JP2003093085A - METHOD FOR PRODUCING POLYESTER, METHOD FOR PRODUCING SUBSTITUTED alpha-HYDROXY ACID AND CLOSTRIDIUM BEIJERINCKII STRAIN HICA 432 - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a useful industrial product from waste such as pulp, paper, waste starch, to avoid exhaustion of oil resources and to solve the environmental pollution by waste. SOLUTION: The method for producing a polyester comprises (1) a process for obtaining one or more kinds of substituted α-hydroxy acids represented by the formula HO-CHR-COOH (R is a >=1C and <=10C hydrocarbon group) by fermenting a saccharide with a bacterium and (2) a process for polymerizing the substituted 2-hydroxy acid or its derivative.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing polyester, and more particularly to a method for recycling sugars, specifically starch, hemicellulose and cellulose.

[0002]

2. Description of the Related Art Conventional general-purpose plastic products are polymer compounds mainly synthesized from petroleum resources. That is, in most cases, polymer compounds such as polyester, polystyrene, nylon, polyethylene, polyvinyl chloride, polyimide and polycarbonate are produced from monomers obtained from petroleum.

However, oil is a limited resource,
It is expected to run out soon. Therefore, a technique for producing general-purpose plastic products from a new raw material replacing petroleum, that is, a renewable raw material is strongly desired.

On the other hand, although the recycling of used paper is being promoted with the spread of personal computers and office automation equipment and the increasing demand for paper from paper containers and the like, the number of times of recycling is limited due to the shortening of the paper fiber, and it cannot be recycled. The handling of such waste paper is a big problem. On the other hand, from the viewpoint of effective use of resources, the use of waste paper, etc. other than thermal recycling is required. Paper has cellulose as its main component, and studies are being made to decompose and reuse this cellulose.

Cellulose is produced in an amount of 10 ⁹ to 10 ¹¹ tons per year, and structural materials, fillers, food additives, and
It is used in large quantities in a wide range of applications such as adhesives. Along with this, the amount of waste cellulose is also increasing year by year.

Hemicellulose, which is present in the plant tissue in an amount of 20 to 30%, is not removed in the pulp production process and is mixed in the paper, and when the amount is large, it reaches about 20%, and when considering the effective utilization of waste paper. In addition, effective utilization of hemicellulose components has become a problem that cannot be ignored. In addition, as the demand for paper has increased, the amount of hemicellulose discarded in the pulp manufacturing process has also increased.

On the other hand, starch is a polymer compound in which D-glucose is polymerized dehydratively, and is an important polysaccharide together with cellulose. Starch is potato, sweet potato,
Manufactured from corn and the like, the worldwide output (corn yield) is about 400 to 500 million tons / year, which is the most produced and renewable resource among natural resources. Therefore, if a general-purpose plastic product can be produced from starch, it is a promising new resource to replace petroleum.

As a technique for decomposing and reusing waste cellulose, for example, a method of extracting hydrocarbons such as methane and ethane from cellulose (Japanese Patent Laid-Open No. 213778/1993),
A method of producing alcohol from cellulose by a microorganism (JP-A-11-299479) is known.

Further, as a technique for utilizing waste hemicellulose, food additives (JP-A-5-17503 and JP-A-5-43470) and foaming agents (JP-A-5-24)
4880), a sizing agent for fiber processing (Japanese Patent Laid-Open No. 5-263).
367), cosmetics (JP-A-6-157238), fiber materials (JP-A-6-220213), and the like have been conventionally studied.

Production of a thermosetting resin material from lignocellulose is described in JP-A-6-126715, and it is produced using an adhesive as a physical property.

[0011]

In order to cope with the further increase of these wastes in the future, it is necessary to develop a new technology capable of recycling resources more efficiently. Therefore, the inventors of the present invention focus on the fact that the increasing amount of waste is pulp, paper, waste starch, and the like, and on the other hand, there is a demand for a plastic raw material that replaces petroleum, and produce a plastic from saccharides. It was investigated. If plastics can be made from sugars, useful industrial products can be provided with abundant starting materials, depletion of petroleum resources can be avoided, and environmental pollution due to waste can be eliminated.

The object of the present invention is therefore to provide a practical process for converting sugars into plastics. Specifically, it is an object to provide a method for producing hemicellulose and a plastic material obtained from cellulose as a raw material, and a method for recycling hemicellulose and cellulose. Moreover, it aims at obtaining high quality plastics using glucan such as cellulose and starch as a starting material.

[0013]

The present invention is a method for producing polyester, which comprises the following steps. (1) HO-C by fermenting sugars with microorganisms
HR-COOH (wherein R represents a hydrocarbon group having 1 to 10 carbon atoms), and a step of obtaining one or more kinds of substituted α-hydroxy acid, (2) the substituted α-hydroxy acid or a derivative thereof. The process of polymerizing.

As the saccharide, starch, glucose or xylose can be used.

When the saccharide is glucose or xylose, prior to the steps (1) and (2), (3)
It is preferable to have a step of hydrolyzing a raw material containing cellulose or hemicellulose to obtain glucose or xylose.

When the saccharide is starch or glucose, the one or more substituted α-hydroxy acids include lactic acid (R is a methyl group), β-phenyllactic acid (R is a benzyl group) and 2-hydroxyisolate. It is selected from the group consisting of caproic acid (wherein R is an isobutyl group).

A preferable microorganism in the step (1) is an anaerobic bacterium, and it is more preferable that the fermentation in the step is carried out in the presence of pyruvic acid.

The substituted α-hydroxy acid derivative is cyclic 2
A lactide monomer, and prior to the polymerization step (2) above,
(4) It is preferable to have a step of dehydrating two molecules of the substituted α-hydroxy acid to form a cyclic diester to obtain the cyclic dimer lactide.

Further, the polyester contains at least one of repeating units represented by the following formulas (I) to (III).

[0020]

[Chemical 4]

[0021]

[Chemical 5]

[0022]

[Chemical 6]

Further, the present invention is a method for producing a substituted α-hydroxy acid, which comprises fermenting saccharides with a microorganism.

The above-mentioned microorganism is preferably an anaerobic bacterium, which produces a substituted α-hydroxy acid by fermentation in the presence of pyruvic acid.

[0025]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention (1) is represented by HO-CHR-COOH (wherein R represents a hydrocarbon group having 1 to 10 carbon atoms) by fermenting sugars with a microorganism. A method for producing a polyester, which comprises a step of obtaining at least two kinds of substituted α-hydroxy acid, and a step of (2) polymerizing the substituted α-hydroxy acid or a derivative thereof.

Specifically, the step (1) may include the following four embodiments.

In the first embodiment, glucan is hydrolyzed to obtain glucose, and the obtained glucose is fermented with a microorganism to produce a substituted α-hydroxy acid, and the substituted α
-A method for producing a polyester, which comprises polymerizing a hydroxy acid or the substituted α-hydroxy acid derivative.

The second embodiment is a polyester characterized by producing a substituted α-hydroxy acid by fermenting starch with a microorganism and polymerizing the substituted α-hydroxy acid or the substituted α-hydroxy acid derivative. Is a manufacturing method.

In the third embodiment, xylose is obtained by hydrolyzing a substance containing at least hemicellulose.
A method for producing a polyester, which comprises producing a substituted α-hydroxy acid by fermenting the obtained xylose with a microorganism, and polymerizing the substituted α-hydroxy acid or the substituted α-hydroxy acid derivative.

In a fourth embodiment, hemicellulose and a substance containing cellulose are hydrolyzed to obtain xylose and glucose, and the obtained xylose and glucose are fermented by a microorganism to produce a substituted α-hydroxy acid, A method for producing a polyester, which comprises polymerizing the substituted α-hydroxy acid or the substituted α-hydroxy acid derivative.

Anaerobic bacteria are preferable as the microorganisms used in the above first to fourth embodiments, Clostridium bacteria are preferable as the anaerobic bacteria, and substituted α-hydroxy acid is produced from glucose as the Clostridium bacteria. Clostridium beige rinki HICA432 (Clostridium beijer)
inckii HICA432) FERM BP-811
Seven strains are preferred.

A. Step of Obtaining Substituted α-Hydroxy Acid Hereinafter, by substituting a saccharide with a microorganism,
The steps to obtain the hydroxy acid will be described in order for these four embodiments.

1. First Embodiment In the first embodiment, a step of hydrolyzing a glucan to obtain glucose, and fermenting the glucose by a microorganism to give HO-CHR-COOH (where R is a carbon number of 1 to 10). A hydrocarbon group)) to obtain one or more substituted α-hydroxy acids.

Each step will be described below.

(Glucan to Glucose) The glucan as a starting material is not particularly limited as long as it is hydrolyzed to produce glucose. However, from the viewpoint of recycling glucan, cellulose and starch, which have a large production amount and have not yet been effectively recycled, are preferable.

The conversion of cellulose into glucose includes, for example, a decomposition method with an enzyme such as cellulase, a decomposition method with an acid such as sulfuric acid or hydrochloric acid, or a decomposition method with supercritical water.

As cellulose as a raw material for obtaining glucose from cellulose, not only commercially available cellulose can be used, but also waste cellulose obtained by appropriately treating wood such as waste paper and waste wood is used. be able to. Therefore, according to the present invention, it is possible to open a new route to the recycling of waste cellulose.

Examples of the conversion of starch into glucose include a hydrolysis method with a dilute acid such as sulfuric acid, a hydrolysis method with an enzyme such as amylase and maltase, and a hydrolysis method with supercritical water.

As starch as a raw material for obtaining glucose from starch, not only commercially available starch can be used, but also waste starch obtained by appropriately treating potato, sweet potato, corn and the like can be used. it can. Therefore, according to the present invention, a new path for recycling waste starch can be opened.

(Substituted α-hydroxy acid from glucose)
The conversion of glucose to substituted α-hydroxy acids is carried out by anaerobic fermentation by microorganisms. The microbial production of hydroxy acid is described in JP-A-5-336981 and JP-A-7-327692, but specific examples of the microorganism producing 2-hydroxyisocaproic acid from glucose are described below. Not listed in.

As a microorganism, α-substituted from glucose
Since few strains have been isolated and identified that produce hydroxyacids, and the production efficiency is low, we diligently screened a new strain. For example, VL medium containing 2-hydroxyisocaproic acid as a substituted α-hydroxy acid and added with 5 g / l glucose and 5 g / l calcium carbonate (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l) 1, NaCl 5 g / l, cysteine hydrochloride 2 g / l) were used to screen strains having the ability to produce 2-hydroxyisocaproic acid under anaerobic conditions at 30 ° C.

[0042] As a result, Clostridium Beiji prn (Clostridiumbeijerinckii)
As a strain capable of producing 2-hydroxyisocaproic acid from glucose in a bacterium belonging to the genus Clostridium such as Clostridium basilica HICA432 ( Clost
ridiumbeijerinckii HICA43
2) The strain was successfully obtained.

The above-mentioned strains were registered at the Japan Patent Organism Depositary Center, National Institute of Advanced Industrial Science and Technology, June 6, 2001.
Deposited on the date (Accession number: FERM P-183
73). After that, it was transferred from the original deposit to the international deposit on July 15, 2002 based on the Budapest Treaty (accession number: FERM BP-8117).

Furthermore, the obtained Clostridium bezierinki HICA432 (Clostridium beij)
erinckii HICA432) FERM BP-8
The 117 strain was analyzed for the product from glucose and found to contain the substitution α shown by HO-CHR-COOH.
It was found that in addition to 2-hydroxyisocaproic acid (R is an isobutyl group) as a hydroxy acid, at least lactic acid (R is a methyl group) and β-phenyllactic acid (R is a benzyl group) are produced. Therefore, these substitutions α-
Polyesters can be made from at least one of the hydroxy acids.

Specifically, when a substituted α-hydroxy acid produced from glucose by HICA432 is used, the obtained polyester has the above formulas (I) to (III).
Having at least one of the repeating units represented by.

Further, the mixture of substituted α-hydroxy acids produced from glucose by HICA432 is purified, if necessary, to contain substantially only β-phenyl lactic acid, substantially only 2-hydroxyisocaproic acid. , Those containing at least β-phenyllactic acid, those containing at least 2-hydroxyisocaproic acid, and the like can be obtained.

When the one or more substituted α-hydroxy acids are purified to contain substantially only β-phenyllactic acid, the resulting polyester mainly contains the structure represented by the formula (II). In this case, since the obtained polyester has a benzyl group in the side chain, it is presumed that it has excellent heat resistance. Moreover, since β-phenyllactic acid has a high polymerization activity, a sufficient degree of polymerization of the obtained polyester can be secured.

The term "mainly contains" means preferably 60 mol% or more, more preferably 70 mol% or more, further preferably 80 mol% or more, and most preferably 90 mol% or more based on the entire repeating unit. Means to occupy.

If the one or more substituted α-hydroxy acids are purified to contain essentially only 2-hydroxyisocaproic acid, the resulting polyester will predominantly contain the structure of formula (III). In this case, since the obtained polyester has an isobutyl group in the side chain, it is presumed that the glass transition temperature is low and the mechanical properties are excellent. Moreover, since 2-hydroxyisocaproic acid has a high polymerization activity, a sufficient degree of polymerization of the obtained polyester can be secured.

When the one or more substituted α-hydroxy acids are purified to contain at least β-phenyl lactic acid, for example lactic acid and β-phenyl lactic acid, the formula (I) and A polyester mainly containing the structure represented by the formula (II) can be produced. In this case, since the obtained polyester has a methyl group and a benzyl group in the side chain, it is presumed that it has an excellent balance of moldability and heat resistance.

When one or more substituted α-hydroxy acids are purified to contain at least 2-hydroxyisocaproic acid, for example lactic acid and 2-hydroxyisocaproic acid, the formula: (I) and formula (I
A polyester mainly containing the structure represented by II) can be produced. In this case, since the obtained polyester has a methyl group and an isobutyl group in the side chain, it is presumed that it has an excellent balance of moldability and mechanical properties.

When the one or more substituted α-hydroxy acids are purified to contain at least β-phenyllactic acid and 2-hydroxyisocaproic acid, for example, structures of formula (II) and formula (III) A polyester containing mainly can be produced. In this case, since the obtained polyester has a benzyl group and an isobutyl group in the side chain, it is presumed that it has an excellent balance of heat resistance and mechanical properties.
Moreover, since β-phenyllactic acid and 2-hydroxyisocaproic acid have high polymerization activity, a sufficient degree of polymerization of the obtained polyester can be secured.

It is also possible to purify each of the substituted α-hydroxy acids and mix a predetermined amount to produce a polyester. In this case, a wide range of physical properties of polyester can be realized.

Clostridium beige rinki HICA4
32 (Clostridium beijerincki
iHICA432) FERM BP-8117 strain is
High production rate of substituted α-hydroxy acids. In particular,
When compared under the same growth conditions, HICA432 shows the amount of substituted α-hydroxy acid produced by an absolutely anaerobic bacterium which is generally known and has the ability to produce substituted α-hydroxy acid in several weeks to several months. Can be achieved. Therefore, sufficient production efficiency of the substituted α-hydroxy acid can be realized.

Also, Clostridium basilica HI
CA432 (Clostridium beijerin
ckiiHICA432) FERM BP-8117 strain has a high growth rate. Specifically, general absolute anaerobic bacteria reach the initial logarithmic growth phase in about one month after subculture, whereas HICA432 reaches the initial logarithmic growth phase in about several days. Therefore, it is possible to easily secure the amount of bacterial cells required to produce a sufficient amount of the substituted α-hydroxy acid.

In the substitution α-hydroxy acid fermentation from glucose, strains are added to a culture solution containing glucose as a main carbon source and nitrogen, phosphorus, minerals, vitamins, etc. as nutrients, and kept under anaerobic conditions. It is done by that.

Since the above strains are absolutely anaerobic bacteria that are killed by oxygen, they strictly realize anaerobic conditions. After injecting the medium, replace the gas phase part of the culture container sealed with a gas-impermeable butyl rubber stopper etc. with nitrogen, carbon dioxide, hydrogen, etc., and also oxygen in the medium such as cysteine, sodium thioglycolate, sodium sulfide, etc. Reducing agent and heat treatment,
It is necessary to remove it by nitrogen aeration.

The glucose concentration in the culture broth may be set according to the type of microorganism used, but is usually 0.1% by mass in order to realize a sufficient amount of the substituted α-hydroxy acid produced.
The amount is preferably 0.5% by mass or more. On the other hand, in order to reduce by-products, it is usually 10% by mass or less and 5% by mass
The following are preferred.

A supplementary carbon source is not necessary and glucose may be used as the single carbon source. As the nitrogen source, organic or inorganic nitrogen sources such as urea, ammonia, ammonium sulfate and ammonium nitrate, and natural nitrogen sources such as corn steep liquor, peptone, meat extract and yeast extract are used. As the inorganic salt, potassium phosphate, sodium phosphate, magnesium sulfate, manganese sulfate, ferrous sulfate, potassium chloride, sodium chloride and the like are used.

The fermentation temperature is usually 15 ° C. or higher and 20 ° C. in order to realize a sufficient amount of the substituted α-hydroxy acid produced.
The above is preferable. On the other hand, in order to reduce by-products and prevent the bacterial strain from dying, the temperature is usually 38 ° C or lower, preferably 37 ° C or lower.

The pH of the culture broth is usually about 5 to 7 and may be appropriately set depending on the nature of the microorganism.

Further, the fermentation time depends on the type of microorganism,
The HICA432 strain can be fermented in 3 days to 1 week, and in 1 to 3 days if pre-culture conditions are selected.

It has been found that the HICA432 strain changes the ratio of the substituted α-hydroxy acid produced with the progress of fermentation. For example, when fermentation is carried out under the conditions described above, lactic acid, β-phenyllactic acid and 2-hydroxyisocaproic acid are produced in almost the same number of moles in the first half of fermentation (for example, up to 3 days after the start of fermentation). As a result, the molar concentration in the medium increases similarly. On the other hand, in the latter half of fermentation (for example, 3 days after the start of fermentation), β-
The molar concentrations of phenyllactic acid and 2-hydroxyisocaproic acid in the medium are constant, and only lactic acid increases.

By utilizing such characteristics, the substituted α-hydroxy acid can be purified more efficiently. For example, if .beta.-phenyl lactic acid and 2-hydroxyisocaproic acid are primarily needed, the mixture of substituted .alpha.-hydroxy acids produced in the early stages of fermentation is purified. on the other hand,
If lactic acid is primarily required, the mixture of substituted α-hydroxy acids produced late in fermentation is purified.

Further, it was also found that the above fermentation characteristics can be controlled by adding pyruvic acid to the medium. For example, when fermented by adding pyruvic acid to the medium,
In the first half of fermentation (for example, until the third day after the start of fermentation), lactic acid, β-phenyllactic acid and 2-hydroxyisocaproic acid are produced in approximately the same molar number, and the molar concentration in the medium is
Each will increase similarly. On the other hand, in the latter half of fermentation (for example, 3 days after the start of fermentation), the molar concentration of 2-hydroxyisocaproic acid in the medium becomes constant, whereas
The molar concentration of lactic acid decreases and the molar concentration of β-phenyllactic acid increases.

By utilizing such characteristics, the substituted α-hydroxy acid can be purified more efficiently. For example, if .beta.-phenyl lactic acid and 2-hydroxyisocaproic acid are primarily needed, the mixture of substituted .alpha.-hydroxy acids produced in the early stages of fermentation is purified. on the other hand,
If β-phenyl lactic acid is primarily required, the mixture of substituted α-hydroxy acids produced later in fermentation is purified.

The concentration of pyruvic acid is preferably 0.1% by mass or more in order to sufficiently suppress the production of lactic acid.
It is more preferably 0.5% by mass or more. On the other hand, in order to reduce by-products, it is preferably 10% by mass or less, more preferably 5% by mass or less.

2. Second Embodiment A second embodiment is one or more kinds represented by HO-CHR-COOH (wherein R represents a hydrocarbon group having 1 to 10 carbon atoms) by fermenting starch with a microorganism. Obtaining a substituted α-hydroxy acid.

Each step will be described below.

(Starch-substituted α-hydroxy acid) As the starch as a raw material, not only commercially available starch can be used but also waste starch obtained by appropriately treating potato, sweet potato, corn and the like. You can Therefore, according to the present invention, a new path for recycling waste starch can be opened.

The step of directly obtaining a substituted α-hydroxy acid from starch without passing through glucose is carried out by anaerobic fermentation by a microorganism. Regarding the microbial production of hydroxy acid, JP-A-5-336981, JP-A-7-327692 and JP-A-2000-300.
Although it is described in Japanese Patent No. 284, etc., it does not specifically describe a microorganism that produces a substituted α-hydroxy acid from starch.

Since few microorganisms have been isolated and identified as strains producing a substituted α-hydroxy acid from starch, and the production efficiency is low, we diligently screened a new strain. At this time, for reasons such as experimental efficiency, instead of directly screening a strain that can convert starch into a substituted α-hydroxy acid, first screen a strain that can convert glucose into a substituted α-hydroxy acid, and obtain the obtained strain. Among them, a strain capable of converting starch into a substituted α-hydroxy acid was further screened.

For example, as the substituted α-hydroxy acid, 2-
VL medium containing hydroxyisocaproic acid and added with 5 g / l glucose and 5 g / l calcium carbonate (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g /
1) was used to screen a strain having the ability to produce 2-hydroxyisocaproic acid under anaerobic conditions at 30 ° C.

Then, the ability of the obtained strain to produce a substituted α-hydroxy acid from starch was examined.

[0075] As a result, Clostridium Beiji prn (Clostridiumbeijerinckii)
As a strain capable of producing 2-hydroxyisocaproic acid from starch in a bacterium belonging to the genus Clostridium, such as Clostridium basilica HICA432 ( Clostr
idiumbeijerinckii HICA432)
Succeeded in acquiring the strain.

Further, the obtained Clostridium bezierinki HICA432 (Clostridium beij)
erinckii HICA432) FERM BP-8
The 117 strain was analyzed for the product from starch, and as with the product from glucose described in the first embodiment, 2-hydroxyisocaproic acid as the substituted α-hydroxy acid represented by HO-CHR-COOH was analyzed. (R is an isobutyl group), at least lactic acid (R
Is a methyl group) and β-phenyllactic acid (R is a benzyl group)
Was found to be produced. Thus, a polyester can be produced from at least one of these substituted α-hydroxy acids.

Specifically, even when a substituted α-hydroxy acid produced from starch by HICA432 is used, the obtained polyester has the above formulas (I) to (I).
It has at least any one of the repeating units represented by II).

The mixture of substituted α-hydroxy acids produced from starch by HICA432 can also be purified, if desired, as in the first embodiment.

The HICA432 used in this embodiment is also used.
Growth rate and the production rate of substituted α-hydroxy acids are also
It was similar to the strain used in the first embodiment.

In the substitution α-hydroxy acid fermentation from starch, starch is used as a main carbon source, and other nutrients such as nitrogen, phosphorus, minerals and vitamins are added to the culture solution.
It is carried out by adding a strain and keeping it under anaerobic conditions.

Since the above strains are absolutely anaerobic bacteria that are killed by oxygen, they strictly realize anaerobic conditions. After injecting the medium, replace the gas phase part of the culture container sealed with a gas-impermeable butyl rubber stopper etc. with nitrogen, carbon dioxide, hydrogen, etc., and also oxygen in the medium such as cysteine, sodium thioglycolate, sodium sulfide, etc. Reducing agent and heat treatment,
It is necessary to remove it by nitrogen aeration.

The starch concentration in the culture broth may be set according to the type of microorganism used, but in order to realize a sufficient amount of the substituted α-hydroxy acid produced, it is usually 0.1% by mass or more, It is preferably 5% by mass or more. On the other hand, in order to reduce by-products, it is usually 10% by mass or less and preferably 5% by mass or less.

A supplementary carbon source is not necessary, and starch may be the single carbon source. As the nitrogen source, organic or inorganic nitrogen sources such as urea, ammonia, ammonium sulfate and ammonium nitrate, corn steep liquor, peptone,
Natural nitrogen sources such as meat extract and yeast extract are used. As the inorganic salt, potassium phosphate, sodium phosphate, magnesium sulfate, manganese sulfate, ferrous sulfate, potassium chloride, sodium chloride and the like are used.

The fermentation temperature is usually 15 ° C. or higher and 20 ° C. in order to realize a sufficient amount of the substituted α-hydroxy acid produced.
The above is preferable. On the other hand, in order to reduce by-products and prevent the bacterial strain from dying, the temperature is usually 38 ° C or lower, preferably 37 ° C or lower.

The pH of the culture broth is usually about 5 to 7 and may be appropriately set depending on the nature of the microorganism.

Further, the fermentation time depends on the type of microorganism,
The HICA432 strain can be fermented in 3 days to 1 week, and in 1 to 3 days if pre-culture conditions are selected.

It has been found that the HICA432 strain changes the ratio of the substituted α-hydroxy acid produced with the progress of fermentation. For example, when fermentation is carried out under the conditions described above, lactic acid, β-phenyllactic acid and 2-hydroxyisocaproic acid are produced in almost the same number of moles in the first half of fermentation (for example, up to 3 days after the start of fermentation). As a result, the molar concentration in the medium increases similarly. On the other hand, lactic acid decreases in the latter half of fermentation (for example, 3rd day after the start of fermentation).

By utilizing such characteristics, the substituted α-hydroxy acid can be purified more efficiently. For example, if .beta.-phenyl lactic acid and 2-hydroxyisocaproic acid are primarily needed, the mixture of substituted .alpha.-hydroxy acids produced late in fermentation is purified.

3. Third Embodiment In a third embodiment of the present invention, a step of hydrolyzing at least hemicellulose-containing material to obtain xylose, and fermenting the xylose with a microorganism to give HO-CHR-COOH (however, , R has 1 to 1 carbon atoms
0 represents a hydrocarbon group) and one or more types of substituted α-
The step of obtaining a hydroxy acid is included.

Each step will be described below.

As the raw material (hemicellulose to xylose) containing hemicellulose, not only commercially available hemicellulose can be used, but also waste paper obtained by appropriately treating waste paper, wood such as waste wood, pulp waste liquid, etc. Hemicellulose can also be used. By establishing a synthetic method of polyester from hemicellulose,
It can open a new route to the recycling of waste hemicellulose. In that respect, the method for producing the polyester of the present invention is also a useful method for recycling hemicellulose.

Conversion of hemicellulose to xylose can be performed, for example, by a method of decomposing with an enzyme such as xylanase,
Examples thereof include a decomposition method with an acid such as sulfuric acid and hydrochloric acid, or a decomposition method with supercritical water.

(Xylose-substituted α-hydroxy acid)
The conversion of xylose to substituted α-hydroxy acids is carried out by anaerobic fermentation by microorganisms. Regarding the microbial production of hydroxy acid, JP-A-5-336981, JP-A-7-327692 and JP-A-2000 are available.
No. 3,00284, etc., but no specific description is given regarding a microorganism that produces a substituted α-hydroxy acid from xylose.

As the microorganism, α-substituted from xylose
Since few strains have been isolated and identified that produce hydroxyacids, and the production efficiency is low, we diligently screened a new strain. At this time, for reasons of experimental efficiency and the like, rather than directly screening for a strain capable of converting xylose into a substituted α-hydroxy acid, the first
Substitute α for glucose similar to that performed for the embodiment of
First, a strain that can be converted into a hydroxy acid was screened, and from the obtained strains, a strain that can also convert xylose into a substituted α-hydroxy acid was further screened.

For example, as the substituted α-hydroxy acid, 2-
VL medium containing hydroxyisocaproic acid and added with 5 g / l glucose and 5 g / l calcium carbonate (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g /
1) was used to screen a strain having the ability to produce 2-hydroxyisocaproic acid under anaerobic conditions at 30 ° C.

Then, the ability of the obtained strain to produce a substituted α-hydroxy acid from xylose was examined.

[0097] As a result, Clostridium Beiji prn (Clostridiumbeijerinckii)
As a strain capable of producing 2-hydroxyisocaproic acid from xylose in a bacterium belonging to the genus Clostridium, such as Clostridium basilica HICA432 ( Clost
ridiumbeijerinckii HICA43
2) The strain was successfully obtained.

Furthermore, the obtained Clostridium bezierinki HICA432 (Clostridium beij)
erinckii HICA432) FERM BP-8
The 117 strain was analyzed for the product from xylose and showed that the substitution α shown by HO-CHR-COOH
It was found that at least β-phenyllactic acid (R is a benzyl group) was produced in addition to 2-hydroxyisocaproic acid (R is an isobutyl group) as a hydroxy acid. Therefore, a polyester can be produced from at least one of these substituted α-hydroxy acids.

Thus, the HICA432 strain was
Unlike the case where glucose or starch is used as the carbon source described in the second embodiment, when xylose is used as the single carbon source, lactic acid is not substantially produced.

Therefore, when using a substituted α-hydroxy acid produced from xylose by HICA432, the resulting polyester has the above formulas (II) and (II
It has at least one of the repeating units represented by I).

Further, 2-hydroxyisocaproic acid and β-produced from xylose by HICA432.
A mixture of phenyl lactic acids can be purified as necessary to obtain those containing substantially only β-phenyl lactic acid, those containing substantially only 2-hydroxyisocaproic acid, and the like. The structure, physical properties and the like of the obtained purified product are the same as those of the substituted α-hydroxy acid purified product obtained from glucose described in the first embodiment.

It is also possible to purify β-phenyllactic acid and 2-hydroxyisocaproic acid, and mix them in predetermined amounts to produce a polyester. in this case,
A wide range of physical properties of polyester can be realized.

In the substitution α-hydroxy acid fermentation from xylose, a strain is added to a culture solution containing xylose as a main carbon source and nitrogen, phosphorus, minerals, vitamins, etc. as nutrients and kept under anaerobic conditions. It is done by that.

Since the above strains are absolutely anaerobic bacteria that are killed by oxygen, they strictly realize anaerobic conditions. After injecting the medium, replace the gas phase part of the culture container sealed with a gas-impermeable butyl rubber stopper etc. with nitrogen, carbon dioxide, hydrogen, etc., and also oxygen in the medium such as cysteine, sodium thioglycolate, sodium sulfide, etc. Reducing agent and heat treatment,
It is necessary to remove it by nitrogen aeration.

The xylose concentration in the culture broth may be set according to the type of microorganism used, but is usually 0.1% by mass in order to realize a sufficient amount of the substituted α-hydroxy acid produced.
The amount is preferably 0.5% by mass or more. On the other hand, in order to reduce by-products, it is usually 10% by mass or less and 5% by mass
The following are preferred.

No auxiliary carbon source is required, and xylose may be used as a single carbon source. As the nitrogen source, organic or inorganic nitrogen sources such as urea, ammonia, ammonium sulfate and ammonium nitrate, and natural nitrogen sources such as corn steep liquor, peptone, meat extract and yeast extract are used. As the inorganic salt, potassium phosphate, sodium phosphate, magnesium sulfate, manganese sulfate, ferrous sulfate, potassium chloride, sodium chloride and the like are used.

The fermentation temperature is usually 15 ° C. or higher and 20 ° C. in order to realize a sufficient amount of the substituted α-hydroxy acid produced.
The above is preferable. On the other hand, in order to reduce byproducts and prevent the bacterial strain from dying, the temperature is usually 37 ° C or lower, preferably 35 ° C or lower.

The pH of the culture broth is usually about 5 to 8, but it may be appropriately set depending on the properties of the microorganism.

Furthermore, the fermentation time depends on the type of microorganism,
The HICA432 strain can be fermented in 3 days to 1 week, and in 1 to 3 days if pre-culture conditions are selected.

As mentioned above, the HICA432 strain produces substantially no lactic acid when xylose is the sole carbon source. Therefore, it is advantageous when at least one of β-phenyllactic acid and 2-hydroxyisocaproic acid is mainly required.

4. Fourth Embodiment In a fourth embodiment, a step of hydrolyzing hemicellulose and a material containing cellulose to obtain xylose and glucose, and fermenting the xylose and glucose with a microorganism to give HO-CHR-COOH. (However, R
Represents a hydrocarbon group having 1 to 10 carbon atoms).

Each step will be described below.

(Hemicellulose and Cellulose to Xylose and Glucose) As the raw material containing hemicellulose and cellulose, not only commercially available hemicellulose and cellulose can be used, but also used papers, woods such as waste materials, pulp waste liquid, etc. Waste hemicellulose and waste cellulose obtained by appropriate treatment can also be used. By establishing a method for synthesizing polyester from hemicellulose and cellulose, it is possible to open a new route to the recycling of waste hemicellulose and waste cellulose. In that respect, the method for producing the polyester of the present invention is also a useful method for recycling hemicellulose and cellulose at the same time.

Conversion from hemicellulose to xylose and from cellulose to glucose can be carried out, for example, by a decomposition method with an enzyme such as xylanase or cellulase, a decomposition method with an acid such as sulfuric acid or hydrochloric acid, or a decomposition method with supercritical water. Is mentioned. Hemicellulose and cellulose can be simultaneously decomposed by a method utilizing a plurality of enzymes or a decomposition method using an acid or supercritical water.

(Substitution from xylose and glucose α-
Substituted from (hydroxy acid) xylose and glucose α-
Conversion to hydroxy acids is carried out by anaerobic fermentation by microorganisms. Regarding the microbial production of hydroxy acid,
JP-A-5-336981, JP-A-7-32769
Although it is described in JP-A No. 2 and JP-A No. 2000-300284, it does not specifically describe a microorganism that produces a substituted α-hydroxy acid from xylose and glucose.

As the microorganism of this embodiment, the same one as in the third embodiment was used. That is, instead of directly screening a strain capable of simultaneously converting both xylose and glucose into a substituted α-hydroxy acid, a strain capable of converting glucose into a substituted α-hydroxy acid is first analyzed as described in the first embodiment. From the thus obtained strains, a strain capable of converting xylose into a substituted α-hydroxy acid was further screened.

Then, the ability of the obtained strain to produce a substituted α-hydroxy acid from xylose was examined.

[0118] As a result, Clostridium Beiji prn (Clostridiumbeijerinckii)
As a strain capable of producing 2-hydroxyisocaproic acid from xylose and glucose in a bacterium belonging to the genus Clostridium, such as Clostridium basilica HICA43
2 ( Clostridium beijerinckii
HICA432) was successfully obtained.

Obtained Clostridium basilica H
ICA432 (Clostridium beijeri)
nckii HICA432) FERM BP-8117
When the product product from xylose and glucose was analyzed with respect to the strain, 2-hydroxyisocaproic acid (R is an isobutyl group) was added as a substituted α-hydroxy acid represented by HO-CHR-COOH, and at least lactic acid (R was methyl was used). Group) and β-phenyllactic acid (R is a benzyl group) were found to be produced. Thus, a polyester can be produced from at least one of these substituted α-hydroxy acids.

Specifically, when a substituted α-hydroxy acid produced from xylose and glucose by HICA432 is used, the obtained polyester has the above formula (I).
To (III) at least one of the repeating units.

In the present embodiment, the substituted α-hydroxy acid produced by HICA432 from xylose and glucose can be purified in the same manner as in the above-described embodiments, and the structure and physical properties of the obtained purified product are also shown in Examples. It is the same as that obtained by the purification of 1.

Substitution α-from xylose and glucose
Hydroxy acid fermentation uses xylose and glucose as main carbon sources, and other nutrients such as nitrogen, phosphorus, minerals,
It is carried out by adding a strain to a culture solution containing vitamins and the like and maintaining it under anaerobic conditions.

Since the above strains are absolutely anaerobic bacteria that are killed by oxygen, they strictly realize anaerobic conditions. After injecting the medium, replace the gas phase part of the culture container sealed with a gas-impermeable butyl rubber stopper etc. with nitrogen, carbon dioxide, hydrogen, etc., and also oxygen in the medium such as cysteine, sodium thioglycolate, sodium sulfide, etc. Reducing agent and heat treatment,
It is necessary to remove it by nitrogen aeration.

The concentrations of xylose and glucose in the culture solution may be set according to the type of microorganism used,
In order to achieve a sufficient amount of substituted α-hydroxy acid produced,
The concentration of at least one of xylose and glucose, or the total concentration of xylose and glucose,
Usually, it is 0.1% by mass or more, preferably 0.5% by mass or more. On the other hand, in order to reduce by-products, it is usually 10% by mass or less and preferably 5% by mass or less.

No auxiliary carbon source is required, and only xylose and glucose may be used as carbon sources. As the nitrogen source, organic or inorganic nitrogen sources such as urea, ammonia, ammonium sulfate and ammonium nitrate, and natural nitrogen sources such as corn steep liquor, peptone, meat extract and yeast extract are used. As the inorganic salt, potassium phosphate, sodium phosphate, magnesium sulfate, manganese sulfate, ferrous sulfate, potassium chloride, sodium chloride and the like are used.

The fermentation temperature is usually 15 ° C. or higher and 20 ° C. in order to realize a sufficient amount of the substituted α-hydroxy acid produced.
The above is preferable. On the other hand, in order to reduce byproducts and prevent the bacterial strain from dying, the temperature is usually 37 ° C or lower, preferably 35 ° C or lower.

The pH of the culture broth is usually about 5 to 8, but it may be appropriately set depending on the properties of the microorganism.

Furthermore, the fermentation time depends on the type of microorganism,
The HICA432 strain can be fermented in 3 days to 1 week, and in 1 to 3 days if pre-culture conditions are selected.

B. Step of obtaining cyclic dimeric lactide from substituted α-hydroxy acid From the substituted α-hydroxy acid obtained in the steps described in the first to fourth embodiments,
Although the polyester can be directly produced, the substituted α-hydroxy acid can be converted into a derivative having high polymerization activity prior to the polymerization step.

For example, two molecules of the substituted α-hydroxy acid are dehydrated to form a cyclic diester, a cyclic dimer lactide is prepared as a derivative of the substituted α-hydroxy acid, and then the cyclic dimer lactide is subjected to ring-opening polymerization. Thereby, polyester can be manufactured. Ring-opening polymerization generally has a high polymerization rate and can produce a polyester having a high degree of polymerization.

As a method for dehydrating two molecules of a substituted α-hydroxy acid to form a cyclic diester, for example, DeanS is used.
Using a reactor equipped with a tarktrap, a substituted α-hydroxy acid and a condensation catalyst such as p-toluenesulfonic acid are subjected to azeotropic dehydration in toluene under a nitrogen atmosphere for 30 hours to appropriately remove water accumulated in the DeanStarktrap. By doing so, the cyclic dimer lactide can be obtained in high yield.

When two molecules of lactic acid are dehydrated to undergo cyclic diesterification under the above conditions, the resulting cyclic dimer lactide is represented by the following formula (I ').

[0133]

[Chemical 7]

Under the above conditions, when two molecules of β-phenyllactic acid are dehydrated for cyclic diesterification, the resulting cyclic dimer lactide is represented by the following formula (II ').

[0135]

[Chemical 8]

Under the above conditions, when two molecules of 2-hydroxyisocaproic acid are dehydrated to form a cyclic diester,
The resulting cyclic dimer lactide is represented by the following formula (III ′).

[0137]

[Chemical 9]

C. Step of Obtaining Polyester from Substituted α-Hydroxy Acid The objective polyester is condensation polymerization by refluxing a substituted α-hydroxy acid and a polymerization catalyst in an organic solvent and removing water generated in the polymerization process to the outside of the reaction system. Can be advanced.

(A) Polymerization catalyst In the condensation polymerization of the substituted α-hydroxy acid, examples of the polymerization catalyst include metals such as tin powder and zinc powder, tin oxide, zinc oxide, magnesium oxide, titanium oxide and aluminum oxide. Metal oxide, tin dichloride, tin tetrachloride,
Metal halides such as tin dibromide, tin tetrabromide, zinc chloride, magnesium chloride and aluminum chloride, tetraphenyltin, tin octylate, p-toluenesulfonic acid and the like can be used.

The amount of the polymerization catalyst used is 0.001 to 10% by mass, preferably 0.1% by weight based on the substituted α-hydroxy acid.
It is 01 to 5 mass%.

(A) Polymerization solvent In the condensation polymerization of the substituted α-hydroxy acid, a polymerization solvent which can be easily separated from water by liquid separation is preferable. For example, toluene, xylene, mesitylene, 1, 2, 3, 5
-Tetramethylbenzene, chlorobenzene, 1,2-dichlorobenzene, 1,3-dichlorobenzene, bromobenzene, 1,2-dibromobenzene, 1,3-dibromobenzene, iodobenzene, 1,2-diiodobenzene, diphenyl ether Solvents such as dibenzyl ether and dibenzyl ether may be used, and these may be used as a mixture.

The amount of the polymerization solvent used is preferably such that the concentration of the substituted α-hydroxy acid is 5 to 50% by mass.

(C) Polymerization conditions In the condensation polymerization of the substituted α-hydroxy acid, the polymerization temperature is
Considering the production rate of the polymer and the thermal decomposition rate of the produced polymer, 50 to 200 ° C., preferably 110 to 18
It is 0 ° C. The condensation polymerization reaction is usually carried out at the distillation temperature of the organic solvent used under normal pressure. When using an organic solvent having a high boiling point, it may be carried out under reduced pressure.

In the condensation polymerization of substituted α-hydroxy acid,
It is preferable to carry out under an inert gas atmosphere, and it may be carried out while replacing the reaction apparatus with an inert gas or while bubbling with an inert gas. Further, water generated in the polymerization reaction process is appropriately removed from the reactor.

The weight average molecular weight of the polyester obtained by the polymerization is the kind of the polymerization solvent, the kind and the amount of the polymerization catalyst,
Various molecular weights can be obtained by changing the conditions such as polymerization temperature and polymerization time.
It is preferably 000 to 1,000,000.

The polyester thus obtained contains a heat-resistant stabilizer, a weather-resistant stabilizer, an antioxidant, an antistatic agent, a flame retardant, a lubricant, a dye, a pigment, an ultraviolet absorber, and a crosslinker, as long as the object of the present invention is not impaired. Agents and the like can be added. The mixing ratio is not particularly limited and may be appropriately determined.

When the polyester is produced by condensation polymerization in an organic solvent at a temperature of less than 200 ° C., it may be colored, or
Impurities due to thermal decomposition are suppressed. Therefore, when it is used for applications such as an adhesive or a coating agent, a desired dyeing can be obtained, which is advantageous in appearance.

The polyester obtained as described above has good melt moldability, and the polyester having a weight average molecular weight of more than 50,000 has sufficient mechanical strength and can be used for many purposes. For example, packaging films, bottles, cups, trays, spoons, knives, forks, garbage bags and the like can be mentioned.

Further, the substituted α-hydroxy acid is an optical isomer (chiral), and the polyester obtained by the direct dehydration condensation polymerization of the substituted α-hydroxy acid also has a chiral structure, so that it is also an optical material. Can be used. However, it is not limited to these applications.

D. Step of Obtaining Polyester from Cyclic Dimer Lactide The method for producing polyester described in the previous section is a method of obtaining a polyester by polymerizing a substituted α-hydroxy acid. The target polyester is a cyclic dimer lactide and a polymerization catalyst. And ring-opening polymerization in an inert gas atmosphere.

(A) Polymerization catalyst In the ring-opening polymerization of the cyclic dimer lactide, examples of the polymerization catalyst include metals such as tin powder and zinc powder, tin oxide,
Metal oxides such as zinc oxide, magnesium oxide, titanium oxide and aluminum oxide, tin halides such as tin dichloride, tin chloride, tin dibromide, tin tetrabromide, zinc chloride, magnesium chloride and aluminum chloride, tetra Phenyltin, tin octylate, etc. can be used. Among these, tin or tin compounds are particularly preferable because they have excellent catalytic activity.

The amount of the polymerization catalyst used is 0.001 to 10% by mass, preferably 0.0 to 10% by mass, based on the cyclic dimer lactide.
It is 1 to 5 mass%.

(A) Polymerization conditions In the ring-opening polymerization of the cyclic dimer lactide, the polymerization temperature is 100 to 200 ° C., preferably 120 to 200 ° C., in consideration of the production rate of the polymer and the thermal decomposition rate of the produced polymer. 18
It is 0 ° C.

Ring-opening polymerization of the cyclic dimer lactide is preferably carried out in an inert gas atmosphere. As the inert gas, for example, nitrogen gas or argon gas can be used.

Regarding the weight average molecular weight of the polyester obtained by the polymerization, various molecular weights can be obtained by changing the conditions such as the type and amount of the polymerization catalyst, the polymerization temperature and the polymerization time.
000 is preferable.

The obtained polyester may be a heat-resistant stabilizer, a weather-resistant stabilizer, an antioxidant, an antistatic agent, a flame retardant, a lubricant, a dye, a pigment, an ultraviolet absorber, a cross-linker, as long as the object of the present invention is not impaired. Agents and the like can be added. The mixing ratio is not particularly limited and may be appropriately determined.

When the polyester is produced by ring-opening polymerization of a cyclic dimer lactide at less than 200 ° C., it is colored or
Alternatively, impurities due to thermal decomposition are suppressed. Therefore,
When it is used for applications such as adhesives and coatings, it has the advantage of appearance because it gives the desired dyeing.

The polyester obtained as described above has good melt moldability, and the polyester having a weight average molecular weight of more than 50,000 has sufficient mechanical strength and therefore can be used for many purposes. For example, packaging films, bottles, cups, trays, spoons, knives, forks, garbage bags and the like can be mentioned.

Hereinafter, the present invention will be described more specifically with reference to Examples. However, the present invention is not limited to these examples. Unless otherwise specified, commercially available high-purity reagents were used.

First, a method for screening a microorganism used in the step of obtaining a substituted α-hydroxy acid by microbial fermentation of glucose described in the first embodiment will be described.

(Screening of 2-Hydroxyisocaproic Acid-Producing Bacteria) Sampling of reduced soil located a few centimeters below the surface layer of the swamp in the infiltrated state, microbial membrane on the surface of the drain, and scum floating on the still water surface 5g
VL medium supplemented with glucose / l glucose and 5 g / l calcium carbonate (peptone 10 g / l, yeast extract 5 g / l
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l).

The medium was injected into a vial to make it anaerobic, then the gas phase was replaced with nitrogen gas, sealed and autoclaved (121 ° C., 98 kPa pressurization, 10 minutes), and the sample was added. I opened it. After adding the sample, the gas phase portion of the vial was replaced with nitrogen gas again, and the vial was sealed with a butyl rubber stopper. After that, static culture was carried out under anaerobic conditions at 30 ° C. for 2 weeks with occasional agitation, and the preculture solution was added to the medium in the above-mentioned medium maintained under anaerobic conditions in the same manner as above.
Volume% was added.

Then, after statically culturing at 30 ° C. for 1 week with occasional agitation, 2-hydroxyisocaproic acid produced in the culture solution was subjected to methylation treatment, and 2-hydroxyisocaproic acid methyl ester was subjected to GC- It was evaluated by MS. Next, the culture solution in which the production of 2-hydroxyisocaproic acid was observed was sterilized, diluted several steps with a cysteine hydrochloride 2 g / l aqueous solution whose pH was adjusted to 7.0, and the roll tube method was used for each dilution step It was cultured in. The medium is agar 2 on the above medium
The thing which added the mass% was used.

Then, the cells were cultured at 30 ° C. for 1 week, and the resulting colonies were suspended in the above medium kept under anaerobic conditions in the same manner as above in a glove box filled with nitrogen gas.
Then, after statically culturing at 30 ° C. for 1 week with occasional stirring, 2-hydroxyisocaproic acid in the culture solution was subjected to methylation treatment and evaluated by GC-MS. A culture solution in which a large amount of 2-hydroxyisocaproic acid was produced was cultured by the roll tube method in the same manner as above, colonies were separated, and a 2-hydroxyisocaproic acid-producing strain was isolated. It isolates Clostridium Beiji of opportunity (Clostridiumbei
jeranckii) and was named HICA432 strain (FERM BP-8117).

It was confirmed that the HICA432 strain produces not only 2-hydroxyisocaproic acid but also lactic acid and phenyllactic acid. Therefore, in the following, H
The ICA432 strain may also be referred to as a 2-hydroxyisocaproic acid producing strain, a lactic acid producing strain, and a phenyllactic acid producing strain.

[0166]

Examples Examples 1-48 below are methods for producing polyesters, which include the step of obtaining a substituted α-hydroxy acid by microbial fermentation of glucose described in the first embodiment. As the microorganism, the 2-hydroxyisocaproic acid producing strain HICA432 strain obtained by the above screening was used.

(Example 1) Cellulose (manufactured by Nippon Paper Industries, K
C Flock W-100) 370 parts by mass of enzyme solution 110
It was put into 50 parts by mass and stirred at 45 ° C. for 8 hours. The enzyme solution was cellulase (Meiji Seika, Meiserase TP6
0) 37 parts by mass of acetic acid / sodium acetate aqueous solution (pH
4.5) What was melt | dissolved in 11000 mass parts was used. After the reaction, 735 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (manufactured by Organo,
Amberlite IR-120B) was passed through and the solvent was distilled off. The reaction mixture was separated and purified to obtain 220 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, internal standard: TMS (tetramethylsilane), D
MSO-d ₆ ) was measured. The chemical shift δ (pp
m) was as follows; α type: 92.12, 73.
04, 72.29, 71.80, 70.58, 61.2
0, β type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

220 parts by weight of this glucose was added to 12100
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and the preculture liquid 1 of the 2-hydroxyisocaproic acid producing strain HICA432 strain isolated by the above-mentioned method
21 parts by mass was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and the remaining solid content was heated under reduced pressure. The sublimated gas was cooled and the precipitated solid was collected to obtain 10 parts by mass of 2-hydroxyisocaproic acid.

Of the resulting 2-hydroxyisocaproic acid
The measurement result of ¹³ C-NMR (100 MHz, TMS, CDCl ₃ ) was as follows (δ / ppm); 180.2
5, 68.95, 43.16, 24.45, 23.2
0, 21.42. From this measurement result, it was confirmed that the desired 2-hydroxyisocaproic acid was obtained.

Also, an optically active column (J & WSCIEN
As a result of evaluating the three-dimensional structure using Cylodex-B manufactured by TIFIC, it was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 100 parts by mass of mesitylene was added to 10 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of tin dichloride, and azeotropic dehydration was carried out under a nitrogen atmosphere. Dean
Water accumulated in the Stark trap was appropriately removed. The polymerization time was 40 hours, and the aliphatic polyester obtained had a weight average molecular weight of 90,000.

Moreover, ¹³ C-NMR (100 MHz, TM
S, CDCl ₃ ) measurement result (δ / ppm) is 16
9.78, 71.38, 39.33, 24.53, 2
It was 3.00 and 21.43, and it was confirmed that the desired aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 90,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 38 to 39 μm, tensile strength: 48 MPa (yield), 48 MPa (breakage), elongation: 5%.

(Example 2) An aliphatic polyester was polymerized using 2-hydroxyisocaproic acid obtained by performing the same operation as in Example 1. Dean Stark tra
Using a reactor equipped with p, 100 parts by mass of 1,3-dichlorobenzene was added to 10 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of zinc chloride, and azeotropic dehydration was carried out under a nitrogen atmosphere. Water accumulated in the Stark trap was appropriately removed. The polymerization time is 45 hours,
The weight average molecular weight of the obtained aliphatic polyester was 140,000.

The aliphatic polyester having a weight average molecular weight of 140,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 57 MPa (yield), 56 MPa (breaking), elongation: 9%.

Example 3 Recycled paper for PPC (Canon Sales, EN-500, A4) used (copied on one side with a copying machine) was cut into 5 mm squares, and 380 parts by mass of the enzyme solution 11300. It was put into a mass part and stirred at 45 ° C. for 10 hours. As the enzyme solution, 40 parts by mass of cellulase (Meijise TP60 manufactured by Meiji Seika Co., Ltd.) was dissolved in 11200 parts by mass of acetic acid / sodium acetate aqueous solution (pH 4.5). After the reaction, 750 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo) to distill off the solvent. The reaction mixture was separated and purified to obtain 210 parts by mass of glucose.

210 parts by weight of this glucose was added to 11600
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and then butyl rubber stopper Seal it with an autoclave (121 ° C,
Pressurized at 98 kPa, 10 minutes). Then, add 3 media
Pre-culture liquid 116 of 2-hydroxyisocaproic acid producing strain HICA432 strain cooled to 0 ° C. and isolated by the above method
Add parts by mass using a syringe with the bottle sealed,
Static culture was carried out at 30 ° C. for 3 days. Then, 9 parts by mass of 2-hydroxyisocaproic acid was recovered in the same manner as in Example 1.

When ¹³ C-NMR was measured, Example 1 was obtained.
A spectrum similar to that of was obtained, and it was confirmed that the desired 2-hydroxyisocaproic acid was obtained.

Using a reactor equipped with a Dean Stark trap, 9 parts by mass of 2-hydroxyisocaproic acid, 0.018 parts by mass of tin octylate and 90% of mesitylene were used.
Add parts by mass, perform azeotropic dehydration under a nitrogen atmosphere, and
Water accumulated in the n Stark trap was appropriately removed. The polymerization time was 40 hours, and the aliphatic polyester obtained had a weight average molecular weight of 50,000.

When ¹³ C-NMR was measured, the same spectrum as in Example 1 was obtained, and it was confirmed that the desired aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 50,000 thus obtained was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 20 MPa (yield), 20 MPa (breakage), elongation: 3%.

Example 4 An aliphatic polyester was polymerized using 2-hydroxyisocaproic acid obtained by performing the same operation as in Example 3. Dean Stark tra
Using a reactor equipped with p, 100 parts by mass of diphenyl ether was added to 9 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of tin powder, and azeotropic dehydration was performed at 130 ° C./12 mmHg, and then in a Dean Stark trap. The accumulated water was appropriately removed. The polymerization time was 48 hours, and the weight average molecular weight of the obtained aliphatic polyester was 1
It was 90,000.

The aliphatic polyester having a weight average molecular weight of 190,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 69 MPa (yield), 67 MPa (breaking), elongation: 13%.

(Example 5) Cellulose (manufactured by Nippon Paper Industries, K
C Flock W-100) 370 parts by mass of enzyme solution 110
It was put into 50 parts by mass and stirred at 45 ° C. for 8 hours. The enzyme solution was cellulase (Meiji Seika, Meiserase TP6
0) 37 parts by mass of acetic acid / sodium acetate aqueous solution (pH
4.5) What was melt | dissolved in 11000 mass parts was used. After the reaction, 735 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (manufactured by Organo,
Amberlite IR-120B) was passed through and the solvent was distilled off. The reaction mixture was separated and purified to obtain 220 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

220 parts by mass of this glucose was added to 12100
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and the preculture liquid 1 of the 2-hydroxyisocaproic acid producing strain HICA432 strain isolated by the above-mentioned method
21 parts by mass was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and the remaining solid content was heated under reduced pressure. The sublimated gas was cooled and the precipitated solid was collected to obtain 10 parts by mass of 2-hydroxyisocaproic acid.

¹³ C of this 2-hydroxyisocaproic acid
_{-NMR (100MHz, TMS, CDCl 3} ) measurement results ([delta] / ppm) is 180.25,68.95,4
It was 3.16, 24.45, 23.20, 21.42, and it was confirmed that the desired 2-hydroxyisocaproic acid was obtained.

Using a reaction apparatus equipped with a Dean Stark trap, 600 parts by mass of toluene was added to 10 parts by mass of 2-hydroxyisocaproic acid and 0.2 parts by mass of p-toluenesulfonic acid, and the mixture was azeotropically distilled for 30 hours in a nitrogen atmosphere. Dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 8.2 parts by mass of cyclic dimer lactide. ¹³ C-NMR of the obtained cyclic dimer lactide (100 MHz,
TMS, CDCl ₃ ) measurement result (δ / ppm) is 1
67.33, 74.14, 38.87, 23.89, 2
It was 3.03 and 21.33, and it was confirmed that the target cyclic dimer lactide was obtained.

The ring-opening polymerization was carried out by heating 8.2 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of tin chloride at 150 ° C. for 12 hours in a nitrogen atmosphere. The weight average molecular weight of the obtained aliphatic polyester was 180,000.

¹³ C-NMR of this aliphatic polyester
(100 MHz, TMS, CDCl ₃ ) measurement result (δ
/ Ppm) is 169.78, 71.38, 39.33,
24.53, 23.00, 21.43, and it was confirmed that the target aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 180,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 68 MPa (yield), 68 MPa (breakage), elongation: 13%.

Example 6 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 5. The ring-opening polymerization was carried out by heating 8.2 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of zinc chloride in a nitrogen atmosphere at 120 ° C. for 5 hours. The weight average molecular weight of the obtained aliphatic polyester was 50,000. ^{When 13} C-NMR was measured, the same spectrum as in Example 5 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 50,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 29 MPa yield), 29 MPa (break), elongation: 4%.

(Example 7) Recycled paper for PPC (Canon Sales, EN-500, A4) used (copied on one side with a copying machine) was cut into 5 mm square, and 380 parts by mass of the enzyme solution 11300 was cut. It was put into a mass part and stirred at 45 ° C. for 10 hours. As the enzyme solution, 40 parts by mass of cellulase (Meijise TP60 manufactured by Meiji Seika Co., Ltd.) was dissolved in 11200 parts by mass of acetic acid / sodium acetate aqueous solution (pH 4.5). After the reaction, 750 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo) to distill off the solvent. The reaction mixture was separated and purified to obtain 210 parts by mass of glucose.

210 parts by weight of this glucose was added to 11600
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and then butyl rubber stopper Seal it with an autoclave (121 ° C,
Pressurized at 98 kPa, 10 minutes). Then, add 3 media
Pre-culture liquid 116 of 2-hydroxyisocaproic acid producing strain HICA432 strain cooled to 0 ° C. and isolated by the above method
Add parts by mass using a syringe with the bottle sealed,
Static culture was carried out at 30 ° C. for 3 days. Then, 9 parts by mass of 2-hydroxyisocaproic acid was recovered in the same manner as in Example 5.

When ¹³ C-NMR was measured, Example 5
A spectrum similar to that of was obtained, and it was confirmed that the desired 2-hydroxyisocaproic acid was obtained. Obtained 2-
By performing the same operation as in Example 5 with 9 parts by mass of hydroxyisocaproic acid, 7.4 parts by mass of a cyclic dimer lactide was obtained.

The ring-opening polymerization was carried out by heating 7.4 parts by mass of the cyclic dimer lactide and 0.008 parts by mass of tin octylate under nitrogen atmosphere at 180 ° C. for 30 hours. The weight average molecular weight of the obtained aliphatic polyester was 680,000. ¹³ C-
When NMR was measured, the same spectrum as in Example 5 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 680,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 30%.

Example 8 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 7. The ring-opening polymerization was carried out by heating 7.4 parts by mass of the cyclic dimer lactide and 0.007 parts by mass of the tin powder at 160 ° C. for 18 hours in a nitrogen atmosphere. The weight average molecular weight of the obtained aliphatic polyester was 390,000. ¹³ C-NMR
As a result, the same spectrum as in Example 5 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 390,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 88 MPa (yield), 87 MPa (breaking), elongation: 22%.

(Example 9) Cellulose (manufactured by Nippon Paper Industries, K
C Flock W-100) 370 parts by mass of enzyme solution 110
It was put into 50 parts by mass and stirred at 45 ° C. for 8 hours. The enzyme solution was cellulase (Meiji Seika, Meiserase TP6
0) 37 parts by mass of acetic acid / sodium acetate aqueous solution (pH
4.5) What was melt | dissolved in 11000 mass parts was used. After the reaction, 735 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (manufactured by Organo,
Amberlite IR-120B) was passed through and the solvent was distilled off. The reaction mixture was separated and purified to obtain 220 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

220 parts by weight of this glucose was added to 12100
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., 121 parts by mass of the preculture liquid of the phenyllactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe with the bottle sealed, and the mixture was allowed to stand at 30 ° C. for 3 days. The culture was performed.

After that, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and then purified to obtain 10 parts by mass of phenyllactic acid. ¹³ C-N of the obtained phenyllactic acid
The measurement results (δ / ppm) of MR (100 MHz, TMS, CDCl ₃ ) were 177.92, 137.63, 13
0.30, 129.46, 127.82, 72.05,
It was 40.38, and it was confirmed that the desired phenyllactic acid was obtained.

Also, an optically active column (J & WSCIEN
As a result of evaluating the three-dimensional structure using TIFIC's Cyclodex-B), it was confirmed that the obtained phenyllactic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 100 parts by mass of mesitylene was added to 10 parts by mass of phenyllactic acid and 0.018 parts by mass of tin dichloride, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark.
Water accumulated in the trap was appropriately removed. Polymerization time is 4
It was 4 hours, and the weight average molecular weight of the obtained aromatic polyester was 180,000.

The aromatic polyester having a weight average molecular weight of 180,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 38 to 39 μm, tensile strength: 69 MPa (yield), 69 MPa (breakage), elongation: 12%.

Example 10 An aromatic polyester was polymerized using phenyllactic acid obtained by the same procedure as in Example 9. Using a reactor equipped with Dean Stark trap, 10 parts by mass of phenyllactic acid and 0.05 of zinc chloride are used.
100 parts by mass of 1,3-dichlorobenzene was added to 2 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Sta.
Water accumulated in the rk trap was appropriately removed. The polymerization time was 16 hours, and the aromatic polyester obtained had a weight average molecular weight of 30,000.

The aromatic polyester having a weight average molecular weight of 30,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 20 MPa (yield), 19 MPa (break), elongation: 2%.

Example 11 Recycled paper for PPC (Canon Sales, EN-500, A4) used (copied on one side with a copying machine) was cut into 5 mm square, and 380 parts by mass of the enzyme solution 11300 was cut. Add to the mass part, 10 at 45 ℃
Stir for hours. The enzyme solution is a cellulase (made by Meiji Seika,
40 parts by mass of macerase TP60) dissolved in 11200 parts by mass of acetic acid / sodium acetate aqueous solution (pH 4.5) was used. After the reaction, 750 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo) to distill off the solvent. The reaction mixture was separated and purified to obtain 210 parts by mass of glucose.

210 parts by weight of this glucose was added to 11600
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and then butyl rubber stopper Seal it with an autoclave (121 ° C,
Pressurized at 98 kPa, 10 minutes). Then, add 3 media
The mixture was cooled to 0 ° C., 116 parts by mass of a preculture liquid of the phenyllactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe with the bottle sealed, and static culture was carried out at 30 ° C. for 3 days. Then, 9 parts by mass of phenyl lactic acid was recovered in the same manner as in Example 1.

As a result of measurement of ¹³ C-NMR, Example 9 was performed.
A spectrum similar to that of was obtained, and it was confirmed that the target phenyllactic acid was obtained.

Using a reactor equipped with a Dean Stark trap, 9 parts by mass of phenyl lactic acid and 0.
90 parts by mass of mesitylene was added to 016 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark tra
Water accumulated in p was appropriately removed. The polymerization time was 40 hours, and the aromatic polyester obtained had a weight average molecular weight of 90,000.

The aromatic polyester having a weight average molecular weight of 90,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 49 MPa (yield), 49 MPa (breaking), elongation: 5%.

Example 12 An aromatic polyester was polymerized by using phenyllactic acid obtained by the same operation as in Example 11. Using a reactor equipped with a Dean Stark trap, 9 parts by mass of phenyl lactic acid and 0.0 of tin powder were used.
100 parts by mass of diphenyl ether is added to 15 parts by mass,
Azeotropic dehydration is performed at 130 ° C / 12 mmHg, and Dean
Water accumulated in the Starktrap was appropriately removed. The polymerization time was 48 hours, and the weight average molecular weight of the obtained aromatic polyester was 250,000.

The aromatic polyester having a weight average molecular weight of 250,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 82 MPa (yield), 81 MPa (breaking), elongation: 18%.

Example 13 Cellulose (manufactured by Nippon Paper Industries,
KC Flock W-100) 370 parts by mass of enzyme solution 11
It was added to 050 parts by mass and stirred at 45 ° C. for 8 hours. The enzyme solution was cellulase (Meiji Seika, Meiserase TP6
0) 37 parts by mass of acetic acid / sodium acetate aqueous solution (pH
4.5) What was melt | dissolved in 11000 mass parts was used. After the reaction, 735 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (manufactured by Organo,
Amberlite IR-120B) was passed through and the solvent was distilled off. The reaction mixture was separated and purified to obtain 220 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

220 parts by mass of this glucose was added to 12100
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., 121 parts by mass of the preculture liquid of the phenyllactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe with the bottle sealed, and the mixture was allowed to stand at 30 ° C. for 3 days. The culture was performed. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and then purified to obtain 10 parts by mass of phenyllactic acid.

¹³ C-NMR (1
00MHz, TMS, CDCl ₃ ) measurement result (δ / p
pm) is 177.92, 137.63, 130.30,
129.46, 127.82, 72.05, 40.38
It was confirmed that the desired phenyllactic acid was obtained.

Using a reactor equipped with DeanStarktrap, 600 parts by mass of toluene was added to 10 parts by mass of phenyllactic acid and 0.2 parts by mass of p-toluenesulfonic acid.
Azeotropic dehydration is performed for 30 hours under a nitrogen atmosphere, and DeanSt
Water accumulated in the arktrap was appropriately removed. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 8.7 parts by mass of cyclic dimer lactide.

The ring-opening polymerization was carried out by heating 8.7 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of tin chloride at 160 ° C. for 10 hours in a nitrogen atmosphere. The weight average molecular weight of the obtained aromatic polyester was 190,000.

The aromatic polyester having a weight average molecular weight of 190,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 69 MPa (yield), 69 MPa (breakage), elongation: 12%.

Example 14 An aromatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 13. The ring-opening polymerization was carried out by heating 8.7 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of zinc chloride at 120 ° C. for 5 hours in a nitrogen atmosphere. The weight average molecular weight of the obtained aromatic polyester was 50,000.

The aromatic polyester having a weight average molecular weight of 50,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 40 to 41 μm tensile strength: 31 MPa (yield), 30 MPa (breaking), elongation: 3%.

(Example 15) Recycled paper for PPC (Canon Sales, EN-500, A4) used (copied on one side with a copying machine) was cut into 5 mm square, and 380 parts by mass of the enzyme solution 11300 was cut. Add to the mass part, 10 at 45 ℃
Stir for hours. The enzyme solution is a cellulase (made by Meiji Seika,
40 parts by mass of macerase TP60) dissolved in 11200 parts by mass of acetic acid / sodium acetate aqueous solution (pH 4.5) was used. After the reaction, 750 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo) to distill off the solvent. The reaction mixture was separated and purified to obtain 210 parts by mass of glucose.

210 parts by weight of this glucose was added to 11600
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and then butyl rubber stopper Seal it with an autoclave (121 ° C,
Pressurized at 98 kPa, 10 minutes). Then, add 3 media
The mixture was cooled to 0 ° C., 116 parts by mass of a preculture liquid of the phenyllactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe with the bottle sealed, and static culture was carried out at 30 ° C. for 3 days. Then, 9 parts by mass of phenyllactic acid was recovered in the same manner as in Example 13.

When ¹³ C-NMR was measured, Example 1
A spectrum similar to that of No. 3 was obtained, and it was confirmed that the target phenyllactic acid was obtained.

9 parts by mass of the obtained phenyllactic acid was used in Example 1
7.8 parts by mass of the cyclic dimer lactide was obtained by performing the same operation as described above.

Ring-opening polymerization was carried out by heating 7.8 parts by mass of the cyclic dimer lactide and 0.007 parts by mass of tin octylate under nitrogen atmosphere at 180 ° C. for 33 hours. The weight average molecular weight of the obtained aromatic polyester was 700,000.

The aromatic polyester having a weight average molecular weight of 700,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 25%.

Example 16 An aromatic polyester was polymerized using the cyclic dimer lactide obtained by the same procedure as in Example 15. 7.8 parts by mass of the cyclic dimer lactide and 0.006 parts by mass of tin powder at 160 ° C. under nitrogen atmosphere
Ring-opening polymerization was performed by heating for 8 hours. The weight average molecular weight of the obtained aliphatic polyester was 380,000.

The aromatic polyester having a weight average molecular weight of 380,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 87 MPa (yield), 86 MPa (breakage), elongation: 16%.

Example 17 Cellulose (manufactured by Nippon Paper Industries,
KC Flock W-100) 340 parts by mass of enzyme solution 10
It was put into 250 parts by mass and stirred at 45 ° C. for 8 hours. The enzyme solution was cellulase (Meiji Seika, Meiserase TP6
0) 34 parts by mass of acetic acid / sodium acetate aqueous solution (pH
4.5) What was melt | dissolved in 10000 mass parts was used. After the reaction, 670 parts by mass of methanol was added, the water-soluble residue was filtered off, and the ion-exchange resin column (manufactured by Organo,
Amberlite IR-120B) was passed through and the solvent was distilled off. The reaction mixture was separated and purified to obtain 200 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

200 parts by mass of this glucose was added to 11000
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and 110 parts by mass of a preculture liquid of the 2-hydroxyisocaproic acid / lactic acid producing strain HICA432 strain isolated by the above method was added using a syringe in a state in which the bottle was sealed, Static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, a mixture of 10 parts by mass of 2-hydroxyisocaproic acid and 12 parts by mass of lactic acid was obtained. I confirmed that.

¹ H-NMR (400 MHz, TM
S, DMSO-d ₆ ) measurement results (δ / ppm) are shown; 2-hydroxyisocaproic acid: 0.88 (t, 6)
H), 1.37 to 1.48 (m, 2H), 1.72 to
1.82 (m, 1H), 3.94 (dd, 1H), Lactic acid: 1.25 (d, J = 6.8Hz, 3H), 4.0
7 (q, J = 6.8 Hz, 1H).

Also, ¹³ C-NMR (100 MHz, TM
S, DMSO-d ₆ ) measurement results (δ / ppm) are shown; 2-hydroxyisocaproic acid: 21.43, 23.1
2, 23.84, 42.84, 68.07, 176.2
3, Lactic acid: 20.39, 65.82, 176.42.

Furthermore, an optically active column (J & WSCIEN
2-using TIFIC's Cyclodex-B)
As a result of evaluating the three-dimensional structure of hydroxyisocaproic acid,
It was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

On the other hand, as a result of evaluating the three-dimensional structure of lactic acid using an optically active column (Restek column 13113, manufactured by UNIFLEX), it was confirmed that the obtained lactic acid was DL-form.

Using a reactor equipped with a Dean Stark trap, 220 parts by mass of mesitylene was added to 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of lactic acid, and 0.044 parts by mass of tin dichloride under a nitrogen atmosphere. Azeotropic dehydration was performed to appropriately remove water accumulated in the DeanStark trap. The polymerization time is 40 hours,
The weight average molecular weight of the obtained aliphatic polyester copolymer was 170,000.

¹³ C-NMR (100 MHz, TMS, C
The measurement result (δ / ppm) of DCl ₃ ) is 16.66, 2
1.43, 22.99, 24.55, 39.30, 6
It was 8.97, 71.37, 169.51, 169.76, and it was confirmed that the target aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 170,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 61 MPa (yield), 59 MPa (breaking), elongation: 14%.

Example 18 An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained by performing the same operation as in Example 17. Dean
Using a reactor equipped with a Stark trap, 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of lactic acid,
Further, 200 parts by mass of 1,3-dichlorobenzene was added to 0.04 parts by mass of zinc powder, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 38 hours, and the weight average molecular weight of the obtained aliphatic polyester copolymer was 100,000.

When ¹³ C-NMR was measured, the same spectrum as in Example 17 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 100,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 49 MPa (yield), 49 MPa (breakage), elongation: 6%.

(Example 19) Recycled paper for PPC (Canon Sales, EN-500, A4) used (copied on one side with a copying machine) was cut into 5 mm square, and 500 parts by mass of the enzyme solution 15050 was cut. Add to the mass part, 10 at 45 ℃
Stir for hours. The enzyme solution is a cellulase (made by Meiji Seika,
50 parts by mass of macerase TP60) dissolved in 15,000 parts by mass of acetic acid / sodium acetate aqueous solution (pH 4.5) was used. After the reaction, 1000 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo) to distill off the solvent. The reaction mixture was separated and purified to obtain 280 parts by mass of glucose.

280 parts by mass of this glucose were added to 15400
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., and 154 parts by mass of a preculture liquid of the 2-hydroxyisocaproic acid / lactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe in a state in which the bottle was sealed, Static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, a mixture of 13 parts by mass of 2-hydroxyisocaproic acid and 16 parts by mass of lactic acid was obtained. I confirmed that.

Using a reactor equipped with Dean Stark trap, 13 parts by mass of 2-hydroxyisocaproic acid, 16 parts by mass of lactic acid, and 0.05 parts of tin octylate were used.
290 parts by mass of mesitylene was added to 8 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark trap.
The water accumulated inside was appropriately removed. The polymerization time was 36 hours, and the aliphatic polyester copolymer obtained had a weight average molecular weight of 60,000.

When ¹³ C-NMR was measured, the same spectrum as in Example 17 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 60,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 22 MPa (yield), 22 MPa (breakage), elongation: 3%.

(Example 20) An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained in the same manner as in Example 19. Dean
Using a reaction apparatus equipped with a Stark trap, 13 parts by mass of 2-hydroxyisocaproic acid, 16 parts by mass of lactic acid,
Further, 320 parts by mass of diphenyl ether was added to 0.052 parts by mass of tin powder, and azeotropic dehydration was carried out at 130 ° C./12 mmHg to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 48 hours, and the weight average molecular weight of the obtained aliphatic polyester copolymer was 2
It was 90,000.

When ¹³ C-NMR was measured, the same spectrum as in Example 17 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 290,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 74 MPa (yield), 73 MPa (breaking), elongation: 23%.

(Example 21) 200 parts by mass of glucose obtained by the same procedure as in Example 17 was added to 11,000 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l, NaCl 5 g / l). 1, cysteine hydrochloride (2 g / l, calcium carbonate 5 g / l) and mixed into a pressure bottle, and the gas phase in the bottle was replaced with a mixed gas of 98% nitrogen and 2% oxygen. After setting to slightly aerobic conditions, seal with a butyl rubber stopper and autoclave (121 ° C, 98k
Pa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and the preculture liquid 110 of the 2-hydroxyisocaproic acid / lactic acid producing strain HICA432 strain isolated by the above-mentioned method
Add parts by mass using a syringe with the bottle sealed,
Static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, a mixture of 4 parts by mass of 2-hydroxyisocaproic acid and 18 parts by mass of lactic acid was obtained. I confirmed that.

Using a reactor equipped with a Dean Stark trap, 210 parts by mass of bromobenzene was added to 4 parts by mass of 2-hydroxyisocaproic acid, 18 parts by mass of lactic acid, and 0.04 parts by mass of tin oxide, and the mixture was added under a nitrogen atmosphere. Azeotropic dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. The polymerization time is 45 hours,
The weight average molecular weight of the obtained aliphatic polyester copolymer was 220,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 17 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 220,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 37 to 38 μm, tensile strength: 69 MPa (yield), 67 MPa (breakage), elongation: 18%.

Example 22 280 parts by mass of glucose obtained by performing the same operation as in Example 19 was cultured in the same manner as in Example 21. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, 5 parts by mass of 2-hydroxyisocaproic acid,
It was confirmed that a mixture of and 16 parts by mass of lactic acid was obtained.

Using a reactor equipped with a Dean Stark trap, 5 parts by mass of 2-hydroxyisocaproic acid, 16 parts by mass of lactic acid, and 200 parts by mass of 1,2-dichlorobenzene in 0.038 parts by mass of p-toluenesulfonic acid. And azeotropic dehydration under a nitrogen atmosphere.
Water accumulated in the Stark trap was appropriately removed. The polymerization time was 32 hours, and the weight average molecular weight of the obtained aliphatic polyester copolymer was 40,000.

When ¹³ C-NMR was measured, the same spectrum as in Example 17 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 40,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 40 to 41 μm, tensile strength: 19 MPa (yield), 19 MPa (breakage), elongation: 2%.

(Example 23) Cellulose (manufactured by Nippon Paper Industries,
KC Flock W-100) 340 parts by mass of enzyme solution 10
It was put into 250 parts by mass and stirred at 45 ° C. for 8 hours. The enzyme solution was cellulase (Meiji Seika, Meiserase TP6
0) 34 parts by mass of acetic acid / sodium acetate aqueous solution (pH
4.5) What was melt | dissolved in 10000 mass parts was used. After the reaction, 670 parts by mass of methanol was added, the water-soluble residue was filtered off, and the ion-exchange resin column (manufactured by Organo,
Amberlite IR-120B) was passed through and the solvent was distilled off. The reaction mixture was separated and purified to obtain 200 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

200 parts by mass of this glucose was added to 11000
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., and 2-hydroxyisocaproic acid / phenyllactic acid producing strain HICA432 isolated by the above-mentioned method was used.
110 parts by mass of the pre-culture liquid of the strain was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, 10 parts by mass of 2-hydroxyisocaproic acid and 1 of phenyllactic acid 1 were obtained.
It was confirmed that 2 parts by mass of the mixture was obtained.

¹ H-NMR (400 MHz, TM
S, DMSO-d ₆ ) measurement results (δ / ppm) are shown; 2-hydroxyisocaproic acid: 0.88 (t, 6)
H), 1.37 to 1.48 (m, 2H), 1.72 to
1.82 (m, 1H), 3.94 (dd, 1H), Phenyl lactic acid: 2.82, 2.95, 4.16, 7.2
5.

Furthermore, an optically active column (J & WSCIEN
2-using TIFIC's Cyclodex-B)
As a result of evaluating the three-dimensional structure of hydroxyisocaproic acid,
It was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

On the other hand, as a result of evaluating the three-dimensional structure of phenyl lactic acid using an optically active column (Restek column 13113, manufactured by UNIFLEX), it was confirmed that the obtained phenyl lactic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of phenyllactic acid, and tin dichloride were added.
220 parts by mass of mesitylene was added to 044 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark tr.
The water accumulated in the ap was appropriately removed. The polymerization time was 40 hours, and the weight average molecular weight of the obtained polyester copolymer was 160,000.

The polyester copolymer having a weight average molecular weight of 160,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 37 to 38 μm, tensile strength: 60 MPa (yield), 60 MPa (breakage), elongation: 13%.

(Example 24) A polyester copolymer was synthesized using 2-hydroxyisocaproic acid and phenyllactic acid obtained by the same procedure as in Example 23. Dean
Using a reactor equipped with a Stark trap, 2-
10 parts by mass of hydroxyisocaproic acid, 1 phenyllactic acid
200 parts by mass of 1,3-dichlorobenzene was added to 2 parts by mass and 0.04 parts by mass of zinc powder, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Starktrap. The polymerization time was 38 hours, and the weight average molecular weight of the obtained polyester copolymer was 110,000.

The polyester copolymer having a weight average molecular weight of 110,000 thus obtained was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 50 MPa (yield), 49 MPa (breaking), elongation: 7%.

(Example 25) Recycled paper for PPC (Canon Sales, EN-500, A4) used (copied on one side with a copying machine) was cut into 5 mm square, and 500 parts by mass of the enzyme solution 15050 was cut. Add to the mass part, 10 at 45 ℃
Stir for hours. The enzyme solution is a cellulase (made by Meiji Seika,
50 parts by mass of macerase TP60) dissolved in 15,000 parts by mass of acetic acid / sodium acetate aqueous solution (pH 4.5) was used. After the reaction, 1000 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo) to distill off the solvent. The reaction mixture was separated and purified to obtain 280 parts by mass of glucose.

280 parts by mass of this glucose were added to 15400
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., and 2-hydroxyisocaproic acid / phenyllactic acid producing strain HICA432 isolated by the above-mentioned method was used.
154 parts by mass of the preculture liquid of the strain was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, 13 parts by mass of 2-hydroxyisocaproic acid and 1 of phenyllactic acid 1 were obtained.
It was confirmed that 6 parts by mass of the mixture was obtained.

Using a reactor equipped with DeanStarktrap, 290 parts by mass of mesitylene was added to 13 parts by mass of 2-hydroxyisocaproic acid, 16 parts by mass of phenyllactic acid, and 0.058 parts by mass of tin octylate, and the mixture was mixed under a nitrogen atmosphere. After boiling dehydration, DeanStarktr
The water accumulated in the ap was appropriately removed. The polymerization time was 36 hours, and the weight average molecular weight of the obtained polyester copolymer was 70,000.

The polyester copolymer having a weight average molecular weight of 70,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 40 to 41 μm, tensile strength: 22 MPa (yield), 21 MPa (breakage), elongation: 4%.

(Example 26) A polyester copolymer was synthesized using 2-hydroxyisocaproic acid and phenyllactic acid obtained by the same procedure as in Example 25. Dean
Using a reactor equipped with Starktrap, 13 parts by mass of 2-hydroxyisocaproic acid, 16 parts by mass of phenyl lactic acid, and 320 parts by mass of diphenyl ether were added to 0.052 parts by mass of tin powder, and azeotropic dehydration was performed at 130 ° C / 12 mmHg. Then, the water accumulated in the DeanStarktrap was appropriately removed. The polymerization time was 48 hours, and the weight average molecular weight of the obtained polyester copolymer was 290,000.

The polyester copolymer having a weight average molecular weight of 290,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 38 to 39 μm, tensile strength: 78 MPa (yield), 77 MPa (breakage), elongation: 23%.

Example 27 Cellulose (manufactured by Nippon Paper Industries,
KC Flock W-100) 340 parts by mass of enzyme solution 10
It was put into 250 parts by mass and stirred at 45 ° C. for 8 hours. The enzyme solution was cellulase (Meiji Seika, Meiserase TP6
0) 34 parts by mass of acetic acid / sodium acetate aqueous solution (pH
4.5) What was melt | dissolved in 10000 mass parts was used. After the reaction, 670 parts by mass of methanol was added, the water-soluble residue was filtered off, and the ion-exchange resin column (manufactured by Organo,
Amberlite IR-120B) was passed through and the solvent was distilled off. The reaction mixture was separated and purified to obtain 200 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

200 parts by mass of this glucose was added to 11000
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., 110 parts by mass of the preculture liquid of the lactic acid / phenyl lactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe in a state in which the bottle was sealed, and 30 ° C.
The cells were statically cultivated for 3 days. Then, the culture solution is filtered, and water and volatile components of the filtrate are distilled off at room temperature under reduced pressure.
As a result of R analysis, it was confirmed that a mixture of 12 parts by mass of lactic acid and 10 parts by mass of phenyllactic acid was obtained.

The analysis results of lactic acid are shown below: ¹ H-NMR (400 MHz, TMS, DMSO-d ₆ ).
δ / ppm: 1.25 (d, J = 6.8 Hz, 3H),
4.07 (q, J = 6.8 Hz, 1H), and the analysis results of phenyllactic acid are shown below: ¹ H-NMR (400 MHz, TMS, DMSO-d ₆ ).
δ / ppm: 2.82, 2.95, 4.16, 7.2
5. Further, an optically active column (manufactured by UNIFLEX, Rest
As a result of evaluating the three-dimensional structure of lactic acid using ek column 13113), it was confirmed that the obtained lactic acid was DL-form.

On the other hand, as a result of evaluating the three-dimensional structure of phenyl lactic acid using an optically active column (Restek column 13113 manufactured by UNIFLEX), it was confirmed that the obtained phenyl lactic acid was L-form.

Using a reaction device equipped with a Dean Stark trap, 220 parts by mass of mesitylene was added to 12 parts by mass of lactic acid, 10 parts by mass of phenyllactic acid, and 0.044 parts by mass of tin dichloride, and azeotropic dehydration was performed under a nitrogen atmosphere. Done,
Water accumulated in the Dean Stark trap was appropriately removed. The polymerization time was 40 hours, and the weight average molecular weight of the obtained polyester copolymer was 150,000.

The polyester copolymer having a weight average molecular weight of 150,000 thus obtained was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 40 to 41 μm, tensile strength: 63 MPa (yield), 61 MPa (breaking), elongation: 7%.

Example 28 A polyester copolymer was synthesized using lactic acid and phenyllactic acid obtained by the same procedure as in Example 27. Using a reactor equipped with a Dean Stark trap, 12 parts by mass of lactic acid, 10 parts by mass of phenyl lactic acid, and 0.04 parts by mass of zinc powder were added to 1,3-
200 parts by mass of dichlorobenzene was added, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 38 hours, and the weight average molecular weight of the obtained polyester copolymer was 100,000.

The polyester copolymer having a weight average molecular weight of 100,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 51 MPa (yield), 49 MPa (breaking), elongation: 6%.

(Example 29) Recycled paper for PPC (Canon Sales, EN-500, A4) used (copied on one side with a copying machine) was cut into 5 mm squares, and 500 parts by mass of the enzyme solution 15050 was cut. Add to the mass part, 10 at 45 ℃
Stir for hours. The enzyme solution is a cellulase (made by Meiji Seika,
50 parts by mass of macerase TP60) dissolved in 15,000 parts by mass of acetic acid / sodium acetate aqueous solution (pH 4.5) was used. After the reaction, 1000 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo) to distill off the solvent. The reaction mixture was separated and purified to obtain 280 parts by mass of glucose.

280 parts by mass of this glucose were added to 15400
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., and 154 parts by mass of the pre-culture liquid of the lactic acid / phenyl lactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe in a state in which the bottle was sealed, and 30 ° C.
The cells were statically cultivated for 3 days. Then, the culture solution is filtered, and water and volatile components of the filtrate are distilled off at room temperature under reduced pressure.
As a result of R analysis, it was confirmed that a mixture of 18 parts by mass of lactic acid and 4 parts by mass of phenyllactic acid was obtained.

Using a reactor equipped with a Dean Stark trap, 210 parts by mass of bromobenzene was added to 18 parts by mass of lactic acid, 4 parts by mass of phenyllactic acid, and 0.04 parts by mass of tin oxide, and azeotropic dehydration was performed under a nitrogen atmosphere. Then, the water accumulated in the Dean Starktrap was appropriately removed. The polymerization time was 45 hours, and the weight average molecular weight of the obtained polyester copolymer was 200,000.

The polyester copolymer having a weight average molecular weight of 200,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 67 MPa (yield), 66 MPa (breaking), elongation: 11%.

Example 30 A polyester copolymer was synthesized by using lactic acid and phenyl lactic acid obtained by performing the same operation as in Example 29. Using a reactor equipped with Dean Stark trap, 18 parts by mass of lactic acid, 4 parts by mass of phenyl lactic acid, and p-toluenesulfonic acid 0.0
200 parts by mass of 1,2-dichlorobenzene was added to 36 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean St.
Water accumulated in the ark trap was appropriately removed. The polymerization time was 30 hours, and the weight average molecular weight of the obtained polyester copolymer was 40,000.

The polyester copolymer having a weight average molecular weight of 40,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 20 MPa (yield), 19 MPa (break), elongation: 2%.

(Example 31) 370 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) was added to 3300 parts by mass of water and dissolved while heating. To this, 3700 parts by mass of 3 mol / l sulfuric acid was added and reacted at 80 ° C. for 5 hours with stirring. After completion of the reaction, anhydrous sodium carbonate was added to neutralize the aqueous solution, and then the solution was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo), and the solvent was distilled off. Then, the reaction mixture was separated and purified to obtain 220 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

220 parts by mass of this glucose was added to 12100
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and the preculture liquid 1 of the 2-hydroxyisocaproic acid producing strain HICA432 strain isolated by the above-mentioned method
21 parts by mass was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and the remaining solid content was heated under reduced pressure. The sublimated gas was cooled and the precipitated solid was collected to obtain 10 parts by mass of 2-hydroxyisocaproic acid.

Of the resulting 2-hydroxyisocaproic acid
¹³ C-NMR (100 MHz, TMS, CDCl ₃ ) measurement results (δ / ppm) were 180.25, 68.95,
It was 43.16, 24.45, 23.20 and 21.42, and it was confirmed that the target 2-hydroxyisocaproic acid was obtained.

Using a reactor equipped with a Dean Stark trap, 100 parts by mass of mesitylene was added to 10 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of tin dichloride, and azeotropic dehydration was carried out under a nitrogen atmosphere. Dean
Water accumulated in the Stark trap was appropriately removed. The polymerization time was 40 hours, and the aliphatic polyester obtained had a weight average molecular weight of 90,000.

¹³ C-NMR (100 MHz, TMS, C
DCl ₃ ) measurement result (δ / ppm) was 169.78,
71.38, 39.33, 24.53, 23.00, 2
It was 1.43, and it was confirmed that the desired aliphatic polyester was synthesized.

Further, an optically active column (J & WSCIEN
As a result of evaluating the three-dimensional structure using Cylodex-B manufactured by TIFIC, it was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

The aliphatic polyester having a weight average molecular weight of 90,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 38 to 39 μm, tensile strength: 48 MPa (yield), 48 MPa (breakage), elongation: 5%.

(Example 32) An aliphatic polyester was polymerized using 2-hydroxyisocaproic acid obtained by performing the same operation as in Example 31. Dean Stark t
Using a reactor equipped with a rap, 10 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of zinc chloride were added to 1, 3 and 2.
-100 parts by mass of dichlorobenzene was added, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 45 hours, and the weight average molecular weight of the obtained aliphatic polyester was 1
It was 40,000.

The aliphatic polyester having a weight average molecular weight of 140,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 57 MPa (yield), 56 MPa (breaking), elongation: 9%.

(Example 33) An aliphatic polyester was polymerized using 2-hydroxyisocaproic acid obtained by performing the same operation as in Example 31. Dean Stark t
Using a reactor equipped with a rap, 90 parts by mass of mesitylene was added to 9 parts by mass of 2-hydroxyisocaproic acid and 0.018 parts by mass of tin octylate, and azeotropic dehydration was carried out under a nitrogen atmosphere to obtain Dean Stark trap. The accumulated water was appropriately removed. The polymerization time was 40 hours, and the aliphatic polyester obtained had a weight average molecular weight of 50,000.

Also, ¹³ C-NMR was measured, and the same spectrum as in Example 31 was obtained, confirming that the desired aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 50,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 20 MPa (yield), 20 MPa (breakage), elongation: 3%.

(Example 34) An aliphatic polyester was polymerized using 2-hydroxyisocaproic acid obtained by performing the same operation as in Example 31. Dean Stark t
Using a reactor equipped with rap, 100 parts by mass of diphenyl ether was added to 9 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of tin powder, and 130 ° C / 12 mmH.
azeotropic dehydration with Dean Stark trap
The water accumulated inside was appropriately removed. The polymerization time was 48 hours, and the aliphatic polyester obtained had a weight average molecular weight of 190,000.

The aliphatic polyester having a weight average molecular weight of 190,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 69 MPa (yield), 67 MPa (breaking), elongation: 13%.

(Example 35) 370 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) was added to 3300 parts by mass of water and dissolved while heating. To this, 3700 parts by mass of 3 mol / l sulfuric acid was added and reacted at 80 ° C. for 5 hours with stirring. After completion of the reaction, anhydrous sodium carbonate was added to neutralize the aqueous solution, and then the solution was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo), and the solvent was distilled off. Then, the reaction mixture was separated and purified to obtain 220 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

220 parts by weight of this glucose was added to 12100
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and the preculture liquid 1 of the 2-hydroxyisocaproic acid producing strain HICA432 strain isolated by the above-mentioned method
21 parts by mass was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and the remaining solid content was heated under reduced pressure. The sublimated gas was cooled and the precipitated solid was collected to obtain 10 parts by mass of 2-hydroxyisocaproic acid.

Of the resulting 2-hydroxyisocaproic acid
¹³ C-NMR (100 MHz, TMS, CDCl ₃ ) measurement results (δ / ppm) were 180.25, 68.95,
It was 43.16, 24.45, 23.20 and 21.42, and it was confirmed that the target 2-hydroxyisocaproic acid was obtained.

Using a reactor equipped with a Dean Stark trap, 600 parts by mass of toluene was added to 10 parts by mass of 2-hydroxyisocaproic acid and 0.2 parts by mass of p-toluenesulfonic acid, and the mixture was azeotropically distilled for 30 hours under a nitrogen atmosphere. Dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 8.2 parts by mass of cyclic dimer lactide.

¹³ C-NMR of a cyclic dimer lactide (10
Measurement result of 0 MHz, TMS, CDCl ₃ (δ / pp
m) is 167.33, 74.14, 38.87, 23.
89, 23.03, 21.33, and it was confirmed that the target cyclic dimer lactide was obtained.

The ring-opening polymerization was carried out by heating 8.2 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of tin chloride in a nitrogen atmosphere at 150 ° C. for 12 hours. The weight average molecular weight of the obtained aliphatic polyester was 180,000.

¹³ C-NMR (10
Measurement result of 0MHz, TMS, CDCl ₃ (δ / p
p) is 169.78, 71.38, 39.33, 24.
53, 23.00, 21.43, and it was confirmed that the desired aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 180,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 68 MPa (yield), 68 MPa (breakage), elongation: 13%.

Example 36 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 35. The ring-opening polymerization was carried out by heating 8.2 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of zinc chloride in a nitrogen atmosphere at 120 ° C. for 5 hours. The weight average molecular weight of the obtained aliphatic polyester was 50,000. ^{When 13} C-NMR was measured, the same spectrum as in Example 35 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 50,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 29 MPa (yield), 29 MPa (breaking), elongation: 4%.

Example 37 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 35. A cyclic dimer lactide (7.4 parts by mass) and tin octylate (0.008 parts by mass) were added under nitrogen atmosphere to 180
The ring-opening polymerization was performed by heating at 30 ° C. for 30 hours. The weight average molecular weight of the obtained aliphatic polyester was 680,000. ¹³
When C-NMR was measured, the same spectrum as in Example 35 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 680,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 30%.

Example 38 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 35. The cyclic dimer lactide (7.4 parts by mass) and the tin powder (0.007 parts by mass) were mixed at 160 ° C. in a nitrogen atmosphere at 1
Ring-opening polymerization was performed by heating for 8 hours. The weight average molecular weight of the obtained aliphatic polyester was 390,000. ¹³ C-N
When the MR was measured, the same spectrum as in Example 35 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 390,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm; tensile strength: 88 MPa (yield), 87 MPa (breaking), elongation: 22%.

(Example 39) 370 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) was added to 3300 parts by mass of water and dissolved while heating. To this, 3700 parts by mass of 3 mol / l sulfuric acid was added and reacted at 80 ° C. for 5 hours with stirring. After completion of the reaction, anhydrous sodium carbonate was added to neutralize the aqueous solution, and then the solution was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo), and the solvent was distilled off. Then, the reaction mixture was separated and purified to obtain 220 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

220 parts by mass of this glucose was added to 12100
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., 121 parts by mass of the preculture liquid of the phenyllactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe with the bottle sealed, and the mixture was allowed to stand at 30 ° C. for 3 days. The culture was performed. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and then purified to obtain 10 parts by mass of phenyllactic acid.

¹³ C-NMR (1
00MHz, TMS, CDCl ₃ ) measurement result (δ / p
pm) is 177.92, 137.63, 130.30,
129.46, 127.82, 72.05, 40.38
It was confirmed that the desired phenyllactic acid was obtained.

Using a reaction apparatus equipped with a Dean Stark trap, 600 parts by mass of toluene was added to 10 parts by mass of phenyllactic acid and 0.2 parts by mass of p-toluenesulfonic acid, and azeotropic dehydration was carried out for 30 hours in a nitrogen atmosphere. Dean
Water accumulated in the Stark trap was appropriately removed. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 8.7 parts by mass of cyclic dimer lactide.

Ring-opening polymerization was carried out by heating 8.7 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of tin chloride at 160 ° C. for 10 hours in a nitrogen atmosphere. The weight average molecular weight of the obtained aromatic polyester was 190,000.

The aromatic polyester having a weight average molecular weight of 190,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 69 MPa (yield), 69 MPa (breakage), elongation: 12%.

Example 40 An aromatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 39. The ring-opening polymerization was carried out by heating 8.7 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of zinc chloride at 120 ° C. for 5 hours in a nitrogen atmosphere. The mass average molecular weight of the obtained aromatic polyester was 50,000.

The aromatic polyester having a weight average molecular weight of 50,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 40 to 41 μm, tensile strength: 31 MPa (yield), 30 MPa (breakage), elongation: 3%.

(Example 41) An aromatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 39. 7.8 parts by mass of the cyclic dimer lactide and 0.007 parts by mass of tin octylate were added under nitrogen atmosphere for 180
The ring-opening polymerization was carried out by heating at 0 ° C for 33 hours. The weight average molecular weight of the obtained aromatic polyester was 700,000.

The aromatic polyester having a weight average molecular weight of 700,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 25%.

(Example 42) An aromatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 39. 7.8 parts by mass of the cyclic dimer lactide and 0.006 parts by mass of tin powder at 160 ° C. under nitrogen atmosphere
Ring-opening polymerization was performed by heating for 8 hours. The weight average molecular weight of the obtained aromatic polyester was 380,000.

The aromatic polyester having a weight average molecular weight of 380,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 87 MPa (yield), 86 MPa (breakage), elongation: 16%.

(Example 43) 340 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) was added to 3000 parts by mass of water and dissolved while heating. To this, 3400 parts by mass of 3 mol / l sulfuric acid was added and reacted at 80 ° C. for 5 hours with stirring. After completion of the reaction, anhydrous sodium carbonate was added to neutralize the aqueous solution, and then the solution was passed through an ion exchange resin column (Amberlite IR-120B, manufactured by Organo), and the solvent was distilled off. Then, the reaction mixture was separated and purified to obtain 200 parts by mass of glucose.

FT-NMR DPX400 manufactured by Bruker
¹³ C-NMR (100
MHz, TMS, DMSO-d ₆ ) were measured. The chemical shift δ (ppm) was as follows: α type: 92.12, 73.04, 72.29, 71.8.
0, 70.58, 61.20, β-type: 96.79, 76.70, 76.59, 74.7.
8, 70.30, 61.00.

From the above measurement results, it was confirmed that the desired glucose was obtained.

200 parts by mass of this glucose was added to 11000
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and 110 parts by mass of a preculture liquid of the 2-hydroxyisocaproic acid / lactic acid producing strain HICA432 strain isolated by the above method was added using a syringe in a state in which the bottle was sealed, Static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, a mixture of 10 parts by mass of 2-hydroxyisocaproic acid and 12 parts by mass of lactic acid was obtained. I confirmed that.

¹ H-NMR (400 MHz, TMS, D
The measurement result (δ / ppm) of MSO-d ₆ ) is shown below; 2-hydroxyisocaproic acid: 0.88 (t, 6)
H), 1.37 to 1.48 (m, 2H), 1.72 to
1.82 (m, 1H), 3.94 (dd, 1H), Lactic acid: 1.25 (d, J = 6.8Hz, 3H), 4.0
7 (q, J = 6.8 Hz, 1H).

In addition, ¹³ C-NMR (100 MHz, TM
The measurement results (δ / ppm) of S, DMSO-d ₆ ) are shown below; 2-hydroxyisocaproic acid: 21.43, 23.1
2, 23.84, 42.84, 68.07, 176.2
3, Lactic acid: 20.39, 65.82, 176.42.

Furthermore, an optically active column (J & WSCIEN
2-using TIFIC's Cyclodex-B)
As a result of evaluating the three-dimensional structure of hydroxyisocaproic acid,
It was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

On the other hand, as a result of evaluating the three-dimensional structure of lactic acid using an optically active column (Restek column 13113, manufactured by UNIFLEX), it was confirmed that the obtained lactic acid was DL-form.

Using a reactor equipped with a Dean Stark trap, 220 parts by mass of mesitylene was added to 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of lactic acid, and 0.044 parts by mass of tin dichloride under a nitrogen atmosphere. Azeotropic dehydration was performed to appropriately remove water accumulated in the DeanStark trap. The polymerization time is 40 hours,
The weight average molecular weight of the obtained aliphatic polyester copolymer was 170,000.

In addition, ¹³ C-NMR (100 MHz, TM
S, CDCl ₃ ) measurement result (δ / ppm) is 16.6
6, 21.43, 22.99, 24.55, 39.3
0, 68.97, 71.37, 169.51, 169.
It was 76, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 170,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 61 MPa (yield), 59 MPa (breaking), elongation: 14%.

Example 44 An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained in the same manner as in Example 43. Dean
Using a reactor equipped with a Stark trap, 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of lactic acid,
Further, 200 parts by mass of 1,3-dichlorobenzene was added to 0.04 parts by mass of zinc powder, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 38 hours, and the weight average molecular weight of the obtained aliphatic polyester copolymer was 100,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 43 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 100,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 49 MPa (yield), 49 MPa (breakage), elongation: 6%.

(Example 45) 200 parts by mass of glucose obtained by the same procedure as in Example 43 was used as 11,000 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l, NaCl 5 g / l). 1, cysteine hydrochloride (2 g / l, calcium carbonate 5 g / l) and injected into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the container was sealed with a butyl rubber stopper, and then autoclaved (121 ° C, 98
kPa pressurization, 10 minutes). Then, culture medium at 30 ℃
Preculture liquid 11 of 2-hydroxyisocaproic acid / lactic acid producing strain HICA432 strain cooled by
0 part by mass was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, a mixture of 9 parts by mass of 2-hydroxyisocaproic acid and 11 parts by mass of lactic acid was obtained. I confirmed that.

Using a reactor equipped with a Dean Stark trap, 210 parts by mass of mesitylene was added to 9 parts by mass of 2-hydroxyisocaproic acid, 11 parts by mass of lactic acid, and 0.04 parts by mass of tin octylate under a nitrogen atmosphere. Azeotropic dehydration was performed to appropriately remove water accumulated in the DeanStark trap. The polymerization time is 36 hours,
The weight average molecular weight of the obtained aliphatic polyester copolymer was 60,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 43 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 60,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 22 MPa (yield), 22 MPa (breakage), elongation: 3%.

(Example 46) An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained in the same manner as in Example 45. Dean
Using a reactor equipped with a Stark trap, 9 parts by mass of 2-hydroxyisocaproic acid, 11 parts by mass of lactic acid, and 0.036 parts by mass of tin powder were added to diphenyl ether 2.
60 parts by mass was added, and azeotropic dehydration was performed at 130 ° C./12 mmHg to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 48 hours, and the aliphatic polyester copolymer obtained had a weight average molecular weight of 29.
It was good.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 43 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 290,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 74 MPa (yield), 73 MPa (breaking), elongation: 23%.

(Example 47) 200 parts by mass of glucose obtained by the same procedure as in Example 43 was used in 11,000 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l, NaCl 5 g / l). 1, cysteine hydrochloride (2 g / l, calcium carbonate 5 g / l) and mixed into a pressure bottle, and the gas phase in the bottle was replaced with a mixed gas of 98% nitrogen and 2% oxygen. After setting to slightly aerobic conditions, seal with a butyl rubber stopper and autoclave (121 ° C, 98k
Pa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and the preculture liquid 110 of the 2-hydroxyisocaproic acid / lactic acid producing strain HICA432 strain isolated by the above-mentioned method 110
Add parts by mass using a syringe with the bottle sealed,
Static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, a mixture of 4 parts by mass of 2-hydroxyisocaproic acid and 18 parts by mass of lactic acid was obtained. I confirmed that.

Using a reactor equipped with a Dean Stark trap, 210 parts by mass of bromobenzene was added to 4 parts by mass of 2-hydroxyisocaproic acid, 18 parts by mass of lactic acid, and 0.04 parts by mass of tin oxide, and the mixture was added under a nitrogen atmosphere. Azeotropic dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. The polymerization time is 45 hours,
The weight average molecular weight of the obtained aliphatic polyester copolymer was 220,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 43 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 220,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 37 to 38 μm, tensile strength: 69 MPa (yield), 67 MPa (breakage), elongation: 18%.

Example 48 An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained by the same procedure as in Example 47. Dean
Using a reactor equipped with a Stark trap, 1 part or 2 parts were added to 4 parts by mass of 2-hydroxyisocaproic acid, 18 parts by mass of lactic acid, and 0.036 parts by mass of p-toluenesulfonic acid.
-200 parts by mass of dichlorobenzene was added, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 30 hours, and the aliphatic polyester copolymer obtained had a weight average molecular weight of 40,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 43 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 40,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 40 to 41 μm, tensile strength: 20 MPa (yield), 20 MPa (breakage), elongation: 2%.

The following Examples 49-74 are methods for producing polyesters which include the step of obtaining a substituted α-hydroxy acid by microbial fermentation of starch described in the second embodiment. Microorganisms, Example 2-hydroxyisocaproic acid-producing strains HICA432 strain used in 1-48, further Clostridium Beiji prn obtained by considering the ability to produce a substituted α- hydroxy acid from starch (Clo
stridembeijerinckii ) HICA
An isolated strain of 432 strain (FERM BP-8117) was used.

It was confirmed that the HICA432 strain produces not only 2-hydroxyisocaproic acid but also lactic acid and phenyllactic acid. Therefore, in the following, H
The ICA432 strain may also be referred to as a 2-hydroxyisocaproic acid producing strain, a lactic acid producing strain, and a phenyllactic acid producing strain.

(Example 49) 250 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) and 12100 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l,
NaCI 5g / l, cysteine hydrochloride 2g / l, calcium carbonate 5g / l) were mixed and injected into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper, and the autoclave (121 ℃, pressurize 98kPa, 10
Min) processed. Then, the medium was cooled to 30 ° C., and the 2-hydroxyisocaproic acid producing strain H isolated by the above method was used.
121 parts by mass of the preculture liquid of the ICA432 strain was added using a syringe with the bottle sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and the remaining solid content was heated under reduced pressure. The sublimated gas was cooled and the precipitated solid was collected to obtain 10 parts by mass of 2-hydroxyisocaproic acid.

Of the resulting 2-hydroxyisocaproic acid
¹³ C-NMR (100 MHz, TMS, CDCl ₃ ) measurement results (δ / ppm) are 180.25 and 68.9.
5, 43.16, 24.45, 23.20, 21.42
It was confirmed that the desired 2-hydroxyisocaproic acid was obtained.

Further, an optically active column (J & WSCIEN
As a result of evaluating the three-dimensional structure using Cylodex-B manufactured by TIFIC, it was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

Next, using a reactor equipped with a Dean Stark trap, 10 parts by mass of 2-hydroxyisocaproic acid and 0.02 part by mass of tin dichloride were added to 1 part of mesitylene.
00 parts by mass was added, and azeotropic dehydration was performed under a nitrogen atmosphere.
Water accumulated in the ean Stark trap was appropriately removed. The polymerization time was 40 hours, and the aliphatic polyester obtained had a weight average molecular weight of 90,000.

¹³ C-NMR (100 MHz, CDC
l ₃ ) measurement result (δ / ppm) is 169.78, 7
1.38, 39.33, 24.53, 23.00, 2
It was 1.43, and it was confirmed that the desired aliphatic polyester was synthesized. The aliphatic polyester having a weight average molecular weight of 90,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 38 to 39 μm, tensile strength: 48 MPa (yield), 48 MPa (breakage), elongation: 5%.

Example 50 An aliphatic polyester was polymerized using 2-hydroxyisocaproic acid obtained by performing the same operation as in Example 49. Dean Stark t
Using a reactor equipped with a rap, 10 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of zinc chloride were added to 1, 3
-100 parts by mass of dichlorobenzene was added, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 45 hours, and the weight average molecular weight of the obtained aliphatic polyester was 1
It was 40,000.

The aliphatic polyester having a weight average molecular weight of 140,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 57 MPa (yield), 56 MPa (breaking), elongation: 9%.

(Example 51) An aliphatic polyester was polymerized using 2-hydroxyisocaproic acid obtained by performing the same operation as in Example 49. Dean Stark t
Using a reactor equipped with a rap, 90 parts by mass of mesitylene was added to 9 parts by mass of 2-hydroxyisocaproic acid and 0.018 parts by mass of tin octylate, and azeotropic dehydration was carried out under a nitrogen atmosphere to obtain Dean Stark trap. The accumulated water was appropriately removed. The polymerization time was 40 hours, and the aliphatic polyester obtained had a weight average molecular weight of 50,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 49 was obtained, and it was confirmed that the desired aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 50,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 20 MPa (yield), 20 MPa (breakage), elongation: 3%.

(Example 52) An aliphatic polyester was polymerized using 2-hydroxyisocaproic acid obtained in the same manner as in Example 49. Dean Stark t
Using a reactor equipped with rap, 100 parts by mass of diphenyl ether was added to 9 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of tin powder, and 130 ° C / 12 mmH.
azeotropic dehydration with Dean Stark trap
The water accumulated inside was appropriately removed. The polymerization time was 48 hours, and the aliphatic polyester obtained had a weight average molecular weight of 190,000.

The aliphatic polyester having a weight average molecular weight of 190,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 69 MPa (yield), 67 MPa (breaking), elongation: 13%.

Example 53 2-Hydroxyisocaproic acid obtained in the same manner as in Example 49 was used to convert it into a cyclic dimer lactide by the following method.

Using a reactor equipped with a Dean Stark trap, 540 parts by mass of toluene was added to 9 parts by mass of 2-hydroxyisocaproic acid and 0.18 parts by mass of p-toluenesulfonic acid, and azeotropically distilled for 30 hours under a nitrogen atmosphere. Dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 7.4 parts by mass of cyclic dimer lactide. ¹³ C-NMR of the obtained cyclic dimer lactide (100 MHz,
TMS, CDCl ₃ ) measurement result (δ / ppm) is 1
67.33, 74.14, 38.87, 23.89, 2
It was 3.03 and 21.33, and it was confirmed that the target cyclic dimer lactide was obtained.

The ring-opening polymerization was carried out by heating 7.4 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of tin chloride at 150 ° C. for 12 hours in a nitrogen atmosphere. The weight average molecular weight of the obtained aliphatic polyester was 180,000.

¹³ C-NMR of this aliphatic polyester
(100 MHz, TMS, CDCl ₃ ) measurement result (δ
/ Ppm) is 169.78, 71.38, 39.33,
24.53, 23.00, 21.43, and it was confirmed that the target aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 180,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 68 MPa (yield), 68 MPa (breakage), elongation: 13%.

Example 54 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 53. The cyclic dimer lactide (7.4 parts by mass) and the tin powder (0.007 parts by mass) were mixed at 160 ° C. in a nitrogen atmosphere at 1
Ring-opening polymerization was performed by heating for 8 hours. The weight average molecular weight of the obtained aliphatic polyester was 390,000. ¹³ C-N
When the MR was measured, the same spectrum as in Example 53 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 390,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 88 MPa (yield), 87 MPa (breaking), elongation: 22%.

Example 55 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 53. A cyclic dimer lactide (7.4 parts by mass) and tin octylate (0.008 parts by mass) were added under nitrogen atmosphere to 180
The ring-opening polymerization was performed by heating at 30 ° C. for 30 hours. The weight average molecular weight of the obtained aliphatic polyester was 680,000. ¹³
When C-NMR was measured, the same spectrum as in Example 53 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 680,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 30%.

(Example 56) 250 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) and 12100 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l,
NaCI 5g / l, cysteine hydrochloride 2g / l, calcium carbonate 5g / l) were mixed and injected into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper, and the autoclave (121 ℃, pressurize 98kPa, 10
Min) processed. Then, the medium was cooled to 30 ° C., 121 parts by mass of the preculture liquid of the phenyllactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe with the bottle sealed, and the mixture was allowed to stand at 30 ° C. for 3 days. The culture was performed. afterwards,
The culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and then purified to obtain 10 parts by mass of phenyllactic acid.

¹³ C-NMR (1
00MHz, TMS, CDCl ₃ ) measurement result (δ / p
pm) is 177.92, 137.63, 130.3
0, 129.46, 127.82, 72.05, 40.
It was 38 and it was confirmed that the desired phenyllactic acid was obtained.

Also, an optically active column (J & WSCIEN
As a result of evaluating the three-dimensional structure using TIFIC's Cyclodex-B), it was confirmed that the obtained phenyllactic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 100 parts by mass of mesitylene was added to 10 parts by mass of phenyllactic acid and 0.018 parts by mass of tin dichloride, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark.
Water accumulated in the trap was appropriately removed. Polymerization time is 4
It was 4 hours, and the weight average molecular weight of the obtained aromatic polyester was 180,000.

The aromatic polyester having a weight average molecular weight of 180,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 38 to 39 μm, tensile strength: 69 MPa (yield), 69 MPa (breakage), elongation: 12%.

Example 57 An aromatic polyester was polymerized using phenyllactic acid obtained by the same procedure as in Example 56. Using a reactor equipped with Dean Stark trap, 10 parts by mass of phenyl lactic acid and 0.
100 parts by mass of 1,3-dichlorobenzene was added to 02 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean St.
Water accumulated in the ark trap was appropriately removed. The polymerization time was 16 hours, and the aromatic polyester obtained had a weight average molecular weight of 30,000.

The aromatic polyester having a weight average molecular weight of 30,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 20 MPa (yield), 19 MPa (break), elongation: 2%.

Example 58 An aromatic polyester was polymerized using phenyllactic acid obtained by the same procedure as in Example 56. Using a reactor equipped with Dean Stark trap, 9 parts by mass of phenyl lactic acid and 0.0
90 parts by mass of mesitylene was added to 15 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain a Dean Stark trap.
The water accumulated inside was appropriately removed. The polymerization time was 40 hours, and the aromatic polyester obtained had a weight average molecular weight of 90,000.

The aromatic polyester having a weight average molecular weight of 90,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 49 MPa (yield), 49 MPa (breaking), elongation: 5%.

(Example 59) Using phenyllactic acid obtained by the same procedure as in Example 56, an aromatic polyester was polymerized. Using a reactor equipped with Dean Stark trap, 9 parts by mass of phenyl lactic acid and 0.01 parts of tin powder were used.
Add 100 parts by mass of diphenyl ether to 5 parts by mass, and
Azeotropic dehydration is performed at 30 ° C / 12 mmHg, and Dean S
Water accumulated in the tarktrap was appropriately removed. The polymerization time was 48 hours, and the weight average molecular weight of the obtained aromatic polyester was 250,000.

The aromatic polyester having a weight average molecular weight of 250,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 82 MPa (yield), 81 MPa (breaking), elongation: 18%.

(Example 60) Using phenyllactic acid obtained by performing the same operation as in Example 56, it was converted into a cyclic dimer lactide by the following method.

Using a reactor equipped with a Dean Stark trap, 540 parts by mass of toluene was added to 9 parts by mass of phenyllactic acid and 0.18 parts by mass of p-toluenesulfonic acid, and azeotropic dehydration was performed for 30 hours in a nitrogen atmosphere. Dean
Water accumulated in the Stark trap was appropriately removed. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 7.8 parts by mass of cyclic dimer lactide.

Ring-opening polymerization was carried out by heating 7.8 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of tin chloride in a nitrogen atmosphere at 160 ° C. for 10 hours. The weight average molecular weight of the obtained aromatic polyester was 190,000.

The aromatic polyester having a weight average molecular weight of 190,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 69 MPa (yield), 69 MPa (breakage), elongation: 12%.

Example 61 An aromatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 60. 7.8 parts by mass of the cyclic dimer lactide and 0.006 parts by mass of tin powder at 160 ° C. under nitrogen atmosphere
Ring-opening polymerization was performed by heating for 8 hours. The weight average molecular weight of the obtained aromatic polyester was 380,000.

The aromatic polyester having a weight average molecular weight of 380,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 87 MPa (yield), 86 MPa (breakage), elongation: 16%.

Example 62 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 60. 7.8 parts by mass of the cyclic dimer lactide and 0.007 parts by mass of tin octylate were added under nitrogen atmosphere for 180
The ring-opening polymerization was carried out by heating at 0 ° C for 33 hours. The weight average molecular weight of the obtained aromatic polyester was 700,000.

The aromatic polyester having a weight average molecular weight of 700,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 25%.

(Example 63) 400 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) and 11000 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l,
NaCI 5g / l, cysteine hydrochloride 2g / l, calcium carbonate 5g / l) were mixed and injected into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper, and the autoclave (121 ℃, pressurize 98kPa, 10
Min) processed. Thereafter, the medium was cooled to 30 ° C., and 110 parts by mass of a preculture liquid of 2-hydroxyisocaproic acid and a lactic acid producing strain HICA432 strain isolated by the above-mentioned method were added using a syringe in a state in which the bottle was sealed, 4 at 30 ° C
The culture was allowed to stand for a day. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, a mixture of 10 parts by mass of 2-hydroxyisocaproic acid and 12 parts by mass of lactic acid was obtained. I confirmed that.

Incidentally, ¹ H-NMR (400 MHz, TM
The measurement results (δ / ppm) of S, DMSO-d ₆ ) are as follows; 2-hydroxyisocaproic acid: 0.88 (t, 6)
H), 1.37 to 1.48 (m, 2H), 1.72 to
1.82 (m, 1H), 3.94 (dd, 1H), Lactic acid: 1.25 (d, J = 6.8Hz, 3H), 4.0
7 (q, J = 6.8 Hz, 1H).

Also, ¹³ C-NMR (100 MHz, TM
S, DMSO-d ₆ ) measurement results (δ / ppm) are as follows: 2-hydroxyisocaproic acid: 21.43, 23.1
2, 23.84, 42.84, 68.07, 176.2
3, Lactic acid: 20.39, 65.82, 176.42.

Furthermore, an optically active column (J & WSCIEN
2-using TIFIC's Cyclodex-B)
As a result of evaluating the three-dimensional structure of hydroxyisocaproic acid,
It was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

On the other hand, as a result of evaluating the three-dimensional structure of lactic acid using an optically active column (Restek column 13113, manufactured by UNIFLEX), it was confirmed that the obtained lactic acid was DL-form.

Using a reactor equipped with a Dean Stark trap, 220 parts by mass of mesitylene was added to 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of lactic acid, and 0.044 parts by mass of tin dichloride under a nitrogen atmosphere. Azeotropic dehydration was performed to appropriately remove water accumulated in the DeanStark trap. The polymerization time is 40 hours,
The weight average molecular weight of the obtained aliphatic polyester copolymer was 170,000.

¹³ C-NMR (100 MHz, TMS, C
The measurement result (δ / ppm) of DCl ₃ ) is 16.66,
21.43, 22.99, 24.55, 39.30, 6
It was 8.97, 71.37, 169.51, 169.76, and it was confirmed that the target aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 170,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 61 MPa (yield), 59 MPa (breaking), elongation: 14%.

Example 64 An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained by performing the same operation as in Example 63. Dean
Using a reactor equipped with a Stark trap, 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of lactic acid,
Further, 200 parts by mass of 1,3-dichlorobenzene was added to 0.04 parts by mass of zinc powder, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 38 hours, and the weight average molecular weight of the obtained aliphatic polyester copolymer was 100,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 63 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 100,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 49 MPa (yield), 49 MPa (breakage), elongation: 6%.

Example 65 An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained by performing the same operation as in Example 63. Dean
Using a reactor equipped with a Stark trap, 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of lactic acid,
And 0.04 parts by weight of tin octylate to mesitylene 21
Add 0 parts by mass and perform azeotropic dehydration in a nitrogen atmosphere to remove De
Water accumulated in the an Stark trap was appropriately removed. The polymerization time was 36 hours, and the aliphatic polyester copolymer obtained had a weight average molecular weight of 60,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 63 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 60,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 22 MPa (yield), 22 MPa (breakage), elongation: 3%.

Example 66 An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained by performing the same operation as in Example 63. Dean
Using a reactor equipped with a Stark trap, 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of lactic acid,
Further, 260 parts by mass of diphenyl ether was added to 0.036 parts by mass of tin powder, and azeotropic dehydration was carried out at 130 ° C./12 mmHg to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 48 hours, and the weight average molecular weight of the obtained aliphatic polyester copolymer was 2
It was 90,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 63 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 290,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 74 MPa (yield), 73 MPa (breaking), elongation: 23%.

Example 67 400 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) and 11000 parts by mass of medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l,
NaCI 5g / l, cysteine hydrochloride 2g / l, calcium carbonate 5g / l) were mixed and injected into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper, and the autoclave (121 ℃, pressurize 98kPa, 10
Min) processed. Thereafter, the medium was cooled to 30 ° C., and 110 parts by mass of a preculture liquid of 2-hydroxyisocaproic acid and a lactic acid producing strain HICA432 strain isolated by the above-mentioned method were added using a syringe in a state in which the bottle was sealed, 3 at 30 ° C
The culture was allowed to stand for a day. Then, the culture solution was filtered, and the water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and as a result of NMR analysis, 4 parts by mass of 2-hydroxyisocaproic acid,
It was confirmed that a mixture of and 18 parts by mass of lactic acid was obtained.

Using a reactor equipped with a Dean Stark trap, 210 parts by mass of bromobenzene was added to 4 parts by mass of 2-hydroxyisocaproic acid, 18 parts by mass of lactic acid, and 0.04 parts by mass of tin oxide, and under a nitrogen atmosphere. Azeotropic dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. The polymerization time is 45 hours,
The weight average molecular weight of the obtained aliphatic polyester copolymer was 220,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 63 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 220,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 37 to 38 μm, tensile strength: 69 MPa (yield), 67 MPa (breakage), elongation: 18%.

(Example 68) An aliphatic polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and lactic acid obtained in the same manner as in Example 67. Dean
Using a reactor equipped with a Stark trap, 1 part or 2 parts were added to 4 parts by mass of 2-hydroxyisocaproic acid, 18 parts by mass of lactic acid, and 0.036 parts by mass of p-toluenesulfonic acid.
-200 parts by mass of dichlorobenzene was added, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 30 hours, and the aliphatic polyester copolymer obtained had a weight average molecular weight of 40,000.

Further, ¹³ C-NMR was measured to find that the same spectrum as in Example 63 was obtained, and it was confirmed that the desired aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 40,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 40 to 41 μm, tensile strength: 20 MPa (yield), 20 MPa (breakage), elongation: 2%.

Example 69 400 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) and 11000 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l,
NaCI 5g / l, cysteine hydrochloride 2g / l, calcium carbonate 5g / l) were mixed and injected into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper, and the autoclave (121 ℃, pressurize 98kPa, 10
Min) processed. Then, the medium was cooled to 30 ° C., and 110 parts by mass of a preculture liquid of the 2-hydroxyisocaproic acid- and phenyllactic acid-producing strain HICA432 strain isolated by the above method was added using a syringe in a state where the bottle was sealed. Three
Static culture was carried out at 0 ° C. for 6 days. Then, the culture solution is filtered,
After distilling off the water and volatile components of the filtrate at room temperature under reduced pressure,
As a result of NMR analysis, 2-hydroxyisocaproic acid 1
It was confirmed that a mixture of 0 parts by mass and 12 parts by mass of phenyllactic acid was obtained.

¹ H-NMR (400 MHz, TM
The measurement results (δ / ppm) of S, DMSO-d ₆ ) are as follows; 2-hydroxyisocaproic acid: 0.88 (t, 6)
H), 1.37 to 1.48 (m, 2H), 1.72 to
1.82 (m, 1H), 3.94 (dd, 1H), Phenyl lactic acid: 2.82, 2.95, 4.16, 7.2
5.

Furthermore, an optically active column (J & WSCIEN
2-using TIFIC's Cyclodex-B)
As a result of evaluating the three-dimensional structure of hydroxyisocaproic acid,
It was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

On the other hand, as a result of evaluating the three-dimensional structure of phenyl lactic acid using an optically active column (Restek column 13113, manufactured by UNIFLEX), it was confirmed that the obtained phenyl lactic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 10 parts by mass of 2-hydroxyisocaproic acid, 12 parts by mass of phenyl lactic acid, and tin dichloride were added.
220 parts by mass of mesitylene was added to 044 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark tr.
The water accumulated in the ap was appropriately removed. The polymerization time was 40 hours, and the weight average molecular weight of the obtained polyester copolymer was 160,000.

The polyester copolymer having a weight average molecular weight of 160,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 37 to 38 μm, tensile strength: 60 MPa (yield), 60 MPa (breaking), elongation: 13%.

Example 70 A polyester copolymer was synthesized by using 2-hydroxyisocaproic acid and phenyllactic acid obtained by the same procedure as in Example 69. Dean
Using a reactor equipped with a Stark trap, 2-
10 parts by mass of hydroxyisocaproic acid, 1 phenyllactic acid
200 parts by mass of 1,3-dichlorobenzene was added to 2 parts by mass and 0.04 parts by mass of zinc powder, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Starktrap. The polymerization time was 38 hours, and the weight average molecular weight of the obtained polyester copolymer was 110,000.

The polyester copolymer having a weight average molecular weight of 110,000 thus obtained was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 50 MPa (yield), 49 MPa (breaking), elongation: 7%.

(Example 71) 400 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) and 11,000 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l,
NaCI 5g / l, cysteine hydrochloride 2g / l, calcium carbonate 5g / l) were mixed and injected into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper, and the autoclave (121 ℃, pressurize 98kPa, 10
Min) processed. Then, the medium was cooled to 30 ° C., and the lactic acid- and phenyllactic acid-producing strain HICA isolated by the above-mentioned method was used.
110 parts by mass of the preculture liquid of the 432 strain was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 4 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and NMR analysis confirmed that a mixture of 12 parts by mass of lactic acid and 10 parts by mass of phenyllactic acid was obtained. did.

The analysis results of lactic acid are as follows: ¹ H-NMR (400 MHz, TMS, DMSO-d ₆ ).
δ / ppm: 1.25 (d, J = 6.8 Hz, 3H),
4.07 (q, J = 6.8 Hz, 1H), and the analysis results of phenyllactic acid are as follows: ¹ H-NMR (400 MHz, TMS, DMSO-d ₆ ).
δ / ppm: 2.82, 2.95, 4.16, 7.2
5. Further, an optically active column (manufactured by UNIFLEX, Rest
As a result of evaluating the three-dimensional structure of lactic acid using ek column 13113), it was confirmed that the obtained lactic acid was DL-form.

On the other hand, as a result of evaluating the three-dimensional structure of phenyllactic acid using an optically active column (Restek column 13113 manufactured by UNIFLEX), it was confirmed that the obtained phenyllactic acid was L-form.

Using a reaction apparatus equipped with a Dean Stark trap, 220 parts by mass of mesitylene was added to 12 parts by mass of lactic acid, 10 parts by mass of phenyllactic acid, and 0.044 parts by mass of tin dichloride, and azeotropic dehydration was performed under a nitrogen atmosphere. Done,
Water accumulated in the Dean Stark trap was appropriately removed. The polymerization time was 40 hours, and the weight average molecular weight of the obtained polyester copolymer was 130,000.

The polyester copolymer having a weight average molecular weight of 130,000 thus obtained was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 61 MPa (yield), 60 MPa (breaking), elongation: 6%.

Example 72 A polyester copolymer was synthesized using lactic acid and phenyllactic acid obtained by the same procedure as in Example 71. Using a reactor equipped with a Dean Stark trap, 12 parts by mass of lactic acid, 10 parts by mass of phenyl lactic acid, and 0.04 parts by mass of zinc powder were added to 1,3-
200 parts by mass of dichlorobenzene was added, and azeotropic dehydration was performed under a nitrogen atmosphere to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 38 hours, and the weight average molecular weight of the obtained polyester copolymer was 80,000.

The polyester copolymer having a weight average molecular weight of 80,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 49 MPa (yield), 49 MPa (breakage), elongation: 6%.

(Example 73) 400 parts by mass of starch (manufactured by Wako Pure Chemical Industries, Ltd.) and 11,000 parts by mass of a medium (peptone 10 g / l, yeast extract 5 g / l, meat extract 2 g / l,
NaCI 5g / l, cysteine hydrochloride 2g / l, calcium carbonate 5g / l) were mixed and injected into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper, and the autoclave (121 ℃, pressurize 98kPa, 10
Min) processed. Then, the medium was cooled to 30 ° C., and the lactic acid- and phenyllactic acid-producing strain HICA isolated by the above-mentioned method was used.
110 parts by mass of the preculture liquid of 432 strain was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and NMR analysis confirmed that a mixture of 18 parts by mass of lactic acid and 4 parts by mass of phenyllactic acid was obtained. did.

Using a reaction apparatus equipped with a Dean Stark trap, 210 parts by mass of bromobenzene was added to 18 parts by mass of lactic acid, 4 parts by mass of phenyllactic acid, and 0.04 parts by mass of tin oxide, and azeotropic dehydration was performed under a nitrogen atmosphere. Then, the water accumulated in the Dean Starktrap was appropriately removed. The polymerization time was 45 hours, and the aliphatic polyester copolymer obtained had a weight average molecular weight of 180,000.

The polyester copolymer having a weight average molecular weight of 180,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 67 MPa (yield), 64 MPa (breakage), elongation: 10%.

Example 74 A polyester copolymer was synthesized using lactic acid and phenyllactic acid obtained by the same procedure as in Example 73. Using a reactor equipped with Dean Stark trap, 18 parts by mass of lactic acid, 4 parts by mass of phenyl lactic acid, and p-toluenesulfonic acid 0.0
200 parts by mass of 1,2-dichlorobenzene was added to 36 parts by mass, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean St.
Water accumulated in the ark trap was appropriately removed. The polymerization time was 30 hours, and the weight average molecular weight of the obtained polyester copolymer was 30,000.

The polyester copolymer having a weight average molecular weight of 30,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 19 MPa (yield), 19 MPa (breakage), elongation: 2%.

The following Examples 75 to 82 are substituted α-by microbial fermentation of xylose described in the third embodiment.
A method for producing a polyester, which comprises the step of obtaining a hydroxy acid.

[0463] Microorganisms, Example 2-hydroxyisocaproic acid-producing strains HICA432 strain used in 1-48, further Clostridium Beiji prn obtained by considering the ability to produce a substituted α- hydroxy acid from xylose (Clostridiumbeijerinckii )
An isolated strain of HICA432 strain (FERM BP-8117) was used.

It was confirmed that HICA432 strain produces not only 2-hydroxyisocaproic acid but also phenyllactic acid from xylose. Therefore, hereinafter, the HICA432 strain may also be referred to as a 2-hydroxyisocaproic acid-producing strain and a phenyllactic acid-producing strain.

(Example 75) 750 parts by mass of hemicellulose was added to 3000 parts by mass of 1% hydrochloric acid, and the mixture was stirred at 130 ° C for 8 hours. After the reaction, the mixture was neutralized with a 10% sodium hydroxide solution, 1800 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B manufactured by Organo). Then, the solvent was distilled off. The reaction mixture is separated and purified to give xylose 55
0 part by mass was obtained.

The obtained xylose was subjected to ion exchange chromatography (TSKgelSugarA manufactured by Tosoh Corporation).
XG column) for identification and confirmation.

550 parts by weight of this xylose were added to 27,000
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., 270 parts by mass of the preculture liquid of the phenyllactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe in a state where the bottle was sealed, and the mixture was allowed to stand at 30 ° C. for 6 days. The culture was performed. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and then purified to obtain 10 parts by mass of phenyllactic acid.

¹³ C-NMR (1
00MHz, TMS, CDCl ₃ ) measurement result (δ / p
pm) is 177.92, 137.63, 130.3
0, 129.46, 127.82, 72.05, 40.
It was 38 and it was confirmed that the desired phenyllactic acid was obtained.

Further, an optically active column (J & WSCIEN
As a result of evaluating the three-dimensional structure using Cylodex-B manufactured by TIFIC, it was confirmed that the obtained phenyllactic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 100 parts by mass of mesitylene was added to 10 parts by mass of phenyllactic acid and 0.018 parts by mass of tin dichloride, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark.
Water accumulated in the trap was appropriately removed. Polymerization time is 4
It was 4 hours, and the weight average molecular weight of the obtained polyester was 200,000.

The polyester having a weight average molecular weight of 200,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 38 to 39 μm, tensile strength: 70 MPa (yield), 69 MPa (break), elongation: 13%.

Example 76 A polyester was polymerized using phenyllactic acid obtained by performing the same operation as in Example 75. Using a reactor equipped with Dean Stark trap, 100 parts by mass of 1,3-dichlorobenzene was added to 10 parts by mass of phenyllactic acid and 0.02 parts by mass of zinc chloride,
Azeotropic dehydration is performed under a nitrogen atmosphere, and Dean Stark is used.
Water accumulated in the trap was appropriately removed. The polymerization time was 16 hours, and the weight average molecular weight of the obtained polyester was 50,000.

The polyester having a weight average molecular weight of 50,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 23 MPa (yield), 20 MPa (breaking), elongation: 3%.

(Example 77) Using phenyllactic acid obtained by performing the same operation as in Example 75, it was converted into a cyclic dimer lactide by the following method.

Using a reactor equipped with a Dean Stark trap, 600 parts by mass of toluene was added to 10 parts by mass of phenyllactic acid and 0.20 parts by mass of p-toluenesulfonic acid, and azeotropic dehydration was carried out for 30 hours in a nitrogen atmosphere. Dea
Water accumulated in the nStark trap was appropriately removed. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 8.7 parts by mass of cyclic dimer lactide.

The ring-opening polymerization was carried out by heating 8.7 parts by mass of the cyclic dimer lactide and 0.01 parts by mass of tin chloride at 160 ° C. for 10 hours in a nitrogen atmosphere. The weight average molecular weight of the obtained aromatic polyester was 190,000.

The aromatic polyester having a weight average molecular weight of 190,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 69 MPa (yield), 69 MPa (breakage), elongation: 12%.

Example 78 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 77. Cyclic dimer lactide (8.7 parts by mass) and tin octylate (0.007 parts by mass) were added under nitrogen atmosphere for 180 times.
The ring-opening polymerization was carried out by heating at 0 ° C for 33 hours. The weight average molecular weight of the obtained aromatic polyester was 700,000. The aromatic polyester having a weight average molecular weight of 700,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 25%.

(Example 79) 750 parts by mass of hemicellulose was added to 3000 parts by mass of 1% hydrochloric acid, and the mixture was stirred at 130 ° C for 8 hours. After the reaction, the mixture was neutralized with a 10% sodium hydroxide solution, 1800 parts by mass of methanol was added, the water-soluble residue was filtered off, and the mixture was passed through an ion exchange resin column (Amberlite IR-120B manufactured by Organo). Then, the solvent was distilled off. The reaction mixture is separated and purified to give xylose 55
0 part by mass was obtained.

The obtained xylose was subjected to ion exchange chromatography (TSKgelSugarA manufactured by Tosoh Corporation).
XG column) for identification and confirmation.

550 parts by weight of this xylose were added to 27,000
Part by weight of medium (peptone 10 g / l, yeast extract 5 g /
1, meat extract 2 g / l, NaCl 5 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, and the gas phase in the bottle was replaced with nitrogen gas, then butyl rubber Seal with a stopper and autoclave (121
C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and the preculture liquid 2 of the 2-hydroxyisocaproic acid producing strain HICA432 strain isolated by the above-mentioned method 2
70 parts by mass was added using a syringe while the bottle was sealed, and static culture was carried out at 30 ° C. for 6 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and then purified to obtain 12 parts by mass of 2-hydroxyisocaproic acid.

Of the resulting 2-hydroxyisocaproic acid
¹³ C-NMR (100 MHz, TMS, CDCl ₃ ) measurement results (δ / ppm) are 180.25 and 68.9.
5, 43.16, 24.45, 23.20, 21.42
It was confirmed that the desired 2-hydroxyisocaproic acid was obtained.

In addition, an optically active column (J & WSCIEN
As a result of evaluation of the three-dimensional structure using Cyclodex-B manufactured by TIFIC, it was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 12 parts by mass of 2-hydroxyisocaproic acid and 0.018 parts by mass of tin dichloride were added to 100 parts of mesitylene.
Add parts by mass, perform azeotropic dehydration under a nitrogen atmosphere, and
Water accumulated in the n Stark trap was appropriately removed. The polymerization time was 44 hours, and the polyester obtained had a weight average molecular weight of 220,000.

The polyester having a weight average molecular weight of 220,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 67 MPa (yield), 67 MPa (breakage), elongation: 16%.

Example 80 A polyester was polymerized using 2-hydroxyisocaproic acid obtained by the same procedure as in Example 79. Dean Stark trap
12 parts by mass of 2-hydroxyisocaproic acid and 0.02 parts by mass of zinc chloride were added with 100 parts by mass of 1,3-dichlorobenzene, and azeotropic dehydration was performed under a nitrogen atmosphere to prepare Dean Stark. Water accumulated in the trap was appropriately removed. The polymerization time was 16 hours, and the polyester obtained had a weight average molecular weight of 60,000.

The polyester having a weight average molecular weight of 60,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 23 MPa (yield), 21 MPa (breakage), elongation: 3%.

Example 81 2-Hydroxyisocaproic acid obtained in the same manner as in Example 79 was used to convert it into a cyclic dimer lactide by the following method.

Using a reactor equipped with a Dean Stark trap, 600 parts by mass of toluene was added to 12 parts by mass of 2-hydroxyisocaproic acid and 0.24 parts by mass of p-toluenesulfonic acid, and the mixture was azeotropically distilled for 30 hours under a nitrogen atmosphere. Dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 9.8 parts by mass of cyclic dimer lactide. ¹³ C-NMR (100 MH of the obtained cyclic dimer lactide
z, TMS, CDCl ₃ ) measurement results (δ / ppm)
Is 167.33, 74.14, 38.87, 23.8.
9, 23.03, 21.33, and it was confirmed that the target cyclic dimer lactide was obtained.

Ring-opening polymerization was carried out by heating 9.8 parts by mass of the cyclic dimer lactide and 0.013 parts by mass of tin chloride in a nitrogen atmosphere at 150 ° C. for 12 hours. The weight average molecular weight of the obtained aliphatic polyester was 180,000.

¹³ C-NMR of this aliphatic polyester
(100 MHz, TMS, CDCl ₃ ) measurement result (δ
/ Ppm) is 169.78, 71.38, 39.33,
24.53, 23.00, 21.43, and it was confirmed that the target aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 180,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 68 MPa (yield), 68 MPa (breakage), elongation: 13%.

Example 82 An aliphatic polyester was polymerized using the cyclic dimer lactide obtained by performing the same operation as in Example 81. The cyclic dimer lactide (7.4 parts by mass) and the tin octylate (0.011 parts by mass) were added under nitrogen atmosphere for 180 times.
The ring-opening polymerization was performed by heating at 30 ° C. for 30 hours. The weight average molecular weight of the obtained aliphatic polyester was 680,000. ¹³
When C-NMR was measured, the same spectrum as in Example 81 was obtained, and it was confirmed that the desired aliphatic polyester was obtained.

The aliphatic polyester having a weight average molecular weight of 680,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 30%.

The following Examples 83-91 are methods for producing polyesters which include the step of obtaining a substituted α-hydroxy acid by microbial fermentation of cellulose and xylose described in the fourth embodiment.

The microorganism was prepared by examining the ability of the 2-hydroxyisocaproic acid-producing strain HICA432 used in Example 1-48 to further produce a substituted α-hydroxy acid from cellulose and xylose. ( Clostridiumbeige
rinckii ) HICA432 strain (FERM BP-
8117) isolates were used.

It was confirmed that the HICA432 strain produced not only 2-hydroxyisocaproic acid but also lactic acid and phenyllactic acid from xylose and glucose. Therefore, in the following, the HICA432 strain is
-Hydroxyisocaproic acid-producing strain, lactic acid-producing strain, and phenyllactic acid-producing strain may also be referred to.

(Example 83) Newspaper was cut into 10 mm square pieces, 500 parts by mass of which was added to 2500 parts by mass of 20% sulfuric acid, and the mixture was stirred at 180 ° C for 12 hours. After the reaction, the mixture was neutralized with a 20% sodium hydroxide solution, 300 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (Amberlite IR-120 manufactured by Organo Corporation) was further added.
It was passed through B) and the solvent was distilled off. The reaction mixture was separated and purified to obtain 25 parts by mass of xylose and 180 parts by mass of glucose.

The xylose and glucose thus obtained were subjected to ion exchange chromatography (TSKge manufactured by Tosoh Corporation).
ISugar AXG column).

25 parts by mass of this xylose and 1 part of glucose
80 parts by mass of 10000 parts by mass of medium (peptone 10 g
/ L, yeast extract 5g / l, meat extract 2g / l, NaC
15 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper and then autoclaved (121 C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., 100 parts by mass of the preculture liquid of the phenyllactic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe in a state where the bottle was sealed, and the mixture was allowed to stand at 30 ° C. for 6 days. The culture was performed. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and then purified to obtain 9 parts by mass of phenyllactic acid.

¹³ C-NMR (1
00MHz, TMS, CDCl ₃ ) measurement result (δ / p
pm) is 177.92, 137.63, 130.3
0, 129.46, 127.82, 72.05, 40.
It was 38 and it was confirmed that the desired phenyllactic acid was obtained.

In addition, an optically active column (J & WSCIEN
As a result of evaluating the three-dimensional structure using Cylodex-B manufactured by TIFIC, it was confirmed that the obtained phenyllactic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 100 parts by mass of mesitylene was added to 9 parts by mass of phenyllactic acid and 0.018 parts by mass of tin dichloride, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark.
Water accumulated in the trap was appropriately removed. Polymerization time is 4
It was 4 hours, and the weight average molecular weight of the obtained polyester was 120,000.

The polyester having a weight average molecular weight of 120,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 50 MPa (yield), 49 MPa (breaking), elongation: 5%.

Example 84 A polyester was polymerized using phenyllactic acid obtained by performing the same operation as in Example 83. Using a reactor equipped with Dean Starktrap, 100 parts by mass of diphenyl ether was added to 9 parts by mass of phenyllactic acid and 0.015 parts by mass of tin powder, and the temperature was adjusted to 130 ° C /
Azeotropic dehydration is performed at 12 mmHg, and DeanStarkt is used.
Water accumulated in the rap was appropriately removed. Polymerization time is 48
It was time, and the weight average molecular weight of the obtained polyester was 220,000.

The polyester having a weight average molecular weight of 220,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 39 to 40 μm, tensile strength: 74 MPa (yield), 73 MPa (breakage), elongation: 14%.

(Example 85) Using phenyllactic acid obtained by performing the same operation as in Example 83, it was converted into a cyclic dimer lactide by the following method.

Using a reactor equipped with a Dean Stark trap, 540 parts by mass of toluene was added to 9 parts by mass of phenyllactic acid and 0.18 parts by mass of p-toluenesulfonic acid, and azeotropic dehydration was performed for 30 hours in a nitrogen atmosphere. Dean
Water accumulated in the Stark trap was appropriately removed. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 7.8 parts by mass of cyclic dimer lactide.

Ring-opening polymerization was carried out by heating 7.8 parts by mass of cyclic dimer lactide and 0.007 parts by mass of tin octylate under nitrogen atmosphere at 180 ° C. for 33 hours. The weight average molecular weight of the obtained aromatic polyester was 700,000.

The aromatic polyester having a weight average molecular weight of 700,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 25%.

(Example 86) Newspaper was cut into 10 mm squares, 500 parts by mass of which were added to 2500 parts by mass of 20% sulfuric acid, and the mixture was stirred at 180 ° C for 12 hours. After the reaction, the mixture was neutralized with a 20% sodium hydroxide solution, 300 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (Amberlite IR-120 manufactured by Organo Corporation) was further added.
It was passed through B) and the solvent was distilled off. The reaction mixture was separated and purified to obtain 25 parts by mass of xylose and 180 parts by mass of glucose.

The xylose and glucose thus obtained were subjected to ion exchange chromatography (TSKge manufactured by Tosoh Corporation).
ISugar AXG column).

25 parts by mass of xylose and 1 part of glucose
80 parts by mass of 10000 parts by mass of medium (peptone 10 g
/ L, yeast extract 5g / l, meat extract 2g / l, NaC
15 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper and then autoclaved (121 C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and 100 parts by mass of a preculture liquid of phenyllactic acid and 2-hydroxyisocaproic acid producing strain HICA432 strain isolated by the above-mentioned method was added using a syringe in a state where the bottle was sealed. The cells were statically cultured at 30 ° C for 6 days. Then, the culture solution is filtered, water and volatile components of the filtrate are distilled off at room temperature under reduced pressure, and then purified to NM.
As a result of R analysis, a mixture of 8.5 parts by mass of phenyllactic acid and 8.0 parts by mass of 2-hydroxyisocaproic acid was obtained.

Incidentally, ¹ H-NMR (400 MHz, TM
The measurement results (δ / ppm) of S, DMSO-d ₆ ) are as follows; 2-hydroxyisocaproic acid: 0.88 (t, 6)
H), 1.37 to 1.48 (m, 2H), 1.72 to
1.82 (m, 1H), 3.94 (dd, 1H), Phenyl lactic acid: 2.82, 2.95, 4.16, 7.2
5.

Furthermore, an optically active column (J & WSCIEN
2-using TIFIC's Cyclodex-B)
As a result of evaluating the three-dimensional structure of hydroxyisocaproic acid,
It was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

On the other hand, as a result of evaluating the three-dimensional structure of phenyl lactic acid using an optically active column (Restek column 13113, manufactured by UNIFLEX), it was confirmed that the obtained phenyl lactic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 2-hydroxyisocaproic acid 8.0
100 parts by mass of mesitylene was added to 0.01 part by mass of tin dichloride, 8.5 parts by mass of phenyl lactic acid, and azeotropic dehydration was performed under a nitrogen atmosphere to obtain Dean Stark.
Water accumulated in the trap was appropriately removed. Polymerization time is 4
It was 0 hours, and the weight average molecular weight of the obtained polyester copolymer was 160,000.

The polyester copolymer having a weight average molecular weight of 160,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 37 to 38 μm, tensile strength: 60 MPa (yield), 60 MPa (breaking), elongation: 13%.

(Example 87) Newspaper was cut into 10 mm square pieces, 500 parts by mass of which was added to 2500 parts by mass of 20% sulfuric acid, and the mixture was stirred at 180 ° C for 12 hours. After the reaction, the mixture was neutralized with a 20% sodium hydroxide solution, 300 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (Amberlite IR-120 manufactured by Organo Corporation) was further added.
It was passed through B) and the solvent was distilled off. The reaction mixture was separated and purified to obtain 25 parts by mass of xylose and 180 parts by mass of glucose.

The xylose and glucose thus obtained were subjected to ion exchange chromatography (TSKge manufactured by Tosoh Corporation).
ISugar AXG column).

25 parts by mass of xylose and 1 part of glucose
80 parts by mass of 10000 parts by mass of medium (peptone 10 g
/ L, yeast extract 5g / l, meat extract 2g / l, NaC
15 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper and then autoclaved (121 C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C. and the phenyllactic acid and lactic acid producing strain HICA432 isolated by the above-mentioned method was used.
100 parts by mass of the preculture liquid of the strain was added using a syringe in a state where the bottle was sealed, and static culture was carried out at 30 ° C. for 4 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and purified and analyzed by NMR. As a result, a mixture of 4 parts by mass of phenyllactic acid and 5 parts by mass of lactic acid was obtained.

The analysis results of lactic acid are as follows: ¹ H-NMR (400 MHz, TMS, DMSO-d ₆ ).
δ / ppm: 1.25 (d, J = 6.8 Hz, 3H),
4.07 (q, J = 6.8 Hz, 1H), and the analysis results of phenyllactic acid are as follows: ¹ H-NMR (400 MHz, TMS, DMSO-d ₆ ).
δ / ppm: 2.82, 2.95, 4.16, 7.2
5. Further, an optically active column (manufactured by UNIFLEX, Rest
As a result of evaluating the three-dimensional structure of lactic acid using ek column 13113), it was confirmed that the obtained lactic acid was DL-form.

On the other hand, as a result of evaluating the three-dimensional structure of phenyl lactic acid using an optically active column (Restek column 13113, manufactured by UNIFLEX), it was confirmed that the obtained phenyl lactic acid was L-form.

Using a reactor equipped with a Dean Stark trap, 5 parts by mass of lactic acid, 4 parts by mass of phenyl lactic acid, and 0.018 parts by mass of tin dichloride were added to 1 part of mesitylene.
00 parts by mass was added, and azeotropic dehydration was performed under a nitrogen atmosphere.
Water accumulated in the ean Stark trap was appropriately removed. The polymerization time was 40 hours, and the weight average molecular weight of the obtained polyester copolymer was 130,000.

The polyester copolymer having a weight average molecular weight of 130,000 thus obtained was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 61 MPa (yield), 60 MPa (breaking), elongation: 6%.

(Example 88) Newspaper was cut into 10 mm square pieces, 500 parts by mass of which was added to 2500 parts by mass of 20% sulfuric acid, and the mixture was stirred at 180 ° C for 12 hours. After the reaction, the mixture was neutralized with a 20% sodium hydroxide solution, 300 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (Amberlite IR-120 manufactured by Organo Corporation) was further added.
It was passed through B) and the solvent was distilled off. The reaction mixture was separated and purified to obtain 22 parts by mass of xylose and 170 parts by mass of glucose.

The xylose and glucose thus obtained were subjected to ion exchange chromatography (TSKge manufactured by Tosoh Corporation).
ISugar AXG column).

22 parts by mass of xylose and 1 part of glucose
70 parts by mass of 10,000 parts by mass of medium (peptone 10 g
/ L, yeast extract 5g / l, meat extract 2g / l, NaC
15 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper and then autoclaved (121 C., 98 kPa pressurization, 10 minutes). Then, the medium was cooled to 30 ° C., and the 2-hydroxyisocaproic acid-producing strain HICA isolated by the above-mentioned method was used.
100 parts by mass of the preculture liquid of 432 strain was added using a syringe while the bottle was sealed, and static culture was performed at 30 ° C. for 6 days. Then, the culture solution was filtered, and water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, followed by purification to obtain 8 parts by mass of 2-hydroxyisocaproic acid.

Of the resulting 2-hydroxyisocaproic acid
¹³ C-NMR (100 MHz, TMS, CDCl ₃ ) measurement results (δ / ppm) are 180.25 and 68.9.
5, 43.16, 24.45, 23.20, 21.42
It was confirmed that the desired 2-hydroxyisocaproic acid was obtained.

Also, an optically active column (J & WSCIEN
As a result of evaluation of the three-dimensional structure using Cyclodex-B manufactured by TIFIC, it was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

[0531] Using a reactor equipped with a Dean Stark trap, 100 parts by mass of mesitylene was added to 8 parts by mass of 2-hydroxyisocaproic acid and 0.018 parts by mass of tin dichloride, and azeotropic dehydration was performed under a nitrogen atmosphere. Dean
Water accumulated in the Stark trap was appropriately removed. The polymerization time was 44 hours, and the weight average molecular weight of the obtained polyester was 100,000.

The polyester having a weight average molecular weight of 100,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 51 MPa (yield), 50 MPa (breakage), elongation: 6%.

(Example 89) Polyester was polymerized using 2-hydroxyisocaproic acid obtained by performing the same operation as in Example 88. Dean Stark trap
Using 2 parts of 2-hydroxyisocaproic acid and 0.015 parts by mass of tin powder, 100 parts by mass of diphenyl ether was added, and azeotropic dehydration was carried out at 130 ° C / 12 mmHg to collect in the Dean Stark trap. Water was removed as appropriate. The polymerization time was 48 hours, and the polyester obtained had a weight average molecular weight of 200,000.

The polyester having a weight average molecular weight of 200,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties; thickness: 39 to 40 μm, tensile strength: 62 MPa (yield), 59 MPa (breaking), elongation: 18%.

Example 90 2-Hydroxyisocaproic acid obtained in the same manner as in Example 88 was used to convert it into a cyclic dimer lactide by the following method.

Using a reactor equipped with a Dean Stark trap, 400 parts by mass of toluene was added to 8 parts by mass of 2-hydroxyisocaproic acid and 0.16 parts by mass of p-toluenesulfonic acid, and the mixture was azeotropically distilled for 30 hours under a nitrogen atmosphere. Dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. After completion of the reaction, the mixture was washed with sodium hydrogen carbonate and toluene was distilled off. The residue was recrystallized from ether to obtain 6.5 parts by mass of cyclic dimer lactide. ¹³ C-NMR of the obtained cyclic dimer lactide (100 MHz,
TMS, CDCl ₃ ) measurement result (δ / ppm) is 1
67.33, 74.14, 38.87, 23.89, 2
It was 3.03 and 21.33, and it was confirmed that the target cyclic dimer lactide was obtained.

Ring-opening polymerization was carried out by heating 6.5 parts by mass of a cyclic dimer lactide and 0.010 parts by mass of tin octylate at 180 ° C. for 30 hours in a nitrogen atmosphere. The weight average molecular weight of the obtained aliphatic polyester was 680,000.

¹³ C-NMR of this aliphatic polyester
(100 MHz, TMS, CDCl ₃ ) measurement result (δ
/ Ppm) is 169.78, 71.38, 39.33,
24.53, 23.00, 21.43, and it was confirmed that the target aliphatic polyester was synthesized.

The aliphatic polyester having a weight average molecular weight of 680,000 obtained above was dissolved in chloroform, and a film was produced from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 98 MPa (yield), 98 MPa (breaking), elongation: 30%.

(Example 91) Newspaper was cut into 10 mm square pieces, 500 parts by mass of which was added to 2500 parts by mass of 20% sulfuric acid, and the mixture was stirred at 180 ° C for 12 hours. After the reaction, the mixture was neutralized with a 20% sodium hydroxide solution, 300 parts by mass of methanol was added, the water-soluble residue was filtered off, and an ion exchange resin column (Amberlite IR-120 manufactured by Organo Corporation) was further added.
It was passed through B) and the solvent was distilled off. The reaction mixture was separated and purified to obtain 25 parts by mass of xylose and 180 parts by mass of glucose.

The obtained xylose and glucose were subjected to ion exchange chromatography (TSKge manufactured by Tosoh Corporation).
ISugar AXG column).

25 parts by mass of xylose and 1 part of glucose
80 parts by mass of 10000 parts by mass of medium (peptone 10 g
/ L, yeast extract 5g / l, meat extract 2g / l, NaC
15 g / l, cysteine hydrochloride 2 g / l, calcium carbonate 5 g / l) and mixed into a pressure resistant bottle, the gas phase in the bottle was replaced with nitrogen gas, and the bottle was sealed with a butyl rubber stopper and then autoclaved (121 C., 98 kPa pressurization, 10 minutes). Thereafter, the medium was cooled to 30 ° C., and 100 parts by mass of a preculture liquid of 2-hydroxyisocaproic acid and a lactic acid producing strain HICA432 strain isolated by the above-described method were added using a syringe in a state where the bottle was sealed, Static culture was carried out at 30 ° C. for 3 days. Then, the culture solution was filtered, water and volatile components of the filtrate were distilled off at room temperature under reduced pressure, and purified and analyzed by NMR. As a result, a mixture of 7 parts by mass of 2-hydroxyisocaproic acid and 6 parts by mass of lactic acid was obtained. .

¹ H-NMR (400 MHz, TM
The measurement results (δ / ppm) of S, DMSO-d ₆ ) are as follows; 2-hydroxyisocaproic acid: 0.88 (t, 6)
H), 1.37 to 1.48 (m, 2H), 1.72 to
1.82 (m, 1H), 3.94 (dd, 1H), Lactic acid: 1.25 (d, J = 6.8Hz, 3H), 4.0
7 (q, J = 6.8 Hz, 1H).

In addition, ¹³ C-NMR (100 MHz, TM
S, DMSO-d ₆ ) measurement results (δ / ppm) are as follows: 2-hydroxyisocaproic acid: 21.43, 23.1
2, 23.84, 42.84, 68.07, 176.2
3, Lactic acid: 20.39, 65.82, 176.42.

Furthermore, an optically active column (J & WSCIEN
2-using TIFIC's Cyclodex-B)
As a result of evaluating the three-dimensional structure of hydroxyisocaproic acid,
It was confirmed that the obtained 2-hydroxyisocaproic acid was L-form.

On the other hand, as a result of evaluating the steric structure of lactic acid using an optically active column (Restek column 13113, manufactured by UNIFLEX), it was confirmed that the obtained lactic acid was DL-form.

Using a reactor equipped with a Dean Stark trap, 100 parts by mass of mesitylene was added to 7 parts by mass of 2-hydroxyisocaproic acid, 6 parts by mass of lactic acid, and 0.018 parts by mass of tin dichloride, and the mixture was added under a nitrogen atmosphere. Azeotropic dehydration was performed to appropriately remove water accumulated in the Dean Stark trap. The polymerization time was 40 hours, and the weight average molecular weight of the obtained aliphatic polyester copolymer was 170,000.

¹³ C-NMR (100 MHz, TMS, C
The measurement result (δ / ppm) of DCl ₃ ) is 16.66,
21.43, 22.99, 24.55, 39.30, 6
It was 8.97, 71.37, 169.51, 169.76, and it was confirmed that the target aliphatic polyester copolymer was synthesized.

The aliphatic polyester copolymer having a weight average molecular weight of 170,000 obtained above was dissolved in chloroform, and a film was prepared from the solution by a casting method. The produced film is colorless and has the following physical properties: thickness: 38 to 39 μm, tensile strength: 61 MPa (yield), 59 MPa (breaking), elongation: 14%.

[0550]

INDUSTRIAL APPLICABILITY In the above-described production method of the present invention, by utilizing a microorganism, a substituted α-hydroxy acid having a high purity to the extent that it can be used as a monomer of polyester is converted into starch, glucose, xylose and glucose. It can be efficiently produced from sugars such as a mixture of xylose. As a result, it becomes possible to obtain a high-quality plastic by using glucan such as cellulose or starch, cellulose, hemicellulose, or a mixture of cellulose and hemicellulose as a starting material. Therefore, it becomes possible to manufacture polyester obtained from waste cellulose, waste hemicellulose, waste starch, etc. as raw materials, and these wastes can be recycled.

   ─────────────────────────────────────────────────── ─── Continued front page    (31) Priority claim number Japanese Patent Application No. 2001-215428 (P2001-215428) (32) Priority date July 16, 2001 (July 16, 2001) (33) Priority claiming country Japan (JP) F term (reference) 4B064 AD32 AD83 CA02 CB07 CD19                       DA16                 4B065 AA23 AC14 BB08 BB18 CA10                       CA12 CA55

Claims (21)

[Claims] 1. A method for producing polyester having the following steps: (1) HO-C by fermenting sugars with a microorganism.
HR-COOH (wherein R represents a hydrocarbon group having 1 to 10 carbon atoms), and a step of obtaining one or more kinds of substituted α-hydroxy acid, (2) the substituted α-hydroxy acid or a derivative thereof. The process of polymerizing. 2. The method for producing a polyester according to claim 1, wherein the saccharide is glucose or xylose. 3. Prior to the steps (1) and (2),
The method for producing a polyester according to claim 2, further comprising (3) a step of hydrolyzing a raw material containing cellulose or hemicellulose to obtain glucose or xylose. 4. The method for producing a polyester according to claim 1, wherein the saccharide is starch. 5. The sugar is starch or glucose, and the one or more substituted α-hydroxy acids are lactic acid (R is a methyl group), β-phenyllactic acid (R is a benzyl group) and 2-hydroxy. The method for producing a polyester according to claim 1, wherein the polyester is selected from the group consisting of isocaproic acid (wherein R is an isobutyl group). 6. The method for producing a polyester according to claim 1, wherein the at least one substituted α-hydroxy acid contains at least β-phenyllactic acid (wherein R is a benzyl group). 7. The method for producing a polyester according to claim 1, wherein the at least one substituted α-hydroxy acid contains at least 2-hydroxyisocapnic acid (wherein R is an isobutyl group). 8. The saccharide is xylose, and the one or more substituted α-hydroxy acids are β-phenyllactic acid (where R is a benzyl group) and 2-hydroxyisocaproic acid (where R is an isobutyl group). The method for producing a polyester according to claim 1, wherein the polyester is at least one of them. 9. The method for producing a polyester according to claim 1, wherein the microorganism in the step (1) is an anaerobic bacterium. 10. The method for producing a polyester according to claim 8, wherein the fermentation in the step (1) is performed in the presence of pyruvic acid. 11. The method for producing a polyester according to claim 8, wherein the anaerobic bacterium is a bacterium belonging to the genus Clostridium. 12. The bacterium belonging to the genus Clostridium is Clostridium bayerikinki HICA432 (Clo).
stridium beijerinckii HIC
A432) FERM BP-8117 strain, The manufacturing method of polyester of Claim 10 characterized by the above-mentioned. 13. The substituted α-hydroxy acid derivative is a cyclic dimer lactide, and prior to the polymerization step (2), (4) two molecules of the substituted α-hydroxy acid are dehydrated to form a cyclic diester. The method for producing a polyester according to claim 1, further comprising a step of performing the same to obtain the cyclic dimer lactide. 14. The polyester has the following formula (I):
The method for producing a polyester according to claim 1, comprising at least one of the repeating units represented by (III). [Chemical 1] [Chemical 2] [Chemical 3] 15. A method for producing a substituted α-hydroxy acid, which comprises fermenting sugars with a microorganism. 16. The method for producing a substituted α-hydroxy acid according to claim 15, wherein the microorganism is an anaerobic bacterium. 17. The method for producing a substituted α-hydroxy acid according to claim 16, wherein the step of obtaining the substituted α-hydroxy acid is performed by fermentation with an anaerobic bacterium in the presence of pyruvic acid. 18. The substituted α according to claim 16, wherein the anaerobic bacterium is a bacterium belonging to the genus Clostridium.
-A method for producing a hydroxy acid. 19. The bacterium belonging to the genus Clostridium is Clostridium vaginii HICA432 (Clo).
stridium beijerinckii HIC
A432) FERM BP-8117 strain, The method for producing a substituted α-hydroxy acid according to claim 18, which is characterized in that 20. Clostridium beyzirinki H
ICA432 (Clostridium beijer
inckii HICA432) FERMBP-811
7 strains. 21. Clostridium, which is used for producing a substituted α-hydroxy acid from a saccharide.
Basirinki HICA432 (Clostridiu
m beijerinckii HICA432) FE
RM BP-8117 strain.