JP2018109103A - Block copolymer and resin composition containing the same, and method for producing block copolymer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new block copolymer applicable as a compatibilizer of a resin.SOLUTION: There is provided a block copolymer in which a polymer block A and a polymer block B are bonded to each other through a connection group X, where the polymer block A is composed of a repeating unit represented by the following general formula (1): [-CHR-CH-CO-O-], the polymer block B has at least one polymer unit selected from the group consisting of aliphatic polyester and polyalkylene glycol, and the connection group X is a group formed by an alkyne-azide reaction, a group formed by a thio-ene reaction or a group formed by a thio-in reaction. In the formula (1), Ris an alkyl group represented by CH; n is an integer of 1 to 15; and * is asymmetric carbon being an R arrangement.SELECTED DRAWING: None

Description

  The present invention relates to a block copolymer having a poly [(R) -3-hydroxyalkanoic acid] block, a resin composition containing the block copolymer, and a method for producing the block copolymer.

  Poly [(R) -3-hydroxyalkanoic acid] (hereinafter abbreviated as “PHA”) is a thermoplastic polyester that is produced and accumulated as an energy storage material in the cells of many microbial species. PHA produced from various natural carbon sources by microorganisms is completely biodegraded by microorganisms in soil and water, and thus is incorporated into the natural carbon cycle process. Therefore, it can be said that PHA is an environmentally friendly plastic that has almost no adverse effects on the ecosystem. In recent years, synthetic plastics have become a serious social problem from the viewpoints of environmental pollution, waste disposal, and petroleum resources, and PHA has attracted attention as an environmentally friendly green plastic, and its practical application is eagerly desired.

  PHA first discovered in microorganisms is poly [(R) -3-hydroxybutyrate] (hereinafter abbreviated as “PHB”), which is a homopolymer of 3-hydroxybutyric acid (hereinafter abbreviated as “3HB”). It is. PHB has high crystallinity, is hard and brittle because of its high degree of crystallinity, and rapidly decomposes at a temperature near the melting point (180 ° C.), so that melt workability is low and the practical range is extremely limited. doing.

  Therefore, in order to improve brittleness by reducing the crystallinity of PHB, an attempt was made to introduce other 3-hydroxyalkanoic acid into the PHB skeleton. For example, side chains such as 3-hydroxypropionic acid (hereinafter abbreviated as “3HP”), 4-hydroxybutyric acid (hereinafter abbreviated as “4HB”), 5-hydroxyvaleric acid (hereinafter abbreviated as “5HV”), etc. PHB skeleton for linear monomers having no side chain and monomers having side chains such as lactic acid, 3-hydroxyvaleric acid (hereinafter abbreviated as “3HV”), 3-hydroxyhexanoic acid (hereinafter abbreviated as “3HHx”) The introduction to has been reported so far. The physical properties of PHA obtained vary greatly depending on the type of monomer to be introduced and the copolymerization ratio, but basically any monomer is introduced, so that the crystallinity of PHB is lowered, so that the melt processability is higher than that of PHB. Improve. Among them, poly (3HB-co-3HHx) (hereinafter abbreviated as “PHBH”) can express a wide range of physical properties from hard to soft by changing the copolymerization ratio of 3HHx, and thus its practical application is strongly desired.

  On the other hand, attempts have been made to change physical properties by chemically linking PHAs having different properties or PHA and other biodegradable resins or the like to form a block copolymer. For example, in Patent Document 1, PHB and PCL are synthesized by synthesizing polycaprolactone (hereinafter abbreviated as “PCL”) or polylactic acid (hereinafter abbreviated as “PLA”) by ring-opening polymerization from the end using PHB as a polymer initiator. Alternatively, a method for producing a diblock copolymer composed of PHB and PLA has been reported. In Non-Patent Document 1, PHB, poly (3HB-co-3HV) (hereinafter abbreviated as “PHBV”), or poly [(R) -3-hydroxyoctanoic acid] (hereinafter abbreviated as “PHO”). A block copolymer composed of PHAs obtained by synthesizing atactic PHB by ring-opening polymerization from the terminal using PHA as a polymer initiator has been reported. Non-Patent Document 2 reports a triblock copolymer composed of PHB and polyethylene glycol (hereinafter abbreviated as “PEG”).

  The block copolymer thus obtained is expected to be developed for drug delivery as described in Non-Patent Document 3. As summarized in Non-Patent Document 4, some biodegradable block copolymers that function as compatibilizers in biodegradable polymer blends have been reported.

US Patent No. 5,439,985

Grazyna Adamus, Wanda Sikorska, Henryk Janeczek, Michal Kwiecien, Michal Sobota, Marek Kowalzuku, European Polymer Jour. 48, pp. 621-631, 2013 Jun Li, Xu Li, Xipping Ni, Kam W. et al. Leon, Macromolecules, vol. 36, pp. 2661-2667, 2003 Neeraj Kumar, Majeti N .; V. Ravikumar, A.R. J. et al. Domb, Advanced Drug Delivery Review, vol. 53 pp. 23-44, 2001 B. Imre, B.M. Pukanzsky, European Polymer Journal, vol. 49, pp. 1215-12333, 2013

  As described above, in order to expand the use of PHA, it is important to improve the physical properties and processability of PHA. For example, the development of a new biodegradable block copolymer that can function as a compatibilizer for other resins, etc. It was desired. In particular, as described above, development of a new block copolymer that can be used as a compatibilizer for PHBH has been strongly desired for the practical use of PHBH. In particular, it is desirable to provide an environmentally friendly block copolymer that uses a PHA block that can be click-reacted as another block polymer as a microbial production block.

  Accordingly, an object of the present invention is to provide a novel block copolymer using an environmentally friendly click-reactive PHA block.

  As a result of intensive studies to solve the above problems, the present inventors have found that a novel block copolymer (biodegradable block copolymer) can be efficiently obtained according to a specific production method. The invention has been completed.

The details of the present invention are as follows.
1. A block copolymer in which a polymer block A and a polymer block B are bonded via a linking group X, and the polymer block A has the following general formula (1)
[—C ^* HR ¹ —CH ₂ —CO—O—] (1)
(In the formula, R ¹ is an alkyl group represented by C _n H _{2n + 1} , n is an integer of 1 to 15, and * indicates an asymmetric carbon in R configuration.)
Consisting of repeating units represented by
The polymer block B has at least one polymer unit selected from the group consisting of aliphatic polyester and polyalkylene glycol,
The linking group X is
A group formed by an alkyne-azide reaction,
A block copolymer which is a group formed by a thiol-ene reaction or a group formed by a thiol-in reaction.
2. 2. The block copolymer according to 1 above, wherein the block copolymer is AB type or ABA type.
3. 3. The block copolymer according to 1 or 2, wherein the aliphatic polyester is a ring-opening polymer of caprolactone or polylactic acid, and the polyalkylene glycol is polyethylene glycol.
4). The group formed by the alkyne-azide reaction is a divalent group having a triazole ring, and the group formed by the thiol-ene reaction is an —S—CR ² ₂ —CR ² ₂ — group (wherein R ² represents a hydrogen atom or an organic group), and the group formed by the thiol-in reaction is an —S—CR ³ ₂ ═CR ³ ₂ — group (wherein R ^3. The block copolymer according to any one of 1 to 3 above, which is a divalent group represented by 3) represents a hydrogen atom or an organic group.
5. 5. The block copolymer according to any one of 1 to 4 above, wherein the number average molecular weight of the polymer block B is 1,000 to 1,000,000.
6). 6. The block copolymer according to any one of 1 to 5, wherein the polymer block A contains (R) -3-hydroxybutyric acid as a monomer unit.
7). 7. The block copolymer according to 6 above, wherein the polymer block A further contains (R) -3-hydroxyhexanoic acid as a monomer unit.
8). 8. The block copolymer according to any one of 1 to 7 above, wherein the number average molecular weight of the polymer block A is 1,000 to 1,000,000.
9. The resin composition containing the block copolymer in any one of said 1-8.
10. 10. The resin composition as described in 9 above, which is a molded product.
11. It is a manufacturing method of the block copolymer of any one of said 1-8,
A reactive functional group having a repeating unit represented by the general formula (1) and selected from an alkynyl group, an alkenyl group, a mercaptoalkyl group, an azidoalkyl group, or an allyl (poly) oxyalkyl group at the carboxy terminus. Polymer A to which group A is bonded;
A process for producing a block copolymer in which at least one polymer B selected from the group consisting of an aliphatic polyester having a reactive functional group B capable of forming a chemical bond with the reactive functional group A and a polyalkylene glycol is reacted. .
12 Culturing a microorganism capable of producing polyhydroxyalkanoic acid in a culture solution in which the polymer A contains an alcohol having an alkynyl group, an alkenyl group, a mercaptoalkyl group, an azidoalkyl group, or an allyl (poly) oxyalkyl group. 12. The production method according to 11 above, produced by

  According to the present invention, for example, a novel block copolymer having a PHA block that can be preferably used as a compatibilizing agent for a resin can be obtained.

It is a figure which shows the structural formula of PHBH which the ethynyl group was introduce | transduced into the one terminal obtained in manufacture example 1. FIG. It is a figure which shows the structural formula of PCL in which the azide group was introduce | transduced into the one terminal obtained in manufacture example 2. It is a figure which shows the structural formula of PCL in which the azide group was introduce | transduced into the both terminal obtained in manufacture example 5. 1 is a diagram showing a structural formula of a block copolymer (PHBH-b-PCL) obtained in Example 1. FIG. FIG. 6 is a schematic view showing a structural formula of a block copolymer (PHBH-b-PCL-PHBH) obtained in Example 4. It is a figure which shows the structural formula of PEG in which the azide group was introduce | transduced into the one terminal obtained in manufacture example 6. It is the schematic which shows the structural formula of the block copolymer (PHBH-b-PEG) obtained in Example 5. FIG. Shows the structure analysis of N _3-PCL obtained in Preparation Example 2 (1), end ethynylation PHBH obtained in Production Example 1, and obtained in Example 1 PHBH-b-PCL (1) It is. 6 is a diagram showing the structural analysis result of N ₃ -PEG obtained in Production Example 6. FIG. It is the schematic which shows the structural-analysis result of the block copolymer (PHBH-b-PEG) obtained in Example 5. It is a figure which shows the structural formula of PLLA by which the azide group was introduce | transduced into the one terminal obtained in manufacture example 7. FIG. It is a figure which shows the structural formula of PDLA in which the azide group was introduce | transduced into the one terminal obtained in manufacture example 8. It is the schematic which shows the structural formula of the block copolymer (PHBH-b-PLLA) obtained in Example 6. FIG. It is the schematic which shows the structural formula of the block copolymer (PHBH-b-PDLA) obtained in Example 7. FIG. It is a figure which shows the structural-analysis result of the block copolymer (PHBH-b-PLLA) obtained in Example 6.

  Hereinafter, the present invention will be described in more detail.

The block copolymer of the present invention is a block copolymer in which a polymer block A and a polymer block B are bonded via a linking group X, and the polymer block A has the following general formula (1)
[—C ^* HR ¹ —CH ₂ —CO—O—] (1)
(In the formula, R ¹ is an alkyl group represented by C _n H _{2n + 1} , n is an integer of 1 to 15, and * indicates an asymmetric carbon in R configuration.)
Consisting of repeating units represented by
The polymer block B has at least one polymer unit selected from the group consisting of aliphatic polyester and polyalkylene glycol,
The linking group X is
A group formed by an alkyne-azide reaction,
A group formed by a thiol-ene reaction or a group formed by a thiol-in reaction.

  The polymer block A in the block copolymer of the present invention is a polymer block of (R) -3-hydroxyalkanoic acid produced from a microorganism, and is composed of a repeating unit represented by the general formula (1). Anything can be used. For example, it may be a polymer block of one or two or more (R) -3-hydroxyalkanoic acids satisfying the general formula (1), or may be a polymer block with other polymer [one or two. Polymer block of one or more kinds of (R) -3-hydroxyalkanoic acid and one or more kinds of other (R) -hydroxyalkanoic acids (for example, (R) -4-hydroxyalkanoic acid)] Can be mentioned.

  Examples of the monomer unit constituting the polymer block A include 3HB, 3HP, 4HB, 3HV, 5HV, 3HHx, 6-hydroxyhexanoic acid (hereinafter abbreviated as “6HHx”), and 3-hydroxyheptanoic acid. , 3-hydroxyoctanoic acid, 3-hydroxynonanoic acid, 3-hydroxydecanoic acid, 3-hydroxyundecanoic acid, 3-hydroxydodecanoic acid and the like. The polymer block A may be composed of a single type of monomer unit or may be composed of a plurality of types of monomer units. When the polymer block A is composed of a plurality of types of monomer units, it preferably contains 3HB. The copolymerization ratio of each monomer unit of the polymer block A is not particularly limited, but when 3HB is contained as a monomer unit, the copolymerization ratio is more preferably 50 mol% or more, and 60 mol% or more. More preferably, it is more preferably 70 mol% or more, and particularly preferably 80 mol% or more. The polymer block A is more preferably poly (3HB-co-3HHx) (hereinafter abbreviated as “PHBH”) containing 3HB and 3HHx. When the polymer block A is PHBH, the lower limit of the copolymerization ratio of 3HHx is preferably 1 mol%, more preferably 2 mol%, and even more preferably 3 mol%. The upper limit is preferably 45 mol%, more preferably 42 mol%, and even more preferably 40 mol%. The polymer block A may further contain monomer units other than the above-described 3HB and 3HHx.

  The lower limit of the number average molecular weight of the polymer block A is preferably 1,000, more preferably 2,000, still more preferably 3,000, and particularly preferably 5,000. The upper limit of the number average molecular weight of the polymer block A is preferably 1,000,000, more preferably 800,000, still more preferably 600,000, and particularly preferably 500,000. The number average molecular weight was calculated by standard polystyrene conversion or NMR by size exclusion chromatography SEC (GPC) as described in detail in the Examples section below.

  The polymer block B in the block copolymer of the present invention has at least one polymer unit selected from the group consisting of aliphatic polyesters and polyalkylene glycols. However, the polymer block B is a polymer block different from the polymer block A.

  Examples of the aliphatic polyester include a ring-opening polymer of caprolactone, polybutylene succinate, polyhydroxyalkanoic acid (eg, polylactic acid, poly-3-hydroxyalkanoic acid, etc.), and the like. When the polymer block B is a polylactic acid block, the monomer unit may be only D-lactic acid, L-lactic acid alone, or both D-lactic acid and L-lactic acid. May be. When both D-lactic acid and L-lactic acid are included as monomer units, the polylactic acid block may be a random copolymer of D-lactic acid and L-lactic acid or a block copolymer. When it is a block copolymer, it may be any of a diblock, a triblock, and a multiblock having 4 or more polymer blocks. Of these, the polylactic acid block is preferably a random copolymer from the viewpoint of low crystallinity.

  Examples of the polyalkylene glycol include polyethylene glycol, polypropylene glycol, and polytetramethylene glycol.

  The lower limit of the number average molecular weight of the polymer block B is preferably 1,000, more preferably 2,000, still more preferably 3,000, and particularly preferably 5,000. The upper limit of the number average molecular weight of the polymer block B is preferably 1,000,000, more preferably 800,000, still more preferably 600,000, and particularly preferably 500,000. The number average molecular weight was calculated by standard polystyrene conversion or NMR by size exclusion chromatography SEC (GPC) as described in detail in the Examples section below.

  When the block copolymer of the present invention is used as, for example, a compatibilizing agent, the molecular weights of the polymer block A and the polymer block B are important for the function expression. When the molecular weight of each block is too small, the entanglement of the polymer molecular chain is lowered and the effectiveness as a compatibilizing agent is impaired. On the other hand, when the molecular weight of each block is too large, the dispersibility in the polymer is lowered, and the effectiveness as a compatibilizing agent is impaired. Furthermore, when the molecular weight of each block is too large, the reaction efficiency by the manufacturing method of this invention mentioned later may fall.

The linking group X that binds the polymer block A and the polymer block B is represented by an alkyne-azide reaction, a thiol-ene reaction, a thiol-in reaction, etc. between the polymer block A and the polymer block B. And a group formed by an alkyne-azide reaction, a group formed by a thiol-ene reaction, or a group formed by a thiol-in reaction. As will be described in detail later, the block copolymer of the present invention comprises a polymer A having a specific reactive functional group A bonded to its carboxy terminus, and a reactivity capable of forming a chemical bond with the reactive functional group A. The polymer is produced by reacting a polymer B having a functional group B, and the reactive functional group A and the reactive functional group B are bonded by a click reaction to introduce the linking group X. As a specific example of the linking group X, for example, when a polymer A or a polymer B having an alkynyl group at one terminal (molecular chain terminal) and an azide at the other terminal (molecular chain terminal) is reacted. A triazole ring is introduced by a cyclization reaction between an azide and an alkyne, and a divalent group having a triazole ring is introduced. In addition, when a polymer A or a polymer B having a thiol at one terminal and an alkenyl group at the other terminal is reacted, a —S—CR ² ₂ —CR ² ₂ — group is obtained by a thiol-ene reaction. (In the formula, two R ^{2 s} are the same or different and each represents a hydrogen atom or an organic group). Here, the organic group is, for example, an methylidene group, an ethylidene group, an isopropylidene group, a sec-butylidene group, a benzylidene group, an o-hydroxybenzylidene group or the like, excluding two hydrogen atoms on the same carbon of the hydrocarbon) And a tetramethylene group, a pentamethylene group, etc., which are obtained by removing one hydrogen atom on two different carbons in a hydrocarbon. These organic groups may be further substituted with one or more substituents. Preferably it is a hydrogen atom. In the thiol-in reaction in which a polymer having a thiol at one terminal and a polymer having an alkynyl group at the other terminal is reacted, a —S—CR ³ ₂ = CR ³ ₂ — group (formula Among these, two R ^{3 s} are the same or different and each represents a hydrogen atom or an organic group). What was mentioned above is mentioned as said organic group. Preferably it is a hydrogen atom.

  As described above, the block copolymer of the present invention is obtained by covalently bonding the polymer block A and the polymer block B via the linking group X. In order to exhibit good physical properties, an AB type block copolymer in which the polymer block A and the polymer block B are combined, or an ABA type in which the polymer block A, the polymer block B, and the polymer block A are combined. The block copolymer (the polymer block A is located at the terminal) is preferred. When the latter ABA type block copolymer has two or more polymer blocks A, these polymer blocks A may be the same or different.

  Next, the manufacturing method of the block copolymer of this invention is demonstrated. The block copolymer of the present invention has a repeating unit represented by the general formula (1) and has an alkynyl group, an alkenyl group, a mercaptoalkyl group, an azide alkyl group, or an allyl (poly) at the carboxy terminus. A polymer A to which a reactive functional group A selected from oxyalkyl groups is bonded; an aliphatic polyester having a reactive functional group B capable of forming a chemical bond with the reactive functional group A, and a group consisting of a polyalkylene glycol It is produced by reacting with at least one polymer B selected from

  The alkynyl group is preferably an alkynyl group having 3 to 8 carbon atoms, and examples thereof include a propynyl group, a butynyl group, a pentynyl group, a hexynyl group, a heptynyl group, and an octynyl group. The alkynyl group may be linear or branched, but is preferably linear. The number of carbon atoms is more preferably 3-6, and among them, a propynyl group, a butynyl group, and a hexynyl group are particularly preferable.

  The alkenyl group is preferably an alkenyl group having 3 to 8 carbon atoms, and examples thereof include a propenyl group, a butenyl group, a pentenyl group, a hexenyl group, a heptenyl group, and an octenyl group. The alkenyl group may be linear or branched, but is preferably linear. The number of carbon atoms is more preferably 3 to 6, and among them, a propenyl group, a butenyl group, and a hexenyl group are particularly preferable.

  The mercaptoalkyl group is preferably a group having 2 to 8 carbon atoms in the alkyl group, such as a mercaptoethyl group, a mercaptopropyl group, a mercaptobutyl group, a mercaptopentyl group, a mercaptohexyl group, a mercaptoheptyl group, a mercapto group. An octyl group is mentioned. The alkyl group may be linear or branched, but is preferably linear. As for carbon number, it is more preferred that it is 2-6, and it is still more preferred that it is 3-6. In particular, a mercaptoethyl group and a mercaptopropyl group are preferable.

  The azide alkyl group is preferably a group having 3 to 8 carbon atoms in the alkyl group, for example, azido propyl group, azido butyl group, azide pentyl group, azide hexyl group, azide heptyl group. Is mentioned. The alkyl group may be linear or branched, but is preferably linear. The number of carbon atoms is more preferably 3-6, and particularly preferably an azidopropyl group and an azidobutyl group.

  As the allyl (poly) oxyalkyl group, an alkyl group having 2 to 6 carbon atoms and having 1 to 3 oxyalkyl is preferably used. Oxyalkyl is more preferably 1. Examples of the oxyalkyl group include an oxyethyl group, an oxypropyl group, an oxybutyl group, an oxypentyl group, and an oxyhexyl group. Among them, preferred examples include an oxyethyl group, an oxypropyl group, and an oxybutyl group. The alkyl group may be linear or branched, but is preferably linear. The total carbon number of the allyl (poly) oxyalkyl group is preferably 2 to 6, and examples thereof include an allyloxyethyl group, an allyloxypropyl group, and an allyloxybutyl group.

As described above, the present invention uses a click reaction to introduce a linking group X into a block copolymer, and when the linking group X is a divalent group having a triazole ring introduced by an alkyne-azide reaction, Polymer A and polymer B, which are raw materials, have an alkynyl group at one of these terminals and an azide alkyl group at the other terminal. Specifically, for example, a PHA having an alkyne group at the carboxy terminus (hereinafter sometimes referred to as “terminal alkynylated PHA”) and a caprolactone ring-opening polymer having an azide group at one or both ends, polylactic acid , Polyalkylene glycol and the like can be reacted. In the case of a —S—CR ² ₂ —CR ² ₂ — group in which the linking group X is introduced by a thiol-ene reaction (wherein R ² is as described above), the starting polymer A and polymer B are One having a thiol at one end and an alkenyl group at the other end is used. In the case of a —S—CR ³ ₂ ═CR ³ ₂ — group in which the linking group X is introduced by a thiol-in reaction (wherein R ³ is as described above), the raw materials, polymer A and polymer B, Any of these having a thiol at one end and an alkynyl group at the other end is used. The conditions for introducing the linking group X by the click reaction are not particularly limited, and a known method can be employed.

  The polymer A may be obtained directly by a method using microbial fermentation, or may be obtained by chemically modifying PHA obtained using microbial fermentation. Those obtained directly using are preferred.

  In the case of microbial fermentation, the polymer A can produce polyhydroxyalkanoic acid in a culture solution containing an alcohol having an alkynyl group, an alkenyl group, a mercaptoalkyl group, an azide alkyl group, or an allyl (poly) oxyalkyl group. It is preferably produced by culturing microorganisms. The alcohol is preferably a primary alcohol.

  Examples of such an alcohol include 2-propyn-1-ol, 3-butyn-1-ol, 4-pentyn-1-ol, 5-hexyn-1-ol; 2-propen-1-ol, 3- Buten-1-ol, 4-penten-1-ol, 5-hexen-1-ol; 2-mercaptoethanol, 3-mercaptopropanol, 4-mercaptobutanol, 5-mercaptopentanol, 6-mercaptohexanol; 4- Azidobutan-1-ol, 5-azidopentan-1-ol, 6-azidohexane-1-ol; ethylene glycol monoallyl ether, propylene glycol monoallyl ether, tetramethylene glycol monoallyl ether, pentamethylene glycol monoallyl ether, etc. Is mentioned. Among them, 2-propen-1-ol, 3-buten-1-ol, 5-hexen-1-ol, 2-propyn-1-ol, 3-butyn-1-ol, 5-hexyn-1-ol Preferred examples include 2-mercaptoethanol, 3-mercaptopropanol, and ethylene glycol monoallyl ether. In addition to this, an alcohol having a branched structure in the alkyl chain portion, an alcohol having a plurality of hydroxy groups, or an alcohol having a plurality of alkynyl groups, alkenyl groups, thiol groups, azide groups, or allyl groups may be used. These alcohols may be used alone or in combination of two or more.

  The production method of the polymer A applies a chain transfer reaction in PHA synthesis in microbial cells, and therefore the alcohol used in the production method functions as a chain transfer agent. Therefore, what alcohol functions as a chain transfer agent depends on the substrate specificity of PHA synthase for alcohol. Therefore, the gene encoding the PHA synthase (hereinafter abbreviated as “PHA synthase gene”) possessed by the microorganism used in the method of the present invention is preferably a PHA synthase gene derived from the genus Aeromonas. As an example, a PHA synthase comprising the amino acid sequence described in SEQ ID NO: 1 and derived from Aeromonas caviae, wherein the 149th asparagine is artificially replaced with serine and the 171st aspartic acid is replaced with glycine. Genes can be used. Alternatively, the PHA synthase gene possessed by the microorganism of the present invention is preferably a PHA synthase gene derived from the genus Ralstonia. As an example, a PHA synthase gene derived from Ralstonia eutropha (Ralstonia eutropha) consisting of the amino acid sequence shown in SEQ ID NO: 2 can be used. Alternatively, the PHA synthase gene possessed by the microorganism of the present invention is preferably a PHA synthase gene derived from the genus Pseudomonas. As an example, Pseudomonas Sp. Consisting of the amino acid sequence set forth in SEQ ID NO: 3 is used. A PHA synthase gene derived from 61-3, in which the 325th serine is artificially replaced with threonine, the 477th serine with arginine, and the 481st glutamine with arginine, can be used.

  In the above microorganism, the host is not particularly limited, and may be either a bacterium or a fungus. For example, Acinetobacter (Acinetobacter), Aeromonas (Aeromonas), Alcaligenes (Alkalinegenes), Allochromatium (Arozobium), Azotobacter (Azotobacter) Deria) genus, Candida genus, Caurobacter genus, Chromobacterium genus, Comamonas genus, Cupriavidus genus, Ectothiorhodospira genus bsiella (Klebsiella), Methylobacterium (Methylobacterium), Nocardia (Nocardia), Paracoccus, Pseudomonas, Ralstonia, Rhizobium, Rhizobium, Rhizobium, Rhizobium Genus, Rhodococcus genus, Rhodospirillum genus, Rickettsia genus, Saccharomyces genus, Sinorhobium genus, S cus (Chiokokkasu) genus, Thiocystis (Chiokisuchisu) genus Vibrio (Vibrio) genus, Wautersia (Woterushia) genus, or Zoog / Loea (Zoogu / Roea) include microorganisms belonging to the genus. Of these, microorganisms belonging to the genus Aeromonas, Alcaligenes, Cupriavidus, Escherichia, Pseudomonas, Ralstonia, etc. are preferred, and microorganisms belonging to the genus Cupriavidus, Escherichia, Ralstonia are more preferred, and the microorganism belonging to the genus Cu belonging to the genus Cu is more preferred than the genus Cu belonging to the genus Curiavidus, Escherichia and Ralstonia. Particularly preferred as the host of the microorganism of the present invention is Cupriavidus necator.

  When the host of the microorganism does not have a PHA synthase gene, or when the PHA synthase gene possessed by the host is not the preferred PHA synthase gene, a preferred PHA synthase gene is introduced into the host by gene recombination. Also good. The method for introducing the PHA synthase gene into the host may be a format in which the gene is held in a plasmid or a format in which it is introduced at an arbitrary position on the chromosome. At this time, the PHA synthase gene originally possessed by the host is preferably lost its function. Examples of a method of deleting the function of the PHA synthase gene include, as an example, a method of deleting the PHA synthase gene over its entire length or partially, or addition or deletion of a base to the PHA synthase gene. Alternatively, a method in which the function of the produced PHA synthase is lost by substitution, and the like can be mentioned. For specific methods of inserting or replacing DNA, see, for example, Green, M. et al. R. and Sambrook, J. et al. , 2012, Molecular Cloning: A Laboratory Manual Fourth Ed. , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

  When the polymer A is produced using the microorganism, any raw material can be used as a carbon source at the time of cultivation as long as the microorganism can assimilate the microorganism. Such a carbon source is not particularly limited, but is preferably sugars such as glucose, fructose, sucrose, palm oil, palm kernel oil (hereinafter abbreviated as “PKO”), corn oil, palm Preferred are oils such as oil, olive oil, soybean oil, rapeseed oil and jatropha oil, fractionated oils or refined by-products thereof, or fatty acids such as lauric acid, oleic acid, stearic acid, palmitic acid and myristic acid, and derivatives thereof. . In addition, yeast extract and polypeptone can also be used. More preferred are vegetable oils such as palm oil and palm kernel oil, or palm olein, palm double olein or palm kernel oil olein which is a low melting point fraction obtained by fractionating palm oil or palm kernel oil. From the viewpoint of avoiding competition with food, oil by-products such as PFAD (palm oil fatty acid distillate), PKFAD (palm kernel oil fatty acid distillate), or rapeseed oil fatty acid distillate are particularly preferred.

  In the production of terminal ethynylated PHA by the microorganism, the microorganism may be cultured using a medium containing the carbon source, a nitrogen source that is a nutrient source other than the carbon source, inorganic salts, and other organic nutrient sources. preferable. Examples of the nitrogen source include ammonia, ammonium chloride, urea, ammonium sulfate, ammonium phosphate and other ammonium salts, as well as peptone, meat extract, yeast extract and the like. Examples of inorganic salts include potassium dihydrogen phosphate, disodium hydrogen phosphate, magnesium phosphate, magnesium sulfate, sodium chloride and the like. Examples of other organic nutrient sources include amino acids such as glycine, alanine, serine, threonine, and proline, and vitamins such as vitamin B1, vitamin B12, and vitamin C.

  Conditions such as culture temperature, culture time, culture pH, medium, etc. may be those normally used in the above microorganisms.

  In the present invention, the method for recovering the polymer A from the microbial cells is not particularly limited. For example, the following method can be used. After completion of the culture, the cells are separated from the culture solution with a centrifuge, and the cells are washed with distilled water, methanol, or the like and dried. The polymer A is extracted from the dried cells using an organic solvent such as chloroform. Cellular components are removed from the organic solvent solution containing the polymer A by filtration or the like, and a poor solvent such as methanol or hexane is added to the filtrate to precipitate the polymer A. Further, the supernatant is removed by filtration or centrifugation, and dried to recover the polymer A.

  In the production method of the present invention, for example, the polymer block (PHA) A and the polymer block B are composed of a caprolactone ring-opening polymer block (hereinafter sometimes abbreviated as polycaprolactone block (B1)). In the case of producing an AB-type or ABA-type block copolymer, a raw material (precursor) of the polycaprolactone block (B1) is a ring-opened polymer of caprolactone (hereinafter referred to as azido capodized at one end or both ends). , “One-end azide PCL” or “both-end azide PCL” is preferably used. Of these, the method for producing the former one-terminal azide PCL is not particularly limited. For example, it can be produced by ring-opening polymerization of ε-caprolactone using 6-azido-1-hexanol as an initiator. it can. In these polymerization reactions, it is preferable to use diphenyl phosphate as a catalyst. After the polymerization reaction, the one-terminal azide PCL can be purified by reprecipitation in a methanol / hexane mixture cooled with liquid nitrogen. Moreover, although the manufacturing method of the latter both terminal azide PCL is not specifically limited, As an example, it can manufacture by carrying out the condensation reaction of one terminal azide PCL and 6-azido-1-hexanoic acid. After the polymerization reaction, both terminal azidated PCL can be purified by reprecipitation in methanol cooled with liquid nitrogen. The molecular weights of the one-end azide PCL and the both-end azide PCL can be adjusted by the amount of ε-caprolactone added to the initiator.

  In the production method of the present invention, for example, when producing an AB type or ABA type block copolymer comprising a polymer block (PHA) A and a polylactic acid block (B2) as the polymer block B, As a raw material for the lactic acid block (B2), it is preferable to use polylactic acid in which one end or both ends are azide (hereinafter, abbreviated as “one end azide PLA” or “both ends azide PLA”). Here, the PLA may be any of a homopolymer of L-lactic acid, a homopolymer of D-lactic acid, and a copolymer of L-lactic acid and D-lactic acid (random copolymer, block copolymer). Good. The former method for producing the one-terminal azide PLA is not particularly limited, but as an example, it can be produced by ring-opening polymerization of lactide using 6-azido-1-hexanol as an initiator. In these polymerization reactions, diazabicycloundecene is preferably used as a catalyst. After the polymerization reaction, the one-terminal azide PLA can be purified by reprecipitation in a methanol / hexane mixture cooled with liquid nitrogen. Moreover, although the manufacturing method of the latter both terminal azide PLA is not specifically limited, As an example, it can manufacture by carrying out the condensation reaction of one terminal azide PLA and 6-azido-1-hexanoic acid. After the polymerization reaction, both terminal azide PLA can be purified by reprecipitation in methanol cooled with liquid nitrogen. In addition, the molecular weights of the one-end azide PLA and the both-end azide PLA can be adjusted by the amount of lactide added to the initiator.

  In the production method of the present invention, for example, when producing an AB type or ABA type block copolymer composed of a polymer block (PHA) A and a polyethylene glycol block (B3) as the polymer block B, polyethylene is used. As a raw material for the glycol block (B3), it is preferable to use polyethylene glycol having one or both ends azide (hereinafter abbreviated as “one-end azide PEG” or “both ends azide PEG”). Of these, the production method of the former one-terminal azido PEG is not particularly limited, but as an example, it can be produced by a condensation reaction of polyethylene glycol monomethyl ether having an appropriate molecular weight and 6-azido-1-hexanoic acid. . Further, the method for producing the latter both-end azide PEG is not particularly limited, but as an example, it can be produced by a condensation reaction of polyethylene glycol having an appropriate molecular weight and 6-azido-1-hexanoic acid. One-end azide PEG and both-end azide PEG can be purified by reprecipitation in diethyl ether cooled with liquid nitrogen.

  The block copolymer of the present invention includes, for example, an antioxidant, an ultraviolet absorber, a colorant such as a dye / pigment, a plasticizer, a lubricant, an inorganic filler, an antistatic agent, an antifungal agent, an antibacterial agent, A foaming agent, a flame retardant, etc. can be contained as needed. Moreover, you may contain another crystal nucleating agent. Moreover, this invention is also a resin composition obtained by blending the said block copolymer with other resin.

  The resin composition obtained as described above may be a molded body (form of a molded body). Conventionally known methods can be applied as a molding method of the resin composition for obtaining the molded body, and examples thereof include injection molding, film molding, blow molding, fiber spinning, extrusion foaming, and bead foaming.

  The molded body can be used for various containers, packaging materials, agricultural and horticultural films, medical materials, and the like.

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples. In addition, breeding of strains, analysis method of monomer unit copolymerization ratio contained in PHA, analysis method of molecular weight of block copolymer and raw material polymer thereof (hereinafter sometimes collectively referred to as polymer), polymer structure The analysis method is as follows.

(Breeding of strains)
The genetic manipulations in the examples herein are described in Green, M. et al. R. and Sambrook, J. et al. (2012). In addition, enzymes, cloning hosts, etc. used for gene manipulation can be purchased from market suppliers and used in accordance with the instruction manual. In addition, the enzyme used for an Example etc. will not be specifically limited if it can be used for gene manipulation.

(Analyzing method of copolymerization ratio of monomer units contained in PHA)
The monomer unit copolymerization ratio contained in PHA was analyzed using ¹ H-NMR. 2 mg of the obtained PHA was dissolved in 2 mL of deuterated chloroform and transferred to a sample tube for measurement. From the area of each detected peak, the copolymerization ratio of the monomer units was calculated.

(Method for analyzing molecular weight of polymer)
The number average molecular weight, weight average molecular weight, and molecular weight dispersion of the polymer were determined by size exclusion chromatography (Size Exclusion Chromatography; hereinafter referred to as SEC measurement). About 1 mg of the polymer was dissolved in 1 mL of tetrahydrofuran and passed through a 0.45 μm membrane filter, and then SEC measurement was performed with an eluent: tetrahydrofuran, a flow rate: 1.0 mL / min, and a column temperature: 40 ° C. Polystyrene was used as a standard substance.
However, as an exception, N ₃ -PEG produced in Production Example 6 and PHBH-b-PEG produced in Example 5 were subjected to SEC measurement under the following conditions. First, 1 mg of polymer was dissolved in 1 mL of dimethylformamide, passed through a 0.45 μm membrane filter, and SEC measurement was performed with eluent: dimethylformamide containing 10 mM lithium chloride, flow rate: 0.6 mL / min, column temperature: 40 ° C. It was. Polystyrene was used as a standard substance.

(Method for analyzing PHBH structure and method for calculating NMR estimated molecular weight)
The structural analysis of PHBH was performed by ¹ H-NMR, ¹ H- ¹ H COSY, and HMQC measurements. From the measurement results, detailed peaks of the backbone of the PHBH main chain and the terminal ethynyl group were assigned, and the copolymer composition ratio of the monomer units contained in PHBH was calculated from the integration ratio.
The absolute molecular weight of PHBH was calculated by ¹ H-NMR measurement. As measurement conditions, about 1 mg of PHBH was dissolved in 600 μL of a CDCl ₃ solution, and measurement was performed with 128 integrations. Since a peak derived from the methylene proton adjacent to the ethynyl group was observed at 4.67 ppm, the integrated value of this peak was set to 2, and the proton (5.2 to 5.2) bonded to the carbon adjacent to the alkyl group of the main chain. The degree of polymerization of the PHBH main chain skeleton was calculated from the ratio with the integrated value of the peak at around 5.3 ppm. Since the molecular weight of the main chain skeleton is randomly copolymerized with molecules with different side chains, after calculating the copolymer composition ratio, add the product of each composition ratio and the molecular weight of the P3HB part and P3HHx part. The molecular weight of the main chain skeleton was calculated. Finally, the absolute molecular weight of PHBH was calculated from ¹ H-NMR by adding the molecular weight of the terminal portion to the product of the molecular weight of the main chain skeleton portion and the degree of polymerization.

(Method for analyzing terminal azide polymer structure and method for calculating NMR estimated molecular weight)
The structural analysis of the terminal azidated polymer was performed by ¹ H-NMR. As measurement conditions, about 1 mg of a polymer was dissolved in 600 μL of a CDCl ₃ solution, and measurement was performed with 128 integrations. Since a peak derived from the methylene proton adjacent to the azide group was observed at 3.25 ppm, the integrated value of this peak was set to 2, and the polymer main component was calculated from the ratio of the integrated value of the peak derived from the main chain structure of each polymer. The degree of polymerization of the chain skeleton was calculated. Finally, the absolute molecular weight of the terminal azide polymer was calculated from ¹ H-NMR by adding the molecular weight of the terminal portion to the product of the molecular weight of the main chain skeleton portion and the degree of polymerization.
However, as an exception, only when calculating the molecular weight of N ₃ -PCL-N ₃ prepared in Production Example 5, the integrated value of the peak derived from the methylene proton adjacent to the azide group was set to 4.

(Block copolymer structure analysis method and NMR estimated molecular weight calculation method)
The structural analysis of the block copolymer was performed by ¹ H-NMR. As measurement conditions, about 1 mg of a polymer was dissolved in 600 μL of a CDCl ₃ solution, and measurement was performed with 128 integrations.
First, a copolymer composition was calculated for PHBH in the block copolymer by the same method as described above, and based on this, the molecular weight (1) of the main chain portion of PHBH was calculated. For the terminal azide polymer of the block copolymer, the molecular weight (2) of the main chain portion was calculated in the same manner as described above.
Next, the integral value of the proton peak derived from the formed triazole ring is set to 1, the integral ratio of the peak derived from the main chain of PHBH (3), and the integral ratio of the peak derived from the main chain of the terminal azide polymer (4) was obtained. Using the above values, the molecular weight of the block copolymer was calculated by the following formula.
(Molecular weight of block copolymer) = (1) × (3) / 2 + (2) × (4) / 2 + (molecular weight in the vicinity of the linked structure)
However, as an exception, only when calculating the molecular weight of PCL-b-PHBH-b-PCL prepared in Example 4, the integral value of the proton peak derived from the triazole ring was set to 2.

<Production Example 1 Production of Terminal Ethynylated PHBH>
As the microorganism, KNK-005 trc-phaJ4b / ΔphaZ1,2,6 strain described in WO2015 / 115619 was used. The KNK-005 trc-phaJ4b / ΔphaZ1,2,6 strain has a full length deletion of the phaZ1 gene and phaZ6 gene on the chromosome, a deletion from the 16th codon to the stop codon of the phaZ2 gene, and SEQ ID NO: on the chromosome. 4. A strain having the PHA synthase gene described in 4 and having an expression regulatory sequence described in SEQ ID NO: 5 inserted upstream of the phaJ4b gene.

(culture)
The strain was cultured as follows.
The composition of the seed medium is 10 g / L meat extract, 10 g / L bactotryptone, 2 g / L yeast extract, 9 g / L disodium hydrogen phosphate 12 hydrate, 1.5 g / L potassium dihydrogen phosphate, pH 6 .8.
The composition of the preculture medium is 5.5 g / L disodium hydrogen phosphate 12 hydrate, 0.95 g / L potassium dihydrogen phosphate, 6.44 g / L ammonium sulfate, 1 g / L magnesium sulfate heptahydrate, 5 mL / L trace metal salt solution (16 g / L iron (III) chloride hexahydrate in 0.1 N hydrochloric acid, 10 g / L calcium chloride dihydrate, 0.2 g / L cobalt chloride hexahydrate, 0. 16 g / L copper sulfate pentahydrate and 0.12 g / L nickel chloride hexahydrate). As the carbon source, palm single olein oil was used at a concentration of 30 g / L.
The composition of the PHA production medium is 5.5 g / L disodium hydrogen phosphate 12 hydrate, 0.95 g / L potassium dihydrogen phosphate, 6.44 g / L ammonium sulfate, 1 g / L magnesium sulfate heptahydrate, 5 mL / L trace metal salt solution (16 g / L iron (III) chloride hexahydrate in 0.1 N hydrochloric acid, 10 g / L calcium chloride dihydrate, 0.2 g / L cobalt chloride hexahydrate, 0. 16 g / L copper sulfate pentahydrate and 0.12 g / L nickel chloride hexahydrate). As the carbon source, palm single olein oil was used at a concentration of 30 g / L.

  Inoculate 50 mL of the glycerol stock of each strain into 10 mL of seed medium and incubate for 24 hours, and add 1.0% to 3 L jar fermenter (Bioneer-300L type manufactured by Marubishi Bioengineer) containing 1.8 L of preculture medium. v / v) Inoculated. The operating conditions were a culture temperature of 34 ° C., a stirring speed of 500 rpm, an aeration rate of 1.8 L / min, and culturing for 28 hours while controlling the pH between 6.7 and 6.8. A 14% aqueous ammonium hydroxide solution was used for pH control.

  PHA production culture was performed as follows. First, 2-propyn-1-ol was added at a concentration of 0.1% (w / v) to a 10 L jar fermenter (Bioneer-1000L type manufactured by Maruhishi Bio-Engineering Co., Ltd.) containing 4 L of PHA production medium. 25% (v / v) of the preculture seed mother was inoculated. The operating conditions were a culture temperature of 34 ° C., a stirring speed of 600 rpm, an aeration rate of 6.0 L / min, and a pH controlled between 6.7 and 6.8. A 25% aqueous ammonium hydroxide solution was used for pH control. The culture was performed for 45 to 54 hours.

(Purification)
Purification was performed by partially modifying the method described in International Publication No. WO2010 / 066753.
First, the culture solution obtained in the above culture step was sterilized by heating and stirring at an internal temperature of 60 to 70 ° C. for 20 minutes. Thereafter, 0.2% by weight sodium dodecyl sulfate was added to the culture medium. Furthermore, after adding sodium hydroxide so that pH might be set to 11.0, it heat-retained at 50 degreeC for 1 hour. Thereafter, high-pressure crushing was performed at a pressure of 450 to 550 kgf / cm ² with a high-pressure crusher (high-pressure homogenizer model PA2K manufactured by Niroso Avi).
An equal amount of distilled water was added to the crushed liquid after high-pressure crushing. After further centrifugation, the supernatant was removed (3-fold concentration). To the aqueous suspension of PHA concentrated 3 times, the same amount of water as the excluded supernatant was added and suspended. 0.2% by weight sodium dodecyl sulfate and 1/100 weight protease of PHA (Novozyme) Product, Esperase) was added, and the mixture was stirred for 2 hours while maintaining the pH at 50 ° C.
After the PHA was centrifuged from the enzyme-treated solution after stirring to remove the supernatant, the same amount of water as the removed supernatant was added to wash the PHA. After performing this operation three times, the precipitate obtained after centrifugation was collected and dried under a vacuum condition at 60 ° C. until there was no change in weight to obtain crude purified PHA.

70 g of the obtained PHA was dissolved in 450 mL of dichloromethane, added to 2 L of ice-cold methanol, and reprecipitated to obtain a white powder. Further, 30 g of the obtained white powder was dissolved in 90 mL of dichloromethane, and added to 900 mL of normal temperature methanol for reprecipitation. The obtained precipitate was dried under reduced pressure to obtain terminal alkyne PHA.
When the structure of the obtained terminal alkynylated PHA was identified by ¹ H-NMR spectrum measurement, it was PHBH in which an ethynyl group was introduced at the terminal shown in FIG. 1 (sometimes referred to as “terminal ethynylated PHBH”). It was confirmed. The 3HHx copolymerization ratio of the obtained terminal ethynylated PHBH was 38 mol%. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

<Production Example 2 Synthesis of N ₃ -PCL (1)>
In a dry box, 50 mmol of ε-caprolactone, 1 mmol of 6-azido-1-hexanol and 0.05 mmol of diphenylphosphoric acid were added to a 15 mL capacity vial, and polymerization was performed at 80 ° C. After 6 hours, an excessive amount of anion exchange resin Amberlyst A21 (manufactured by Organo) was added to terminate the polymerization. Thereafter, purified by reprecipitation into liquid nitrogen cooled methanol / hexane (90:10) mixed solvent gave N _3-PCL (1) of a white powder.
When the structure of the obtained N ₃ -PCL (1) was identified by ¹ H-NMR spectrum measurement, it was confirmed to be PCL in which an azide group was introduced at one end shown in FIG. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

<Production Example 3 Synthesis of N ₃ -PCL (2)>
In a dry box, 100 mmol of ε-caprolactone, 1 mmol of 6-azido-1-hexanol and 1 mmol of diphenylphosphoric acid were added to a 15 mL capacity vial, and polymerization was performed in a toluene solution at room temperature. After 21 hours, an excessive amount of anion exchange resin Amberlyst A21 was added to terminate the polymerization. Thereafter, purified by reprecipitation into liquid nitrogen cooled methanol / hexane (90:10) mixed solvent, to obtain a white powder of N _3-PCL (2).
When the structure of the obtained N ₃ -PCL (2) was identified by ¹ H-NMR spectrum measurement, it was confirmed to be PCL in which an azido group was introduced at one end shown in FIG. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

<Production Example 4 Synthesis of N ₃ -PCL (3)>
In a dry box, 200 mmol of ε-caprolactone, 1 mmol of 6-azido-1-hexanol and 1 mmol of diphenylphosphoric acid were added to a 15 mL capacity vial, and polymerization was performed in a toluene solution at room temperature. After 21 hours, an excessive amount of anion exchange resin Amberlyst A21 was added to terminate the polymerization. Thereafter, purified by reprecipitation into liquid nitrogen cooled methanol / hexane (90:10) mixed solvent, to obtain a white powder of N _3-PCL (3).
When the structure of the obtained N ₃ -PCL (3) was identified by ¹ H-NMR spectrum measurement, it was confirmed to be PCL having an azido group introduced at one end shown in FIG. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

<Production Example 5 Synthesis of N ₃ -PCL-N ₃ >
Schlenk tube containing 5 g of one-terminal azide-modified PCL (1) produced in Production Example 2, 454 mg of N, N-dimethyl-4-aminopyrrolidine, and 776 mg of 1-ethyl-3- (3-dimethylamidopropyl) carboxamide hydrochloride 20 mL of a dichloromethane solution in which 424 mg of 6-azido-1-hexanoic acid was dissolved was added after argon bubbling, and a condensation reaction was performed. The reaction was carried out at 25 ° C. for 20 hours under an argon atmosphere. The solvent was distilled off under reduced pressure to obtain a crude product, which was then added to methanol cooled with liquid nitrogen and reprecipitated to obtain N ₃ -PCL-N ₃ as a white powder.
When the structure of the obtained N ₃ -PCL-N ₃ was identified by ¹ H-NMR spectrum measurement, it was confirmed to be PCL having an azido group introduced at both ends shown in FIG. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

<Synthesis of Example 1 PHBH-b-PCL (1)>
In a Schlenk tube containing 4 g of terminal ethynylated PHBH obtained in Production Example 1, 2.05 g of N ₃ -PCL (1) obtained in Production Example 2, and 69.4 mg of copper bromide, N, N, N ′, Click reaction was performed by adding 20 mL of dichloromethane in which 0.10 mL of N ″, N ″ -pentamethyldiethylenetriamine was dissolved after argon bubbling. The reaction was carried out at 25 ° C. for 48 hours under an argon atmosphere. After confirming the progress of the reaction by ¹ H-NMR, in order to remove excess N ₃ -PCL (1), a polystyrene resin having an ethynyl group was added to the system, followed by a click reaction at 25 ° C. for 72 hours. It was. Thereafter, the reaction solution was diluted with dichloromethane, and after removing polystyrene resin and copper bromide using an alumina column, the solvent was distilled off under reduced pressure to obtain solid PHBH-b-PCL (1).
When the structure of the obtained PHBH-b-PCL (1) was identified by ¹ H-NMR spectrum measurement, it was an AB type block copolymer having a PHBH block and a PCL block (1) shown in FIG. It was confirmed. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

<Synthesis of Example 2 PHBH-b-PCL (2)>
In the same manner as in Example 1, PHBH-b-PCL (2) was obtained using terminal ethynylated PHBH obtained in Production Example 1 and N ₃ -PCL (2) obtained in Production Example 3.
When the structure of the obtained PHBH-b-PCL (2) was identified by ¹ H-NMR spectrum measurement, it was an AB type block copolymer having a PHBH block and a PCL block (2) shown in FIG. It was confirmed. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

<Synthesis of Example 3 PHBH-b-PCL (3)>
In the same manner as in Example 1, PHBH-b-PCL (3) was obtained using the terminal ethynylated PHBH obtained in Production Example 1 and N ₃ -PCL (3) obtained in Production Example 4.
When the structure of the obtained PHBH-b-PCL (3) was identified by ¹ H-NMR spectrum measurement, it was an AB type block copolymer having a PHBH block and a PCL block (3) shown in FIG. It was confirmed. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

<Synthesis of Example 4 PHBH-b-PCL-b-PHBH>
To a Schlenk tube containing 2.07 g of terminal ethynylated PHBH obtained in Production Example 1, 0.50 g of N ₃ -PCL-N ₃ obtained in Production Example 5 and 36.8 mg of copper bromide, N, N, N A click reaction was performed by adding 15 mL of dichloromethane in which 53.5 μL of ', N ″, N ″ -pentamethyldiethylenetriamine was dissolved after argon bubbling. The reaction was carried out at 25 ° C. for 72 hours under an argon atmosphere. After confirming the progress of the reaction, in order to remove the excessive amount of N ₃ -PCL-N ₃ , a polystyrene resin having an ethynyl group was added to the system, followed by a click reaction at 25 ° C. for 72 hours. Thereafter, the reaction solution was diluted with dichloromethane, and after removing polystyrene resin and copper bromide using an alumina column, the solvent was distilled off under reduced pressure to obtain solid PHBH-b-PCL-b-PHBH.
When the structure of the obtained PHBH-b-PCL-b-PHBH was identified by ¹ H-NMR spectrum measurement, it was an ABA type block copolymer having a PHBH block and a PCL block shown in FIG. confirmed. The molecular weights calculated by NMR analysis and SEC measurement are shown in Table 1, respectively.

(Results and discussion)
The structural analysis result of the terminal ethynylated PHBH obtained in Production Example 1 is shown in FIG. Since a peak a (δ = 4.67 ppm) derived from a methylene proton adjacent to the ethynyl group was observed, an ethynyl group at the PHBH terminal was confirmed.
The structural analysis result of N ₃ -PCL (1) obtained in Production Example 2 is shown in FIG. The peak b (δ = 3.25 ppm) derived from the methylene proton adjacent to the azide group was observed, and the molecular weight of Production Example 2 shown in Table 1 was almost the same as the theoretical molecular weight calculated from the monomer addition amount. It was confirmed that N ₃ -PCL (1) was synthesized. By the same method, it was confirmed that the intended polymers were synthesized for the polymers of Production Examples 3 to 5.
The terminal ethynylated PHBH obtained in Production Example 1 and N ₃ -PCL (1) obtained in Production Example 2 were linked by a click reaction according to the method described in Example 1, and the resulting PHBH-b-PCL was obtained. The structural analysis result of (1) is shown in FIG. 8, but the disappearance of the peak a, the shift of the peak b to the peak c (near 4.3 to 4.4 ppm), and the peak e (from the triazole ring formation) (δ = 7.60 ppm) was observed, and the other peaks were assigned consistently. In addition, from the results of Table 1, an increase in molecular weight was observed in Example 1 as compared with Production Example 1 and Production Example 2. From the above results, it was confirmed that the desired PHBH-b-PCL (1) was synthesized. The same procedure was performed for the polymers of Examples 2-4. In Example 2, the SEC trace of the product was shifted to the high molecular weight side while maintaining the unimodality compared to before the reaction. From these results, it was confirmed that the intended polymers were synthesized for the polymers of Examples 2 to 4.

<Synthesis of Production Example 6 N _3-PEG>
To a Schlenk tube containing 1 g of polyethylene glycol monomethyl ether, 52.9 mg of N, N-dimethyl-4-aminopyrrolidine, and 82.9 mg of 1-ethyl-3- (3-dimethylamidopropyl) carboxamide hydrochloride, 6-azido- 15 mL of dichloromethane in which 45.3 mg of 1-hexanoic acid was dissolved was added after argon bubbling to carry out a condensation reaction. The reaction was carried out at 25 ° C. for 72 hours under an argon atmosphere. After confirming the progress of the reaction, impurities were removed by passing through an alumina column. Then, the crude product obtained by distilling off the solvent under reduced pressure was added into diethyl ether cooled with liquid nitrogen and reprecipitated to obtain white powder of N ₃ -PEG.
When the structure of the obtained N ₃ -PEG was identified by ¹ H-NMR spectrum measurement, it was confirmed to be a PEG having an azido group introduced at one end as shown in FIG.

<Synthesis of Example 5 PHBH-b-PEG>
In a Schlenk tube containing 110 mg of terminal ethynylated PHBH obtained in Production Example 1, 100 mg of N ₃ -PEG obtained in Production Example 6 and 1.7 mg of copper bromide, N, N, N ′, N ″, N ′ Dichloromethane (10 mL) in which 2.7 μL of '-pentamethyldiethylenetriamine was dissolved was added after argon bubbling, and a click reaction was performed. The reaction was carried out at 25 ° C. for 48 hours under an argon atmosphere. After confirming the progress of the reaction, a polystyrene resin having an azide group was added to the system in order to remove the excessively added terminal ethynylated PHBH, and then a click reaction was performed at 25 ° C. for 48 hours. Thereafter, the reaction solution was diluted with dichloromethane, and after removing polystyrene resin and copper bromide using an alumina column, the solvent was distilled off under reduced pressure to obtain solid PHBH-b-PEG.
When the structure of the obtained PHBH-b-PEG was identified by ¹ H-NMR spectrum measurement, it was confirmed to be an AB type block copolymer having a PHBH block and a PEG block shown in FIG.

(Results and discussion)
The structural analysis results of N ₃ -PEG obtained in Production Example 6 are shown in FIG. The ¹ H-NMR spectrum of the product after purification is consistently assigned, and the peak d derived from the methyl group proton at the PEG end (δ = 3.38 ppm) and the peak a derived from the methylene proton adjacent to the azide group (δ = From the integration ratio of 3.25 ppm, the quantitative progress of the condensation reaction was confirmed. As a result of the SEC measurement of the product, the trace showed unimodality, and the shape of the peak did not change before and after the reaction. From the above results, it was confirmed that the target N ₃ -PEG was synthesized.
The terminal ethinylated PHBH obtained in Production Example 1 and the N ₃ -PEG obtained in Production Example 6 were linked by a click reaction according to the method described in Example 5, and the structural analysis results of the obtained PHBH-b-PEG were obtained. As shown in FIG. 10, the disappearance of the peak a and the peak e (δ = 7.60 ppm) derived from the formation of the triazole ring were observed, and other peaks were also assigned without contradiction. In addition, from the results of Table 2, an increase in molecular weight was observed in Example 5 as compared with Production Example 1 and Production Example 6. From the above results, it was confirmed that the target PHBH-b-PEG was synthesized.

<Production Example 7 Synthesis of N ₃ -PLLA>
L-(-)-lactide 900 mg, 6-azido-1-hexanol 12.8 mg, and diazabicycloundecene 13.6 mg were added to a 15 mL vial in a dry box, and 25 ° C. in 6.24 mL of dichloromethane. The polymerization was carried out at After 20 minutes, an excess amount of benzoic acid was added to terminate the polymerization. Thereafter, purified by reprecipitation into methanol solution cooled with liquid nitrogen to obtain N ₃ -PLLA white solid.
When the structure of the obtained N ₃ -PLLA was identified by ¹ H-NMR spectrum measurement, it was confirmed that it was poly-L-lactic acid (PLLA) having an azido group introduced at one end as shown in FIG. .

<Production Example 8 Synthesis of N ₃ -PDLA>
N ₃ -PDLA was obtained by using D-(−)-lactide instead of L-(−)-lactide in the same manner as in Production Example 7.
When the structure of the obtained N ₃ -PDLA was identified by ¹ H-NMR spectrum measurement, it was confirmed that it was poly-D-lactic acid (PDLA) having an azido group introduced at one end as shown in FIG. .

<Synthesis of Example 6 PHBH-b-PLLA>
In a Schlenk tube containing 250 mg of terminal ethynylated PHBH obtained in Production Example 1, 200 mg of N ₃ -PLLA obtained in Production Example 7, and 3.87 mg of copper bromide, N, N, N ′, N ″, N ′ A click reaction was performed by adding 5.6 μL of dichloromethane in which 5.6 μL of '-pentamethyldiethylenetriamine was dissolved, after argon bubbling. The reaction was carried out at 25 ° C. under an argon atmosphere for 24 hours. After confirming the progress of the reaction, a polystyrene resin having an azide group was added to the system in order to remove the excessively added terminal ethynylated PHBH, and then click reaction was carried out at 25 ° C. for 96 hours. Thereafter, the reaction solution was diluted with dichloromethane, and after removing the polystyrene resin and copper bromide using an alumina column, the solvent was distilled off under reduced pressure to obtain solid PHBH-b-PLLA.
When the structure of the obtained PHBH-b-PLLA was identified by ¹ H-NMR spectrum measurement, it was confirmed to be an AB type block copolymer having a PHBH block and a PLLA block shown in FIG.

<Synthesis of Example 7 PHBH-b-PDLA>
In the same manner as in Example 6, PHBH-b-PDLA was obtained using the terminal ethynylated PHBH obtained in Production Example 1 and N ₃ -PDLA obtained in Production Example 8.
When the structure of the obtained PHBH-b-PDLA was identified by ¹ H-NMR spectrum measurement, it was confirmed to be an AB type block copolymer having a PHBH block and a PDLA block shown in FIG.

(Results and discussion)
The terminal ethynylated PHBH obtained in Production Example 1 and N ₃ -PLLA obtained in Production Example 7 are linked by a click reaction according to the method described in Example 6, and the resulting PHBH-b-PLLA is structured. The analysis result is shown in FIG. 15, and the disappearance of the peak a (δ = 4.67 ppm) derived from the methylene proton adjacent to the ethynyl group of ethinylated PHBH and the peak j (δ = 7. 60 ppm) was observed, and other peaks were assigned consistently. Further, from the results of Table 3, an increase in molecular weight was observed in Example 6 and Example 7 as compared with Production Example 1, Production Example 7, and Production Example 8. From the above results, it was confirmed that the target PHBH-b-PLLA and PHBH-b-PDLA were synthesized.

Claims (12)

A block copolymer in which a polymer block A and a polymer block B are bonded via a linking group X, and the polymer block A has the following general formula (1)
[—C ^* HR ¹ —CH ₂ —CO—O—] (1)
(In the formula, R ¹ is an alkyl group represented by C _n H _{2n + 1} , n is an integer of 1 to 15, and * indicates an asymmetric carbon in R configuration.)
Consisting of repeating units represented by
The polymer block B has at least one polymer unit selected from the group consisting of aliphatic polyester and polyalkylene glycol,
The linking group X is
A group formed by an alkyne-azide reaction,
A block copolymer which is a group formed by a thiol-ene reaction or a group formed by a thiol-in reaction.   The block copolymer according to claim 1, wherein the block copolymer is an AB type or an ABA type.   The block copolymer according to claim 1 or 2, wherein the aliphatic polyester is a ring-opening polymer of caprolactone or polylactic acid, and the polyalkylene glycol is polyethylene glycol. The group formed by the alkyne-azide reaction is a divalent group having a triazole ring, and the group formed by the thiol-ene reaction is an —S—CR ² ₂ —CR ² ₂ — group (wherein R ² represents a hydrogen atom or an organic group), and the group formed by the thiol-in reaction is an —S—CR ³ ₂ ═CR ³ ₂ — group (wherein R The block copolymer according to any one of claims 1 to ³ , wherein ³ represents a hydrogen atom or an organic group.   The number average molecular weight of the said polymer block B is 1,000-1,000,000, The block copolymer of any one of Claims 1-4.   The block copolymer according to any one of claims 1 to 5, wherein the polymer block A contains (R) -3-hydroxybutyric acid as a monomer unit.   The block copolymer according to claim 6, wherein the polymer block A further contains (R) -3-hydroxyhexanoic acid as a monomer unit.   The block copolymer according to any one of claims 1 to 7, wherein the number average molecular weight of the polymer block A is 1,000 to 1,000,000.   The resin composition containing the block copolymer of any one of Claims 1-8.   The resin composition according to claim 9, which is a molded body. A method for producing a block copolymer according to any one of claims 1 to 8,
A reactive functional group having a repeating unit represented by the general formula (1) and selected from an alkynyl group, an alkenyl group, a mercaptoalkyl group, an azidoalkyl group, or an allyl (poly) oxyalkyl group at the carboxy terminus. Polymer A to which group A is bonded;
A process for producing a block copolymer in which at least one polymer B selected from the group consisting of an aliphatic polyester having a reactive functional group B capable of forming a chemical bond with the reactive functional group A and a polyalkylene glycol is reacted. .   Culturing a microorganism capable of producing polyhydroxyalkanoic acid in a culture solution in which the polymer A contains an alcohol having an alkynyl group, an alkenyl group, a mercaptoalkyl group, an azidoalkyl group, or an allyl (poly) oxyalkyl group. The manufacturing method of Claim 11 manufactured by these.

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