JP2622799B2 - Method of producing useful substances from waste - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing useful substances by microorganisms using food waste and processed wastes as raw materials, that is, a method for recycling wastes. For organic waste generated in large quantities from food factories such as microorganisms, coffee grounds, bran, okara, etc., extract microbial assimilating substances contained in waste in solid or liquid form by specific means. And a method for producing a specific useful substance by a useful microorganism using the same as a culture medium.

[0002]

2. Description of the Related Art Conventionally, organic wastes from food manufacturing factories and processing plants such as beer lees, brewer's yeast, coffee grounds, bran, and okara are used as they are, or after being dried or modified by some microorganisms. And sell it to external users as fertilizer and / or feed.

[0003] However, in fact, feed used for multi-head breeding of fertilizers, livestock, and poultry used for agriculture is,
Low-value-added fertilizers and feeds produced using such wastes as raw materials are produced or imported in Japan and abroad because homogeneous and inexpensive fertilizers and feeds are produced or imported. It cannot compete with fertilizer and feed produced as raw materials in terms of cost.

[0004] For example, at present, a part of surplus brewer's yeast is sold as feed (additive) for livestock, but the cost required for drying is 120 yen / kg, while the selling price of the user is less. It is 60 yen / kg, and a complete deficit production has been forced. In response to this reality, there has been a strong demand from the food industry for the emergence of a technology for extracting or producing high value-added useful substances using organic wastes as raw materials.

[0005]

As described in detail above, the prior art recycling method and / or technology using organic waste directly as raw material, that is, the production of fertilizers, feeds, etc., is an additional process. It is not feasible as a business that has low value and can essentially secure profits. The present invention has solved the above-mentioned problems of the prior art, and the residue of the primary material used for fermentation and other purposes is not organic waste, but the primary material is qualitatively converted. and it was evaluated as a new raw material (secondary raw materials), to produce high-value-added materials primarily using microorganisms function, self-contained microorganisms in consideration as to minimize the material to be disposed of final It is an object of the present invention to provide a method for producing a useful substance according to the present invention.

[0006]

The present invention mainly relates to microorganisms
Producing lytic enzymes or lytic enzymes from organic waste consisting of
Solubilized by the first microorganism having the function of releasing it from the body
To produce and accumulate useful substances in the body using the culture medium obtained
This is a method for producing a useful substance from waste, characterized by culturing two microorganisms (also referred to as useful microorganisms) , whereby the above problem can be solved. TECHNICAL FIELD The present invention relates to a technology for producing a novel useful substance having a technical idea substantially different from that of the conventional technology, and effectively utilizes various organic wastes mainly discharged from food factories in a low-dimensional manner. Alternatively process, regarded as a secondary resource before disposal, produce or qualitatively so-called self-contained you rolling conversion plant and / or substances producing useful substances by use of what is now prepared as culture media useful microorganisms At the same time, an object of the present invention is to provide an innovative technology for producing useful substances by using microorganisms for the purpose of establishing a treatment technique capable of simultaneously achieving waste treatment.

[0007] The culture medium used in the present invention is mainly a microorganism.
It is prepared from organic waste consisting of the body . In the organic waste used in the present invention , the microorganism is preferably a living cell such as brewer's yeast, but may be a dead cell . In the present invention, components that can be contained in organic waste are
In addition, organic solutions, organic solids (for example, beer lees,
Okara, etc.) . Form of organic waste to be made tone is optional, may be a solid in a liquid, extraction of the culture medium may be appropriately selected depending on the separation technique. For example, in the case of a solid, the culture medium may be extracted as it is by pressing or the like. Alternatively, the culture medium may be separated and extracted by converting the solid state into a liquid state or the liquid state into a solid state. As a solvent for obtaining a liquid organic waste, water is usually used, but if desired, another suitable solvent may be used in combination.

[0008] A preferred method of preparing the culture medium is
For example, the following may be mentioned. When the components of the organic waste are microorganisms such as cells, the microorganisms are treated with a lytic enzyme such as lysozyme to decompose (dissolve) the cell wall and cell membrane and to dissolve or disperse the intracellular components. . In place of the lytic enzyme, a microorganism producing the enzyme,
For example, Myxococcus, Myxobacter, Mycobacter, Cystobacter, Stigmatella (St)
igmatella) and the like.

[0009] as a component of organic waste, coffee grounds, if it contains solids such as spent grains, formed extracted by dispersing it should be processed by pressing or the like, or water
How to mix minutes with things . In this case, the solid content may be pulverized to a desired particle size, and the temperature of the water to be used may be selected to obtain a desired medium component. The culture medium is effective for effectively culturing the microorganisms and for facilitating the purification of the useful substances, which is obtained by sufficiently removing water-insoluble substances which cannot be decomposed by the useful microorganisms (here, And produces lytic enzymes for insolubles
As a means for removing such insolubles and extracting a culture medium, conventionally known separation means, for example, filtration (membrane, screen, cloth, etc.), centrifugation, spontaneous sedimentation and the like can be mentioned. .

[0010] Further, in the culture medium, when there is a component whose component is lacking in a desired microorganism culture, the component may be appropriately supplemented. The kind of useful microorganism used in the present invention can be variously selected without particular limitation according to the target useful substance. Useful substances produced inside and outside the microorganism
And utilize useful substances accumulated in the microorganisms.

[0011] Further, an organic waste containing the microorganism newly produced after culturing a useful microorganism producing a useful substance with the prepared culture medium is used as an organic waste as a raw material of the culture medium in the present invention (recycled). Ru can also be included) to.

The useful substance produced according to the present invention is separated from microorganisms and then separated from various microorganisms by various means known in the art.
For example, it can be purified by a precipitation method, a solvent extraction method, various types of chromatography, ultrafiltration and the like. Hereinafter, the present invention will be described in more detail. In the present invention, food production, various useful substances from organic waste discharged from the processing plant,
For example, when producing and manufacturing fine chemicals such as bioactive substances, chemicals, enzymes, and vitamins ,
The organism produces a specific lytic enzyme and / or lytic enzyme.
Depending on the function of the microorganisms produced,
Take out the active ingredient and remove it mainly from solid waste.
The various active ingredients necessary for the growth of microorganisms are at room temperature
Extraction with water, hot water or hot water, and using these as a culture medium for useful microorganisms, culturing specific microorganisms, it is possible to produce specific useful substances with extremely high added value at low cost. This is the gist of the present application.

Heretofore, it has been customary to convert this kind of organic waste into fertilizer and / or feed as it is, by slight drying, or by utilizing the function of microorganisms. Competitive and unprofitable, both qualitatively and costly, hindered the recycling of organic waste as a method of solving global environmental problems.

The novel method for producing useful substances by microorganisms according to the present invention is an innovative recycling method that fundamentally solves the fatal deficiencies of this recycling method, and produces high value-added substances. By doing so, it is economically feasible and the waste can be treated at the same time. As a total process, the present invention is a multifunctional and excellent resource for organic waste that is unprecedented. It is a chemical method.

[0015] The present invention relates to a fermentation plant or food production and production.
And / or surplus microorganisms discharged from processing plants or
Mainly solid waste, such as beer yeast, beer cake,
Coffee grounds, bran, included in which a so-called organic waste your or Rana, can be a growth source for useful microorganisms substances, trace elements, particular means a component consisting trace growth factors such as, in the liquid in the process is taken out, This is an innovative method for producing a useful substance for the purpose of producing a physiologically active substance, an enzyme, or various fine chemicals by using this as a culture medium for the useful microorganism and growing the microorganism.

At present, there are not many food production plants in Japan that discharge this kind of organic waste, but for example, in the case of a beer production plant, for example, there are approximately 40 production plants, and The amount of discharged beer cake is estimated only to be approximately 30,000 tons / year in a dry state, and the excess brewer's yeast amounts to 140,000 tons / year.

Although some of these organic wastes are currently sold to the outside as fertilizer and / or feed, they are practically unprofitable due to the low added value of the products, and the plant is not profitable. It is an economic burden, and the remaining beer cake and / or yeast is incinerated. In view of this, by applying the present invention to the recycling of these organic wastes, these factories will inevitably receive immeasurable benefits.

Next, the excellent functions and effects of the present invention will be described with reference to FIG. 1 by taking an organic waste from a beer manufacturing plant as an example. FIG. 1 shows a flow for treating organic waste according to the method of the present invention. First, FIG.
Moves from the beer lees storage tank 1 storing beer lees, which is an organic solid waste to be effectively used in the above, to the screen 2 and removes so-called barley epidermis that cannot be assimilated by useful microorganisms with a screen having a certain opening. I do. The beer cake that has passed through the screen is pulverized appropriately by a pulverizer 3 and then sieved, if necessary, to remove the sieve residue.
Perform slow stirring for ~ 4 hours to extract the aqueous component in the beer cake to the water side. If necessary, an appropriate amount of a supplemental component 19 such as a nutrient or a trace element is added to the extract, and the extract is once stored in the culture solution storage tank 6.

On the other hand, the surplus yeast from the beer production process is transferred from the surplus yeast storage tank 7 in which the surplus yeast is stored to the yeast dissolution tank 8, and an appropriate amount (5 to 10 times) of water 4 is added. An appropriate amount of commercially available lytic enzyme 9 such as lysozyme is added to the yeast solution, and the yeast and lysozyme are contacted with gentle stirring for about 1 day to dissolve the yeast cell wall and cell membrane, and transfer the yeast content solution to the solution side. .

In addition, for dissolving the yeast in the yeast dissolving tank 8, even if a commercially available expensive enzyme such as lysozyme is not used, an enzyme which is widely distributed in nature and easily available to anyone can be produced. Microbial cells 10 such as slime bacterium are separately concentrated and added to the yeast dissolving tank 8 and mixed and cultured for 2 to 3 days with aeration of sterilized air.
The cell membrane is almost completely dissolved, and the components in the yeast migrate to the liquid side.

In order to ensure the lytic action and function of mucus cells, Myxococcus xanthus (Myxococcus) is used.
xanthus, ATCC 25232), Cystobacter fuscus.
us, ATCC 25194), Stigmatella erecta, ATC
Brewer's yeast Saccharomyces cere.
visiae), there was a slight difference in the ability of each myxobacteria, but at a culture temperature of 25 ° C. (normal temperature) and a culture time of 2.0 days to 5.0 days, the yeast cell wall and membrane were Almost completely dissolved. The step of lysing yeast by myxobacteria, that is, the step of preparing a culture medium in the present invention, is a central part of the present invention. By applying this preparation method, it is possible to produce and produce high-value-added useful substances at extremely low cost. Can be.

After the insoluble residue is removed from the mixed solution in the yeast dissolving tank 8 by the next screen 11, the mixed solution is temporarily stored in the culture solution storage tank 6 and mixed with the extract of beer lees. Beer lees discharged from a beer factory are approximately 4k
g absolute dry matter / kL beer, surplus yeast discharge about 20kg
Absolutely dry matter / kL beer, the emission ratio of both is approximately 1: 5
become.

The following table shows the results obtained by obtaining beer meal and surplus yeast from a beer manufacturing plant.

[0024]

[Table 1]

[0025]

[Table 2]

As can be easily understood from the analysis results in the above table, both beer meal (extract) and brewer's yeast (dissolution) sufficiently contain organic nutrients, vitamins and inorganic salts, and are useful for various useful microorganisms. Has a nearly complete composition and usually requires little supplement 19, such as nutrients for nutrition correction and / or fortification. In this example, if the method is limited to the recycling of organic waste in a beer manufacturing plant, the step of extracting a culture medium from beer cake can be omitted in view of the small amount of discharge.

The mixed solution of the extract of beer lees and / or the solution of surplus bacterial cells (yeast) is transferred from the culture solution storage tank 6 to the culture tank 13 after inoculating various useful microorganisms 12,
Culture is performed by setting culture conditions for the growth characteristics and biological characteristics of various useful microorganisms. The culture tank 13 is of course sterile. If the useful microorganism is anaerobic,
The structure is such that air from outside is mixed in. In the case of aerobic, sterile air 14 is introduced to supply oxygen to the microorganisms.

In the present invention, useful microorganisms that produce useful substances in the culture tank need not be specified .
In a non-hyaluronic acid production, Streptococcus Zuepidemikusu (Streptococcus zooepi
demicus) (Reference Current Microb)
ology, Vol. 1 to 8 [1987
Year]), in the production of γ-linolenic acid, Mortierella elongata (Mortierella erogata)
ngata) (TTCC 16271), for production of eicosapentaenoic acid, Alteromonas putrifaciens.
s) (ATCC 8073), Mortierella spinosa (A)
TCC16272) is selected.

When producing deoxyribonucleic acid (DNA) or adenosine triphosphate (ATP), Rhodotorul is widely distributed as a wild yeast in nature and readily available to anyone.
a sp. ), Torula (Torula sp.), Mycotorula (Mycotorula sp.), Pichia (P
ichia sp. ), Candida
sp. ) Or Trichosporone (Trichosp)
oron sp. ) May be selected.

As described above, one of the useful microorganisms is
After production of useful substances in the microbial bodies in mass culture to,
Extraction of useful substances from microorganisms, ordinary microorganism separation equipment 1
In step 5, the microorganism is separated, and a useful substance 18 is obtained by passing through a separation / purification device 17 in order to purely extract a useful substance of interest from the extracted liquid .

The secondary surplus microorganisms separated by the microorganism separation device 15 may be newly used as organic waste in the method of the present invention, or the water content may be reduced to a certain value or less by ordinary drying means. It can be dehydrated and turned into fertilizer and / or feed 16.

[0032]

EXAMPLES The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. Example 1 As organic wastes to be recycled, beer meal (barley meal) discharged from a beer manufacturing plant and surplus yeast Saccharomyces cerevisiae (Saccharomyces c)
erevisiae).

First, powdery dried beer cake about 100
A 10-fold tap water was added to the g, and a component that could be a nutrient source for useful microorganisms contained in the beer lees was extracted with gentle stirring at room temperature for 5 hours (insoluble components were filtered off). On the other hand, about 5 yeast in a viable bacterial state dehydrated by a dehydrator
About 10 times (about 5 liters (L)) of tap water was added to 00g, and the myxobacterium Myxococcus xanthus was separately cultured and increased in volume.
(ATCC 25232) culture solution 300cc,
After culturing at a water temperature of 25 ° C. for 3 days, the cell wall and cell membrane of the viable yeast cells were almost completely dissolved, and the bacterial cell content solution was eluted to the liquid side (unresolved residues were sieved).

About 1 L of the extract from beer lees was mixed with about 5 L of a brewer's yeast solution, and the mixture was used for culturing useful microorganisms. The composition of this liquid was as shown in Table 3.

[0035]

[Table 3]

This mixture is placed in a standard type jar fermenter having an effective volume of 2 L, and sterilized by an autoclave. A lactic acid bacterium Streptococcus zooepidemics (Streptococcus), which was separately cultured under sterile conditions and pure, was separately added to this mixture.
zooepidemicus) 5cc
The mixture was inoculated at a ratio of / L-mixture, and cultured at a culture temperature of 30 ° C for 48 hours in a batch system. Removed culture was precipitated by addition of A Ruko <br/> Le, the precipitate was dissolved by adding water, repeated four times the operation of precipitation by addition of alcohol again, a white powdery hyaluronic acid Was collected.

Using the above mixture as a culture medium, Streptococcus zooepidemics (Streptococcus)
The amount of hyaluronic acid produced when cultivating S. zooepidemicus is 4 to 5 g / L, and a good result comparable to the amount of hyaluronic acid produced when a standard medium used for culturing this microorganism is usually obtained. Was done. The culture medium used in Example 1 was prepared based on brewer's grains and brewer's yeast, but the quality and quantity of the culture medium components were derived from brewer's yeast. Significant effects can be expected to compensate for the deficiencies.

[0038] In Comparative Examples against Comparative Example 1, bacteria of the active ingredient in powdered spent grains is a step of extracting with water are identical, by myxobacteria respect brewer's yeast as a live bacteria The body was not dissolved but diluted with water, mixed with the extract at a ratio of about 1: 5, and used as a culture medium under the same conditions as in Example 1 for Streptococcus zooepidemics (St.
reptococcus zooepidemicu
Although s) was cultured, the production of hyaluronic acid was extremely lower than that of Example 1, and was only 0.2 to 0.3 g / L.

Furthermore, in the comparative example , not only the amount of production of hyaluronic acid is small, but also the yeast cells are not dissolved, so that a burden is imposed on the cell separation step 15 and the separation / purification step 17 is also subjected to impurities as impurities. Excessive extracellular polymer flowed in, preventing complete separation and purification.

[0040]

According to the present invention, the following effects can be obtained. (1) The organic waste discharged from the food manufacturing plant, the water-soluble extract of solid waste, and the bacterial cell content of bacterial waste by dissolving the bacterial cells with myxobacteria. By taking out the mixture and culturing microorganisms that produce various useful substances by using a mixture of the two as a culture medium, a substance with high added value can be produced, and complete recycling of organic waste can be achieved. (2) By adopting the present invention, it is possible to produce high value-added useful substances from low value-added organic waste, so that a conventional economically unprofitable waste recycling method has been developed. It is possible, and the realization of a self-contained factory can be fully expected. (3) The present invention enables not only the production of organic waste but also the treatment thereof, which contributes significantly to the preservation of the global environment. (4) The so-called final residue after producing useful substances can be effectively used as fertilizer and / or feed.

[Brief description of the drawings]

FIG. 1 shows a flow of treating organic waste from a beer manufacturing plant according to the method of the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Beer lees storage tank 2 Screen 3 Crusher 4 Water 5 Extraction tank 6 Culture storage tank 7 Surplus yeast storage tank 8 Yeast dissolution tank 9 Lysis enzyme 10 Microorganism 11 Screen 12 Useful microorganisms 13 Culture tank 14 Sterilized air 15 Microbial decomposition equipment 16 Fertilizer and // feed 17 Separation / purification equipment 18 Useful substances

Claims (4)

(57) [Claims] (1) dissolving organic waste mainly consisting of microorganisms;
It has the function of producing and releasing bacterial or lytic enzymes from the body.
Useful in culture medium obtained by solubilization by the first microorganism
A method for producing a useful substance from waste, comprising culturing a second microorganism that produces and accumulates a substance in a body . 2. The culture medium is decomposed by a second microorganism.
Water-insoluble substances and / or first microorganisms that cannot
2. The method according to claim 1, wherein said material has been sufficiently removed.
A method for producing a useful substance from the waste described in the above. 3. The second microorganism as organic waste
The method for producing a useful substance from waste according to claim 1, wherein the method is solubilized by a first microorganism . 4. The method according to claim 1, wherein the first microorganism is Myxococcus,
Mixobacter, cystobacter and stigmatella
The method for producing a useful substance from waste according to claim 1, wherein the method is a myxobacterium selected from the group consisting of :