JP5024848B2 - Method of converting biomass into solidified material and method of using the solidified material - Google Patents

Description

  The present invention relates to a method for converting a resource (biomass) generated as waste in a biological industry such as agriculture, forestry and fisheries and food industry into a solidified material, a solidified material produced by the conversion method, and a method for using the solidified material. Is.

While importance is attached to the recycling of biomass aimed at sustainable use of resources, most of the waste biomass produced in biological industries such as agriculture, forestry and fisheries and the food industry is high in perishable organic matter. The processing is a problem.
Examples of these wastes include non-food parts and primary processing residues of crops such as rice straw and rice husks, waste crops during production adjustment, livestock manure and livestock waste during meat processing, Waste from primary processing residue of fishery products, forestry waste such as thinned wood, sawdust, waste fungus bed, etc., and food industry produce waste at the time of food production, unsold goods, etc. . In addition, the garbage as a general waste contains a large amount of organic matter, but the composition of the components is not constant, so some of the resources have been put to practical use, such as gasification and composting by methane fermentation. It is currently being burned directly. In addition, it is desired to develop treatment methods for biomass such as building waste, pruning residue, waste paper, and waste cloth.

By appropriately processing these biomasses and developing their applications, recycling is promoted by their cascade use. For example, a technology for converting into pellets, biodegradable pots, and the like has been developed by injection molding technology using okara or corn-derived biomass that is by-produced in the tofu production process (Patent Document 1).
On the other hand, technologies have been developed and used for silage fermentation, composting, and converting food waste biomass into liquid feed by lactic acid fermentation or the like. In addition, methane fermentation has been used for a long time for decomposition and volume reduction of waste biomass and conversion to gas energy.
In addition, composting technology has been widely used mainly for household waste, and volume reduction has been performed by aerobic decomposition. However, many waste biomass derived from the agriculture, forestry and fisheries industry and the food industry will rot immediately and become sludge that emits a foul odor. Will drop significantly.

One of the most important treatments for preventing the decay of biomass is a reduction in water activity before the decay. It is known that microorganisms such as mold, yeast, and bacteria stop growing at water activities of about 0.7, 0.75, and 0.8 or less, respectively.
Therefore, by drying these biomasses, it becomes possible to store them while preventing microbial degradation of the quality. This is the same for biomass with low uniformity, such as trash, and drying improves storage properties and increases added value by itself.

The purpose of the biomass drying process is to improve storage, but by forming it into a solid material with an appropriate size and shape according to the purpose, not only storage but also transportation and use Workability is dramatically improved. If the dry biomass is in the form of fine powder, it will not only be scattered as dust and difficult to disperse, but there is also a risk that the worker will be sucked and exposed. Importance is great.
Moreover, when performing shaping | molding, it is desirable to convert into the solidification raw material shape | molded by the shape according to a pellet form, a film form, or a use. If biomass rich in high-quality organic matter can be efficiently converted into solidified materials, in addition to biodegradable materials, it can be used for feed, fertilizer, fermentation medium, soil modifier, carbonization raw materials, pyrolysis raw materials, etc. It can be expected to lead to the development of applications.

Patent No. 3697234 Specification

However, with regard to existing technologies related to the conversion of biomass into solidified materials, such as fertilizer and feed pelletization technology, in order to ensure the formability and stability in the process of transporting and working the molded products, However, there is a problem that the cost of the molding process is increased due to the addition of the binder, the pulverization process of biomass, the high temperature / high pressure treatment process, and the like.
In this way, the use of biomass is expected to greatly expand by streamlining the conversion process to solidified materials that are effective for the use of highly perishable biomass, but there are limits to existing technologies. is there. So far, only a limited number of examples of fertilizer and feed have been put to practical use. Under such circumstances, there is a strong demand for breakthroughs relating to the supply of binders and the improvement of the processing characteristics of biomass in the process of converting to solidified materials in order to increase the availability of biomass.

As a result of intensive efforts to solve the above problems, the inventors have found that it is possible to improve binder supply and biomass processing characteristics by devising a fermentation process for biomass, which is a waste resource. The present inventors have succeeded in developing a method for producing a highly useful solidified material while solving various problems.
In the present invention, it is possible to improve the characteristics at the time of molding or pretreatment of biomass for conversion into a solidified material and the characteristics of the solidified material as a molded product by the fermentation process of biomass. In addition to the method of fermenting biomass and converting it into a solidified material alone, we have also developed a method of converting it into a solidified material after mixing with other biomass. It has been found that non-constant biomass and biomass with high water content such as sludge, which are difficult to treat, can be converted to solidified materials by the latter mixing method.
Furthermore, it is possible to prevent spoilage by devising the biomass fermentation process, and it is possible to improve feed characteristics, fertilizer characteristics, characteristics as fermentation media, soil modification characteristics, carbonization characteristics, thermal decomposition characteristics, etc. Become.

The invention provided by the present application is as follows.
[Claim 1]: The following (A) and (B) are performed on a biomass raw material containing at least one or more components belonging to plant cell wall components and protein components. The manufacturing method of the solidification raw material which has the binding property of the components which show the property of.
(A): The process of obtaining the fermented material containing the extracellular component which has a binder characteristic by fermenting the said biomass raw material using the 1 or more types of microorganisms which have the characteristics of (a) and (b) below.
(a): One or more microorganisms belonging to polyglutamic acid-producing bacteria, gellan gum-producing bacteria, polygalactosamine-producing bacteria and curdlan-producing bacteria.
(b): A microorganism having a resolution for at least one component belonging to the plant cell wall component and protein component contained in the raw material.
(B): The process which shape | molds and solidifies the said fermented material or the composition containing the said fermented material.
(C): The property of being difficult to disintegrate under pressure and easily disintegrating in water.
[Claim 2]: The method for producing a solidified material according to claim 1, wherein the composition containing the fermented product contains the unfermented biomass raw material.
[Claim 3] The method for producing a solidified material according to claim 1 or 2, wherein the microorganism is Pseudomonas elodea and / or Bacillus subtilis.
[Claim 4]: The biomass raw material is a non-food part of a crop, a waste crop, a livestock manure, a waste during processing of meat, a waste during processing of marine products, a thinning material, sawdust, a waste fungus bed, The waste at the time of food manufacture, the waste at the time of food distribution, wastes at the time of food distribution, scrap waste, construction waste material, pruning residue, waste paper, and old cloth; Manufacturing method of solidified material.

According to the present invention, there is provided a manufacturing technology of a solidified material characterized by adding a fermentation process of the biomass for effective use of biomass which is an industrial waste resource. Thereby, various characteristics can be imparted to the biomass.
According to the present invention, it becomes possible to increase the added value according to the characteristics of various biomass, and it is considered that the development of various useful materials from biomass utilizing the advanced fermentation engineering technology of Japan will be paved.

The present invention will be described below.
The solidification material manufacturing technology includes biomass drying and heat treatment as pre-treatments, adjusting to the optimal moisture conditions, and using a general granulator, etc., irradiation with gamma rays and enzyme treatment In addition to a solidification method involving polymer cross-linking and the like, a solidification method involving heating and pressurization by a machine is conceivable.
As a molding method using a machine, widely used techniques such as injection, extrusion, and compression (hot pressing) can be used. Examples of the drying step include natural drying, wind drying, and heat drying.

The main factors to be considered in the pre-treatment stage and processing stage until the biomass is formed into a solidified material are: biomass preservability, biomass moisture content, liquid separation from solid biomass, particle size, from outside the system The amount of binder added, the efficiency of mass transfer to the molding apparatus, the efficiency during solidification by heat / pressure treatment, and the like can be mentioned.
In this invention, the characteristic of the biomass which concerns on these factors can be improved by fermenting biomass before a shaping | molding process.
Regarding fermentation of biomass, fermentation is performed using biomass or components decomposed and modified by physical, chemical or biological pretreatment as all or part of the fermentation medium. In that case, as main components, plant storage polysaccharides such as starch, fructan, glucomannan, plant cell wall components such as cellulose, hemicellulose, pectin, lignin, carbohydrate components derived from microorganisms and animals such as chitin, chitosan, glucan, Organic components such as lactose, proteins, and lipids that are by-produced from dairy products, or their degradation / modification products are used.

In the present invention, when determining the fermentation conditions of the target biomass, it is important to make various active adjustments. Active adjustment includes analyzing the composition of biomass and performing screening for searching for known microorganisms or appropriate microorganisms from nature. In addition, the fermentation characteristics of microorganisms are optimized using a known mutation introduction technique, genetic engineering technique, protein engineering technique using genetic manipulation technique, metabolic engineering technique, or the like.
Examples of the fermentation characteristics to be improved in the present invention include biomass utilization characteristics, characteristics relating to growth characteristics, and characteristics of fermentation products. In order to promote the fermentation of the microbial function and appropriately cope with the change in the characteristics of the biomass as the raw material, a subcomponent for adjusting the pH or adjusting the nutrient component is appropriately added. Through these active adjustments, it is possible to determine the strain to be used, the amount of the main component, the subcomponent composition, the water content, the culture temperature / pH, the culture time, the stirring conditions, etc., and the biomass conversion process can be optimized. It becomes possible.

The problem of the storage stability of biomass is a particularly serious problem for biomass that is generally easily degradable and has a high water activity, such as food waste. Until now, in order to improve the storage stability, the biomass had to be bacteriostatic or bacteriostatically dried, but in the present invention, the fermentation process is performed prior to the rot process by the rot bacteria mixed in. To suppress the growth of spoilage bacteria.
In addition, it is known about alcoholic fermentation and organic acid fermentation that fermentation under culture conditions with little growth of spoilage bacteria is possible by using microorganisms with high resistance and growth ability under culture conditions with high selectivity. It is possible to ferment biomass using this method. Common selection pressures include pH, temperature, humidity, alcohol concentration, salt concentration, antibiotics, and the like, and the ability to decompose hardly decomposable biomass.
Furthermore, by inoculating biomass with an overwhelmingly large amount of fermentation bacteria as a starter, the culture conditions become advantageous for the fermentation bacteria. By using these known knowledge related to fermentation production in the pretreatment stage in the formation of biomass, not only is it possible to reduce the labor involved in suppressing the decay of biomass, but it is also possible to perform fermentation with high reproducibility. Become.
Screening for microorganisms exhibiting resistance to selective pressure can be easily found by screening using growth activity in a medium that reproduces an environment with selective pressure as an index, and can be used in the present invention. it can.

Regarding the volume reduction of biomass for improving the molding characteristics, the destructive promotion by the collapse of the structure of the matrix and the granular structure, the formation of flocs, etc. is concerned. For example, dehydration and volume reduction prior to conversion to a solidified material can be achieved by utilizing the substance resolution of microorganisms, the ability to modify pH, and the like. In addition, it has been known for a long time that, when yogurt is used to lower the pH using lactic acid bacteria, protein hydration water is released and protein aggregation is promoted, and water separation occurs. It can also be used in the invention.
Microorganisms that are expected to have floc-forming ability by producing extracellular polysaccharides and cell wall components that are charged in solution include zygotes such as Mucor, Absidia, and Rhizopus that produce chitosan. Aspergillus genus, Paecilomyces genus, Neurospora genus, etc. that produce polygalactosamine are known, and can be used as a technique in the present invention by fermenting biomass.
Microorganisms having the activity of promoting floc formation can be easily screened by a method such as mixing a cell culture solution and a dispersion of an insoluble substance and examining changes in the sedimentation rate of the insoluble substance, and used in the present invention. Is possible.
On the other hand, when considering the economic efficiency of biomass solidification including a dehydration step, it is not always efficient to add extra moisture from the outside. If possible, it is desirable to utilize a solid phase culture technique such as bran culture with reduced amount of water.

  In addition to promoting dehydration, techniques for reducing the amount of biomass by fermentation, such as methane fermentation and composting, are widely known, and can be used as appropriate in the present invention. In the present invention, it is desirable to review and optimize the conditions in consideration of the efficiency of the entire biomass forming process even when using known techniques such as methane fermentation and composting. For example, it is necessary to consider the effect of the loss of useful substances on the added value of the solidified material as the final product when the existing volume reduction technology is used.

Compared with existing methods, when a part of the biomass decomposes due to fermentation of the biomass and the entire structure collapses or softens, resulting in finer particles or improved fluidity and moldability. Therefore, it is possible to carry out operations such as transportation, addition, and mixing with a small amount of power, which is considered to contribute to cost reduction in the molding process.
Due to the action of degrading bacteria such as plant cell wall components and crustacean shell components, the structure of these biomasses in which the hardly decomposable components are associated is dissociated and softened. Many of these fungi and bacteria play the role of biomass decomposers in nature, and they are known to decompose, insolubilize, or assimilate finely-degradable components by single or combined action. It has been.

In the present invention, based on these known findings, it is possible to effectively utilize microorganisms characterized by having degradability of these hardly decomposable components for converting biomass into solidified materials. It becomes a feature. Examples of plant cell wall degrading bacteria include, for example, Acetobacter, Acetovibrio, Acidobacterium, Aeromonas, Bacillus, Cellulomonas, Celluvibrio, Clostridium, Pseudoalteromonas, Pseudomonas, Streptomyces Genus, Thermomonospora, Ruminococcus, Xanthomonas, Aspergillus, Penicillium, Fusarium, Humicola, Hypocrea, Macrophomina, Nectria, Neurospora, Robillarda , Trichoderma, Verticikkium, Agricus, Phanerochaete, Schizophyllum, Sclerotium, Coniophora, Cryptococcus, Filobacidiella, Irpex, Trametes, Ustilago, Dictyostelium A genus or the like can be used.
Examples of the crustacean shell-degrading bacteria include, for example, the chitinase producing bacteria Aeromonas, Alteromonas, Arthrobacter, Aspergillus, Bacillus, Burkholderia, Cellulomonas, Clostridium , Coccidioides spp, Cryptococcus spp, Enterobacter spp, Hypocrea spp, Lactococcus spp, Metarhizium spp, Saccharophagus spp, Pseudoalteromonas spp, Pseudomonas spp, Pyrococcus spp, Rhizopus spp, Serrattomy spp Genus, Thermococcus, Thermomyces, Xanthomonas, Trichoderma, Vibrio, etc. can be used.

  Moreover, about biomass containing many proteins, processing residues of livestock and seafood, microbial cell residues, plant residues and the like are included. For example, okara, a tofu residue, is a biomass rich in protein. Microorganisms with high proteolytic ability show good growth in a medium containing okara, and can improve the fluidity by minimizing okara particles. Examples of microorganisms that produce powerful proteases include Aspergillus, Bacillus, Kluyveromyces, Rhizopus, Rhizomucor, and Streptomyces.

  In addition to those exemplified above, microorganisms that degrade plant cell wall components, crustacean shell components, or protein components have an activity of decomposing those components when cultured in a medium containing these components. Can be easily screened, and the microorganism can be used in the present invention. By appropriately applying selective pressure during growth under the culture conditions in the above medium, it becomes possible to screen for degrading bacteria having high environmental suitability during fermentation of the target biomass and high activity of suppressing spoilage.

Most biomass contains persistent components such as cellulose, chitin, and lignin. In order to make the fermentation process of biomass more efficient, it is not always necessary to set the fermentation stage as one stage. For example, depending on the ability to assimilate biomass and useful bacteria after culturing koji molds to make koji and then fermenting it with yeast, depending on the ability to assimilate biomass and useful bacteria, Added value increases efficiency.
Further, as other important components contained in a large amount in the waste, plant storage polysaccharide components such as starch, lactose, lipids and the like are considered. These components can also easily find a highly assimilating microorganism by screening, and can be utilized in the present invention. In order to confer assimilability for each component to microorganisms that produce useful substances from biomass, genes encoding starch-degrading enzyme systems such as amylase and maltase, genes encoding β-galactosidase, and lipase genes By introducing, the utilization characteristics of the microorganism are improved.

By culturing microorganisms having fermentation ability such as hardly degradable biomass, not only the substrate is dissociated and softened, but also extracellular substances and organic substances accumulate in the culture solution, and the bacterial cells grow. Unless pretreatment such as pulverization is performed, the persistent biomass that is a substrate is large in volume and has low dispersibility and low binder activity, but various organic substances accumulated in fermentation broth after fermentation and microscopic cells It is clear that is highly dispersible and has a high binder activity.
Moreover, it is thought that it contributes to the improvement of the solidification characteristic of the substrate itself by covering the periphery of the substrate with fermenting bacteria or producing a polymer such as a biofilm. Thus, it is clear that fermentation using these organic assimilating bacteria improves the molding characteristics as a solidified material of biomass and the characteristics of the molded article, and the fermentation of biomass is performed in the present invention. This is considered to be a great advantage when converting to a solidified material.

Regarding the improvement of the molding characteristics of the biomass and the properties of the molded product for conversion to a solidified material, the particulates that make up the biomass are bound together and stably exist so that they do not crumble and return to the particulates. For this purpose, a method for producing the binder molecules by fermentation can be considered.
So far, protein-based materials such as corn-derived zein, glycerol, polysaccharides and the like have been used so far, but a method of intentionally producing a binder by fermentation of biomass has not been developed so far.
As binders for fermentation production, components such as microbial cell walls, exopolysaccharides and mucilage, low-molecular fermentation products, or partial structures of plant cell wall components that are by-produced from biomass by the fermentation process are assumed. Is done. Substances derived from microorganisms include, for example, constituents of Ascomycetes such as Neisseria gonorrhoeae and fungi, constituents of Basidiomycetes such as edible mushrooms, constituents of bacteria such as yeast, lactic acid bacteria, and Bacillus natto, A hydrolyzate, autolysate, extracellular polysaccharide component, extracellular polypeptide, etc. can be used.

Examples of extracellular components expected to have good binder properties include curdlan, gellan gum, xanthan gum, hyaluronic acid, pullulan, dextran, succinoglycan, polyglutamic acid, polyhydroxyalkanoic acid, ε-poly-L-lysine, Polygalactosamine and the like, such as the production of these macromolecular substances, for example, Agrobacterium, Alkaligenes, Pseudomonas, Xanthomonas, Streptococcus, Aureobasidium, Leuconostoc, Bacillus, Ralstonia, Azotobacter, Spirillum, Vibrio, Photobacterium, Streptomyces, Aspergillus, and the like can be used.
Many of these components are also known as food gels, and it is possible to develop methods that maximize the binder activity by utilizing known methods. Most of these are dissolved / dispersed at 100 ° C. or lower and gelled by cooling or adding calcium salts or other gels.

In the molding process of converting biomass into solidified material according to the present invention, it is solidified at a much lower temperature than pressing and drying by heat and pressure, so it is possible to greatly reduce the use of energy by heat and pressure. It is. Furthermore, the high elasticity and stability of many gels greatly contribute to the physical stability and chemical stability of the solidified material after molding.
Extracellular components with good binder activity by growing microorganisms in solid or liquid media, collecting extracellular polysaccharides, and searching for microorganisms that produce binders that match the characteristics of the main biomass Can be easily screened and can be utilized in the present invention.

So far, research on the production of exopolysaccharide, a useful substance, from waste biomass has been reported, and olive milling wastewater (Lopez, MJ et al., J. Appl. Microbiol., 90, 829-835 (2001)), chestnut extract (Liakoupoulou-Kyriakiswa, M. et al., Appl. Biochem. Biotechnol., 82, 175-183 (1999)), cassava bagasse hydrolyzate (Woiciechowski, AL et al., Appl. Biochem Biotechnol., 118, 305-312 (2004)) and the like have been tried to produce exopolysaccharides.
However, these research results are aimed at the extraction of exopolysaccharides from biomass, and are not aimed at converting the entire biomass used into solidified materials. It can be utilized in the present invention by optimizing according to the inventive concept.

Furthermore, by modifying the fermentation ability of microorganisms using known mutation introduction techniques, genetic engineering techniques, protein engineering techniques using genetic manipulation techniques, metabolic engineering techniques, etc. Giving microorganisms lacking the ability to produce extracorporeal polysaccharides and mucilages, converting nutritional properties such as carbon sources that can be used during the production of extracorporeal polysaccharides and mucilages, and providing metabolic control mechanisms It is possible to control the structure of the product or to control the structure of the product by modifying the structure of the enzyme.
It is possible to modify the assimilation properties of the fermentation microorganisms, the growth environment characteristics, the structure and physical properties of the products, and to produce useful substances that can improve the characteristics of the solidified material. Clearly, it will be an effective technique to achieve. For example, a biosynthetic gene for xanthan gum possessed by Xanthomonas campestris was introduced into the genus Sphingomonas, and xanthan gum was biosynthesized (Pollock, TJ et al., J. Ind. Microbiol. Biotechnol., 19, 92-97 (1997) ), And an example in which β-galactosidase gene is introduced into X. campestris bacteria that do not have lactose-assimilating ability to impart lactose-assimilating properties (Fu, JF. And Tseng, YH., Appl. Environ. Microbiol. 56 , 919-923 (1990)) can be used in the present invention by optimizing according to the concept of the present invention.

Regarding the improvement of utilization characteristics, plant cell wall components constituting biomass, crustacean shell components, protein components, plant storage polysaccharide components, enzyme genes related to hydrolysis and metabolism of lactose, lipids, etc., receptor genes for uptake By introducing the above, it is possible to impart new biomass utilization to microorganisms used for biomass fermentation.
These genes can be cloned from degrading bacteria for each component, and can also be obtained from microorganisms whose degradation activity is unknown by cloning based on the PCR method using the similarity of gene sequences, Furthermore, it is also possible to obtain a gene sequence by artificially synthesizing part or all of the gene sequence. The protein of interest is biosynthesized by introducing it into a host cell using an appropriate vector and expressing it inside the cell.

  As for genes related to environmental resistance, when the expression of a gene possessed by a microorganism is reduced, environmental resistance can be imparted by introducing a mutation for suppressing the expression of the gene. In addition, it is easy to screen for microorganisms having genes for improving environmental resistance, such as antibiotic inactivation enzymes. The environmental resistance is imparted by introducing and expressing the gene for improving environmental resistance of the microorganism into a biomass-degrading bacterium having no resistance.

  Even when biosynthesis ability of low molecular weight useful substances that provide extracellular components, plasticity and various functionalities is given to microorganisms used for biomass fermentation, genes and gene groups from these microorganisms having biosynthetic activity By acquiring and introducing / expressing, the production of useful substances will be made more efficient. In such cases, use of metabolic engineering technology, ribosome engineering technology, protein engineering technology, etc., as appropriate, increases the amount of precursor and raw material required for biosynthesis of the target substance, and controls metabolism. By promoting or suppressing, or by directing many substances to be used for biosynthesis of the target product, it becomes possible to efficiently produce useful substances while assimilating biomass.

By utilizing protein engineering technology, carbohydrate engineering technology, etc., it becomes possible to modify the structure of useful substances produced by biomass fermentation, and further to the object of the present invention to produce a solidified material with high added value. It becomes possible to produce a material having a suitable structure.
A known technique can be utilized in the present invention to produce a useful substance having a new structure from biomass. For example, it suppresses the action of glycosyltransferases on sugar residues that are the side chains of polymers such as xanthan gum, synthesizes polysaccharides having a structure similar to cellulose, or uses chitin deacetylase to inactivate insoluble polymers such as chitin. It differs by eliminating the side chain N-acetyl group and converting it into a chitosan structure that has a charge and a floc-forming ability in dilute acid solution, or by modifying the substrate recognizability of glycosyltransferase used in polysaccharide synthesis Attempts may be made to modify the polymer structure by acting on monosaccharide derivatives having a structure. In addition to the examples given here, it is possible to convert biomass into a solidified material with higher added value by using a biosynthetic material modified by utilizing known advanced technology in the present invention. Become.

  Regarding the molding using the fermented biomass, there are a method of molding the biomass alone and a method of molding the fermented biomass by mixing with other biomass. The latter method has low uniformity and high septic properties, so it is a biomass that is difficult to ferment. It can be applied to fermentation residues such as soy sauce residue, animal waste such as bone meal, livestock manure, methane fermentation waste and the like. This means that it is possible to produce a solidified material partially mixed with biomass that is extremely difficult to develop.

At the time of molding, it is desirable that the substance having binder activity is uniformly dispersed in the sample. Therefore, it is conceivable to appropriately add a process for solubilizing the active ingredient by adding a solubilizer, mixing / dispersing, or the like. Moreover, in order to adjust the quality of the solidified material or increase its usefulness, it is easily conceived to add subcomponents as appropriate.
For example, in the case of fertilizers, addition of inorganic components that serve as nutrients, in the case of feeds, addition of seasoning substances for improving palatability, adjustment of ingredients to make a highly effective fermentation medium source, etc. Is given as an example.

The physical or chemical stability of the solidified material refers to the property that the shape of the solidified material does not collapse or chemically deteriorate during storage, transportation and use. During the production of the solidified material, it is desirable to appropriately fermentatively produce a component that imparts elasticity and plasticity or to add it as a subcomponent in order to enhance physical stability such as vibration stability and impact stability.
Examples of the component imparting plasticity include polyhydric alcohols such as glycerin, polyethylene glycol, and polypropylene glycol, organic acids such as lactic acid and malic acid, urea, and the like. If a component that imparts plasticity can be produced by fermentation with the production of a substance having good binder activity, addition of subcomponents can be suppressed.

  When using the solidified material as a fertilizer or soil modifier, for example, control of the C / N ratio by decomposing or denitrifying organic matter in the biomass, nitrogen fixation, etc., production of organic nitrogen content that is slow acting nitrogen, The release of ingredients with soil aggregating activity is an advantage. In addition, stabilization of soil pH by a substance having a buffering capacity, provision of proliferation activity of specific useful microorganisms such as actinomycetes by chitin and peptidoglycan derived from microorganisms, provision of elicitor activity by chitin oligosaccharides, and the like. Polyglutamic acid has high water retention and stickiness, and is expected to have a binder activity for soil particles. Moreover, it is thought that it plays a role as slow acting nitrogen as a polypeptide. For example, a solidified material prepared under a mild condition using a fermented biomass produced with polyglutamic acid releases polyglutamic acid easily by soaking in water, so it has a high fertilizer effect and soil improvement effect. it is conceivable that.

Fermentation process that contributes to the improvement of characteristics when using solidified material as feed or as a fermentation medium source includes fermentative synthesis, taste, and texture of nutrient components, digestible components and vitamins using biomass・ Improvement of flavor etc. can be considered. More specifically, it can be a low molecular weight starch and plant cell wall constituents for improving digestibility, fermentative production of essential amino acids, organic acids, alcohol, riboflavin, vitamins, ubiquinones, etc., and a nutrient source for enteric bacteria For fermentation production of functional components and the like, and in the case of feed, fermentation conversion for the purpose of pre-culturing useful intestinal bacteria having a probiotic action can be used.
If the solidified material contains heat-sensitive ingredients such as microorganisms, vitamins, and trace active ingredients, dry at low temperature, put it in a semi-dry state, mix the culture solution immediately before or after completion of drying, etc. It is desirable to devise a processing method. In addition, it is common in all application developments. However, when producing biomass by fermentation, it is necessary to pay attention to various components such as mycotoxins and endotoxins and the toxicity of microorganisms themselves.

Examples of microorganisms that can be expected to exhibit properties for use as solidified fertilizer, soil conditioner and feed include Brevibacterium, Proteus, Rhizopus, Bacillus, Pseudomonas, Alkaligenes, Corynebacterium, Arthrobacter, Saccharomyces, Zymomonas, Lactobacillus, Serratia, Aerobacter, Aspergillus, Penicillium, Acetobacter, Gluconobacter, Streptococcus , Leuconostoc, Pediococcus, Methylobacillus, Methylobacterium, Flavobacterium, Pichia, Rhizobium, Streptomyces, Agrobacterium, Erythrobacter, Candida, Eremothecium, etc. Can be mentioned.
Fermentation can be carried out by directly using the metabolic pathways of these microorganisms, or assimilation can be altered using a known technique such as genetic engineering or metabolic engineering, or a new ability can be imparted to the assimilating bacteria.

  For the utilization of carbonization characteristics and combustion characteristics of solidified materials, fermentation production of binders according to the application becomes important, fermentation conversion for improving shape stability and combustion efficiency of solidified materials during high heat treatment, Fermentation conversion to reduce the production of nitrogen-based compounds and sulfur-based compounds that are by-products due to nitrifying bacteria, desulfurizing bacteria, etc. is important.

  Development of a process centering on the fermentation process in the present invention enables efficient production of a solidified material. By using a known machine manufacturing technique, fermentation conditions determined after examining the biomass conversion method in the present invention, pretreatment of biomass, storage and transport conditions according to the physical properties of the fermentation product, other materials and It is clear that it is possible to develop an apparatus for optimizing the manufacturing process in accordance with the mixing conditions, molding process conditions, molding storage / transport conditions, and the like.

  Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples.

Example 1
It is a gel producing gel such as gellan gum to an agar medium (LB agar medium) prepared by adding agar (2%) to LB liquid medium (composition: 1% Bacto-tryptone, 0.5% Yeast extract, 1% NaCl) After inoculation with Pseudomonas elodea ATCC 31461 and growing at 32 ° C., a single colony was inoculated into 10 ml of LB liquid medium and cultured overnight at 32 ° C. with shaking. Then inoculate 1 ml of the culture solution into a medium containing 2% (air dry weight) okara, 0.3% Yeast extract, 0.5% Peptone, 0.3% NaCl, and shake culture at 32 ° C at 180 rpm. went. As a control, 1 ml of LB liquid medium was inoculated instead of the bacterial culture. From the cell culture and control on day 3, 1.5 ml was taken out and applied to 0.2 g of wood flour (air-dried chopsticks finely ground (using a pencil sharpener)), and then mixed with a spatula. This was shaped by hand into a dumpling having a length of 20 mm, a width of 10 mm, and a height of about 9 mm, and placed on a hot plate and dried at 150 ° C. for 30 minutes. After drying, both were taken out and cooled to room temperature. As a result of performing the operation of pressurizing the central part of this dumpling-shaped pellet with 5 kg / cm ² for 10 seconds (pressurized area approximately 0.20 cm ² ) using FRUIT HARDNESS TESTER 10 times, there were 3 control pellets. However, the cell culture broth pellet did not collapse.

(Example 2)
As in Example 1, after inoculating P. elodea ATCC 31461 on LB agar medium and growing at 32 ° C, single colonies were inoculated into 10 ml of LB liquid medium and shaken overnight at 32 ° C. Cultured. Then, inoculate 1 ml of the culture solution into a medium containing 2% (air-dried weight) plant cell wall component cellulose fine powder, 0.3% Yeast extract, 0.5% Peptone, 0.3% NaCl, and rotate 180 ° C at 32 ° C. Shaking culture was performed at / min. On the other hand, instead of the bacterial cell culture, 1 ml of LB liquid medium was inoculated as a control.
The cell culture on day 3 and the entire control culture were centrifuged (5,500 rpm, 15 minutes, manufactured by Hitachi, Ltd.), and the supernatant was removed except for 1.5 ml of the precipitate. The obtained precipitate was mixed with 0.2 g of wood flour with a spatula. Next, this mixture was shaped by hand into a dumpling having a length of about 20 mm, a width of 10 mm, and a height of about 9 mm, and placed on a hot plate and dried by heating at 150 ° C. for 30 minutes.
After drying, both were taken out and cooled to room temperature. As a result of one operation of pressurizing the central part of this dumpling-shaped pellet at 9 kg / cm ² for 10 seconds (pressurized area of about 0.20 cm ² ) using FRUIT HARDNESS TESTER, the control pellet gradually Although the cellulose fine powder was broken and spattered and disintegrated, the pellet derived from the cell culture broth did not disintegrate at all.

(Example 3)
After inoculating Bacillus subtilis NAFM5, a polyglutamic acid-producing bacterium (natto), on LB agar medium and growing at 37 ° C, inoculate a single colony into 10 ml LB liquid medium and shake overnight at 37 ° C. Cultured at last. Then, inoculate 5 ml (air dry weight) of okara with 5 ml (air dry weight) of okara in a medium containing 15 ml of 1% glucose and 1% sodium glutamate, and then inoculate into a medium sterilized by autoclaving. Incubation was performed. On the other hand, instead of the bacterial cell culture, 1 ml of LB liquid medium was inoculated as a control.
After taking 0.4 g of the medium from the cell culture and control on the 6th day and suspending it in 1.5 ml of water, it was applied to 0.2 g of wood flour described in Example 1, and this was mixed lightly with a spatula. Then, the mixture was shaped by hand into a dumpling having a length of 24 mm, a width of 14 mm, and a height of about 7 mm, and placed on a hot plate and dried by heating at 150 ° C. for 30 minutes.
Then, both were taken out and cooled to room temperature. The control pellet collapsed as a result of one operation of pressing the central part of this dumpling-shaped pellet with a FRUIT HARDNESS TESTER at 2.5 kg / cm ² for 1 second (pressurized area approximately 0.20 cm ² ). Although the wood flour was scattered, the pellet derived from the cell culture broth did not collapse.

Example 4
1.0 g of each of the cell culture and the control sample after completion of the cell culture in Example 3 was taken out, put into a 5 ml plastic syringe (manufactured by Terumo Corporation, without needle, middle mouth), and moved to the tip. . The plunger part of this syringe was pushed using a FRUIT HARDNESS TESTER, and the pressure when the sample was discharged smoothly from the tip (1 mm / sec or more) was measured. As a result, the cell culture was 0.17 kg / cm ² In contrast, the control sample required a pressure of 1.9 kg / cm ² , which was more than 10 times. From this, improvement in operability by culturing biomass was confirmed.

(Example 5)
0.6 g of the cell culture and the control sample after completion of the cell culture in Example 3 were taken out, and 0.6 g of the wood flour prepared in Example 1 and water were mixed together with 0.2 ml. Cut the tip of a 1.0 ml plastic syringe (manufactured by Terumo Corporation), prepare a tip with the same inner diameter as the entire inner diameter of the syringe, and after extruding it after filling the above mixture, diameter 4 mm The pellet was formed in a length of about 12 mm. This was processed on a hot plate at 150 ° C. for 30 minutes and dried to obtain dry pellets. Three dry pellets made from each sample were placed in a 50 ml plastic tube and shaken vigorously up and down for 10 minutes, but both did not collapse. Furthermore, after putting 2 dry pellets made from each sample into a 50 ml plastic tube, 10 ml of water was added and the lid was closed, and this was shaken at 28 ° C. for 120 reciprocations / minute for 12 hours. After standing at room temperature for 7 days and stirring for 10 seconds using a vortex mixer, the pellet derived from the cell culture completely disintegrates and the wood flour disperses, and it is clear that the disintegration in water is high. However, the control pellet hardly disintegrated.

(Example 6)
1.0 g of the wood flour prepared in Example 1 was placed in a 100 ml Erlenmeyer flask, and 1 ml of an aqueous solution containing 2% Peptone, 0.1% Yeast Extract and 0.5% MgSO ₄ .7H ₂ O was added to 0.7% K. After adding 1 ml of an aqueous solution containing ₂ HPO ₄ and 0.3% KH ₂ PO ₄ , the silicon stopper was closed and the mixture was autoclaved (121 ° C., 20 minutes) as the medium. This medium was inoculated with a 5 mm square agar block containing mycelia from a PDA medium slant on which Trichoderma reesei NBRC 31329 was grown, and left to stand at 32 ° C. for 8 days. A medium not inoculated with an agar block with a mycelium was used as a control. After culturing, each culture was taken out and shaped into dumplings of about 24 mm in length, 14 mm in width, and 7 mm in height according to the method of Example 3 and placed on a hot plate at 150 ° C. Heat drying for 30 minutes.
Then, both were taken out and cooled to room temperature. As a result of one operation of pressing the central part of this dumpling-shaped pellet at 5 kg / cm ² for 1 second using a FRUIT HARDNESS TESTER (pressure area approximately 0.20 cm ² ), the control pellet collapsed. Although the wood flour was scattered, the pellet derived from the cell culture broth did not collapse.

The present invention provides a technology for solidifying biomass, which is a waste resource generated in biological industries such as agriculture, forestry and fisheries and food industries. Moreover, the obtained solidified material can be added to high value and effectively used in various fields such as fertilizer and feed.

Claims (4)

The following (A) and (B) steps are performed on a biomass raw material containing at least one or more components belonging to plant cell wall components and protein components. For producing a solidified material having a binding property of.
(A): The process of obtaining the fermented material containing the extracellular component which has a binder characteristic by fermenting the said biomass raw material using the 1 or more types of microorganisms which have the characteristics of (a) and (b) below.
(a): One or more microorganisms belonging to polyglutamic acid-producing bacteria, gellan gum-producing bacteria, polygalactosamine-producing bacteria and curdlan-producing bacteria.
(b): A microorganism having a resolution for at least one component belonging to the plant cell wall component and protein component contained in the raw material.
(B): The process which shape | molds and solidifies the said fermented material or the composition containing the said fermented material.
(C): The property of being difficult to disintegrate under pressure and easily disintegrating in water. The manufacturing method of the solidification raw material of Claim 1 in which the composition containing the said fermented material contains the said unfermented biomass raw material. The method for producing a solidified material according to claim 1, wherein the microorganism is Pseudomonas elodea and / or Bacillus subtilis. Non-food part of crops, waste crops; livestock manure, waste during processing of meat; waste during processing of marine products; thinned wood, sawdust, waste fungus bed; waste during food production A solidified material according to any one of claims 1 to 3, which is one or more of: waste from food distribution; waste from garbage, building waste, pruning residue, waste paper, and waste cloth; Method.