TW200911998A - Method for producing glyceric acid - Google Patents

Abstract

There is provided is a method for producing glyceric acid from glycerol, which comprises culturing a specific bacterium capable of producing glyceric acid in a culture medium containing glycerol, or making cells of the bacterium, processed cells of the bacterium or immobilized product thereof contact with glycerol.

Description

200911998 IX. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates to a method for producing glyceric acid from glycerol using microorganisms. [Prior Art] Biodiesel fuel (BDF) is a carbon-neutral light oil alternative fuel and is currently receiving attention as a fuel to help solve environmental problems such as energy depletion, global warming and air pollution. ζ% Biodiesel fuels are manufactured from vegetable oils, animal fats, waste oils, and the like. Today, waste liquids containing glycerin are manufactured as a by-product. There is no specific use for this waste oil and it can only be discarded today. As for the method of using the biodiesel waste liquid, Japanese Unexamined Patent Publication No. Hei No. 2006-180782 discloses a method of producing hydrogen and ethanol from waste liquid glycerol by Enterobacter bacteria.

Regarding the production of an organic acid, Japanese Unexamined Patent Publication No. 2003-235592 discloses a method of producing an organic acid from glucose as a carbon source G by an aerobic bacterium (especially a rod-shaped bacterium). PCT International Publication WO 02/00846 揭示1 discloses a strain (Mannheimia Sp. 55E) which produces a variety of organic acids from glucose. However, bacteria which can produce glyceric acid from glycerol have not been known to date. SUMMARY OF THE INVENTION In order to utilize resources efficiently, it is desirable to produce useful materials from biodiesel waste liquid. The object of the present invention is to provide a glycerol acid from glycerol in a simple manner. 131377.doc 200911998

a method of cultivating a bacterium belonging to a genus of bacteria belonging to the genus of Methylobacterium, Arthrobacter, and Pseudonocardia in a medium containing glycerin; or Streptomyces globisporus subsp. globisporus, Streptomyces albovinaceus, Streptomyces althioticus, Streptomyces arenae, Streptomyces erythrogriseus, Streptomyces fumigatus (Streptomyces fumanus), Streptomyces gardenri, Streptomyces globosus, Streptomyces longispororuber, Streptomyces moderatus, Streptomyces sclerogranulatus, Streptomyces senoensis, Streptomyces tanashiensis subsp. cephalomyceticus, Streptomyces tuirus, Bacillus subtilis, Brevibacterium buta Bacteria of the species of the bacterium, a group of bacteria of the genus Pseudomonas sp. and Nocardia globerula (hereinafter, these bacteria are collectively referred to as "for the present invention The glycerin-producing bacteria " or simply the "bacteria producing glyceric acid" is particularly cultivating the bacterium under aerobic conditions, or the bacterial cell, the processed cell of the bacterium or the bacterium thereof The immobilized product is contacted with glycerin. That is, the present invention provides the following: 1. A method for producing glyceric acid from glycerol, the method comprising: 131377.doc 200911998 cultivating a glycerol-containing medium having the ability to produce glyceric acid from glycerol and belonging to a bacterium selected from the group consisting of a bacterium of the genus Bacillus belonging to the genus, Arthrobacter, and Pseudonocardia; or contacting the bacterial cell, the processed cell of the bacterium, or the immobilized product thereof with glycerin; 2_1 The method wherein the bacterium belonging to the genus of bacteria belonging to the genus Bacteroides, Arthrobacter, and Pseudonocardia is Methylobacterium extorquens, Rhodesianum methyl bacilli ( Methylobacterium rhodesianum), Methylobacterium zatmani, Roseroparaffinus Arthrobacter roseoparaffinus, Arthrobacter paraffineus, Arthrobacter sp., Arthrobacter globiformis or self a bacterium of Pseudonocardia autotrophyica; 3. the method of 2 above, The bacterium belonging to the genus of the genus of the genus Bacillus, Arthrobacter, and Pseudonocardia is M. exoskeletus JCM 2802 strain, Rhodesianum faecalis JCM 2810 strain, zatmani methyl Bacillus JCM 2819 strain, roseoparaffinus, Arthrobacter sp. ATCC 15584 strain, Arthrobacter sphaeroides ATCC 21535 strain, Arthrobacter sp. ATCC 21908 strain, Arthrobacter sp. NBRC 12137 strain or Nocardia donovani NBRC 12743 strain; A method for producing glyceric acid from glycerol, the method comprising: cultivating a glycerol-containing medium having the ability to produce glyceric acid from glycerol and belonging to a bacterium selected from the group consisting of the genus Coccidella sp., the genus Streptomyces erythraea, the genus 131377.doc 200911998 Streptomyces, Streptomyces cerevisiae, Streptomyces erythrogriseus, Streptomyces faecalis, Streptomyces faecalis, Streptomyces styrofolia, Streptomyces erythraea, Streptomyces moderatus, Streptomyces granulosus, Streptomyces cerevisiae, Streptomyces vaginalis Streptomycin, Streptomyces, Bacillus subtilis, Bacterium butanicum, Pseudomonas and Nocardia - a bacterium of the bacterium of the bacterium population; or contacting the bacterial cell, the processed cell of the bacterium, or the immobilized product thereof with glycerin; Γ 5. The method according to the above 4, wherein the bacterium is selected from the bacterium of the genus Cocci Spores, Streptomyces erythraea, Streptomyces faecalis, Streptomyces faecalis, Streptomyces erythrogriseus, Streptomyces fumigatus, Streptomyces faecalis, Streptomyces styrofolia, Streptomyces faecalis, Streptomyces moderatus, Streptomyces granulosus a strain of a bacterium of the genus Streptomyces cerevisiae, Streptomyces cerevisiae, Streptomyces faecalis, Bacillus subtilis, Brevibacterium butanicum, Pseudomonas spp., and Nocardia The bacterium is Streptomyces cocci, ATCC 21903 strain, Streptomyces erythropolis NBRC 12739 strain, Streptomyces facilis NBRC 15956 strain, Streptomyces cerevisiae NBRC 13016 strain, Streptomyces erythrocriseus NBRC 14601 strain, Streptomyces fumigatus NBRC 13042 strain, Streptomyces natto strain NBRC 12865, Streptomyces cerevisiae NBRC 15874 strain, Streptomyces cerevisiae NBRC 13488 strain, Streptomyces cerevisiae NBRC 13432 Strains, Streptomyces granulosus NBRC 14301 strain, Streptomyces cerevisiae NBRC 13 843 strain, Streptomyces solani sp. NBRC 13929 strain, Streptomyces sp. NBRC 15617 strain, Bacillus subtilis NBRC 3026 strain 131377.doc 200911998 strain, Butanecum sinensis ATCc 21196 strain, Pseudomonas sp. ATCC 53617 strain or Nocardia sphaeroides ATCc 15076 strain; 6. The method according to any one of the above 1 to 5, wherein the bacterium is aerobic The method of any one of the above 1 to 6, further comprising recovering glyceric acid from the culture obtained by the culture; and 8. The method according to any one of the above 1 to 7, wherein the glycerin is From biodiesel waste. ('Embodiment> Examples of the bacterium for producing glyceric acid used in the present invention are the genus, strains and strains described in the above 1 to 5. Among the bacteria producing glyceric acid, preferred bacteria are capable of Glyceric acid is produced from glycerol in a higher yield. Examples of such bacteria include Nocardia globosa ATCC 15076 strain, Bacillus exobacillus] 2] 2802 strain, 1 > 11 〇 (^ 341111111 methyl Bacillus 1 (::) 2810 strain, Artobacter rosette ATCc 15584 strain, Arthrobacter sphaeroides ATCC 21535 strain, Arthrobacter species atcC 21908 strain 'Bacillus cerevisiae NBRC 12137 strain, Nocardia donovani NBRC 12743 strain, Streptomyces sp. sp. variant ATCC 21903 strain, Streptomyces cerevisiae NBRC 13016 halobacterium, erythrogriseus strain NBRC 14601 strain, Streptomyces faecalis NBRC 13042 strain, Streptomyces NBRC 12865 strain, Streptomyces cerevisiae NBRC 15874 strain , Streptomyces cerevisiae NBRC 13488 strain, Streptomyces cerevisiae NBRC 13432 strain, Streptomyces granulosus NBRC 14301 strain, Streptomyces cerevisiae NBRC 13 843 strain, Streptomyces sp. Rc 13929 strain, Paul 131377.doc 200911998 Streptomyces sp. NBRC 15617 strain, Bacillus subtilis NBRC 3026 strain, Brevibacterium butanicum ATCC 21196 strain, Streptomyces erythropolis NBRC 12739 strain, S. isothiocyanum NBRC 15956 strain, zatmani methyl Bacillus strain JCM 2819 and Pseudomonas ATCC 53617 strain, but not limited thereto. Rhodesianum methyl bacillus JCM 2810 strain, 2 & 31 Methyl bacillus 1〇^2819 strain, Arthrobacter VIII 7' (^ 21908 Strains, Streptomyces isothiorum NBRC 15956 strain, Streptomyces erythropolis NBRC 12739 strain and Pseudomonas sp. ATCC 53617 strain, zatmani methyl bacillus JCM 2819 strain, Streptomyces erythropolis NBRC 12739 strain and pseudomonas The strain ATCC 53617 is more preferred. Herein, the production of glyceric acid in "higher yield" means that glyceric acid is accumulated from glycerol in a medium at a yield of about 0.1 g/L or higher. In general, the higher the yield The better, for example, the accumulation of glyceric acid in the medium at a yield of about 0.5 g/L or higher, preferably about 1 g/L or higher. Methyl bacillus JCM 2802 strain, Rhodesianum methyl bacillus J CM 28 1 0 strain zatmani methyl Bacillus strain JCM 2819 available from said present, Saitama 351-0198, Wako, 2-1 Hirosawa, RIKEN BioResource Center "Japan Collection of Microorganisms Depositary / sector microorganisms. Nocardia sphaeroides ATCC 15076 strain, Artobacterium roseoparaffinus ATCC 15584 strain, Arthrobacter sphaeroides ATCC 21535 strain, Arthrobacter sp. ATCC 21908 strain, Streptomyces cocci, ATSC 21903 strain, Brevibacterium butanicum ATCC 211 The % strain and the Pseudomonas sp. ATCC 53617 strain were purchased from VA 20108 USA, Manassas, PO BOX 1549, American Type Culture Center. Arthrobacter globulin NBRC 12137 131377.doc 10· 200911998 strain, Nocardia donovani NBRC 12743 strain, Streptomyces erythraea NBRC 12739 strain, Streptomyces isothiorum NBRC 15956 strain, Streptomyces cerevisiae NBRC 13016 strain, Streptomyces erythrogriseus NBRC 14601 strain, Streptomyces facilis NBRC 13042 strain, Streptomyces nitrite NBRC 12865 strain, Streptomyces cerevisiae NBRC 15874 strain, Streptomyces cerevisiae NBRC 13488 strain 'moderatus Streptomyces NBRC 13432 strain, Streptomyces nucleatum NBRC 14301 Strain, Streptomyces cerevisiae nBRC 13843 strain, Streptomyces acuminata variant NBRC 13929 strain, Bao

Streptomyces NBRC15617 strain and Bacillus subtilis NBRC 3026 strain were purchased from Chiba 292-0818 Japan, Kisarazu-shi, 2-5-8

Kazusakamatari, the merger management agency of the National Institute of Technology and Evaluation, the Biotechnology Division, and the Bioresources Center. The glycerol-producing bacteria used in the present invention may be not only wild-type strains, but also any given naturally occurring or artificial mutant strains, including xenon radiation, ultraviolet radiation or chemical mutagens (for example, N-methyl _ N, _nitro-N_nitrosoguanidine) is a recombinant, or a recombinant obtained by genetic engineering techniques such as cell fusion and genetic recombination. The host of the recombinant may belong to any genus as long as it is a transformable microorganism. However, the host preferably belongs to (4) of the parental strain (4) of the gene in which the target is produced. It is preferred to select a bacterium which produces glyceric acid having an improved ability to convert from glycerol to glyceric acid. The medium of glycerol may be a medium obtained by adding human pure glycerin or a mixture containing glycerin. In the mixture containing glycerol, the group other than glycerin or its content is preferably used in the present invention. The fruit of the glycerin-containing linear acid-containing hydrazine is not particularly limited, but it is preferably used as a bio-diesel waste liquid. One of the methods for producing biodiesel fuel is to produce fatty acid methyl ester (FAME) by alcoholysis of triacetin using a base catalyst. In this method, the glycerin-containing waste liquid is produced as a by-product (referred to as biodiesel waste liquid). The waste liquid is generally contaminated by catalysts, unconverted fatty acids which vary with the oil used, and the like. For example, the group of biodiesel waste liquid used in the following examples is glycerol·51%, decyl alcohol: 11%, potassium hydroxide: 8%, water: 4%, and other glycerides: 26%. It is also possible to use bio-wood with an indefinite composition

Oil waste liquid, for example, as described in s. Papanik〇la〇u et al., Bio-Resource Technology (2002) 82: 43-49: 65%, potassium/sodium salt: 4 to 5%, methanol: 1% Water: 28%, and in M Gonzalez Pajuel〇 et al.;

Glycerol 65%, sodium salt: 5 as described in Ind Microbiol Biotechnol (2004) 31:442-446. /. Or less. When biodiesel waste liquid is added to the medium of the method of the present invention, glyceric acid can be produced in the same or higher yield and conversion rate as in the case of adding pure glycerin. The medium used in the method of the present invention may be of any kind as long as it contains the usual components necessary for cultivating bacteria, and is not limited to a specific medium. In the present invention, glyceric acid is obtained even in a medium having a simple composition containing a carbon source, a nitrogen source, and an inorganic salt. The medium of the method of the present invention contains the H content as a carbon source in the range of about 0.1 to 500 g/L, and preferably from about 0.2 to 500 g/L, for the growth of bacteria and the production of glyceric acid. When the medicinal waste liquid of the medicinal medicinal liquor is used as a glycerin source, depending on the effluent, depending on the degree of aging, the effluent or glycerin may be diluted therein until 131377.doc 200911998 The glycerin content in the medium falls within the above range . The medium may contain substances other than glycerin as a carbon source, but the amount thereof should be limited to such an extent that it does not affect the production of glyceric acid from glycerin. Examples of the carbon source used for the hairpin are glucose, fructose, starch, lactose, arabinose, xylose, dextrin, syrup and malt extract, but are not limited thereto. The other carbon source content is preferably about 10% by weight or less of glycerin, and more preferably about! % by weight or less. The medium preferably contains glycerol as a single carbon source.

Examples of the nitrogen source include inorganic nitrogen compounds such as ammonia, ammonium sulfate, ammonium chloride, and succinic acid, urea, and the like. Organic nitrogen sources such as tantalum powder, cottonseed meal, soy flour, corn syrup, dry yeast, yeast extract, peptone, meat extract and casein amino acid may also be added to the culture medium. It is advantageous to use a combination of a carbon source and a nitrogen source. Since low-purity sources containing trace growth factors and large amounts of inorganic nutrients are also suitable, it is not necessary to use them in a pure state. If necessary, inorganic salts such as dihydrogen hydride "dipotassium hydrogen hydride, sodium chloride, magnesium sulfate, manganese sulfate, calcium carbonate, calcium carbonate, sodium iodide, potassium iodide, and cobaltated cobalt may be used. Defoaming agents such as liquid paraffin, decyl alcohol, vegetable oil, mineral oil and anthrone can also be added as needed, especially when the medium is significantly foamed. Other components such as multivitamins may be added to the medium as needed. Nitrogen sources, inorganic salts and other components are known to those skilled in the art. In the present invention, bacteria producing glyceric acid can be cultured under anaerobic conditions, but it is preferably carried out under aerobic conditions. Aerobic conditions refer to culture in the presence of molecular oxygen. Ventilation, agitation and shaking can be performed to provide oxygen. Culture can be used 131377.doc -13- 200911998: any common device for living organisms. The method of culturing under aerobic conditions τ can be used to culture bacteria and produce glyceric acid in any convenient manner necessary to produce anaerobic conditions. 0 Bacteria culture under surface oxygen conditions can be carried out by introducing carbon dioxide or an inert gas (milk, argon, etc.) or without aeration.

The mass production of glyceric acid is preferably carried out under liquid submerged culture conditions. When breeding in a large tank, it is preferred to inoculate the bacteria into the second & tank during the growing period to avoid delayed propagation during the process of producing glyceric acid. That is, the bacteria are preferably first inoculated into a relatively small amount of medium, and cultured in the growing season: to produce the strain 'and then aseptically transfer the strains to the large tank culture solution (4) and the aeration can be carried out in various ways. . (d) It can be carried out by means of a spiral or a mechanical agitation device similar to a spiral, a rotation or vibration of the fermentation tank, or a pumping device. Ventilation allows the sterilized air to be passed through the culture solution and is dispensed <in the operating towel, the aeration operation can also produce a mixing effect. In the case of culturing in a liquid submerged medium, a culture method such as batch culture, bed batch culture, and continuous culture can be appropriately selected and used. The culture conditions are suitable for cultivating the glycerol producing bacteria of the present invention. For example, the culture temperature is from about 4 to about 15, preferably from about 20 to about. The pH of the medium is from about 5 to 9, and preferably from about 6 to 8. When the pH of the culture medium decreases with the production of glyceric acid, a base (e.g., aqueous ammonia solution, calcium carbonate, sodium hydroxide, and potassium hydroxide) may be added to the culture system as needed to adjust the pH so as to fall within the above range. The composition of the medium and other culture conditions can be appropriately adjusted by those skilled in the art. 131377.doc -14- 200911998. Adjustment conditions will also be considered to further increase the yield of glyceric acid. The bacteria used in the method of the present invention may be bacterial cells, processed bacterial cells or immobilized products thereof. Herein, the processed bacterial cells represent ruptured bacteria, 'field cells' or enzymes extracted from cultured substances (including bacterial cells and culture supernatant). Examples of the processed bacterial cells include those obtained by culturing bacterial cells with an organic acid (for example, C and ethanol), lyophilization or alkali treatment, those obtained by physical or enzymatic chemical disruption of bacterial cells, or crude enzymes isolated or extracted therefrom. . Specifically, the cultured fine g is centrifuged, and the cells to be collected are subjected to physical grinding methods (for example, ultrasonic, Dyno_mill and French compression treatment) or chemistry using a surfactant or a decomposing enzyme (for example, lysozyme). The rupture method is broken. The resulting solution is centrifuged or membraned to remove insolubles, and the resulting cell-free extract is subjected to separation/purification methods such as cation exchange chromatography, anion exchange chromatography, hydrophobic chromatography, gel filtration chromatography. Method and metal chelate chromatography to fractionate and purify the enzyme. Examples of the carrier used in the chromatography include an insoluble polymer carrier such as cellulose, dextroethylidene (DEAE), phenyl or butyl, dextrin and cyclosporin. Commercially available columns filled with a carrier can also be used. The disruption of the bacterial cells and the extraction of the enzyme can be carried out by a method known to those skilled in the art and the above methods. / Examples of contacting bacterial cells, incubated bacterial cells or their immobilized products with glycerol are as follows. An example of a method for producing glyceric acid from glycerol using bacterial cells or processed bacterial cells is a method of suspending and culturing bacterial cells in a glycerol-containing substrate solution and reacting 131377.doc 200911998. Bacterial cells are cultured to produce glyceric acid bacteria, followed by centrifugation to prepare a gluten-free glycerin concentration of about 〇1 to 50% by weight. The reaction temperature is generally about 4 to 4 Torr. Oh, and preferably from about 2 to 37. Hey. The pH of the reaction solution is generally from about 5 to 9, and preferably from about 6 to 8. When the pH of the medium is lowered as the production of the glycerol I, a base (e.g., ammonia aqueous solution of carbonic acid, sodium hydroxide, and hydroxide) can be added to the culture system as needed to adjust the pH to fall within the above range. The method for producing glyceric acid from glycerol using immobilized bacterial cells or immobilized processed bacterial cells is, for example, filling the fixed bacterial cells or immobilized processed bacterial cells into a column, and passing the glycerol-containing receiving solution The method of the column. Bacterial cells or processed bacterial cell lines are obtained by culturing bacteria producing glyceric acid and then centrifuging. The method of immobilizing bacterial cells is, for example, a method of extensively immobilizing a gel, and a method of immobilizing an ion exchange material. Examples of gels that can be used include carrageenan, agar, mannan, PVA, and polypropylene guanamine gel. The suitable particle size of the gel is about 10 mm in diameter, but the size varies with the type of gel. Examples of the ion exchange material include a cellulose-based material, a styrene-divinylbenzene-based material, and an ion exchange material mainly composed of benzophenone. The glycerin wave in the substrate is preferably from about 0.01 to 50% by weight. SH compounds (such as mercaptoethanol, cysteine, and glutathione), reducing agents (such as sulfites), and enzyme activators (such as magnesium ions and manganese ions) may also be added. It varies with the size of the column and the quality of the fixture. As the speed index of the treatment solution, it is preferred that the space velocity (ml/ml resin.hr) is from 〇5 to 1 〇. The separation and purification of glyceric acid is carried out according to a conventionally known method. For example, 131377.doc -16 - 200911998 After the completion of the culture, the culture solution is filtered or separated to obtain an upper layer liquid. From the supernatant liquid, for example, glyceric acid can be isolated, for example, by concentrated crystallization, but the separation and purification of glyceric acid are not limited thereto. Specifically, glyceric acid may be isolated from the supernatant by, for example, solvent extraction, or may be isolated and purified by, for example, ion exchange chromatography (in which glycerate is adsorbed and dissolved after ion exchange resin). And separation), separation of 'insoluble treatment by fractional precipitation, fractional crystallization by crystallization, membrane separation by reverse osmosis membrane, and concentrated crystallization by separation of metal salts such as calcium ions. Specifically, for example, glyceric acid can be isolated and purified according to the method described in Japanese Unexamined Patent Publication No. Hei. The invention is further illustrated by the following examples. It is to be understood that the invention is not limited to the examples, and various modifications may be made within the scope of the invention. EXAMPLES Example 1 The rhodesianum methylbacterium JCM 281 strain was coated on agar medium a for plate culture (composition: 3 g potassium dihydrogen phosphate, 6 § disodium hydrogen phosphate, 0.5 g sodium chloride, 8 Ammonium chloride, 492 mg of magnesium sulfate heptahydrate, 147 mg of calcium carbonate dihydrate, 1 mg of yeast extract, lyoglycerol, 20 g of agar and 1 l of distilled water (final pH 7.4) and The cells were allowed to stand at 3 (rc for 4 days). The strain grown on the above plate was inoculated into 3 mL of medium B by a platinum loop, which was used for tube culture (composition: the same composition as agar medium A, but not Containing calcium carbonate, yeast extract and agar) 'Culturing (pre-culture) with shaking at 200 rpm for 24 hours at 30 C. 30 μί The above-mentioned grown strain culture medium was transferred to 3 mL of medium, 131377.doc • 17- 200911998 B was used for tube culture and cultured at 30 rpm with shaking at 200 rpm (main culture). After 4 days from the start of the reaction, 〇·4 g of glycerol was consumed and 〇lg of glycerol was accumulated per liter. 2 Wang 1; 111311 Thiobacillus <1 € 1 ^ 2819 strain is the same as in Example 1 The reaction was carried out. As a result, 0.3 g of glycerol was consumed and 0.3 g of glyceric acid was accumulated per 1 liter. Example 3 1'11〇(163131111111) Bacillus licheniformis>^1^2810 strain was applied to agar medium VIII. Culture (composition: 3 g potassium dihydrogen phosphate, 6 g phosphoric acid sodium, 0.5 g gasification sodium, 1 g gasification money, 492 mg sulfuric acid sulfate, 147 mg dihydrate gasification mother, 1 mg mg yeast Extract, 1 layer of glycerin, 20 g of agar and 1 L of distilled water (final pH 7.4)), and allowed to stand at 3 〇 ac for 4 days. The strain grown on the above plate was inoculated to 3 by recording ring. In the medium C, the medium c was used for test tube culture (composition: the same composition as agar medium A, but no agar), and shake culture (preculture) with 2 〇 () 卬 melon at 3 〇t for 24 hours. The culture medium of the above-mentioned growth strain was transferred to 3 mL of medium C, which was used for test tube culture, and cultured at 200 rpm with shaking at 30 rpm (main culture). After starting the reaction for 4 days, 0_9 g of glycerol was consumed. 9 g of glycerol was accumulated per 1 liter. Example 4 zatmani methyl bacillus jCM 2819 strain was as in Example 3 Mode reaction 'but the pre-culture period was changed from 24 hours to 6 days, and the main culture period was changed from 4 days to 6 days. Results 'Ug glycerol was consumed and 1.3 g of glyceric acid was accumulated per liter. Example 5 131377.doc -18- 200911998 The B. mobilis strain JCM 2802 was reacted in the same manner as in Example 3, but the period from pre-culture was changed from 24 hours to 6 days, and the period from main culture to 4 days was changed to 6 days. As a result, i.4 g of glycerin was consumed and 4 g of glyceric acid was accumulated per 1 liter. Example 6 Nocardia sphaeroides ATCC 15076 strain was reacted in the same manner as in Example 3. As a result, 5.8 g of glycerin was consumed and 〇 2 g of glyceric acid was accumulated per 1 liter. Example 7 Streptomyces coelis sp. variant ATCC 21903 strain was reacted in the same manner as in Example 3. As a result, 7.3 g of glycerin was consumed and accumulated per liter of glycerol. Example 8 S. erythraea NBRC 12739 strain was reacted in the same manner as in Example 3. As a result, 10.3 g of glycerol was consumed and 丨.1 g of glyceric acid was accumulated per liter. Example 9 The strain Streptomyces isothiocyanum NBRC 15956 was reacted in the same manner as in Example 3. As a result, 2.2 g of glycerin was consumed and 5 g of glyceric acid was accumulated per 1 liter. Example 10 Roseoparaffinus Arthrobacter ATCC 15584 strain as in the example! The same way of reacting. As a result, g 6 g of glycerin was consumed and 0.2 g of glycerol was per 1 well. Example 11 Nocardia pseudobacteria NBRC 12743 strain was reacted in the same manner as in Example 3. As a result, 3.7 g of glycerin was consumed and 1 § of glyceric acid was accumulated per 1 liter. Example 12 131377.doc -19- 200911998 Dendrobium MATCC 21535 strain was reacted in the same manner as in Example 3, but the pre-culture period was changed from 24 hours to 6 days, and the main culture period was changed from 4 days to 6 days. As a result, 4.3 g of glycerin is consumed and each! Literated 〇 & glyceric acid. Example 13 Abacillus sp. ATCC 2 strain was reacted in the same manner as in Example 3. As a result, 5.7 g of glycerin was consumed and 7 g of glyceric acid was accumulated per liter. Example 14

The butanicum bacillus ATCC 2丨i 96 strain was reacted in the same manner as in the case of hydrazine. As a result, 〇·5 g of glycerin was consumed and 2 g of glyceric acid was accumulated per liter. Example 15 S. cerevisiae NBRC 13016 strain was reacted in the same manner as in Example 3. As a result, 4.8 g of glycerin was consumed and 2 g of glyceric acid was accumulated per liter. Example 16

The Whrogriseus sclerotium NBRC 146〇1 strain was reacted in the same manner as in Example 3, but the pre-culture period was changed from 24 hours to 6 days and the main culture period.

The time was changed from 4 days to 6 days. As a result, 3 7 g of glycerol was consumed and ^ liter accumulated glyceric acid. S Example 1 7 Streptomyces facilis NBRC 13042 strain reacted in the same manner as in Example 3 'but the period from pre-culture was changed from 24 hours to 6 days, and during the main culture period from 4 days to 6 days. Results 8.7 § glycerol was consumed And accumulated 2 g of glyceric acid per liter. Example 1 8 Streptomyces NBRC 12865 strain was reacted in the same manner as in Example 3 'but the period from pre-culture was changed from 24 hours to 6 days, and during the main culture period from \ days 131377.doc •20- 200911998 to 6 days . As a result, 3 3 g of glycerin was consumed and 〇i g of glyceric acid was accumulated per 1 liter. Example 19 S. sphaeroides NBRC 15874 strain was reacted in the same manner as in Example 3, except that the pre-culture period was changed from 24 hours to 6 days, and the main culture period was changed from 4 days to 6 days. As a result, 4 g g glycerol was consumed and 〇丨g glyceric acid was accumulated per liter. Example 20 S. chrysogenum NBRC 13488 strain was spread on agar medium A for plate culture (composition: 3 g potassium dihydrogen phosphate, 6 § disodium hydrogen phosphate, 0.5 g sodium chloride, 1 g gas) Ammonium, 492 mg magnesium sulfate heptahydrate, 147 mg monohydrate gasification mom, 1 mg mg yeast extract, layer glycerin, 20 g agar and 1 L steamed water (final pH 7.4)) Allow to stand for 4 days at 3 °C. The strain grown on the above plate was inoculated into 3 mL of medium C by a platinum ring, which was used for test tube culture (composition: same composition as agar medium A, but no agar), at 3 °c The cells were shaken and cultured (precultured) at 2 rpm for 6 days. 30 pL of the above-mentioned grown strain culture medium was transferred to 3 mL of medium D for use in test tube culture (composition: the same composition as the above-mentioned medium C for test tube culture, but instead of 10 g of glycerol containing 19.6 g for manufacture) The glycerin fraction of the by-product of the biodiesel fuel (glycerol·51%, methylation: 11%, oxidative osmolarity: 8%, water: 4%, and others containing glycerides: 26%)) was at 3 〇. The armpit was shaken at 200 rpm (main culture). The reaction took 6 days to consume 11 g of glycerol and accumulated 3 g of glyceric acid per liter. Example 21 The moderatus Streptomyces NBRC 13432 strain reacted in the same manner as in Example 3 'but changed from 24 hours to 6 days during pre-culture period' and during the main culture period from 4 131377.doc 21 200911998 liters accumulated 0.2 g glycerol days to 6 day. As a result, 5.3 g of glycerin is consumed and each! acid. Example 22 The moderatus Streptomyces NBRC 13432 strain was reacted in the same manner as in Example 20. As a result, 6.3 g of glycerin was consumed and 1 liter per liter of glycerol was used. Example 23 Bacillus subtilis NBRC 3〇26 strain was reversed from the same manner as in Example 3. As a result, 1.0 g of glycerin was consumed and 1 g of glyceric acid was accumulated per 1 liter. Example 24 Pseudomonas sp. ATCC 53617 strain was reacted in the same manner as in Example i. As a result, 5. g g glycerol was consumed and accumulated per liter of glycerol. Example 25 Pseudomonas sp. ATCC 53617 strain was reacted in the same manner as in Example 3. As a result, 5.8 g of glycerin was consumed and 6 g of glyceric acid was accumulated per liter. Example 26 Streptomyces nucleatum NBRC 143 〇 lg strain was reacted in the same manner as in Example 2 (). As a result, 5. sg of glycerol was consumed and 〇ig glyceric acid was accumulated per liter. Example 27 S. cerevisiae gNBRC 13843 g strain was reacted in the same manner as the example paste. As a result, 9.2 g of glycine* was consumed and 2. 2 g of glyceric acid was accumulated per 1 liter. Example 28 Field Streptomyces sp. No. NBRC 13929 strain was reacted in the same manner as in Example 3, but changed from 24 hours to 6 days during pre-culture and from 4 days to 6 days during the main pull-up period. 74 g of glycerol and accumulate 131377.doc •22·200911998 ο.1 g of glyceric acid per liter. Example 29 Streptomyces NBRc 15617 strain should be pre-stretched as in the same manner as in Example 3, from 24 hours to 6 days, to δ days.蛀Special and main culture period from 4 days Example 3 母 Mother liter accumulated 〇.4g glyceric acid. The S. globosa NBRC m37 strain was reacted in the same manner as in the Example 丄. As a result, 4.5 g of glycerin was consumed and 〇·2 § glyceric acid was accumulated per liter. Ο Industrial Applicability According to the present invention, glyceric acid is produced from glycerol (biodiesel waste liquid) in a simple manner by microorganisms. Glyceric acid is used as an additive or a resin intermediate or lubricant, and the like. Accordingly, the present invention is used to make useful materials from waste materials. 131377.doc •23·

Claims (1)

200911998 X. Patent Application Range: 1. A method for producing glyceric acid from glycerol, the method comprising: cultivating a glycerol-containing medium having the ability to produce glyceric acid from glycerol and belonging to a group selected from the group consisting of Methylobacterium a bacterium belonging to the genus of the genus Arthrobacter and Pseudonocardia, or a bacterium of the genus of the genus; or the bacterial cell, the processed cell of the bacterium or the immobilized product thereof, is contacted with glycerin. (2) The method according to claim 1, wherein the bacterium belonging to the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus Extorquens), rhodesianum Methylobacterium rhodesianum, zatmani Methylobacterium zatmani, roseoparaffinus Arthrobacter roseoparaffinus, Arthrobacter paraffineus, Arthrobacter sp., sphere A bacterium of Arthrobacter O globiformis or Pseudonocardia autotrophica. The method of claim 2, wherein the method is selected from the group consisting of genus Bacillus, Arthrobacter, and Pseudonocardia. The bacterium belonging to the genus Bacteroides is a strain of Methyl bacillus JCM 2802, a strain of Rhodesianum methyl bacillus JCM 2810, a strain of M. zatmani JCM 2819, a strain of Arthrobacter roseoparaffinus ATCC 15584, and an Arthrobacter bacterium ATCC 21535 Strains, Arthrobacter sp. ATCC 21908 strain, ball 131377.doc 200911998 Arthrobacter NBRC 12137 Strain or autotrophic Nocardia NBRC 12743 strain. A method for producing glyceric acid from glycerol, the method comprising: cultivating a glycerol-containing medium having the ability to produce glyceric acid from glycerol and belonging to a strain selected from the group consisting of Streptomyces globisporus subsp. globisporus, white wine Streptomyces albovinaceus, Streptomyces althioticus, Streptomyces arenae, Streptomyces erythrogriseus, Streptomyces fumanus, Streptomyces gardenri, globular chains Streptomyces globosus, Streptomyces longispororuber, Streptomyces moderatus, Streptomyces sclerogranulatus, Streptomyces senoensis, Streptomyces senoensis Variant (Streptomyces tanashiensis subsp. cephalomyceticus), Streptomyces tuirus, Bacillus subtilis, Brevibacterium butanicum, Pseudomonas sp. The group consisting of bacterial species in the species Nocardia globerula (Nocardia globerula); or the bacterial cells, the bacterial cells or the processed product is contacted with the immobilized glycerol. 5. The method of claim 4, wherein the method is selected from the group consisting of Coccidella sp., Streptomyces erythropolis, Streptomyces faecalis, Streptomyces faecalis, Streptomyces faecalis, 131377.doc 200911998 erythrogriseus Streptomyces, Streptomyces fumigatus, Jiade Streptomyces, Streptomyces styrofolia, Rhizoctonia solani, Streptomyces moderatus, Streptomyces granulosus, Streptomyces cerevisiae, Streptomyces cerevisiae, Protecting Bacillus, Bacillus subtilis, Brevibacterium butanicum, The bacterium of the genus Pseudomonas sp. and the Nocardia sphaeroides is a strain of Streptomyces cocci, ATCC 21903, S. erythropolis NBRC 12739, Streptomyces isothiomas NBRC 15956 strain, Bacillus subtilis NBRC 13016 strain, erythrogriseus bacillus NBRC 14601 strain, Streptomyces fumigatus NBRC 13042 strain, Streptomyces NBRC 12865 strain, Streptomyces sp. NBRC 15874 strain, Keyius red bond NBRC 13488 strain, Moderatus sclerotium NBRC 13432 strain, Streptomyces nucleatum NBRC 14301 strain, Streptomyces cerevisiae NBRC 13843 strain, Streptomyces auratus Species NBRC 13929 strain, the protective strain NBRC 15617 Streptomyces, Bacillus subtilis strain NBRC 3026, butanicum Brevibacterium strain ATCC 21196, Pseudomonas sp. Strain ATCC 53617 or pellets strain Nocardia sp ATCC 15076. 6. The method of any one of claims 1 to 5, wherein the bacterium is cultured under aerobic conditions. 7. The method of any one of claims 1 to 6, further comprising recovering glyceric acid from the culture obtained from the culture. 8. The method of any one of claims 1 to 7, wherein the glycerol is derived from biodiesel waste. 131377.doc 200911998 VII. Designated representative map: (1) The representative representative of the case is: (none) (2) The symbol of the symbol of the representative figure is simple: 8. If there is a chemical formula in this case, please reveal the best indication of the characteristics of the invention. Chemical formula: (none) 131377.doc