CN102277319A - Rhizobium radiobacter and method for producing curdian fermentation liquor by using strain - Google Patents

Rhizobium radiobacter and method for producing curdian fermentation liquor by using strain Download PDF

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CN102277319A
CN102277319A CN 201110219188 CN201110219188A CN102277319A CN 102277319 A CN102277319 A CN 102277319A CN 201110219188 CN201110219188 CN 201110219188 CN 201110219188 A CN201110219188 A CN 201110219188A CN 102277319 A CN102277319 A CN 102277319A
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culture
fermentation
temperature
seed
ventilation
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CN102277319B (en
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张禹
张国佩
张少华
刘学珍
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Hebei Xinhe Biochemical Co ltd
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Hebei Xinhe Biochemical Co Ltd
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Abstract

The invention belongs to microbial fermentation, and particularly discloses Rhizobium radiobacter and a method for producing curdian fermentation liquor by using the strain. The method for producing the curdian fermentation liquor by using the strain comprises the following steps of: inoculating seed liquid which contains Rhizobium radiobacter CGMCC No.5031 serving as the strain after amplification culture into culture solution of a fermentation tank according to the inoculation amount of between 5 and 10 mass percent, wherein the fermentation culture solution contains a carbon source, an essential nitrogen source and nutrient substances; regulating the pH value of the fermentation culture solution, and performing ventilation fermentation; and supplementing a carbon source, a nitrogen source, nutrient substances and water in the fermentation process, and performing ventilation fermentation for not less than 90 hours to obtain fermentation liquid containing curdian. By the method, the defect that the content of solid matter in the curdian fermentation liquor is low in the prior art is overcome; and the method has the advantages that: the carbon source in the fermentation liquor can be better utilized by the strain, the transformation rate is high in the fermentation process, the content of the solid matter in the fermentation liquor is high, the production cost is greatly reduced and the like.

Description

Radiation root nodule bacterium and adopt this bacterial classification to produce the method for curdlan fermented liquid
Technical field
The invention belongs to microbial fermentation, be meant the radiation root nodule bacterium especially and adopt this bacterial classification to produce the method for curdlan fermented liquid.
Background technology
Curdlan is a kind of new microbial exocellular polysaccharide, because this polysaccharide has the peculiar property that forms gel under heating condition, so be called hot gel again, English name Curdian can be widely used in industries such as food, medicine.The traditional technology of curdlan preparation is that (bacterial classification uses Agrobacterium with the production bacterial classification Agrobacterium biovar1) to be inoculated in sucrose be to ferment in the aseptic culture medium of main carbon source, processing condition such as controlled temperature, PH, fermentation ends obtain the curdlan fermented liquid, and above-mentioned prior art exists the low defective of solid content in the prepared curdlan product.
Summary of the invention
One of purpose of the present invention is to provide a kind of radiation root nodule bacterium that can produce curdlan, promptly the radiation root nodule bacterium ( Rhizobium radiobacter) CGMCC NO.5031.
Two of purpose of the present invention is to provide the method for utilizing radiation root nodule bacterium CGMCC NO.5031 to produce the curdlan fermented liquid, makes solid content index in the fermented liquid higher and be better than currently available products.
Overall technology design of the present invention is:
Be used among the present invention to produce curdlan bacterial classification radiation root nodule bacterium ( Rhizobium radiobacter) submit China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation on July 12nd, 2011, deposit number is: CGMCC NO.5031.
The determined dna sequence result of this bacterial classification is:
GGGCTACCATGCAGTCGACGCCCCGCAAGGGGAGTGGCAGACGGGTGAGTAACGCGTGGGAATCTACCCATCTCTGCGGAATAGCTCTGGGAAACTGGAATTAAGACCGCGTACGCCCTACGGGGGAAAGATTTATCGGGGATGGATGAGCCCGCGTTGGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCCATAGCTGGTCTGAGAGGATGATCAGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGAGTGATGAAGGCCTTAGGGTTGTAAAGCTCTTTCACCGATGAAGATAATGACGGTAGTCGGAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGGGCTAGCGTTGTTCGGAATTACTGGGCGTAAAGCGCACGTAGGCGGATATTTAAGTCAGGGGTGAAATCCCGCAGCTCAACTGCGGAACTGCCTTTGATACTGGGTATCTTGAGTATGGAAGAGGTAAGTGGAATTACCGAGTGTAGAGGTGAAATTCGTAGATATTCGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGTCCATTACTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATGTTAGCCGTCGGGCAGTATACTGTTCGGTGGCGCAGCTAACGCATTAAACATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGCAGAACCTTACCAGCTCTTGACATTCGGGGTATGGGCATTGGAGACGATGTCCTTCAGTTAGGCTGGCCCCAGAACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCTTAGTTGCCAGCATTTAGTTGGGCACTCTAAGGGGACTGCCGGTGATAAGCCGAGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTACGGGCTGGGCTACACACGTGCTACAATGGTGGTGACAGTGGGCAGCGAGACAGCGATGTCGAGCTAATCTCCAAAAGCCATCTCAGTTCGGATTGCACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGCAGATCAGCATGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTTGGTTTTACCCGAAGGTAGTGCGCTAACCGCAAGGAGGCAGCTACC。
Above-mentioned bacterial classification has as cellular form and physicochemical property described in the following table:
Test subject The result Test subject The result Test subject The result Test subject The result
Cellular form Shaft-like Gramstaining Negative Catalase + Oxydase +
Beta-galactosidase enzymes + Arginine dihydrolase - Lysine decarboxylase - Ornithine decarboxylase -
Urase + The tryptophane desaminase - Citrate trianion utilizes - V.P. reaction -
Indoles produces - H2S produces - Gelatine liquefication - Nitrate reduction -
The starch hydrolysis - The casein hydrolysis - The polychrom hydrolysis + Glucose produces acid +
Seminose produces acid + Sucrose produces acid + Pectinose produces acid + Wood sugar produces acid +
Rhamnosyl produces acid + Melibiose produces acid + Cellobiose produces acid + Lactose produces acid +
Contrast - The D-melibiose + The p-phenylglycollic acid - The L-Histidine +
The a-cyclodextrin - Methyl glucoside + Methylene-succinic acid - Oxyproline +
Dextrin - The D-psicose + The a-ketobutyric acid - The L-leucine -
Glycogen - The D-raffinose + The a-keto-glutaric acid - The L-ornithine +
Tween40 - The L-rhamnosyl + The a-oxopentanoic acid - The L-phenylalanine -
Tween80 - The D-sorbyl alcohol + D, L-lactic acid + The L-proline(Pro) +
N-acetylgalactosamine - Sucrose + Propanedioic acid - The L-Pyrrolidonecarboxylic acid +
The N-acetylglucosamine + The D-trehalose + Propionic acid - The D-Serine -
Ribitol + Turanose + Quinic acid + The L-Serine +
L-arabinose + Xylitol + The D-saccharinic acid - The L-Threonine +
The D-arabitol + Pyruvic Acid Methyl ester + The certain herbaceous plants with big flowers diacid - D, the L-carnitine -
The D-cellobiose + Succinic Acid one methyl esters + Succinic Acid + The g-aminobutyric acid +
The i-tetrahydroxybutane - Acetate + Bromosuccinic acid + Urocanic acid +
D-fructose + Suitable-aconic acid + Succinamic acid - Inosine -
The L-Fucose + Citric acid - Glucuronamide b Uridine +
The D-semi-lactosi + Formic acid - The L alanimamides - Thymidine -
Gentiobiose + The D-galactonolactone + The D-L-Ala b Phenyl-ethyl amine -
A-D-glucose + The D-galacturonic acid - The L-L-Ala + Putrescine -
The m-inositol + The D-glyconic acid + The L-alanyl-glycine + The 2-monoethanolamine -
The a-D-lactose + The b-methyl glucoside - Altheine + 2, the 3-butyleneglycol -
Lactulose + The D-glucuronic acid + The L-aspartic acid + Glycerine +
Maltose + The a-hydroxybutyric acid - L-L-glutamic acid + D, the L-glycerophosphate -
D-N.F,USP MANNITOL + The b-hydroxybutyric acid - The glycyl aspartic acid + The D-glucose-1-phosphate +
The D-seminose + The g-hydroxybutyric acid - Glycyl L-glutamic acid + The D-Robison ester +
With the radiation root nodule bacterium is the method that bacterial classification is produced the curdlan fermented liquid, and this method comprises following processing step:
A, the seed liquor that contains radiation root nodule bacterium CGMCC NO.5031 after will spreading cultivation are that the inoculum size of 5%-10% is linked in the nutrient solution of fermentor tank according to mass percent, contain carbon source, necessary nitrogenous source and nutritive substance in the fermentation culture;
The pH of B, adjusting fermentation culture carries out aerobic fermentation;
C, mend carbon source, nitrogenous source, nutritive substance and water during the fermentation, the aerobic fermentation through being not less than 90 hours obtains containing the fermented liquid of curdlan.
Concrete technical solution of the present invention also has:
Fermentation culture in the described steps A is made up of the component of following mass percent:
Glucose 6%-8%, SODIUMNITRATE 0.3%-0.45%, dipotassium hydrogen phosphate 0.1%-0.2%, potassium primary phosphate 0.15%-0.3%, FeSO 410ppm-15ppm, vitamin H 20ppm-30ppm, polyether antifoam agent 0.03%-0.1%, surplus are sterilized water; The pH=6-7 of fermentation culture.
Fermentation condition among the described step C is:
C 1, in 10 hours: 30 ℃-32 ℃ of temperature, pressure 0.04 MPa-0.06MPa, pH value 6-7, ventilation 0.3vvm-0.4vvm;
C 2, 10-20 hour: 30 ℃-32 ℃ of temperature, pressure 0.04 MPa-0.06 MPa, pH value 6-7, ventilation 0.4vvm-0.5vvm;
C 3, 20-30 hour: temperature 30-32 ℃, pressure 0.04MPa-0.06MPa, pH value 6-6.5, ventilation 0.6vvm-0.7vvm;
C 4, 30-40 hour: 28 ℃-30 ℃ of temperature, pressure 0.04MPa-0.06MPa, pH value 6-6.5, ventilation 0.7vvm-0.8vvm; Press 1/4 feed supplement of fermentating liquid volume; Benefit into material form by the component of following mass percent:
Glucose 10%-12%, SODIUMNITRATE 0.3%-0.45%, dipotassium hydrogen phosphate 0.1%-0.2%, potassium primary phosphate 0.15%-0.3%, FeSO 410ppm-15ppm, vitamin H 20ppm-30ppm, polyether antifoam agent 0.03%-0.1%, surplus are sterilized water;
C 5, 40-60 hour: 28 ℃-30 ℃ of temperature, pressure 0.04MPa-0.06MPa, ventilation 0.5vvm-0.6vvm, mended the sterilized water of fermented liquid cumulative volume 1%-1.5% in 50 hours;
C 6, 60-70 hour: 29 ℃ of temperature, pressure 0.04MPa, pH value 6-7, ventilation 0.5vvm-0.6vvm, mended the sterilized water of fermented liquid cumulative volume 1.5%-2% in 65 hours;
C 7, 70-80 hour: 28 ℃-32 ℃ of temperature, pressure 0.04MPa-0.06MPa, pH value 6-7, ventilation 0.4vvm-0.5vvm;
C 8, 80-90 hour: 28 ℃-32 ℃ of temperature, pressure 0.04MPa-0.06MPa, ventilation 0.5vvm-0.6vvm;
C 9, 90-120 hour: 28 ℃-32 ℃ of temperature, pressure 0.04MPa, pH value 6-7, ventilation 0.4vvm-0.5vvm.
The process of spreading cultivation comprises in the described steps A:
A 1, slant strains is made shake-flask seed;
A 2, shake-flask seed is made first order seed;
A 3, first order seed is made secondary seed, secondary seed is inserted nutrient solution in the fermentor tank, ferment.
Steps A 1Be with making shake-flask seed through cultivation in the shake-flask seed nutrient solution after the former bacterium access sterilization in 2 square centimeters of slant strains of inoculation shovel picking, the culture condition of shake-flask seed is that culture temperature is that 28 ℃-32 ℃, rotating speed are that 160-200 rev/min, culture cycle are shaking culture under 24-36 hour the condition, and the shake-flask seed nutrient solution is made up of the component of following massfraction:
Glucose 0.8%-1.5%, fish meal 0.5%-0.15%, Tryptones 0.2%-0.3%, soybean protein powder 0.1%-0.19%, soya-bean oil 0.01%-0.03%, surplus are sterilized water.
Steps A 2In be to be to make first order seed through aerated culture in the first order seed nutrient solution after the inoculum size of 0.2%-0.5% inserts sterilization according to mass percent with shake-flask seed, the culture condition of first order seed is that culture temperature is that 28 ℃-32 ℃, pressure are that 0.04MPa-0.06MPa, ventilation 0.3vvm-0.4vvm, culture cycle are aerated culture under 26-48 hour the condition, and the first order seed nutrient solution is made up of the component of following massfraction:
Glucose 2%-2.5%, ammonium chloride 0.3%-0.5%, ammonium sulfate 0.3%-0.5%, sal epsom 0.1%-0.2%, ferrous sulfate 10ppm-20ppm, polyether antifoam agent 0.05%-0.1%, surplus are sterilized water.
Steps A 3Be to be to make secondary seed through aerated culture in the secondary seed nutrient solution after the inoculum size of 5%-10% inserts sterilization according to mass percent with first order seed, the culture condition of secondary seed is that culture temperature is that 28 ℃-32 ℃, pressure are that 0.04MPa-0.06MPa, ventilation 0.5 vvm-0.7vvm, culture cycle are aerated culture under 24 hours the condition, and the secondary seed nutrient solution is made up of the component of following massfraction:
Glucose 2%-2.5%, ammonium chloride 0.4%-0.6%, ammonium sulfate 0.3%-0.5%, sal epsom 0.1%-0.2%, ferrous sulfate 10ppm-20ppm, polyether antifoam agent 0.05%-0.1%, surplus are sterilized water.
Steps A 1In the sterilising conditions of shake-flask seed nutrient solution be vapor pressure 0.1 Mpa-0.12Mpa, 121 ℃-125 ℃ of temperature, time 30-35 minute.
The sterilising conditions of first order seed nutrient solution and secondary seed nutrient solution is 121 ℃-125 ℃ of temperature among steps A 2, the A3, time 25-30 minute.
The detection method of product discrimination method and solid substance is among the present invention:
One, the mensuration of solid content:
(1) fermented liquid is poured in the small beaker of certain volume, stirred with glass stick and make its no bubble, floating the surface with glass stick, the fermented liquid outer cup cleans up.Fermented liquid is extracted with extraction using alcohol, sedimentation, the filter cloth of 90-95%, all thoroughly is moved into after it is torn up in the culture dish, the little fire baking of microwave oven 3 minutes, puts into 100 then +Dry to constant weight in 5 ℃ of baking ovens (about 4 hours), take out and weigh.Actual detected fixture content has as a result reached solid content in every liter of fermented liquid of 78g/L() more than.
(2) calculate:
Solid content (g/L)=(claiming sample gram number/fermentating liquid volume) * 100%
Two, product is differentiated: because of the food security standard (declaration original text) of the curdlan product of China still is in public notification period, issuing and implementation.According to JECFA(2001) in the record discrimination method fermented liquid is detected, detected result is: all meet following standard, then result of determination is a curdlan.
Detection method and result:
(1) solubleness: water insoluble and ethanol (standard).Actual detected result: water insoluble and ethanol.
(2) solubleness in alkali: get the 0.2g sample and add in the 5ml water, add the NaOH solution of 1ml3N, vibration, sample dissolution.Actual detected result: sample dissolution.
(3) form gel: to mass percent is 2% suspension boiling water bath heating 10 minutes, is cooled to room temperature, forms solid gel.Actual detected result: form solid gel.
(4) cupric tartrate precipitin reaction: to the 10ml mass percent concentration is to add the 5ml analytical pure vitriol oil in 2% the sample suspension, and 30 minutes postcooling of boiling water bath are used BaCO 3Neutralization, 900 rev/mins centrifugal 10 minutes, get the 1ml supernatant liquor, add in the basic copper tartrate solution of 5ml heat, generate a large amount of red copper oxidule precipitation.Actual detected result: generate a large amount of red copper oxidule precipitation.
Substantive distinguishing features that the present invention is obtained and significant technical progress are:
1, because bacterial classification disclosed in this invention can utilize the carbon source in the fermented liquid preferably, transformation efficiency height in the fermenting process, production cost significantly reduces.After testing, the solid content in the fermented liquid reaches the solid content in every liter of fermented liquid of 78g/L(at least) more than.
2, prepared curdlan product meets FCC V and JECFA(2001 after testing) standard.
3, adopt moisturizing, supplying technics in the fermenting process, help the synthetic curdlan of fast and stable.
Embodiment
Below in conjunction with embodiment the present invention is further described; but it is not as a limitation of the invention; protection scope of the present invention is as the criterion with the content of claim record, and any equivalence techniques means of having done according to this specification sheets are replaced, and all do not break away from protection scope of the present invention.
Embodiment 1
Present embodiment comprises following processing step:
A, the seed liquor that contains radiation root nodule bacterium CGMCC NO.5031 after will spreading cultivation are that 5% inoculum size is linked in the nutrient solution of fermentor tank according to mass percent, contain carbon source, necessary nitrogenous source and nutritive substance in the fermentation culture;
The pH of B, adjusting fermentation culture carries out aerobic fermentation;
C, mend carbon source, nitrogenous source, nutritive substance and water during the fermentation, the aerobic fermentation through being not less than 90 hours obtains containing the fermented liquid of curdlan.
Fermentation culture in the described steps A is made up of the component of following mass percent:
Glucose 6%, SODIUMNITRATE 0.3%, dipotassium hydrogen phosphate 0.1%, potassium primary phosphate 0.15%, FeSO 410ppm, vitamin H 20ppm, polyether antifoam agent 0.03%, surplus are sterilized water; The pH=6 of fermentation culture.
Fermentation condition among the described step C is:
C 1, in 10 hours: 30 ℃ of temperature, pressure 0.04 MPa, pH value 6, ventilation 0.3vvm;
C 2, 10-20 hour: 30 ℃ of temperature, pressure 0.04 MPa, pH value 6, ventilation 0.4vvm;
C 3, 20-30 hour: 30 ℃ of temperature, pressure 0.04MPa, pH value 6, ventilation 0.6vvm;
C 4, 30-40 hour: 28 ℃ of temperature, pressure 0.04MPa, pH value 6, ventilation 0.7vvm; Press 1/4 feed supplement of fermentating liquid volume; Benefit into material form by the component of following mass percent:
Glucose 12%, SODIUMNITRATE 0.3%, dipotassium hydrogen phosphate 0.1%, potassium primary phosphate 0.15%, FeSO 410ppm, vitamin H 20ppm, polyether antifoam agent 0.03%, surplus are sterilized water;
C 5, 40-60 hour: 28 ℃ of temperature, pressure 0.04MPa, ventilation 0.5vvm, mended the sterilized water of fermented liquid cumulative volume 1% in 50 hours;
C 6, 60-70 hour: 29 ℃ of temperature, pressure 0.04MPa, pH value 6, ventilation 0.5vvm, mended the sterilized water of fermented liquid cumulative volume 1.5% in 65 hours;
C 7, 70-80 hour: 28 ℃ of temperature, pressure 0.04MPa, pH value 6, ventilation 0.4vvm;
C 8, 80-90 hour: 28 ℃ of temperature, pressure 0.04MPa, ventilation 0.5vvm;
C 9, 90-120 hour: 28 ℃ of temperature, pressure 0.04MPa, pH value 6, ventilation 0.4vvm.
The process of spreading cultivation comprises in the described steps A:
A 1, slant strains is made shake-flask seed;
A 2, shake-flask seed is made first order seed;
A 3, first order seed is made secondary seed, secondary seed is inserted nutrient solution in the fermentor tank, ferment.
Steps A 1Be with making shake-flask seed through cultivation in the shake-flask seed nutrient solution after the former bacterium access sterilization in 2 square centimeters of slant strains of inoculation shovel picking, the culture condition of shake-flask seed is that culture temperature is that 28 ℃, rotating speed are that 160 rev/mins, culture cycle are shaking culture under 24 hours the condition, and the shake-flask seed nutrient solution is made up of the component of following massfraction:
Glucose 0.8%, fish meal 0.5%, Tryptones 0.2%, soybean protein powder 0.1%, soya-bean oil 0.01%, surplus are sterilized water.
Steps A 2In be to be to make first order seed through aerated culture in the first order seed nutrient solution after 0.2% inoculum size inserts sterilization according to mass percent with shake-flask seed, the culture condition of first order seed is that culture temperature is that 28 ℃, pressure are that 0.04MPa, ventilation 0.3vvm, culture cycle are aerated culture under 26 hours the condition, and the first order seed nutrient solution is made up of the component of following massfraction:
Glucose 2%, ammonium chloride 0.3%, ammonium sulfate 0.3%, sal epsom 0.1%, ferrous sulfate 10ppm, polyether antifoam agent 0.05%, surplus are sterilized water.
Steps A 3Be to be to make secondary seed through aerated culture in the secondary seed nutrient solution after the inoculum size of 5%-10% inserts sterilization according to mass percent with first order seed, the culture condition of secondary seed is that culture temperature is that 28 ℃, pressure are that 0.04MPa, ventilation 0.5 vvm, culture cycle are aerated culture under 24 hours the condition, and the secondary seed nutrient solution is made up of the component of following massfraction:
Glucose 2%, ammonium chloride 0.5%, ammonium sulfate 0.4%, sal epsom 0.15%, ferrous sulfate 10ppm, polyether antifoam agent 0.06%, surplus are sterilized water.
Steps A 1The sterilising conditions of middle shake-flask seed nutrient solution is vapor pressure 0.1 Mpa, 121 ℃ of temperature, 30 minutes time.
The sterilising conditions of first order seed nutrient solution and secondary seed nutrient solution is 121 ℃ of temperature among steps A 2, the A3,25 minutes time.
Embodiment 2
Present embodiment comprises following processing step:
A, the seed liquor that contains radiation root nodule bacterium CGMCC NO.5031 after will spreading cultivation are that 10% inoculum size is linked in the nutrient solution of fermentor tank according to mass percent, contain carbon source, necessary nitrogenous source and nutritive substance in the fermentation culture;
The pH of B, adjusting fermentation culture carries out aerobic fermentation;
C, mend carbon source, nitrogenous source, nutritive substance and water during the fermentation, the aerobic fermentation through being not less than 90 hours obtains containing the fermented liquid of curdlan.
Concrete technical solution of the present invention also has:
Fermentation culture in the described steps A is made up of the component of following mass percent:
Glucose 8%, SODIUMNITRATE 0.45%, dipotassium hydrogen phosphate 0.2%, potassium primary phosphate 0.3%, FeSO 415ppm, vitamin H 30ppm, polyether antifoam agent 0.1%, surplus are sterilized water; The pH=7 of fermentation culture.
Fermentation condition among the described step C is:
C 1, in 10 hours: 32 ℃ of temperature, pressure 0.06MPa, pH value 7, ventilation 0.4vvm;
C 2, 10-20 hour: 32 ℃ of temperature, pressure 0.06 MPa, pH value 7, ventilation 0.5vvm;
C 3, 20-30 hour: 32 ℃ of temperature, pressure 0.06MPa, pH value 6.5, ventilation 0.7vvm;
C 4, 30-40 hour: 30 ℃ of temperature, pressure 0.06MPa, pH value 6.5, ventilation 0.8vvm; Press 1/4 feed supplement of fermentating liquid volume; Benefit into material form by the component of following mass percent:
Glucose 10%, SODIUMNITRATE 0.45%, dipotassium hydrogen phosphate 0.2%, potassium primary phosphate 0.3%, FeSO 415ppm, vitamin H 30ppm, polyether antifoam agent 0.1%, surplus are sterilized water;
C 5, 40-60 hour: 30 ℃ of temperature, pressure 0.06MPa, ventilation 0.6vvm, mended the sterilized water of fermented liquid cumulative volume 1.5% in 50 hours;
C 6, 60-70 hour: 29 ℃ of temperature, pressure 0.04MPa, pH value 7, ventilation 0.6vvm, mended 2% sterilized water of fermented liquid cumulative volume in 65 hours;
C 7, 70-80 hour: 32 ℃ of temperature, pressure 0.06MPa, pH value 7, ventilation 0.5vvm;
C 8, 80-90 hour: 32 ℃ of temperature, pressure 0.06MPa, ventilation 0.6vvm;
C 9, 90-120 hour: 32 ℃ of temperature, pressure 0.04MPa, pH value 7, ventilation 0.5vvm.
The process of spreading cultivation comprises in the described steps A:
A 1, slant strains is made shake-flask seed;
A 2, shake-flask seed is made first order seed;
A 3, first order seed is made secondary seed, secondary seed is inserted nutrient solution in the fermentor tank, ferment.
Steps A 1Be with making shake-flask seed through cultivation in the shake-flask seed nutrient solution after the former bacterium access sterilization in 2 square centimeters of slant strains of inoculation shovel picking, the culture condition of shake-flask seed is that culture temperature is that 32 ℃, rotating speed are that 200 rev/mins, culture cycle are shaking culture under 36 hours the condition, and the shake-flask seed nutrient solution is made up of the component of following massfraction:
Glucose 1.5%, fish meal 0.15%, Tryptones 0.3%, soybean protein powder 0.19%, soya-bean oil 0.03%, surplus are sterilized water.
Steps A 2In be to be to make first order seed through aerated culture in the first order seed nutrient solution after 0.5% inoculum size inserts sterilization according to mass percent with shake-flask seed, the culture condition of first order seed is that culture temperature is that 32 ℃, pressure are that 0.06MPa, ventilation 0.4vvm, culture cycle are aerated culture under 48 hours the condition, and the first order seed nutrient solution is made up of the component of following massfraction:
Glucose 2.5%, ammonium chloride 0.5%, ammonium sulfate 0.5%, sal epsom 0.2%, ferrous sulfate 20ppm, polyether antifoam agent 0.1%, surplus are sterilized water.
Steps A 3Be to be to make secondary seed through aerated culture in the secondary seed nutrient solution after 10% inoculum size inserts sterilization according to mass percent with first order seed, the culture condition of secondary seed is that culture temperature is that 32 ℃, pressure are that 0.06MPa, ventilation 0.7vvm, culture cycle are aerated culture under 24 hours the condition, and the secondary seed nutrient solution is made up of the component of following massfraction:
Glucose 2.5%, ammonium chloride 0.6%, ammonium sulfate 0.3%, sal epsom 0.2%, ferrous sulfate 15ppm, polyether antifoam agent 0.08%, surplus are sterilized water.
Steps A 1The sterilising conditions of middle shake-flask seed nutrient solution is vapor pressure 0.12Mpa, 125 ℃ of temperature, 35 minutes time.
The sterilising conditions of first order seed nutrient solution and secondary seed nutrient solution is 125 ℃ of temperature among steps A 2, the A3,30 minutes time.
Embodiment 3
Present embodiment comprises following processing step:
A, the seed liquor that contains radiation root nodule bacterium CGMCC NO.5031 after will spreading cultivation are that 8% inoculum size is linked in the nutrient solution of fermentor tank according to mass percent, contain carbon source, necessary nitrogenous source and nutritive substance in the fermentation culture;
The pH of B, adjusting fermentation culture carries out aerobic fermentation;
C, mend carbon source, nitrogenous source, nutritive substance and water during the fermentation, the aerobic fermentation through being not less than 90 hours obtains containing the fermented liquid of curdlan.
Concrete technical solution of the present invention also has:
Fermentation culture in the described steps A is made up of the component of following mass percent:
Glucose 7%, SODIUMNITRATE 0.4%, dipotassium hydrogen phosphate 0.15%, potassium primary phosphate 0.2%, FeSO 412ppm, vitamin H 25ppm, polyether antifoam agent 0.08%, surplus are sterilized water; The pH=7 of fermentation culture.
Fermentation condition among the described step C is:
C 1, in 10 hours: 31 ℃ of temperature, pressure 0.05MPa, pH value 6.5, ventilation 0.35vvm;
C 2, 10-20 hour: 31 ℃ of temperature, pressure 0.05MPa, pH value 6.5, ventilation 0.45vvm;
C 3, 20-30 hour: 31 ℃ of temperature, pressure 0.05MPa, pH value 6, ventilation 0.65vvm;
C 4, 30-40 hour: 29 ℃ of temperature, pressure 0.05MPa, pH value 6, ventilation 0.75vvm; Press 1/4 feed supplement of fermentating liquid volume; Benefit into material form by the component of following mass percent:
Glucose 11%, SODIUMNITRATE 0.4%, dipotassium hydrogen phosphate 0.15%, potassium primary phosphate 0.2%, FeSO 412ppm, vitamin H 25ppm, polyether antifoam agent 0.08%, surplus are sterilized water;
C 5, 40-60 hour: 29 ℃ of temperature, pressure 0.05MPa, ventilation 0.5vvm, mended the sterilized water of fermented liquid cumulative volume 1.2% in 50 hours;
C 6, 60-70 hour: 29 ℃ of temperature, pressure 0.04MPa, pH value 6, ventilation 0.55vvm, mended 1.5% sterilized water of fermented liquid cumulative volume in 65 hours;
C 7, 70-80 hour: 29 ℃ of temperature, pressure 0.05MPa, pH value 6, ventilation 0.45vvm;
C 8, 80-90 hour: 28 ℃-32 ℃ of temperature, pressure 0.05MPa, ventilation 0.55vvm;
C 9, 90-120 hour: 28 ℃-32 ℃ of temperature, pressure 0.04MPa, pH value 6, ventilation 0.45vvm.
The process of spreading cultivation comprises in the described steps A:
A 1, slant strains is made shake-flask seed;
A 2, shake-flask seed is made first order seed;
A 3, first order seed is made secondary seed, secondary seed is inserted nutrient solution in the fermentor tank, ferment.
Steps A 1Be with making shake-flask seed through cultivation in the shake-flask seed nutrient solution after the former bacterium access sterilization in 2 square centimeters of slant strains of inoculation shovel picking, the culture condition of shake-flask seed is that culture temperature is that 30 ℃, rotating speed are that 180 rev/mins, culture cycle are shaking culture under 30 hours the condition, and the shake-flask seed nutrient solution is made up of the component of following massfraction:
Glucose 1%, fish meal 0.4%, Tryptones 0.25%, soybean protein powder 0.15%, soya-bean oil 0.02%, surplus are sterilized water.
Steps A 2In be to be to make first order seed through aerated culture in the first order seed nutrient solution after 0.3% inoculum size inserts sterilization according to mass percent with shake-flask seed, the culture condition of first order seed is that culture temperature is that 30 ℃, pressure are that 0.05MPa, ventilation 0.35vvm, culture cycle are aerated culture under 36 hours the condition, and the first order seed nutrient solution is made up of the component of following massfraction:
Glucose 2.2%, ammonium chloride 0.4%, ammonium sulfate 0.4%, sal epsom 0.15%, ferrous sulfate 15ppm, polyether antifoam agent 0.08%, surplus are sterilized water.
Steps A 3Be to be to make secondary seed through aerated culture in the secondary seed nutrient solution after 8% inoculum size inserts sterilization according to mass percent with first order seed, the culture condition of secondary seed is that culture temperature is that 30 ℃, pressure are that 0.05MPa, ventilation 0.6vvm, culture cycle are aerated culture under 24 hours the condition, and the secondary seed nutrient solution is made up of the component of following massfraction:
Glucose 2.5%, ammonium chloride 0.6%, ammonium sulfate 0.5%, sal epsom 0.2%, ferrous sulfate 20ppm, polyether antifoam agent 0.1%, surplus are sterilized water.
Steps A 1The sterilising conditions of middle shake-flask seed nutrient solution is vapor pressure 0.1Mpa, 121 ℃ of temperature, 30 minutes time.
The sterilising conditions of first order seed nutrient solution and secondary seed nutrient solution is 121 ℃ of temperature among steps A 2, the A3,30 minutes time.
According to JECFA(2001) in the discrimination method of record the fermented liquid of embodiment 1-3 preparation is detected, detected result is: the fermented liquid for preparing among the embodiment 1-3 is a curdlan.The solid content that the detection method of disclosed solid substance is measured fermented liquid among the embodiment 1-3 to specifications respectively is 78g/L, 81g/L, 79g/L.
SEQUENCE LISTING
＜110〉Hebei Xinhe Biologicgal Chemical Co., Ltd
＜120〉radiation root nodule bacterium and adopt this bacterial classification to produce the method for curdlan fermented liquid
<130> none
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1363
<212> DNA
<213> Rhizobium sp.
<400> 1
gggctaccat gcagtcgacg ccccgcaagg ggagtggcag acgggtgagt aacgcgtggg 60
aatctaccca tctctgcgga atagctctgg gaaactggaa ttaagaccgc gtacgcccta 120
cgggggaaag atttatcggg gatggatgag cccgcgttgg attagctagt tggtggggta 180
aaggcctacc aaggcgacga tccatagctg gtctgagagg atgatcagcc acattgggac 240
tgagacacgg cccaaactcc tacgggaggc agcagtgggg aatattggac aatgggcgca 300
agcctgatcc agccatgccg cgtgagtgat gaaggcctta gggttgtaaa gctctttcac 360
cgatgaagat aatgacggta gtcggagaag aagccccggc taacttcgtg ccagcagccg 420
cggtaatacg aagggggcta gcgttgttcg gaattactgg gcgtaaagcg cacgtaggcg 480
gatatttaag tcaggggtga aatcccgcag ctcaactgcg gaactgcctt tgatactggg 540
tatcttgagt atggaagagg taagtggaat taccgagtgt agaggtgaaa ttcgtagata 600
ttcggaggaa caccagtggc gaaggcggct tactggtcca ttactgacgc tgaggtgcga 660
aagcgtgggg agcaaaacag gattagatac cctggtagtc cacgccgtaa acgatgaatg 720
ttagccgtcg ggcagtatac tgttcggtgg cgcagctaac gcattaaaca ttccgcctgg 780
ggagtacggt cgcaagatta aaactcaaag gaattgacgg gggcccgcac aagcggtgga 840
gcatgtggtt taattcgaag caacgcgcag aaccttacca gctcttgaca ttcggggtat 900
gggcattgga gacgatgtcc ttcagttagg ctggccccag aacaggtgct gcatggctgt 960
cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac cctcgccctt 1020
agttgccagc atttagttgg gcactctaag gggactgccg gtgataagcc gagaggaagg 1080
tggggatgac gtcaagtcct catggccctt acgggctggg ctacacacgt gctacaatgg 1140
tggtgacagt gggcagcgag acagcgatgt cgagctaatc tccaaaagcc atctcagttc 1200
ggattgcact ctgcaactcg agtgcatgaa gttggaatcg ctagtaatcg cagatcagca 1260
tgctgcggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca tgggagttgg 1320
ttttacccga aggtagtgcg ctaaccgcaa ggaggcagct acc 1363

Claims (10)

1. radiation root nodule bacterium CGMCC NO.5031 is characterized in that the dna sequence dna of this bacterial classification is:
GGGCTACCATGCAGTCGACGCCCCGCAAGGGGAGTGGCAGACGGGTGAGTAACGCGTGGGAATCTACCCATCTCTGCGGAATAGCTCTGGGAAACTGGAATTAAGACCGCGTACGCCCTACGGGGGAAAGATTTATCGGGGATGGATGAGCCCGCGTTGGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCCATAGCTGGTCTGAGAGGATGATCAGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGAGTGATGAAGGCCTTAGGGTTGTAAAGCTCTTTCACCGATGAAGATAATGACGGTAGTCGGAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGGGCTAGCGTTGTTCGGAATTACTGGGCGTAAAGCGCACGTAGGCGGATATTTAAGTCAGGGGTGAAATCCCGCAGCTCAACTGCGGAACTGCCTTTGATACTGGGTATCTTGAGTATGGAAGAGGTAAGTGGAATTACCGAGTGTAGAGGTGAAATTCGTAGATATTCGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGTCCATTACTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATGTTAGCCGTCGGGCAGTATACTGTTCGGTGGCGCAGCTAACGCATTAAACATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGCAGAACCTTACCAGCTCTTGACATTCGGGGTATGGGCATTGGAGACGATGTCCTTCAGTTAGGCTGGCCCCAGAACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCTTAGTTGCCAGCATTTAGTTGGGCACTCTAAGGGGACTGCCGGTGATAAGCCGAGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTACGGGCTGGGCTACACACGTGCTACAATGGTGGTGACAGTGGGCAGCGAGACAGCGATGTCGAGCTAATCTCCAAAAGCCATCTCAGTTCGGATTGCACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGCAGATCAGCATGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTTGGTTTTACCCGAAGGTAGTGCGCTAACCGCAAGGAGGCAGCTACC。
2. be the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that this method comprises following processing step:
A, the seed liquor that contains radiation root nodule bacterium CGMCC NO.5031 after will spreading cultivation are that the inoculum size of 5%-10% is linked in the nutrient solution of fermentor tank according to mass percent, contain carbon source, necessary nitrogenous source and nutritive substance in the fermentation culture;
The pH of B, adjusting fermentation culture carries out aerobic fermentation;
C, mend carbon source, nitrogenous source, nutritive substance and water during the fermentation, the aerobic fermentation through being not less than 90 hours obtains containing the fermented liquid of curdlan.
3. according to claim 2 is the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that the fermentation culture in the described steps A is made up of the component of following mass percent:
Glucose 6%-8%, SODIUMNITRATE 0.3%-0.45%, dipotassium hydrogen phosphate 0.1%-0.2%, potassium primary phosphate 0.15%-0.3%, FeSO 410ppm-15ppm, vitamin H 20ppm-30ppm, polyether antifoam agent 0.03%-0.1%, surplus are sterilized water; The pH=6-7 of fermentation culture.
4. according to claim 2 is the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that the fermentation condition among the described step C is:
C 1, in 10 hours: 30 ℃-32 ℃ of temperature, pressure 0.04MPa-0.06MPa, pH value 6-7, ventilation 0.3vvm-0.4vvm;
C 2, 10-20 hour: 30 ℃-32 ℃ of temperature, pressure 0.04MPa-0.06MPa, pH value 6-7, ventilation 0.4vvm-0.5vvm;
C 3, 20-30 hour: temperature 30-32 ℃, pressure 0.04MPa-0.06MPa, pH value 6-6.5, ventilation 0.6vvm-0.7vvm;
C 4, 30-40 hour: 28 ℃-30 ℃ of temperature, pressure 0.04MPa-0.06MPa, pH value 6-6.5, ventilation 0.7vvm-0.8vvm; Press 1/4 feed supplement of fermentating liquid volume; Benefit into material form by the component of following mass percent:
Glucose 10%-12%, SODIUMNITRATE 0.3%-0.45%, dipotassium hydrogen phosphate 0.1%-0.2%, potassium primary phosphate 0.15%-0.3%, FeSO 410ppm-15ppm, vitamin H 20ppm-30ppm, polyether antifoam agent 0.03%-0.1%, surplus are sterilized water;
C 5, 40-60 hour: 28 ℃-30 ℃ of temperature, pressure 0.04MPa-0.06MPa, ventilation 0.5 vvm-0.6vvm, mended the sterilized water of fermented liquid cumulative volume 1%-1.5% in 50 hours;
C 6, 60-70 hour: 29 ℃ of temperature, pressure 0.04MPa, pH value 6-7, ventilation 0.5 vvm-0.6vvm, mended the sterilized water of fermented liquid cumulative volume 1.5%-2% in 65 hours;
C 7, 70-80 hour: 28 ℃-32 ℃ of temperature, pressure 0.04MPa-0.06MPa, pH value 6-7, ventilation 0.4vvm-0.5vvm;
C 8, 80-90 hour: 28 ℃-32 ℃ of temperature, pressure 0.04MPa-0.06MPa, ventilation 0.5 vvm-0.6vvm;
C 9, 90-120 hour: 28 ℃-32 ℃ of temperature, pressure 0.04MPa, pH value 6-7, ventilation 0.4vvm-0.5vvm.
5. according to claim 2 is the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that the process of spreading cultivation comprises in the described steps A:
A 1, slant strains is made shake-flask seed;
A 2, shake-flask seed is made first order seed;
A 3, first order seed is made secondary seed, secondary seed is inserted nutrient solution in the fermentor tank, ferment.
6. according to claim 5 is the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that described steps A 1Be with making shake-flask seed through cultivation in the shake-flask seed nutrient solution after the former bacterium access sterilization in 2 square centimeters of slant strains of inoculation shovel picking, the culture condition of shake-flask seed is that culture temperature is that 28 ℃-32 ℃, rotating speed are that 160-200 rev/min, culture cycle are shaking culture under 24-36 hour the condition, and the shake-flask seed nutrient solution is made up of the component of following massfraction:
Glucose 0.8%-1.5%, fish meal 0.5%-0.15%, Tryptones 0.2%-0.3%, soybean protein powder 0.1%-0.19%, soya-bean oil 0.01%-0.03%, surplus are sterilized water.
7. according to claim 5 is the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that described steps A 2In be to be to make first order seed through aerated culture in the first order seed nutrient solution after the inoculum size of 0.2%-0.5% inserts sterilization according to mass percent with shake-flask seed, the culture condition of first order seed is that culture temperature is that 28 ℃-32 ℃, pressure are that 0.04MPa-0.06MPa, ventilation 0.3vvm-0.4vvm, culture cycle are aerated culture under 26-48 hour the condition, and the first order seed nutrient solution is made up of the component of following massfraction:
Glucose 2%-2.5%, ammonium chloride 0.3%-0.5%, ammonium sulfate 0.3%-0.5%, sal epsom 0.1%-0.2%, ferrous sulfate 10ppm-20ppm, polyether antifoam agent 0.05%-0.1%, surplus are sterilized water.
8. according to claim 5 is the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that described steps A 3Be to be to make secondary seed through aerated culture in the secondary seed nutrient solution after the inoculum size of 5%-10% inserts sterilization according to mass percent with first order seed, the culture condition of secondary seed is that culture temperature is that 28 ℃-32 ℃, pressure are that 0.04MPa-0.06MPa, ventilation 0.5 vvm-0.7vvm, culture cycle are aerated culture under 24 hours the condition, and the secondary seed nutrient solution is made up of the component of following massfraction:
Glucose 2%-2.5%, ammonium chloride 0.4%-0.6%, ammonium sulfate 0.3%-0.5%, sal epsom 0.1%-0.2%, ferrous sulfate 10ppm-20ppm, polyether antifoam agent 0.05%-0.1%, surplus are sterilized water.
9. according to claim 6 is the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that described steps A 1In the sterilising conditions of shake-flask seed nutrient solution be vapor pressure 0.1 Mpa-0.12Mpa, 121 ℃-125 ℃ of temperature, time 30-35 minute.
According to claim 7 or 8 described be the method that bacterial classification is produced the curdlan fermented liquid with the radiation root nodule bacterium, it is characterized in that the sterilising conditions of first order seed nutrient solution and secondary seed nutrient solution is 121 ℃-125 ℃ of temperature among described steps A 2, the A3, time 25-30 minute.
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CN105420160A (en) * 2015-12-18 2016-03-23 东华大学 Curdlan producing strain and application thereof
CN106399145A (en) * 2016-05-17 2017-02-15 江南大学 Rhizobium radiobacter and method for producing curdlan gum through fermentation of rhizobium radiobacter
CN106434495A (en) * 2016-12-02 2017-02-22 张星昊 Rhizobium pusense and method for preparing beta-1,3 glucan fermenting liquid by rhizobium pusense
CN108467876A (en) * 2018-03-16 2018-08-31 华东师范大学 A kind of fermentation process improving curdlan yield
CN110982860A (en) * 2019-12-10 2020-04-10 山东天智绿业生物科技有限公司 Fermentation method of curdlan
CN113943761A (en) * 2021-12-22 2022-01-18 山东国力生物科技有限公司 Preparation method of micromolecular beta-1, 3-glucan
CN113957024A (en) * 2021-12-22 2022-01-21 山东国力生物科技有限公司 Rhizobium GL-1803 and application thereof in preparation of insoluble beta-glucan

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Title
《LWT-FOOD SCIENCE AND TECHNOLOGY》 201105 Ben Salah R等 Fermentation of date palm juice by curdlan gum production from Rhizobium radiobacter ATCC 6466 (TM): Purification, rheological and physico-chemical characterization 1026-1034 1-10 第44卷, 第4期 *
《山东食品发酵》 20101020 赵双枝等 放射性土壤杆菌产可得然胶最佳条件的研究 28-32 1-10 , 第3期 *

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CN105420160A (en) * 2015-12-18 2016-03-23 东华大学 Curdlan producing strain and application thereof
CN105420160B (en) * 2015-12-18 2019-03-08 东华大学 A kind of Curdlan producing bacterial strain and its application
CN106399145A (en) * 2016-05-17 2017-02-15 江南大学 Rhizobium radiobacter and method for producing curdlan gum through fermentation of rhizobium radiobacter
CN106434495A (en) * 2016-12-02 2017-02-22 张星昊 Rhizobium pusense and method for preparing beta-1,3 glucan fermenting liquid by rhizobium pusense
CN106434495B (en) * 2016-12-02 2019-05-17 张星昊 A kind of Bodhisattva rhizobium and its β -1 is produced, the method for 3 dextran fermentation liquid
CN108467876A (en) * 2018-03-16 2018-08-31 华东师范大学 A kind of fermentation process improving curdlan yield
CN108467876B (en) * 2018-03-16 2021-12-07 华东师范大学 Fermentation method for increasing yield of curdlan
CN110982860A (en) * 2019-12-10 2020-04-10 山东天智绿业生物科技有限公司 Fermentation method of curdlan
CN113943761A (en) * 2021-12-22 2022-01-18 山东国力生物科技有限公司 Preparation method of micromolecular beta-1, 3-glucan
CN113957024A (en) * 2021-12-22 2022-01-21 山东国力生物科技有限公司 Rhizobium GL-1803 and application thereof in preparation of insoluble beta-glucan
CN113943761B (en) * 2021-12-22 2022-03-22 山东国力生物科技有限公司 Preparation method of micromolecular beta-1, 3-glucan

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