CN102796780A - Method for producing gamma-polyglutamic acid by fermentation of glutamic acid mother liquid - Google Patents
Method for producing gamma-polyglutamic acid by fermentation of glutamic acid mother liquid Download PDFInfo
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- CN102796780A CN102796780A CN2012103276546A CN201210327654A CN102796780A CN 102796780 A CN102796780 A CN 102796780A CN 2012103276546 A CN2012103276546 A CN 2012103276546A CN 201210327654 A CN201210327654 A CN 201210327654A CN 102796780 A CN102796780 A CN 102796780A
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Abstract
The invention relates to a method for producing gamma-polyglutamic acid by fermentation of glutamic acid mother liquid, which belongs to the field of microbial fermentation. The method comprises the following steps of: inoculating 10% of the glutamic acid mother liquid in a fermentation medium to be subjected to fermental cultivation, wherein the cultivation conditions are that the pH ranges from 7.0 to 7.4, the temperature ranges from 30 to 40 DEG C; at an aerobic condition, performing shake cultivation on the mother liquid at 150-250 r/min or performing stirring cultivation on the mother liquid at 100-400 rpm; feeding glucose and the glutamic acid mother liquid into the mother liquid during fermentation to maintain the concentration of the glucose and glutamic acid between 10g/L and 15g/L, wherein the fermentation time is 48-72 hours; centrifuging fermentation liquor to remove thalluses and part of solid contents, thus obtaining the gamma-polyglutamic acid (PGA). The method is simple in technology, and can achieve the efficient use of waster liquid in glutamic acid production, reduce energy consumption and save cost by utilizing the fermentation of the glutamic acid in the glutamic acid mother liquid to produce the gamma-polyglutamic acid.
Description
Technical field
The invention belongs to the microbial fermentation field, be specifically related to a kind of method of utilizing sodium glutamate mother liquid fermentative prodn gamma-polyglutamic acid-.
Background technology
Gamma-polyglutamic acid-(γ-PGA) is a kind of anionic polymer, it by L-L-glutamic acid or/and forms amido linkage generation afterwards through alpha-amino group and γ-carboxyl between the D-L-glutamic acid monomer.γ-PGA can be water-soluble, biodegradable, edible, to human non-toxic property, environmentally safe.It all has very big using value at agricultural, environment and biomedicine field, comprises the slow-releasing system of biodegradable diaper, water reservoir, medicine or fertilizer.
Up to the present, existing much about utilizing microbial fermentation to produce the report of γ-PGA.Different bacterial classifications, the staple of substratum are also identical not to the utmost.Comprise L-glutamic acid 20 g/L in the substratum like B. licheniformis ATCC 9945, glycerine 80 g/L, Hydrocerol A 12 g/L, NH
4Cl 7 g/L, γ-PGA output is 17-23 g/L (Troy, 1973); B. comprise L-glutamic acid 30 g/L in the substratum of subtilis IFO 3335, Hydrocerol A 20 g/L, γ-PGA output is 10-20 g/L (Kunioka and Goto, 1994); B. comprise fructose 75 g/L, NH in the substratum of subtilis TAM-4
4Cl 18g/L, γ-PGA output is 20 g/L (Ito etc., 1973); B. comprise glucose 75 g/L, NH in the substratum of licheniformis A35
4Cl 18g/L, γ-PGA output is 8-12 g/L (Cheng etc., 1989); B. comprise L-glutamic acid 70 g/L in the substratum of subtilis F02-1, glucose 1 g/L, cereal leach liquor 20 g/L, γ-PGA output is 50 g/L (Kubota etc., 1993); B. comprise SANMALT-S 60 g/L in the substratum of subtilis (natto), soy sauce 70 g/L, Sodium Glutamate 30 g/L, γ-PGA output is 35 g/L (Ogawa etc., 1993).
People such as Qiao Changsheng make fermentation production rate reach 1.18 g/Lh, number of patent application: 201110216717.6 through adding the fed-batch medium of being made up of glucose, an ammonium nitrate, calcium chloride and iron(ic)chloride during the fermentation; Utilize Bacillus licheniformis as producing bacterial strain, realize the coproduction of γ-PGA and L-glutamic acid, number of patent application: 201110331788.0; Small molecules in γ-PGA broth extraction process is seen through liquid to add in the fermentor tank and carries out feed supplement, fermentative prodn γ-PGA, number of patent application: 201110426526.2.
Sand is green for a long time to wait the people with Soybean Pretreatment, and inoculation bacillus natto, fixed temperature and humidity 1-2 stir the extraction bacillus natto with saline water and secrete the γ-PGA on the soybean surface, number of patent application: 200410043690.5 round the clock.
Sun Weizhong with soybean cake powder and wheat bran as solids component; Add the inorganic salt that glycerine, Hydrocerol A, L-glutamic acid, ammonium chloride and magnesium, manganese, calcium, iron and potassium are formed; Common fermention medium, fermentative prodn γ-PGA, the number of patent application: 200510091813.7 formed.
People such as Li Xiangye add sugar and cultivate aerobic spore-bearing bacilli in substratum when fed batch cultivation or batch culture, fermentative prodn γ-PGA, number of patent application: 200510091813.7.
People such as Shi Qingshan utilize saccharine material to ferment, and in fermentation raw material, do not add L-L-glutamic acid, number of patent application: 200610122640.5.
People such as Xu Zhinan are with the glutamic acid fermentation intermediate product, and glutami acid fermentation liquor, crystalline mother solution or bran acid are added in the fermention medium, in the fermented liquid content of γ-PGA up to 45-55 g/L, number of patent application: 200610155277.7.
But in existing γ-PGA fermentation manufacturing technique, not about utilizing the report of sodium glutamate mother liquid.
Summary of the invention
The present invention provides a kind of method of utilizing sodium glutamate mother liquid fermentative prodn gamma-polyglutamic acid-, and the more original technology of this method has significantly reduced production cost, in addition, sodium glutamate mother liquid has been carried out efficient utilization.
The technical scheme that the present invention takes is to comprise the following steps:
(1) utilizes activated lichem bacillus strain to carry out fermentation culture, in the fermentation culture process, slant strains is at first carried out liquid seeds cultivate, be inoculated in 10 % again and carry out fermentation culture in the fermention medium; Culture condition; Shaking culture under pH 7.0 ~ 7.4,30 ~ 40 ℃ of temperature, the aerobic conditions: 150-250 r/min or stir culture 100-400 rpm; Stream adds glucose and sodium glutamate mother liquid in the fermenting process; Make the concentration of glucose and L-glutamic acid all maintain 10-15 g/L, fermentation time is 48 ~ 72 h;
(2) in supernatant, add ethanol to remove thalline and part solid substance with fermented liquid is centrifugal, again γ-PGA is redissolved, carry out spraying drying, obtain refining γ-PGA.
Because glutami acid fermentation liquor and bran acid belong to process product, glutamic wherein is very high, and very suitable extraction L-glutamic acid is used for producing monosodium glutamate; And the glutamic in the crystalline mother solution is very low, and 2%-3% is only arranged.The used sodium glutamate mother liquid of the present invention is meant after the L-glutamic acid neutralizer process decolouring deironing that pass through primary crystallization again, uncrystallized Sodium Glutamate solute still is present in the middle of the liquid, and this liquid is referred to as sodium glutamate mother liquid.Sodium Glutamate content wherein is 30%-40%, also has some soluble impurities in addition, evaporative crystallization once more in the industrial production.The present invention originates its precursor as γ-PGA, flows during the fermentation to add.And soluble impurity can be easy in follow-up finished product treating processes, remove.
Technology of the present invention is simple, utilizes the glutamic acid fermentation in the sodium glutamate mother liquid to produce gamma-polyglutamic acid-, can realize to waste liquor from gourmet powder production efficient utilization, cut down the consumption of energy, practice thrift cost.
Embodiment
The used sodium glutamate mother liquid of the present invention is meant after the L-glutamic acid neutralizer process decolouring deironing that pass through primary crystallization again, uncrystallized Sodium Glutamate solute still is present in the middle of the liquid, and this liquid is referred to as sodium glutamate mother liquid.Sodium Glutamate content wherein is 30%-40%, also has some soluble impurities in addition, evaporative crystallization once more in the industrial production.The present invention originates its precursor as γ-PGA, flows during the fermentation to add.This section and repetition before can be removed.
Embodiment 1:
In 500 mL triangular flasks of 90 mL substratum are housed, inoculate the seed culture fluid of 10 mL Bacillus licheniformis, pH 7.0; 30 ℃, shaking culture 150 r/min cultivate 72 h under the aerobic conditions; During the fermentation, stream adds glucose and sodium glutamate mother liquid, makes the concentration of glucose and L-glutamic acid all maintain 10 g/L; After the fermentation ends, be 13.4 g/L through the output that detects γ-PGA;
Centrifugal removal thalline and part solid substance add ethanol in supernatant, γ-PGA is redissolved again, and carry out spraying drying, obtain refining γ-PGA.
Embodiment 2:
In 500 mL triangular flasks of 90 mL substratum are housed; Inoculate the seed culture fluid of 10 mL Bacillus licheniformis, 7.2,35 ℃ of pH; Shaking culture 200 r/min cultivate 60 h under the aerobic conditions; During the fermentation, stream adds glucose and sodium glutamate mother liquid, makes the concentration of glucose and L-glutamic acid all maintain 12.5 g/L; After the fermentation ends, be 14.6 g/L through the output that detects γ-PGA;
Centrifugal removal thalline and part solid substance add ethanol in supernatant, γ-PGA is redissolved again, and carry out spraying drying, obtain refining γ-PGA.
Embodiment 3:
In 500 mL triangular flasks of 90 mL substratum are housed, inoculate the seed culture fluid of 10 mL Bacillus licheniformis, pH 7.4; 40 ℃, 250 r/min cultivate 48h, during the fermentation; Stream adds glucose and sodium glutamate mother liquid, makes the concentration of glucose and L-glutamic acid all maintain 15 g/L.After the fermentation ends, be 13.1 g/L through the output that detects γ-PGA;
Centrifugal removal thalline and part solid substance add ethanol in supernatant, γ-PGA is redissolved again, and carry out spraying drying, obtain refining γ-PGA.
Embodiment 4:
In 500 mL triangular flasks of 90 mL substratum are housed; Inoculate the seed culture fluid of 10 mL Bacillus licheniformis, 7.2,37 ℃ of pH; Shaking culture 180 r/min cultivate 72 h under the aerobic conditions; During the fermentation, stream adds glucose and sodium glutamate mother liquid, makes the concentration of glucose and L-glutamic acid all maintain 12.5 g/L; After the fermentation ends, be 19.8 g/L through the output that detects γ-PGA;
Centrifugal removal thalline and part solid substance add ethanol in supernatant, γ-PGA is redissolved again, and carry out spraying drying, obtain refining γ-PGA.
Embodiment 5:
In 50 L fermentor tanks of 27 L substratum are housed, inoculate the seed culture fluid of 3 L Bacillus licheniformis, pH 7.0; 30 ℃, mixing speed is 200 rpm, cultivates 72 h; During the fermentation, air flow is 1.0 vvm, and continuous feeding stream adds glucose and sodium glutamate mother liquid; Making the concentration of glucose and L-glutamic acid all maintain 10 g/L, after the fermentation ends, is 44.1 g/L through the output that detects γ-PGA;
Centrifugal removal thalline and part solid substance add ethanol in supernatant, γ-PGA is redissolved again, and carry out spraying drying, obtain refining γ-PGA.
Embodiment 6:
In 50 L fermentor tanks of 27 L substratum are housed, inoculate the seed culture fluid of 3 L Bacillus licheniformis, pH 7.2; 35 ℃, mixing speed is 300 rpm, cultivates 60 h; During the fermentation, air flow is 1.0 vvm, and continuous feeding stream adds glucose and sodium glutamate mother liquid; Making the concentration of glucose and L-glutamic acid all maintain 12.5 g/L, after the fermentation ends, is 48.5 g/L through the output that detects γ-PGA;
Centrifugal removal thalline and part solid substance add ethanol in supernatant, γ-PGA is redissolved again, and carry out spraying drying, obtain refining γ-PGA.
Embodiment 7:
In 50 L fermentor tanks of 27 L substratum are housed, inoculate the seed culture fluid of 3 L Bacillus licheniformis, pH 7.4; 40 ℃, mixing speed is 400 rpm, cultivates 48 h; During the fermentation, air flow is 1.0 vvm, and continuous feeding stream adds glucose and sodium glutamate mother liquid; Making the concentration of glucose and L-glutamic acid all maintain 15 g/L, after the fermentation ends, is 43.2 g/L through the output that detects γ-PGA;
Centrifugal removal thalline and part solid substance add ethanol in supernatant, γ-PGA is redissolved again, and carry out spraying drying, obtain refining γ-PGA.
Embodiment 8:
In 50 L fermentor tanks of 27 L substratum are housed, inoculate the seed culture fluid of 3 L Bacillus licheniformis, pH 7.0; 37 ℃, mixing speed is 300 rpm, cultivates 72 h; During the fermentation, air flow is 1.0 vvm, and continuous feeding stream adds glucose and sodium glutamate mother liquid; Making the concentration of glucose and L-glutamic acid all maintain 12.5 g/L, after the fermentation ends, is 57.8 g/L through the output that detects γ-PGA;
Centrifugal removal thalline and part solid substance add ethanol in supernatant, γ-PGA is redissolved again, and carry out spraying drying, obtain refining γ-PGA.
Claims (1)
1. a method of utilizing sodium glutamate mother liquid fermentative prodn gamma-polyglutamic acid-is characterized in that comprising the following steps:
(1) utilizes activated lichem bacillus strain to carry out fermentation culture, in the fermentation culture process, slant strains is at first carried out liquid seeds cultivate, be inoculated in 10 % again and carry out fermentation culture in the fermention medium; Culture condition; Shaking culture under pH 7.0 ~ 7.4,30 ~ 40 ℃ of temperature, the aerobic conditions: 150-250 r/min or stir culture 100-400 rpm; Stream adds glucose and sodium glutamate mother liquid in the fermenting process; Make the concentration of glucose and L-glutamic acid all maintain 10-15 g/L, fermentation time is 48 ~ 72 h;
(2) in supernatant, add ethanol to remove thalline and part solid substance with fermented liquid is centrifugal, again γ-PGA is redissolved, carry out spraying drying, obtain refining γ-PGA.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103667412A (en) * | 2013-12-27 | 2014-03-26 | 天津北洋百川生物技术有限公司 | Method for fermentation production of high molecular weight gamma-polyglutamic acid |
| CN103755470A (en) * | 2013-12-31 | 2014-04-30 | 吉林中粮生化有限公司 | Sustained-release corn specific fertilizer produced from aginomoto mother liquor |
| CN103772013A (en) * | 2013-12-31 | 2014-05-07 | 吉林中粮生化有限公司 | Controlled-release fertilizer special for rice and produced by monosodium glutamate mother solution |
| CN113912464A (en) * | 2021-11-18 | 2022-01-11 | 五家渠慧尔生物科技有限公司 | Preparation method and application of polyglutamic acid particles |
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| CN100999745A (en) * | 2006-12-18 | 2007-07-18 | 浙江大学 | Process of preparing gamma-poly glutaminic acid |
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| CN100999745A (en) * | 2006-12-18 | 2007-07-18 | 浙江大学 | Process of preparing gamma-poly glutaminic acid |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103667412A (en) * | 2013-12-27 | 2014-03-26 | 天津北洋百川生物技术有限公司 | Method for fermentation production of high molecular weight gamma-polyglutamic acid |
| CN103755470A (en) * | 2013-12-31 | 2014-04-30 | 吉林中粮生化有限公司 | Sustained-release corn specific fertilizer produced from aginomoto mother liquor |
| CN103772013A (en) * | 2013-12-31 | 2014-05-07 | 吉林中粮生化有限公司 | Controlled-release fertilizer special for rice and produced by monosodium glutamate mother solution |
| CN113912464A (en) * | 2021-11-18 | 2022-01-11 | 五家渠慧尔生物科技有限公司 | Preparation method and application of polyglutamic acid particles |
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