Method for preparing poly-gamma-glutamic acid by fermenting maize raw material Bacillus subtilis

A technology of Bacillus subtilis and glutamic acid, applied in the field of microbial fermentation, can solve the problems of difficult removal of bacteria, large dosage, and difficulty of increasing polyglutamic acid

Active Publication Date: 2009-08-12
HEILONGJIANG KANPURE BIOTECH
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AI Technical Summary

Problems solved by technology

[0021] Problems in the production of γ-PGA by industrial fermentation γ-PGA fermentation has several factors that are different from other fermentations, resulting in high fermentation costs: 1 Raw material cost: carbon sources such as glucose, citric acid, glycerin, glutamic acid, and peptone are generally added to fermentation Or the amount of raw materials such as yeast extract is relatively large, and the yield of the product is lower than that of other fermented products
Undoubtedly, the cost of raw materials accounts for a large proportion of the production cost; 2. Production cost: γ-PGA is a high-molecular polymer, and as the fermentation time increases, the viscosity of the fermentation liquid increases continuously, and dissolved oxygen becomes a difficult problem in the fermentation. For some measures, the cost of introducing pure oxygen will increase compared with that of passing air. If a special fermenter is used, such as increasing the height-to-diameter ratio, the particularity of the fermenter increases the fixed cost, and the stirring power consumption is also large. These factors are the same as other Compared with fermented products, it undoubtedly increases the cost
The extraction of fermentation products in the later stage is also a problem that is relatively difficult and requires high cost. For example, the viscosity of the fermentation broth is high, and the bacteria in the fermentation broth are not easy to remove. When extracting γ-PGA, organic solvent extraction is generally used. The volume of the organic solvent and the fermentation broth The ratio is generally 3:1, and the consumption of ethanol accounts for a large proportion. This part of the cost will undoubtedly increase the difficulty of industrial production of polyglutamic acid

Method used

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  • Method for preparing poly-gamma-glutamic acid by fermenting maize raw material Bacillus subtilis
  • Method for preparing poly-gamma-glutamic acid by fermenting maize raw material Bacillus subtilis
  • Method for preparing poly-gamma-glutamic acid by fermenting maize raw material Bacillus subtilis

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example 1

[0070] 1. Seed culture preparation of slant seed medium: peptone 5.0g / L, beef extract 3.0g / L, sodium chloride 3.0g / L, agar 20.0g / L, pH value 7.4, sterilized at 121°C for 20min, cooled to room temperature, Inoculate and incubate at a constant temperature of 35°C for 24 hours.

[0071] Inoculate the seeds on the slant medium into the liquid seed medium, the liquid seed medium formula: peptone 5.0g / L, beef extract 3.0g / L, cornstarch 20.0g / L, glutenic acid 20.0g / L, NaCl3.0g / L, pH 7.4. Sterilize at 121°C for 20 minutes, cool to room temperature, inoculate, and culture at 35°C, 200r / min shaker for 24 hours.

[0072] 2 Liquid fermentation Put the mature liquid seeds into the fermentation medium according to the inoculation amount of 8%. The fermentation medium formula: peptone 5.0g / L, beef extract 3.0g / L, corn starch 50.0g / L, gluten acid 50.0g / L L, sodium chloride 3.0g / L, KH 2 PO 40 1.0g / L, MgSO 4 0.5g / L, pH7.4. Sterilize at 121°C for 20 minutes, cool to room temperature, inocul...

example 2

[0075] 1 Seed culture Same as Example 1.

[0076] 2 Liquid fermentation Put the mature liquid seeds into the fermentation medium according to the inoculum amount of 8%. The fermentation medium formula: peptone 5.0g / L, beef extract 3.0g / L, corn dextrin 50.0g / L, glutenic acid 50.0g / L, sodium chloride 3.0g / L, KH 2 PO 40 1.0g / L, MgSO 4 0.5g / L, pH7.4. Sterilize at 121°C for 20 minutes, cool to room temperature, inoculate, and culture at 35°C, 200r / min shaker for 72 hours.

[0077] 3 Extraction of γ-PGA Same as Example 1.

example 3

[0079] 1 Seed culture Same as Example 1.

[0080] 2. Liquid fermentation Put the mature liquid seeds into the fermentation medium according to the inoculum size of 8%. L, gluten acid 50.0g / L, sodium chloride 2.0g / L, KH 2 PO 40 1.0g / L, MgSO 4 0.5g / L, pH7.4. Sterilize at 121°C for 20 minutes, cool to room temperature, inoculate, and culture at 35°C, 200r / min shaker for 72 hours.

[0081] Preparation of corn saccharification solution: Weigh 100g corn flour, add 300mL water, adjust pH to 7.6 with NaOH, heat to 100°C, add 10u α-amylase / g raw material, and stir for 15min. Cool to 62°C, adjust pH to 5.0 with HCl, add 120 u / g of glucoamylase, keep warm for 10 hours, filter to obtain corn saccharification solution, adjust pH to 7.4, and set aside.

[0082] 3 Extraction of γ-PGA Same as Example 1.

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Abstract

The invention relates to a method for preparing a high molecular polymer Poly gamma-glumatic acid gamma-PGA from a corn raw material by the liquid state fermentation of bacillus subtilis. The method is characterized in that: the method for preparing a high molecular polymer Poly gamma-glumatic acid gamma-PGA from corns taken as a basic raw material by the liquid state fermentation of the bacillus subtilis screens Bacillus subtilis HCUL-B115, which is stored in the China Center Of Culture Collection, and the storage number is CGMCC No. 2283. The bacillus subtilis HCUL-B115 uses corn, corn starch, corn dextrin, and corn saccharification hydrolysis solution as basic raw materials to prepare the gamma-PGA by the deep liquid state fermentation. The method has the advantages that: the strain (Bacillus subtillis HCUL-B115) can use the corn, the corn starch, the corn dextrine and the corn saccharification hydrolysis solution as the basic raw materials to prepare the gamma-PGA by the liquid state fermentation, and the raw materials are low in price, so that the cost is lowered, and the industrialized production is easily realized.

Description

technical field [0001] The invention relates to microbial fermentation technology, in particular to a method for preparing polymer polyγ-glutamic acid (Polyγ-glumatic acid γ-PGA) by liquid state fermentation of corn raw material Bacillus subtilis. Background technique [0002] Poly γ-glumatic acid γ-PGA is an anionic polyamino acid produced by microbial fermentation in nature, which is formed by D-glutamic acid and L-glutamic acid through α-amino and γ-carboxyl groups of high molecular weight polymers. [0003] [0004] There is also a free carboxyl group at the α carbon atom of each repeating unit, so the molecular chain of γ-PGA has a large number of highly active free side chain carboxyl groups, which can form hydrogen bonds within or between molecules. γ-PGA is usually composed of about 5,000 glutamic acid monomers, and its relative molecular mass is generally 100-1,000kD. It has no typical peptide chain structure, nor is it a cyclic polypeptide. The basic skeleton i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12N1/20C12R1/125
Inventor 梁金钟刘颖徐伟
Owner HEILONGJIANG KANPURE BIOTECH
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