Process of preparing gamma-polyglutamic acid by bacillus subtilis and glutamic acid bacillus mixed cultivating system

A technology of Corynebacterium glutamicum and Bacillus subtilis, which is applied in the field of preparation of γ-polyglutamic acid, can solve the problems of high production cost, long production cycle, and large consumption of raw materials, and achieve efficient and cheap preparation and good hygroscopic performance and moisturizing properties, and the effect of reducing production costs

Inactive Publication Date: 2010-06-02
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Although the fermentative production of γ-polyglutamic acid has made great progress, there are still shortcomings such as the need for a large amount of exogenous glutamic acid addition, the consumption of more raw materials, the longer production cycle, and low production efficiency. The problem of high production cost has seriously restricted the large-scale industrial application of polyglutamic acid products

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The following medium components are measured by mass volume percentage, and the volume is supplemented with water.

[0041] 1) Preparation of Bacillus subtilis seed liquid:

[0042] Step 1: Bacillus subtills (Bacillus subtills) zju-7 was cultured on a slant at 37°C for 8 hours to prepare mature slant seeds. The slant nutrient medium: glucose 1.5%, peptone 1%, glutamic acid 3%, MgSO 4 0.1%, NaCl 1%, K 2 HPO 4 0.01%, pH 7;

[0043] Step 2: Inoculate the Bacillus subtilis capable of producing polyglutamic acid into the liquid seed medium from the above-mentioned mature solid slant, and culture it in a shaker flask at 37°C for 13 hours at a rotation speed of 200rpm to prepare liquid seeds for mixed strain fermentation , OD 600 For 3, the liquid seed medium: glucose 2%, peptone 1.5%, glutamic acid 3%, MgSO 4 0.1%, NaCl 1%, K 2 HPO 4 0.01%, pH7, filling volume 20ml / 500ml shake flask, sterilized at 115°C for 20 minutes.

[0044] 2) Preparation of Corynebacterium g...

Embodiment 2

[0051] As in Example 1, the glucose in the Corynebacterium glutamicum fermentation medium in the mixed culture system was replaced with sucrose. After the fermentation, the γ-polyglutamic acid was separated and purified as in Example 1, and the yield could reach 18.3 g / L.

Embodiment 3

[0053] 1) Preparation of Bacillus subtilis seed liquid:

[0054] Step 1: Bacillus subtilis (Bacillus subtills) zju-7 was cultured on a slant at 37°C for 8 hours to prepare mature slant seeds. The slant nutrient medium: glucose 1.5%, peptone 1%, glutamic acid 3%, MgSO 4 0.1%, NaCl 1%, K 2 HPO 4 0.01%, pH 7;

[0055] The second step: inoculate the Bacillus subtilis capable of producing polyglutamic acid into the liquid seed medium from the above-mentioned mature solid slant, and culture it in a shaker flask at 37°C for 12 hours at a rotation speed of 200rpm to prepare liquid seeds for mixed strain fermentation , OD 600 0.5, the liquid seed medium: glucose 2%, peptone 1.5%, glutamic acid 3%, MgSO 4 0.1%, NaCl 1%, K 2 HPO 4 0.01%, pH7, filling volume 20ml / 500ml shake flask, sterilized at 115°C for 20 minutes.

[0056] 2) Preparation of Corynebacterium glutamicum seed solution

[0057] In the first step, Corynebacterium glutamicum (Corynebacterium glutamicum) S9114 was...

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Abstract

The present invention is process of preparing poly-gamma-glutamic acid in mixed culture system of Bacillus subtilis and Corynebacterium glutamicum. Bacillus subtilis strain and Corynebacterium glutamicum strain are cultured in culture medium containing carbon source, nitrogen source, inorganic salt and biotin and under optimized condition to obtain poly-gamma-glutamic acid fermenting liquor, whichis treated through deposition in organic solvent, dialysis and drying to obtain poly-gamma-glutamic acid. The process has low production cost and high yield.

Description

[0001] The invention relates to a method for preparing gamma-polyglutamic acid using a mixed culture system of bacillus subtilis and corynebacterium glutamicum. Background technique [0002] γ-polyglutamic acid (poly γ-glutamic acid, γ-PGA) is a homo-amino acid compound that can be synthesized by microorganisms. As a biopolymer material, γ-polyglutamic acid has the advantages of good biodegradability, edible, and non-toxic to human body and environment. Therefore, γ-polyglutamic acid and its derivatives are widely used in food, cosmetics, medicine and water treatment. [0003] At present, the methods for synthesizing γ-polyglutamic acid mainly include chemical synthesis, extraction and microbial synthesis. However, the chemical synthesis method has a long synthesis route, many by-products, low yield, and the product has a small molecular weight, which is difficult to meet the requirements as a new type of drug carrier material, and has no industrial application value. Howeve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P39/00C12P1/00C12R1/15C12R1/125
Inventor 徐志南石峰岑沛霖
Owner ZHEJIANG UNIV
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