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

A technology of Corynebacterium glutamicum and Bacillus subtilis is applied in the field of preparing γ-polyglutamic acid, which can solve the problems of long production cycle, high consumption of raw materials and high production cost, and achieves reduction of production cost, good moisture absorption performance and high production cost. Moisturizing properties, the effect of efficient and inexpensive preparation

Inactive Publication Date: 2007-07-18
ZHEJIANG UNIV
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AI Technical Summary

Benefits of technology

In this patented technology, there are several technical benefits that make Bacilluss Subtilis sporadically producing gamma- polyglucosaminidase A more effective than other methods like enzymes or chemical reactions. These improvements include increased stability against heat stress, reduced sensitivity towards oxidants, improved absorption capabilities when applied topically onto skin, better control over pH levels, ability to formulate into various forms without losing their effectiveness due to dilution caused by sweatiness or dryness, low costs compared to existing techniques, and greater flexibility in adjustments made based upon these factors.

Problems solved by technology

Technological Problem addressed in this patents relates to improving the efficient and economically viable manufacture of α-picolylic acid(α-PA), another key ingredient found naturally occurring within baccharides called beta cyclamsulfuronosporans (BSCNPS). These natural sources consist primarily of various types of organism including yeast cells, but they cannot easily achieve higher yields due to their limited supply compared to other commercially available polysaccharide producers like lycopene monocyanate and nigrosine. To solve these issues, novel techniques involving enzyme catalysis combined with immobilization technique were developed.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The following medium components are measured in percent by mass and volume, and water is used to make up the volume.

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

[0042] The first step: Bacillus subtills (Bacillus subtills) zju-7 was cultured on a slant solid 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%, PH7;

[0043] The second step: inoculate the polyglutamic acid-producing Bacillus subtilis from the above mature solid slant to the liquid seed medium, culture it in a shake flask at 37°C for 13 hours, rotate at 200 rpm, and prepare liquid seeds for the fermentation of mixed strains , OD 600 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, 20ml / 500ml shake flask, sterilized at 115°C for 20 minutes.

[0044] 2) Preparation of Corynebacterium glutamicum seed liquid

[0045] In the f...

Embodiment 2

[0051] As in Example 1, the glucose in the fermentation medium of Corynebacterium glutamicum 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 solution:

[0054] The first step: Bacillus subtills (Bacillus subtills) zju-7 was cultured on a slant solid 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%, pH7;

[0055] Step 2: Inoculate the polyglutamic acid-producing Bacillus subtilis from the mature solid sloping surface in the liquid seed culture medium, culture it in a shake flask at 37°C for 12 hours, rotate at 200 rpm, and prepare liquid seeds for the fermentation of mixed strains , OD 600 Is 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, 20ml / 500ml shake flask, sterilized at 115°C for 20 minutes.

[0056] 2) Preparation of Corynebacterium glutamicum seed liquid

[0057] In the first step, Corynebacterium glutamicum S9114 was cultured on a slope at 37°C for 8 hours to prepare mature slope seeds....

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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, which is 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

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Claims

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Application Information

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Owner ZHEJIANG UNIV
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