Bacillus subtilis and application thereof in preparation of gamma-D-polyglutamic acid

A technology of Bacillus subtilis and polyglutamic acid, applied in the field of fermentation engineering, can solve the problems of industrial production of natural γ-PGA, etc., and achieve the effects of cost reduction, convenient and simple operation, and extensive cultivation conditions

Active Publication Date: 2015-03-25
NANJING SHINEKING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no industrial production of natural γ-PGA with high optical purity

Method used

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  • Bacillus subtilis and application thereof in preparation of gamma-D-polyglutamic acid
  • Bacillus subtilis and application thereof in preparation of gamma-D-polyglutamic acid
  • Bacillus subtilis and application thereof in preparation of gamma-D-polyglutamic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Batch fermentation of B. subtilis PG-8 in a 7.5 L tank to produce γ-D-PGA.

[0043] Seed medium: glucose 5g / L, beef extract 5g / L, peptone 5g / L, MgSO 4 ·7H 2 O0.5g / L, use NaOH solution to adjust pH6.8.

[0044] Fermentation medium: glucose 40g / L, monosodium glutamate 40g / L, yeast extract 5g / L, (NH 4 ) 2 SO 4 5g / L, K 2 HPO 4 ·3H 2O2.5g / L, MgSO 4 ·7H 2 O0.1g / L, use NaOH solution to adjust pH6.8.

[0045] Cultivate Bacillus subtilis B.subtilis PG-8 (CGMCC NO:6324) in the seed medium, 30°C, 200r / min shaker conditions for 15h, and the seed solution is 5% (v / v) of the seed amount Inoculate in a fermenter pre-installed with 4.5L sterilized fermentation medium for cultivation, culture conditions: 30°C, 400r / min, ventilation rate 1.2L / min, turn on the pH automatic control device during the fermentation process, use HCl Or ammonia water is used to control the pH value of the fermentation broth between 6.5-7.5. After 48 hours of fermentation, the concentratio...

Embodiment 2

[0048] Example 2: γ-D-PGA was produced by fermenting B. subtilis PG-8 in a 7.5 L tank with controlled rotational speed in stages.

[0049] Seed medium: glucose 5g / L, beef extract 5g / L, peptone 5g / L, MgSO 4 ·7H 2 O0.5g / L, use NaOH solution to adjust pH7.0.

[0050] Fermentation medium: acid-treated molasses 20g / L, glucose 10g / L, sucrose 10g / L, peptone 2g / L, (NH 4 ) 2 SO 4 5g / L, MSG wastewater 80g / L, K 2 HPO 4 ·3H 2 O2.5g / L, MgSO 4 ·7H 2 O0.1g / L, use NaOH solution to adjust pH7.0.

[0051] Cultivate Bacillus subtilis B.subtilis PG-8 (CGMCC NO:6324) in the seed medium, 32°C, 220r / min shaker conditions for 16h, and the seed solution is 5% (v / v) of the seed amount Inoculate in a fermenter pre-installed with 4.5L sterilized fermentation medium for culture, culture conditions: keep the temperature at 32°C throughout the fermentation process, the ventilation rate is 1.2L / min, open the pH automatic control device during the fermentation process, Utilize HCl or NaOH to contro...

Embodiment 3

[0054] Example 3: 7.5L tank fed-batch fermentation of B. subtilis PG-8 to produce γ-D-PGA.

[0055] Seed medium: glucose 5g / L, beef extract 5g / L, peptone 5g / L, MgSO 4 ·7H 2 O0.5g / L, use NaOH solution to adjust pH6.8.

[0056] Fermentation medium: molasses 90g / L, peptone 5g / L, (NH 4 ) 2 SO 4 5g / L, MSG wastewater 90g / L, K 2 HPO 4 ·3H 2 O2.5g / L, MgSO 4 ·7H 2 O0.1g / L, use NaOH solution to adjust pH6.8.

[0057] Cultivate Bacillus subtilis B.subtilis PG-8 (CGMCC NO:6324) in the seed medium, 32°C, 200r / min shaker conditions for 16h, and the seed solution is 4% (v / v) of the seed amount Inoculate in a fermenter pre-installed with 4.5L sterilized fermentation medium for cultivation, culture conditions: 32°C, 400r / min, ventilation rate 1.4L / min, turn on the pH automatic control device during the fermentation process, use HCl Or NaOH controls the pH value of the fermentation broth between 6.8-7.2. When the residual total sugar in the fermentation broth is 10g / L, turn on the f...

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Abstract

The invention discloses bacillus subtilis. According to the bacillus subtilis, the category name is bacillus subtilis PG-8, the bacillus subtilis is collected at China general microbiological culture collection center (CGMCC), the registered number is CGMCC NO. 6324, and the collection data is 10, July, 2012. The invention further discloses application of the bacillus subtilis in preparation of gamma-D-polyglutamic acid. The bacillus subtilis is cultured in a culture medium containing L-glutamic acid only, and then, the gamma-D-polyglutamic acid of 30-60 g / L can be accumulated under optimized conditions. According to the bacillus subtilis, a variety of carbon and nitrogen sources can be utilized, culture conditions are extensive, the operation is convenient and simple, the yield of the gamma-D-polyglutamic acid is high, and the production cost is low, so that the bacillus subtilis can be widely applied to the fields of plant water conservation, food additives and the like.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering, and relates to a high-yield gamma-D-polyglutamic acid (gamma-D-PGA) microbial strain Bacillus subtilis and its application. Background technique [0002] γ-polyglutamic acid (γ-PGA for short) is an extracellular polymer produced by a variety of bacilli (such as Bacillus anthracis, Bacillus megaterium, Bacillus licheniformis and Bacillus subtilis, etc.). γ-PGA is a homopolyamino acid in which glutamic acid monomers are condensed with γ-carboxyl groups and amino groups, and the main chain is a nylon-4 structure (1). γ-PGA has many unique physical, chemical and biological properties such as excellent biodegradability, film-forming property, fibrosis, plasticity, cohesiveness, moisture retention, etc. Today, γ-PGA is emphasizing environmental protection and sustainable development. -PGA and its derivatives have very broad application prospects. It can be used in cosmetics, food, di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/125
Inventor 徐虹冯小海张丹李莎粱金丰
Owner NANJING SHINEKING BIOTECH CO LTD
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