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A strain of Brevibacterium flavum producing l-proline and its application

A technology of Brevibacterium flavum and proline, which is applied in the field of bioengineering and can solve the problems of inhibiting L-proline synthesis and the like

Active Publication Date: 2020-09-04
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The production of proline by microbial fermentation mainly uses glutamic acid as a precursor to synthesize L-proline. L-glutamic acid is then synthesized into L-proline, but high concentrations of glutamic acid can inhibit the synthesis of L-proline

Method used

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  • A strain of Brevibacterium flavum producing l-proline and its application
  • A strain of Brevibacterium flavum producing l-proline and its application
  • A strain of Brevibacterium flavum producing l-proline and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: the screening of bacterial strain

[0025] Screening liquid medium: sodium glutamate 70g, glucose 5g, beef extract 5g, yeast powder 5g, peptone 10g, sodium chloride 5g, dilute to 1L with distilled water, pH7.0-7.2, sterilize at 121°C for 30min.

[0026] Screening solid medium: sodium glutamate 65g, glucose 5g, beef extract 5g, yeast powder 5g, peptone 10g, sodium chloride 5g, agar 20g, distilled water to 1L, pH7.0~7.2, sterilized at 121℃ 30min.

[0027] Obtain a strain of Brevibacterium flavum through screening and acclimatization of bacterial strain, it can grow on the screening liquid medium that sodium glutamate concentration is 70g / L and the screening solid medium that sodium glutamate concentration is 65g / L, this The strain was named Brevibacterium flavum FMME444.

[0028] Brevibacterium flavum FMME444, named Brevibacterium flavum FMME444 taxonomically, was deposited in the China Center for Type Culture Collection on January 17, 2019, with the depos...

Embodiment 2

[0029] Example 2: Fermentation of Brevibacterium flavum FMME444 to produce L-proline

[0030] Solid plate medium: glucose 5g, beef extract 5g, yeast powder 5g, peptone 10g, sodium chloride 5g, agar 20g, dilute to 1L with distilled water, pH7.0~7.2, sterilize at 121°C for 30min.

[0031] Slant medium: 5g glucose, 5g beef extract, 5g yeast powder, 10g peptone, 5g sodium chloride, 20g agar, dilute to 1L with distilled water, pH7.0-7.2, sterilize at 121°C for 30min.

[0032] Seed medium: glucose 30g, potassium dihydrogen phosphate 1-2g, magnesium sulfate 0.5-1g, corn steep liquor 28-30g, urea 2.5-3g, biotin 1000μg, V B1 1000μg, dilute to 1L with distilled water, adjust the pH to 7.0-7.2 with sodium hydroxide solution, and sterilize at 121°C for 30min.

[0033] Fermentation medium: initial glucose 60, corn steep liquor 10g, sodium glutamate 40g, potassium dihydrogen phosphate 1g, anhydrous magnesium sulfate 0.3g, biotin V H 200 μg, V B1 200 μg, dilute to 1 L with distilled wat...

Embodiment 3

[0038] Example 3: Fermentation of Brevibacterium flavum FMME444 to produce L-proline

[0039] The fermentation process is the same as in Example 2, except that the fermentation temperature is 33°C. After the fermentation, the supernatant was collected and the content of L-proline in the fermentation broth was determined by ninhydrin chromogenic method to be 106.7 g / L.

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Abstract

The invention discloses an L-proline-producing Brevibacterium flavum and its application, belonging to the technical field of bioengineering. The Brevibacterium flavum has been deposited in the China Center for Type Culture Collection on January 17, 2019, the preservation number is CCTCC NO: M2019052, and the preservation address is Wuhan University, Wuhan, China. Brevibacterium flavum (Brevibacterium flavum) FMME444 of the present invention shows the ability of accumulating high-level L-proline, and its tolerance to sodium glutamate reaches 70g / L, which reduces the effect of adding sodium glutamate in the fermentation process. It has toxic effects on cells and further promotes the conversion of glutamate to proline. The yield of L-proline fermented by the bacteria is as high as 100-110g / L, and the yield of glucose is as high as 45%-50%.

Description

technical field [0001] The invention relates to an L-proline-producing Brevibacterium flavum and its application, belonging to the technical field of bioengineering. Background technique [0002] L-proline belongs to the heterocyclic amino acid group and is one of the "non-essential" amino acids in the human body. It has certain physiological activities and is widely used in medicine, food and seasoning, agriculture and organic chemistry, especially in medical research and treatment. role has been increasingly valued. With the deepening of research on L-proline, the scope of application will be further expanded. Therefore, the production of L-proline has very broad application prospects. [0003] At present, the production of L-proline mainly includes chemical synthesis, extraction from natural protein hydrolyzate and biological fermentation. Due to the low cost of raw materials, mild reaction conditions and easy large-scale production, the bio-fermentation method has bec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P13/24C12R1/13
CPCC12N1/20C12P13/24C12N1/205C12R2001/13
Inventor 罗秋玲刘立明季立豪张权刘佳陈修来
Owner JIANGNAN UNIV