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A method for producing phenylalanine by microbial fermentation

A fermentation method, phenylalanine technology, applied in the field of bioengineering, can solve problems such as slow cell growth, achieve the effects of improving economic benefits, reducing the amount of feeding, and reducing the amount of ammonium sulfate

Inactive Publication Date: 2016-11-30
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Excessive addition of L-tyrosine in the fermentation process will cause a large number of cells to grow, so that less carbon flow will flow to the synthesis of L-phenylalanine; too little added L-tyrosine will cause cell growth. Slow or even too little growth, so there is not enough bacteria to produce more L-phenylalanine

Method used

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  • A method for producing phenylalanine by microbial fermentation
  • A method for producing phenylalanine by microbial fermentation
  • A method for producing phenylalanine by microbial fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 This example illustrates the method of producing L-phenylalanine using Escherichia coli BL21, which is preserved by our laboratory. (Escherichia coli BL21 was purchased from Beijing Quanshijin Biotechnology Co., Ltd.)

[0024] Specifically include the following steps:

[0025] (1) Activation of strains: During slant culture, the strains are cultured in a constant temperature incubator after streaking on the slant medium, at a temperature of 30-40°C, and the activation culture time is 1-2d, which is used for seed medium inoculation and strain preservation;

[0026] (2) Seed cultivation: add 50mL of seed culture medium to a 500mL Erlenmeyer flask, sterilize with high-pressure steam at 121°C for 20 minutes, and insert strains on the slant medium after cooling.

[0027] (3) Fermentation culture: the volume of fermentation medium in a 1L fermenter is 0.5-0.8L, the inoculum size is 5%-10% (v / v), the temperature is 30-40°C, the stirring speed is 200-500rpm, and the...

Embodiment 2

[0031] Example 2 This example illustrates the optimization method for the preparation of L-phenylalanine by Escherichia coli BL21 (pET / P-aspC), mainly involving the optimization of ammonium sulfate concentration

[0032] In this embodiment, the bacterial strain used is Escherichia coli BL21 (pET / P-aspC), and the concentration of ammonium sulfate (g / L) in the fermentation medium is 0, 5, 10, 20, 30 respectively. Other cultivation and fermentation methods and implementation Example 1 is the same.

[0033] In this example, different concentrations of ammonium sulfate were used to ferment and prepare L-phenylalanine, and the results were as follows figure 1 shown. Ammonium sulfate contains nitrogen required for bacterial growth and amino acid production, but an excess supply of ammonium ions can lead to competitive inhibition problems. Studies have shown that ammonium concentrations exceeding 3-5g / L tend to cause inhibition. Such as figure 1The ammonium sulfate of initial 10g / ...

Embodiment 3

[0034] Example 3 This example illustrates the optimization method for the preparation of L-phenylalanine by Escherichia coli BL21 (pET / P-aspC), which mainly involves the optimization of the types of organic nitrogen sources

[0035] In this example, the strain used is Escherichia coli BL21 (pET / P-aspC), the type of organic nitrogen source in the fermentation medium is changed, and other cultivation and fermentation methods are the same as in Example 1.

[0036] The above-mentioned organic nitrogen sources are respectively a single organic nitrogen source and a compound organic nitrogen source, and the total nitrogen amount of the added organic nitrogen source is equivalent; in the compound organic nitrogen source, the two single organic nitrogen sources are calculated according to their nitrogen content 1:1 added; the type and concentration of a single organic nitrogen source are yeast powder 10g / L, corn steep liquor dry powder 11.5g / L, cottonseed cake powder 10g / L, bean cake e...

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Abstract

The invention relates to an efficient and safe method for producing L-phenylalanine by microbial fermentation, which belongs to the technical field of bioengineering. The concentration of ammonium sulfate and the type of organic nitrogen source were optimized, and the multi-stage L-tyrosine index feeding method was used to significantly increase the concentration of L-phenylalanine in the fermentation broth. The output of L-phenylalanine reached about 56 g / L. At the same time, the amount of ammonium sulfate, yeast powder and L-tyrosine is significantly reduced compared with common fermentation methods, which greatly reduces the production cost of L-phenylalanine.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a new method for fermenting and preparing L-phenylalanine, specifically for the process of fermenting and producing L-phenylalanine, optimizing the nitrogen source and multi-stage index L-phenylalanine. Proposed amino acid feeding strategy. Background technique [0002] L-phenylalanine, also known as L-phenylalanine, is one of the eight essential amino acids that are necessary for the human body but cannot be synthesized by itself. It is widely used in the fields of medical drugs, cosmetics, food and its additives. In particular, As one of the raw materials for the synthetic sweetener aspartame. The traditional production methods of L-phenylalanine are chemical synthesis method and enzymatic method, but the chemical synthesis method has the disadvantages of environmental pollution, most of the products are racemates, and the subsequent processing technology is complicated; ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/22C12R1/19
Inventor 陈可泉袁佩佩何珣曹伟佳王震肖文赵艳欧阳平凯
Owner NANJING UNIV OF TECH
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