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A strain producing l-phenylalanine and its application

A technology of phenylalanine and Escherichia coli, which is applied in the direction of bacteria, microorganism-based methods, enzymes, etc., can solve the problem of not being able to accumulate a large amount of phenylalanine, and achieve the effect of high capacity and broad application prospects

Inactive Publication Date: 2017-10-20
沙县鑫创生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because end products such as L-phenylalanine have a strong feedback inhibitory effect on the key enzymes in the synthetic and metabolic pathway of aromatic amino acids in wild strains, the cells cannot accumulate phenylalanine in large quantities. Therefore, if the actual fermentation production, it is urgent to obtain strains with high ability to produce L-phenylalanine

Method used

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  • A strain producing l-phenylalanine and its application
  • A strain producing l-phenylalanine and its application
  • A strain producing l-phenylalanine and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1, the preparation of the bacterial strain that produces L-phenylalanine

[0035] 1. Construction of recombinant plasmid pBV220-aroG-pheA

[0036] Genomic DNA of Escherichia coli 10245 was extracted, used as a template, and primers aroG-f and aroG-r were used as primers for PCR amplification to obtain aroG gene.

[0037] Genomic DNA of Escherichia coli 10245 was extracted, used as a template, and primers pheA-f and pheA-r were used as primers for PCR amplification to obtain the pheA gene.

[0038] EcoRI and KpnⅠ double digestion of aroG gene to obtain the gene fragment; EcoRI and KpnⅠ double digestion of pMD-18T to obtain a large vector fragment; the gene fragment was connected to the large vector fragment to obtain a recombinant plasmid, which was named pMD-18T-aroG .

[0039] KpnⅠ and BamHI double-digested the pheA gene to obtain a gene fragment; KpnⅠ and BamHI double-digested pMD-18T to obtain a large vector fragment; the gene fragment was connected to t...

Embodiment 2

[0111] Example 2, the acid production (L-phenylalanine) ability of bacterial strain MD1078 and Escherichia coli 10245

[0112] Acid production capacity of 50L fermenters of Escherichia coli MD1078 and Escherichia coli 10245

[0113] 1. Seed culture (using a 30L automatic fermenter, the liquid volume of the seed medium is 20L): inoculate Escherichia coli MD1078 and Escherichia coli 10245 into the seed medium respectively, and the control parameters of the 30L seed fermentation tank are 36°C, dissolved oxygen (DO) is 40%-60%, the glucose concentration in the seed medium is 30g / L, and the cultivation time is 12 hours, and the seed liquid (seed liquid OD 610 =30).

[0114] 2. Fermentation culture (using a 100L fully automatic fermenter, the liquid volume of the fermentation medium is 50L): Use a 100L fermenter for fermentation. The seed liquid is inoculated into the fermentation medium at a volume of 25%. The control parameters of the fermenter are 36°C, dissolved oxygen (DO) 35...

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Abstract

The invention discloses a bacterial strain for producing L-phenylalanine and an application of the bacterial strain. The recombinant escherichia coli MD1078 has a preservation number of CCTCC NO:M2013509. In a 100L full-automatic fermentation tank, L-phenylalanine yield is 86.5g / L under the conditions with a temperature of 36 DEG C, 35%-50% of dissolved oxygen and fermentation time of 48 hours. The escherichia coli MD1078 is high in capacity of producing L-phenylalanine and has a wide application prospect in the practical production of L-phenylalanine.

Description

technical field [0001] The invention relates to a bacterial strain producing L-phenylalanine and its application. Background technique [0002] L-phenylalanine (L-phenylalanine) is a white crystalline powder. It is one of the essential amino acids that humans and animals cannot synthesize in vivo. It is widely used in food, feed, medicine and cosmetics and other fields. As one of the two raw materials for the synthesis of Aspartame, L-phenylalanine is widely used in the preparation of low-calorie, high-sweet dipeptide sweetener Aspartame. L-phenylalanine The market demand for amino acid is increasing rapidly. In addition, L-phenylalanine is also an essential raw material for antineoplastic drugs and amino acid infusion preparations. With its continuous development in the field of medicine, the demand is also increasing. Therefore, the research on the biosynthetic pathway of L-phenylalanine, the improvement of L-phenylalanine-producing bacteria and the research on its indus...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P13/22C12R1/19
CPCC12N9/1085C12N9/88C12N9/90C12P13/222C12Y205/01054C12Y402/01051C12Y504/99005
Inventor 吴伟斌施巧琴翁雪清黄钦耿赵燕玉陈炳生吴松刚
Owner 沙县鑫创生物科技有限公司