Method for improving fermentation yield and saccharic acid conversion rate of L-phenylalanine

A sugar-acid conversion rate, phenylalanine technology, applied in the field of bioengineering

Active Publication Date: 2021-01-22
乐康珍泰(天津)生物技术有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] With the continuous increase of domestic consumption of L-phenylalanine, the current domestic production of L-Phe can no longer meet the demand, and the import share is increasing. Using Escherichia coli to ferment phenylalanine, the conversion rate of sugar and acid Only about 25%, the output is mostly 50-80 g / L, which has a large room for improvement compared with the theoretical value

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A method for improving L-phenylalanine fermentation yield and sugar-acid conversion rate, the specific steps are as follows:

[0021] Strain activation: take the E. coli strain in the glycerin bacteria preservation tube and place it on the slant medium for passaging activation. The components of the slant medium are glucose 1.5 g / L, yeast powder 6 g / L, peptone 10 g / L, K 2 HPO 4. 3H 2 O 0.8 g / L, tyrosine 1 g / L, agar powder 25 g / L;

[0022]Seed culture: The activated strains were cultured in the seed medium at a temperature of 36±1°C, the pH was controlled at 6.8~7.2 by ammonia water, the composition of the seed medium was glucose 30 g / L, yeast powder 5 g / L, Peptone 1 g / L, citric acid 1.2g / L, MgSO 4 .7H 2 O 2 g / L, KH 2 PO 4 2.0 g / L, ammonium sulfate 2.0 g / L, V B1 1 mg / L, FeSO 4 .7H 2 O 10 mg / L, MnSO 4 .H 2 O 5 mg / L, vitamin H 1 mg / L, tyrosine 3 g / L, kanamycin 20 mg / L;

[0023] Fermentation culture: Inoculate the fermentation medium with 25% bacterial inoculu...

Embodiment 2

[0026] This part of the embodiment does not use amino acid chelate, but adds the metal ion of the mass concentration of amino acid chelate, aims at contrasting with embodiment 6-11 as control group 1.

[0027] Use 5 L fermentors, culture method and control parameter are identical with embodiment 1; Medium is MgSO 4 .7H 2 O 2 g / L, yeast powder 4 g / L, peptone 1.2 g / L, citric acid 1 g / L, K 2 HPO 4 .3H 2 O 7 g / L, tyrosine 1 g / L, glutamic acid 800mg / L, FeSO 4 52 mg / L, MnSO 4 27.5 mg / L, ZnSO 4 30 mg / L, CuSO 4 25mg / , CaCl 2 25 mg / L, V B1 2mg / L, V B 3 2mg / L, V B5 2 mg / L, kanamycin 10 mg / L; sterilization conditions: 0.1 MPa, 121°C, 20 min; fermentation ended after 50 hours. When placed in the tank, the dry weight of the bacteria was 54 g / L, the output of L-phenylalanine was 83.5 g / L, and the sugar-acid conversion rate was 25%.

Embodiment 3

[0029] This part of the embodiment does not use amino acid chelate, but adds the amino acid of mass concentration such as amino acid chelate, aims at contrasting with embodiment 6-11 as control group 2.

[0030] Use 5 L fermentors, culture method and control parameter are identical with embodiment 1; Medium is MgSO 4 .7H 2 O 2 g / L, yeast powder 4 g / L, peptone 1.2 g / L, citric acid 1 g / L, K 2 HPO 4 .3H 2 O 7 g / L, tyrosine 1.039 g / L, glutamic acid 964 mg / L, methionine 100 mg / L, V B1 2mg / L, V B 3 2mg / L, V B5 2 mg / L, kanamycin 10 mg / L; sterilization conditions: 0.1 MPa, 121°C, 20 min; fermentation ended after 50 h. When put into the tank, the dry weight of the bacteria was 46 g / L, the output of L-phenylalanine was 67 g / L, and the sugar-acid conversion rate was 16%.

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Abstract

The invention provides a method for improving the fermentation yield and saccharic acid conversion rate of L-phenylalanine. The method comprises the following steps of fermenting phenylalanine escherichia coli producing bacteria to obtain L-phenylalanine, and feeding amino acid chelated trace elements along with an organic nitrogen source in the fermentation process, wherein the amino acid chelated trace elements are composed of methionine chelated iron, glutamic acid chelated manganese, glutamic acid chelated zinc, tyrosine chelated copper and glutamic acid chelated calcium; and inorganic elements are changed into organic elements through chelation reaction of amino acid and metal cations, and the amino acid chelate with more stable chemical properties is formed. Therefore, trace elementions can be effectively prevented from forming insoluble compounds, the cell growth rate and the cell viability are improved, and then the yield and the saccharic acid conversion efficiency of phenylalanine are improved.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for improving the fermentation yield of L-phenylalanine and the conversion rate of sugar and acid. Background technique [0002] In recent years, with the increasing market demand for the new sweetener aspartame, L-phenylalanine (L-Phenylalanine, L-phe), as the main raw material for the production of aspartame, has rapidly increased its demand. Big. In addition, L-Phe plays an important role in pharmaceutically active compounds such as anti-inflammatory drugs, central nervous system neuropeptides, HIV protease inhibitors and vitamin B 6 It is widely used, and it can also be used as a carrier of anticancer drugs, which can effectively introduce anticancer drugs into the tumor site, while enhancing the therapeutic effect, and greatly reducing the side effects of drugs, which means that the application in the field of medicine is more and more popular. Pay attentio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/22C12N1/20C12N1/38C12R1/19
CPCC12P13/222C12N1/20C12N1/38
Inventor 徐庆阳李志华陈志超张宁杨柳
Owner 乐康珍泰(天津)生物技术有限公司
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