L-tyrosine preparation method through enzymatic conversion

A technology of tyrosinase and tyrosine, applied in the direction of biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., to achieve good economic and social benefits, simple process flow, and mild reaction conditions

Active Publication Date: 2013-07-31
湖北新生源生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the problem of production cost, the enzymatic synthesis of L-tyrosine with L-serine or pure pyruvate has no obvious added value at present, so it has no practical significance to synthesize L-tyrosine with L-serine or pure pyruvate enzymatic method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The amino acid feed solution containing L-serine is derived from keratin hydrolyzate.

[0029] 1. Take Escherichia coli K-12 wet bacteria 10 g and Citrobacter freundii Add 15 g of ATCC 8090 wet bacteria to 1000 ml of mixed amino acid feed solution containing 90g / L L-serine (the total amino acid content in the feed solution is 10%), add 0.2 g pyridoxal phosphate and 0.01 g Tween 80, add 154 ml of 500 g / L phenol aqueous solution, maintain the phenol content in the conversion liquid at 10 g / L, and control the pH value of the conversion liquid with 25% ammonia water by volume, at a temperature of 45°C, After 15 hours of enzymatic reaction, the concentration of L-tyrosine in the conversion solution was 147.3 g / L after the reaction, and the molar conversion rate of L-serine was 95%.

[0030] 2. Put the transformation solution into a centrifuge, centrifuge at a speed of 4000 rpm for 15 minutes, collect 226 g of the wet cell and solid L-tyrosine mixture, dissolve the soli...

Embodiment 2

[0032]The amino acid feed solution containing L-serine is derived from keratin hydrolyzate.

[0033] 1. Take Escherichia coli K-12 wet bacteria 11 g and Citrobacter freundii Add 15 g of ATCC 8090 wet bacteria to 1000 ml of mixed amino acid feed solution containing 80g / L L-serine (the total amino acid content in the feed solution is 20%), add 0.2 g of pyridoxal phosphate and 0.1 g of Tween 80, add 137 ml of 500 g / L phenol aqueous solution, maintain the phenol content in the conversion solution at 10 g / L, control the pH of the conversion solution with 25% volume percent ammonia water, and enzymatically catalyze the reaction at 40°C After 14 hours of reaction, the concentration of L-tyrosine in the conversion solution was 131 g / L, and the molar conversion rate of L-serine was 95%.

[0034] 2. Put the transformation solution into a centrifuge, centrifuge at a speed of 4000 rpm for 15 minutes, collect 202 g of the wet cell and solid L-tyrosine mixture, dissolve the solid mixt...

Embodiment 3

[0036] The amino acid feed solution containing L-serine is derived from keratin hydrolyzate.

[0037] 1. Take Escherichia coli K-12 wet bacteria 10 g and Citrobacter koseri Add 14 g of ATCC BAA-895 wet bacteria to 1000 ml of mixed amino acid feed liquid containing 60 g / L L-serine (the total amino acid content in the feed liquid is 30%), add 0.2 g pyridoxal phosphate and 0.1 g Add 103 ml of 500 g / L phenol aqueous solution to polyethylene glycol octylphenyl ether (OP), maintain the phenol content in the conversion solution at 10 g / L, and control the pH of the conversion solution at 25% by volume with ammonia water , at a temperature of 30°C, the enzymatic reaction was carried out for 18 hours. After the reaction, the concentration of L-tyrosine in the conversion solution was 98.3 g / L, and the molar conversion rate of L-serine was 95%.

[0038] 2. Put the transformation solution into a centrifuge, centrifuge at a speed of 4000 rpm for 15 minutes, collect 158 ​​g of the wet ...

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PUM

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Abstract

The invention provides an L-tyrosine preparation method through enzymatic conversion. The method comprises the following steps: 1, respectively culturing strains having the activities of serine deaminase and strains having the activities of tyrosine phenol-lyase in a culture medium; 2, mixing two cells of the serine deaminase and the tyrosine phenol-lyase with an L-serine-containing amino acid material liquid, adding an aqueous phenol solution having a concentration of 500g/L, pyridoxal phosphate having a concentration of 0.2g/L and a surfactant having a concentration of 0.01-1.0g/L, adjusting the pH value to 6-11 by using ammonia water having a volume percentage of 25%, and carrying out an enzymatic reaction for 6-35h; and 3, centrifuging the obtained conversion liquid for 15min in 4000r/min, collecting the obtained mixture, dissolving the mixture by using 200-7000ml of pure water, adding 10mol/L sodium hydroxide in a dropwise manner to adjust the pH value to 12-13, heating to 80DEG C while stirring, adjusting the pH value of the obtained filtrate to 6 by using 6mol/L hydrochloric acid, and drying to obtain required L-tyrosine.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an L-tyrosinase conversion preparation method. Background technique [0002] L-tyrosine is an aromatic amino acid, one of the important amino acids needed for protein synthesis in animals, and its amino and carboxyl groups have important physiological functions. In the field of medicine, L-tyrosine is the precursor for the synthesis of thyroxine, dopa, and adrenal gland; in the field of food, L-tyrosine is an important additive; clinically, L-tyrosine is also used for reproduction Contraceptive Efficacy Research. According to current literature reports, the preparation methods of L-tyrosine mainly include extraction method, chemical synthesis method, fermentation method and enzymatic conversion method. [0003] 1. Extraction method [0004] The extraction method to produce L-tyrosine is to use natural protein resources such as human hair, pig hair, feathers, pig blood meal, etc. a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P13/22C12R1/19C12R1/01C12R1/18
Inventor 肖国安王先兵焦庆才刘均忠章平
Owner 湖北新生源生物工程有限公司
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