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Preparation process for synthesizing D-phenylalanine by using high-activity bio-enzyme

A technology of phenylalanine and preparation process, applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of small production scale, high cost, low yield and the like

Inactive Publication Date: 2011-05-04
崔传僧
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the production of D-phenylalanine mainly adopts the split process, which has small production scale, low yield and high cost, and is difficult to meet the needs of domestic and foreign markets.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Cultivate the Psudomonas No.1 strain with multiple culture medium to obtain a fermentation broth. Disinfection process of fermentation broth at all levels: disinfection steam temperature 120-130°C, pressure 1.2-1.5 kg / cm 2 , The disinfection time is 30 minutes. The process control conditions of the three-stage fermentation are as follows:

[0018] a, primary fermentation: 2% liquid sugar in the fermentation broth, 2% corn steep liquor, 0.3% sodium chloride, 0.1% ammonium sulfate, and the balance is water. Use hydrochloric acid and liquid caustic soda to adjust the pH value to 6.7-7.0. The fermentation temperature is 35-37°C, the stirring speed is 60 rpm, and the incubation time is 20 hours;

[0019] b, secondary fermentation: 2% liquid sugar in the fermentation broth, 2% corn steep liquor, 0.3% sodium chloride, 0.1% ammonium sulfate, 0.06% magnesium sulfate, and the balance is water. Use hydrochloric acid and liquid caustic soda to adjust the pH value to 6.7-7.0...

Embodiment 2

[0024] (1) Cultivate the Psudomonas No.1 strain with multiple culture medium to obtain a fermentation broth. Disinfection process of fermentation broth at all levels: disinfection steam temperature 120-130°C, pressure 1.2-1.5 kg / cm 2 , The disinfection time is 30 minutes. The process control conditions of the three-stage fermentation are as follows:

[0025] a, primary fermentation: 2% liquid sugar in the fermentation broth, 2% corn steep liquor, 0.3% sodium chloride, 0.1% ammonium sulfate, and the balance is water. Use hydrochloric acid and liquid caustic soda to adjust the pH value to 6.7-7.0. The fermentation temperature is 35-37°C, the stirring speed is 60 rpm, and the incubation time is 20 hours;

[0026] b, secondary fermentation: 2% liquid sugar in the fermentation broth, 2% corn steep liquor, 0.3% sodium chloride, 0.12% ammonium sulfate, 0.05% magnesium sulfate, and the balance is water. Use hydrochloric acid and liquid caustic soda to adjust the pH value to 6.7-7....

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Abstract

The invention discloses a preparation process for synthesizing D-phenylalanine by using high-activity bio-enzyme, comprising the steps of: culturing Psudomonas No.1 strains by using a multivariant culture medium, flocculating and collecting mycelium, putting the collected mycelium in a transformation tank, and putting 5-benzyl hydantoin at the same time, wherein the weight of the 5-benzyl hydantoin is 1 / 5-1 / 6 of the total weight of fermentation liquor before the flocculation; protecting the transformation tank by using nitrogen, wherein the pressure is 0.1-0.2kg, the transformation temperature is 90DEG C, the stirring speed is 120RPM, the pH is kept at 8 and the transformation time is 24h; and conventionally collecting the transformed clear liquor, then refining, concentrating, evaporating, crystallizing, centrifuging and drying the obtain the finished product. In the invention, high-activity D-hydantoinase and N-carbamoyl enzyme can be generated after the fermentation of the strains. Through the transformation at proper transformation temperature, the transformation rate of the enzyme to a substrate is stabilized at 99-100%, so that the product yield of the D-phenylalanine can achieve 84-86%.

Description

technical field [0001] The invention relates to a method for preparing nitrogen-containing organic compounds by using enzymes, in particular to a preparation process for synthesizing D-phenylalanine by using high-activity biological enzymes. Background technique [0002] D-phenylalanine is an important unnatural amino acid, which has important application value in the fields of medicine and food. It is currently the most promising type II diabetes treatment drug in the world, NategliniDe, also known as Starlix. precursor, showing excellent application prospects. It is also an intermediate of drugs for treating tumors. In addition, D-phenylalanine is also used in health care products such as weight loss. At present, the production of D-phenylalanine mainly adopts the split process, which has small production scale, low yield and high cost, and is difficult to meet the needs of domestic and foreign markets. Contents of the invention [0003] The purpose of the present inve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/22C12R1/38
Inventor 崔传僧
Owner 崔传僧
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