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Method for producing L-aminothiopropionic acid by enzyme conversion

A cysteine, enzymatic conversion technology, applied in the field of Pseudomonas putida enzymatic conversion substrate to produce L-cysteine, can solve the problems of high energy consumption, low yield, environmental pollution, etc., to achieve Effects of improving stability and improving utilization efficiency

Inactive Publication Date: 2009-01-21
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the problems of low yield, high energy consumption and serious environmental pollution in the prior art, and provide a method for producing L-cysteine ​​by microbial enzymatic conversion of substrates

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1: Preparation of Pseudomonas putida TS1138 bacterial suspension

[0016] Pick a ring of bacteria lawn of Pseudomonas putida TS-1138 from a fresh slant and insert it into 30mL seed medium (glucose 30g / L, DL-ATC 3H 2 O5g / L, yeast powder 15g / L, peptone 10g / L, urea 3g / L, MnSO 4 ·H 2 O 1g / L, K 2 HPO 4 ·3H 2 O3g / L, MgSO 4 ·7H 2 O 0.5g / L, FeSO 4 ·7H 2 O 0.01g / L, NaCl 1.5g / L, pH 7.5, 0.1MPa 15min) in a 500mL Erlenmeyer flask, sealed with 8 layers of gauze, and cultured in a rotary shaker at 190r / min at 29°C for 14h, then inoculated with 10% The amount of seed culture was inserted into 30mL enzyme-producing medium (glucose 30g / L, DL-ATC 3H 2 O 4g / L, corn steep liquor 3g / L, urea 3g / L, MnSO 4 ·H 2 O 1g / L, K 2 HPO 4 ·3H 2 O 3.0g / L, MgSO 4 ·7H 2 O 0.5g / L, FeSO 4 ·7H 2 O 0.01g / L, NaCl3 g / L, pH 7.5, 0.1MPa 15min) in a 500mL Erlenmeyer flask, sealed with 8 layers of gauze, cultured in a rotary shaker at 170r / min, 29°C for 22h, and centrifuged for 10min (4...

Embodiment 2

[0017] Example 2: Cultivate Pseudomonas putida TS1138 in shake flasks and carry out enzymatic reaction at 42°C to produce L-cysteine

[0018] Add 0.6% DL-ATC solution (containing 0.6% DL-ATC, 0.6% K 2 HPO 4 , pH7.5 to 8.0) 20ml, then add 10ml of the TS-1138 cell suspension of the above-mentioned Example 1, and mix well. After 2.5 hours of reaction at a constant temperature of 42°C, the cysteine ​​content was determined to be 1.65 g / l by the acid ninhydrin method.

Embodiment 3

[0019] Example 3: Cultivate Pseudomonas putida TS1138 in shake flasks and perform enzymatic reaction at 38°C to produce L-cysteine

[0020] Add 0.6% DL-ATC solution (containing 0.6% DL-ATC, 0.6% K 2 HPO 4 , pH7.5 to 8.0) 20ml, then add 10ml of the TS-1138 cell suspension of the above-mentioned Example 1, and mix well. Keep the temperature at 38° C., and react for 3 hours. The content of cysteine ​​is determined to be 1.60 g / l by the acid ninhydrin method.

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Abstract

The invention discloses a technology for transformation production of L-cysteine by the microbial enzyme method. Pseudomonas putida TS-1138 is taken as a strain to be tested; and after amplification culture and zymogencous culture, the Pseudomonas putida TS-1138 is converted into DL-2-amino-Delta2thiazolines-4carboxylic acid (DL-ATC for short) by taking whole cells as an enzyme source, wherein the concentration of substrate solution is between 0.05 and 5.0 percent, the substrate solution is added between 0 and 9 hours, the reaction temperature is between 36 and 42 DEG C, and the reaction time is between 2 and 12 hours. The technology has the advantages of simple process, low cost of raw materials for production, low energy consumption and no pollution.

Description

【Technical field】 [0001] The invention belongs to the technical field of amino acid production, and relates to a method for Pseudomonas putida to produce L-cysteine ​​by enzymatically converting a substrate. 【Background technique】 [0002] L-cysteine ​​(L-cysteine) is an amino acid with active sulfur (sulfhydryl) among more than 20 amino acids that constitute proteins. It has been widely used in medicine, food additives and cosmetics. In medicine, cysteine ​​and its derivatives can be used in liver medicine and antidote, antipyretic and analgesic, ulcer treatment, fatigue recovery, infusion and comprehensive amino acid preparations, especially expectorant. In terms of food, cysteine ​​can be used as a bread fermentation aid. In cosmetics, it is mainly used in perm essence, sunscreen, hair nourishing essence, and hair tonic. Because in most microorganisms, mercapto groups come from sulfate ions, and sulfate ions can be reduced to certain sulfides, it is difficult to prepare...

Claims

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Application Information

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IPC IPC(8): C12P13/12C12R1/40
Inventor 陈宁怀丽华马雷徐庆阳刘淑云
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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