Construction and application of cysteine single-cell biosensor

A biosensor, cysteine ​​technology, applied in microorganism-based methods, microbial determination/inspection, microorganisms, etc., can solve the problem of lack of rapid, sensitive and highly specific detection, achieve real-time detection, enhance downstream genes Expression, strong specificity
CN110283764AInactive Publication Date: 2019-09-27TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
Publication Date
2019-09-27
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a preparation method of a biosensor for detecting L-cysteine and an application of the biosensor in detecting yield of escherichia coli L-cysteine. Main components of the biosensor comprise a transcriptional control gene CcdR coding gene from pantoea ananatis, a promoter region and a terminator region of the transcriptional control gene CcdR coding gene, a promoter region of a gene ccdA from pantoea ananatis, a green fluorescent protein coding gene eGFP and a plasmid framework pTrc-Mob. The biosensor can have a good linear relation under the L-cysteine concentration of 0-32 mmol L<-1> and has the highest fluorescence intensity value and cysteine concentration relation under the L-cysteine concentration of 0.01-8 mmol L<-1>. Through combination with a high-throughput screening system, the biosensor performs real-time detection on the yield of escherichia coli L-cysteine, can screen out high-yield mutants of L-cysteine and mutants for key enzymes in biological synthesis of L-cysteine. Specific, quantitative and real-time detection of L-cysteine can be realized, and the sensitivity is high.
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Description

technical field

[0001] The invention belongs to the technical field of biosensors, and relates to the construction and application of an L-cysteine ​​biosensor. Background technique

[0002] L-cysteine ​​is an important sulfur-containing amino acid, which is widely used in food, medicine and cosmetic industries. Because it contains a highly reducing sulfhydryl group, L-cysteine ​​can be used as a natural reducing agent to break disulfide bonds in proteins. For example, the expectorant liquid with its derivative acetyl L-cysteine ​​as the main component can break the disulfide bond in the sputum to reduce the viscosity of sputum; another example, the perm liquid with L-cysteine ​​as the main component can Break the disulfide bond in hair, so it is widely used in the hairdressing industry; another example, L-cysteine ​​and its derivative glutathione can break the disulfide bond in tyrosinase to inactivate it, It can reduce the melanin produced by tyrosinase, so it is used in...

Claims

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