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β-lactamases and their coding genes and their applications

A lactamase and gene technology, applied in the field of biotechnology, can solve problems such as carbapenem antibiotic pollution and carbapenem antibiotic metabolism, and achieve the effect of removing antibiotic pollution

Active Publication Date: 2021-07-23
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since the human body cannot completely metabolize carbapenem antibiotics, they will eventually be excreted into the environment, resulting in the pollution of carbapenem antibiotics in the environment
In addition, some carbapenem antibiotic fermentation factories also discharge some waste water containing this type of antibiotic, which leads to the pollution of carbapenem antibiotics in the environment
However, there is no effective removal method for the carbapenem antibiotics polluted in these environments

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] This example is used to illustrate the resistance of Escherichia coli transformed with the β-lactamase coding gene provided by the invention to β-lactam antibiotics

[0041] (1) Preparation of competent E.coli BL21 cells

[0042] 1) Streak culture competent Escherichia coli BL21 on LB solid medium, culture at 37°C for 12-16 hours.

[0043] 2) Pick fresh monoclonal colonies, inoculate them into 4ml LB liquid medium, culture with shaking at 37°C for 12-16 hours.

[0044] 3) On the third day, add 1ml of bacterial solution to 100ml of LB liquid medium at a ratio of 1:100, shake at 37°C and 220rpm for about 2 hours, and measure OD every half an hour 600 , when OD 600 When it reaches 0.5, the culture is stopped.

[0045] 4) After 30 minutes of ice bathing the bacterial solution, the bacterial solution was dispensed into 50 ml pre-cooled centrifuge tubes, and centrifuged at 4500 rpm for 10 minutes at 4°C.

[0046] 5) Discard the supernatant, add a small amount of pre-cooled ...

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PUM

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Abstract

The invention relates to the field of biotechnology, in particular to β-lactamases, their coding genes and their applications. The amino acid sequence of the β-lactamase shown is selected from the group consisting of SEQ ID No:6, SEQ ID No:7, SEQ ID No:8, SEQ ID No:9 and SEQ ID No:10. Also provided is a gene encoding β-lactamase, as well as recombinant vectors and transgenic cells containing the gene, and applications of β-lactamase, genes, recombinant vectors and transgenic cells in hydrolyzing β-lactam antibiotics, It also provides a composition containing β-lactamase and / or transgenic cells and a method for hydrolyzing β-lactam antibiotics. The β-lactamase can efficiently hydrolyze β-lactam antibiotics, especially carbapenem antibiotics, so that they can be applied to the environment polluted by corresponding antibiotics to achieve the purpose of removing the pollution of corresponding antibiotics.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a β-lactamase, and also to a gene encoding the β-lactamase, a recombinant vector and a cell containing the gene encoding the β-lactamase, and a β-lactamase Application of amidase, gene encoding the β-lactamase, recombinant vector containing the gene and cells in hydrolyzing β-lactam antibiotics, and a composition and method for hydrolyzing β-lactam antibiotics. Background technique [0002] Carbapenems are new β-lactam antibiotics. Compared with penicillin antibiotics in structure, carbapenem antibiotics have the biggest difference in that the sulfur atom on the thiazole ring is replaced by carbon, and there is an unsaturated diatom between C2 and C3 of carbapenem antibiotics. key. It is precisely because of structural differences that carbapenems exhibit a wider antibacterial spectrum and stronger antibacterial activity. This new type of β-lactam antibiotics mainly inhibi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/86C12N15/55A62D3/35
CPCA62D3/35A62D2101/28C12N9/86C12Y305/02006
Inventor 王超田井井冯婕张刚
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI