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Construction and Application of Cell Lysis Systems in the Expression of Mycotoxin Degrading Enzyme in Escherichia coli |
Cheng-cheng ZHAO,Chang-po SUN,Xiao-jiao CHANG,Song-ling WU,Zhen-quan LIN() |
Academy of National Food and Strategic Reserves Administration, Institute of Cereal Processing Science and Technology, Beijing 100037,China |
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Abstract Objective: Extracellular production of recombinant proteins in Escherichia coli is limited by the inefficiency of inherent secretion system. An inducible cell lysis system was designed and constructed to enhance secretion of intracellular recombinant protein in E.coli.Methods: Considering colicin E7 could promote cell lysis, E.coli cell lysis systems were constructed by co-expressing target protein and colicin E7 lysis to release the recombinant proteins to culture medium.Results: A reporter protein (red fluorescent protein, RFP), as a recombinant protein, was co-expressed with E7 in E.coli to evaluate the cell lysis systems. Expression of recombinant protein was controlled by T7 promoter. While E7 cassette was controlled by two promoters (T7 promoter and araBAD promoter) which determined the expression timing of E7 and therefore determined the timing of cell lysis. Compared to one-step induction by IPTG, the two-step induction by IPTG and L-arabinose was better for the production and secretion of recombinant proteins. The two-step inducible lysis system was also used to express zearalenone (ZEN) degrading enzyme, and high enzyme activity was detectable in the culture supernatant samples. The secreted enzyme could degrade about 5.8μg ZEN in 30minutes at 37℃.Conclusion: The colicin E7 assisted two-step inducible cell lysis system could be potential for expression of recombinant proteins and their secretion to extracellular in E. coli.
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Received: 21 August 2018
Published: 08 May 2019
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Corresponding Authors:
Zhen-quan LIN
E-mail: zhenquanlin09@gmail.com
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