High expression and activity of an L-methionine γ-lyase gene in Escherichia coli

HU Hai-Yan1 DU Shao-Ping1 XIA Feng-Geng1 HUANG Kui-Ying1 ZHOU Shi-Ning2

(1.Guangzhou Institute of Microbiology, Guangzhou, Guangdong, China 510663)
(2.State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, Guangdong, China 510275)

【Abstract】[Background] In order to develop new microbial resources in the ocean, we constructed a deep-sea metagenomic library by adopting the culture-independent metagenomic technology, and carried out studies on the important genes. [Objective] To identify and highly express the methionine γ-lyase gene in Escherichia coli from the DNA library of deep-sea sediments. [Methods] The gene mgl was overexpressed by pET-28a(+) system in E. coli BL21(DE3), which was induced by isopropyl β-D-1-thiogalactopyranoside, and the expression conditions were optimized to obtain the high level of recombinant methionine lyase (rMGL). The recombinant protein was purified by affinity chromatography and the enzyme activity was determined. [Results] The product rMGL had the molecular weight consistent with the predicted 46 kD, with high L-methionine lyase activity. rMGL could use L-methionine or DL-homocysteine as substrate, while had little activity for L-cysteine or L-cystine. Its relative activity for DL-homocysteine was 1.4 times of that for L-methionine. [Conclusion] mgl gene from the deep sea metagenomic library can efficiently express rMGL using pET-28 a(+)/BL21(DE3).

【Keywords】 Metagenomic library; Idiomarina; L-methionine γ-lyase; Induced expression; Purification of recombinant protein;


【Funds】 Special Scientific Research Project of Guangzhou Science and Technology Plan (201607010326)

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(Translated by CHEN T)


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This Article


CN: 11-1996/Q

Vol 46, No. 12, Pages 3225-3232

December 2019


Article Outline


  • 1 Materials and methods
  • 2 Results
  • 3 Discussion
  • 4 Conclusion
  • References