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Efficient production of methionine from 2-amino-4-methylthiobutanenitrile by recombinant Escherichia coli harboring nitrilase

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Methionine as an essential amino acid has been attracting more attention for its important applications in food and feed additives. In this study, for efficient production of methionine from 2-amino-4-methylthiobutanenitrile, a codon-optimized nitrilase gene was newly synthesized and expressed, and the catalytic conditions for methionine production were studied. The optimal temperature and pH for methionine synthesis were 40 °C and 7.5, respectively. The recombinant nitrilase was thermo-stable with half-life of 5.52 h at 40 °C. The substrate loading was optimized in given amount of catalyst and fixed substrate/catalyst ratio mode to achieve higher productivity. Methionine was produced in 100 % conversion within 120 min with a substrate loading of 300 mM. The production of methionine with the immobilized resting cells in packed-bed reactor was investigated. The immobilized nitrilase exhibited good operation stability and retained over 80 % of the initial activity after operating for 100 h. After separation, the purity and the total yield of methionine reached 99.1 and 97 %, respectively. This recombinant nitrilase could be a potential candidate for application in production of methionine.

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Acknowledgments

The authors gratefully acknowledge the National Natural Science Foundation of China (No. 21202150), the Research Program of Science and Technology Department of Zhejiang Province (No. 2011R09043-07) and Natural Science Foundation of Zhejiang Province (No. R3110155).

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Authors and Affiliations

  1. Institute of Bioengineering, Zhejiang, University of Technology, Hangzhou, 310014, People’s Republic of China

    Li-Qun Jin, Zong-Tong Li, Zhi-Qiang Liu, Yu-Guo Zheng & Yin-Chu Shen

  2. Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, People’s Republic of China

    Li-Qun Jin, Zong-Tong Li, Zhi-Qiang Liu, Yu-Guo Zheng & Yin-Chu Shen

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  1. Li-Qun Jin
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  2. Zong-Tong Li
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  3. Zhi-Qiang Liu
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Correspondence to Yu-Guo Zheng.

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Jin, LQ., Li, ZT., Liu, ZQ. et al. Efficient production of methionine from 2-amino-4-methylthiobutanenitrile by recombinant Escherichia coli harboring nitrilase. J Ind Microbiol Biotechnol 41, 1479–1486 (2014). https://doi.org/10.1007/s10295-014-1490-8

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  • DOI: https://doi.org/10.1007/s10295-014-1490-8

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