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Rational enhancement of enzyme-catalyzed enantioselective reaction by construction of recombinant enzymes based on additive strategy

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Abstract

A rational enhancement of kinetic resolution process for producing (S)-N-(2-ethyl-6-methylphenyl) alanine from racemic methyl ester using lipase B from Candida antarctica (CalB) was investigated. With the benefit results that lipase CalB-catalyzed reactions can be effectively regulated using amino acids (such as histidine and lysine) as additives, CalBs modified (mCalBs) by n-histidines at the N terminal and n-lysines at the C terminal were constructed and expressed. The results show that both soluble and precipitated mCalBs can effectively catalyze the hydrolysis reaction without adding any extra additives. The enantioselective ratio (E value) of soluble and precipitated mCalBs could be improved from 12.1 to 20.3, which were higher than that (E value was only 10.2) of commercial Novozym 435 (immobilized CalB). The study indicated that the amino acid-rich molecules introduced on lipase CalB can produce positive effects on enantioselectivity of enzyme. It provides unusual ideas for reasonable regulation of enzyme-catalyzed reactions.

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Acknowledgements

The authors are grateful for the financial support provided by the National Natural Science Foundation of China (Grant Nos. 21672081 and 21372098), Jilin Provincial Science and Technology Sustentation Program (Grant Nos. 20160204004SF and 20190201165JC), Jilin Province Development and Reform Commission (Grant No. 2016C047-1).

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Authors and Affiliations

  1. Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, College of Life Sciences, Jilin University, Changchun, 130012, People’s Republic of China

    Yu Han, Xiaoxue Zhou & Liangyu Zheng

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  1. Yu Han
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  2. Xiaoxue Zhou
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  3. Liangyu Zheng
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Correspondence to Liangyu Zheng.

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Han, Y., Zhou, X. & Zheng, L. Rational enhancement of enzyme-catalyzed enantioselective reaction by construction of recombinant enzymes based on additive strategy. Bioprocess Biosyst Eng 42, 1739–1746 (2019). https://doi.org/10.1007/s00449-019-02170-1

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  • DOI: https://doi.org/10.1007/s00449-019-02170-1

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