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Expression and characterization of a CALB-type lipase from Sporisorium reilianum SRZ2 and its potential in short-chain flavor ester synthesis

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Abstract

A lipase from Sporisorium reilianum SRZ2 (SRL) with 73% amino acid sequence identity to Candida antarctica lipase B (CALB) was cloned and overexpressed in Pichia pastoris. The recombinant SRL showed a preference for short-chain p-nitrophenyl esters. It achieved maximum activity at pH 8.0 and 65°C for p-nitrophenyl hexanoate (C6) with Km and kcat/Km values of 0.14 mmol·L−1 and 1712 min−1 · mmol · L−1 at 30°C, respectively. SRL displayed excellent thermostability and pH stability, retaining more than 79% of its initial activity after incubation at 60°C for 72 h and 75% at pH 3 to 11 for 72 h. It also maintained most of its activity in the presence of inhibitors and detergents except sodium dodecyl sulfate, and it tolerated organic solvents. SRL was covalently immobilized and successfully used for ethyl hexanoate synthesis in cyclohexane or in a solvent-free system with a high conversion yield (> 95%). Furthermore, high conversion yield was also achieved for the synthesis of various short-chain flavor esters when high substrate concentrations of 2 mol · L−1 were applied. This study indicated that a CALB-type lipase from S. reilianum SRZ2 showed great potential in organic ester synthesis.

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Authors and Affiliations

  1. The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou, 310014, China

    Jiang-Wei Shen, Xue Cai, Bao-Juan Dou, Feng-Yu Qi, Xiao-Jian Zhang, Zhi-Qiang Liu & Yu-Guo Zheng

  2. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, China

    Jiang-Wei Shen, Xue Cai, Bao-Juan Dou, Feng-Yu Qi, Xiao-Jian Zhang, Zhi-Qiang Liu & Yu-Guo Zheng

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  1. Jiang-Wei Shen
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Shen, JW., Cai, X., Dou, BJ. et al. Expression and characterization of a CALB-type lipase from Sporisorium reilianum SRZ2 and its potential in short-chain flavor ester synthesis. Front. Chem. Sci. Eng. 14, 868–879 (2020). https://doi.org/10.1007/s11705-019-1889-x

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