Functional characterization of a novel aspartic acid rich lipase, TALipC, from Trichosporon asahii MSR54: solvent-dependent enantio inversion during esterification of 1-phenylethanol

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• 作者：Arti Kumari ; Rani Gupta
• 关键词：Enantioselectivity ; Lipase ; 1 ; Phenylethanol ; Regioselectivity Trichosporon asahii
• 刊名：Biotechnology Letters
• 出版年：2015
• 出版时间：January 2015
• 年：2015
• 卷：37
• 期：1
• 页码：121-130
• 全文大小：1,095 KB
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  3. Gupta N, Rathi P, Gupta R (2002) Simplified / para- nitrophenyl palmitate assay for lipases and esterases. Anal Biochem 31:98-9 CrossRef
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• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

A novel lipase gene TAlipC was isolated from Trichosporon asahii MSR54 and functionally expressed in Pichia pastoris. The protein was His-tagged and purified to homogeneity by affinity chromatography. Sequence comparison revealed a high homology with lipases from Cryptococcus sp. It had a GX type oxyanion hole and a GHSLG-type conserved penta-peptide similar to those in the lipases from Yarrowia lipolytica. The enzyme had optimal activity at pH 8 and 50?°C. It was specific for long chain fatty acid groups of p-nitrophenol esters and triacylglycerols, showing regio- and enantio-selectivity. It was activated by Mg2+ ions (20?mM) and had a predicted Mg-binding domain at the aspartic acid-rich C-terminal. Solvent-based enantio- inversion was the key feature of the enzyme where it showed (S)-selectivity in 1,4-dioxane and 2-propanol and (R)-selectivity in hexane during chiral separation of (±)1-phenylethanol by esterification.