title: Purification and Immobilization of a Novel Enantioselective Lipase from Tsukamurella tyrosinosolvents for Efficient Resolution of Ethyl 2-(2-oxopyrrolidin-1-yl) Butyrate.
abstract: A highly enantioselective lipase from Tsukamurella tyrosinosolvents E105 was purified via ultrasonic extraction, precipitation, and chromatographic steps. The enzyme was purified about 38-fold with the recovery yield of 9 % and was confirmed as a dimer protein consisting of two identical subunits with a molecular mass of 24 kDa. The purified lipase was used to catalyze resolution of racemic ethyl 2-(2-oxopyrrolidin-1-yl) butyrate to (S)-2-(2-oxopyrrolidin-1-yl) butyric acid. The maximum activity of such lipase was obtained at pH 7.5, 35 °C, and the highest relative activity (156.80 %) was observed in the presence of 0.5 mM Co2+. Subsequently, the lipase was encapsulated within a mixture of 3 % sodium alginate and 0.8 % carrageenan, and then cross-linked with 0.6 % glutaraldehyde to enhance its biocatalytic capability and stability. Comparing with 36.9 % product yield and 97.5 % product ee of free lipase, the highest product yield of 46.3 % and ee of 98.5 % for immobilized lipase were achieved with the presence of 20 mM substrate. In addition, the reusability of immobilized lipase was also investigated, which could maintain 63.7 % of its initial conversion yield after seven repeated batch reactions. Thus, the evaluated enantioselective lipase in this work has a good potential for further industrial application.
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Huang
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, China.
authors.forename: Ren
authors.initials: R
authors.lastname: Yan
authors.affiliation: Zhejiang Pharmaceutical College, Ningbo, 315100, China.
authors.forename: Jun-Yao
authors.initials: JY
authors.lastname: He
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, China. wangpu@zjut.edu.cn.
authors.forename: Pu
authors.initials: P
authors.lastname: Wang
mesh_terms: D000192:Actinomycetales / Q000201:enzymology*; D000464:Alginates / Q000494:pharmacology; D055162:Biocatalysis / Q000187:drug effects; D002087:Butyrates / Q000737:chemistry* / Q000378:metabolism*; D002351:Carrageenan / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D004800:Enzymes, Immobilized / Q000378:metabolism*; D020723:Glucuronic Acid / Q000494:pharmacology; D005976:Glutaral / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008049:Lipase / Q000302:isolation & purification* / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D011759:Pyrrolidines / Q000737:chemistry* / Q000378:metabolism*; D013237:Stereoisomerism; D013696:Temperature
chemical_list: D000464:Alginates; D002087:Butyrates; D004800:Enzymes, Immobilized; D006603:Hexuronic Acids; D007477:Ions; D008670:Metals; D011759:Pyrrolidines; C582009:ethyl 2-(2-oxopyrrolidin-1-yl)butyrate; D020723:Glucuronic Acid; D002351:Carrageenan; D008049:Lipase; D005976:Glutaral
keywords: Characterization; Immobilization; Lipase; Purification; Tsukamurella tyrosinosolvents E105