Purification and characterization of the extracellular lipase Lip2 from Yarrowia lipolytica

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• 作者：Mingrui Yu ; Shaowei Qin ; Tianwei Tan
• 关键词：Lipase ; Yarrowia lipolytica ; MALDI-TOF mass spectrometry ; Purification ; Glycosylation
• 刊名：Process Biochemistry
• 出版年：2007
• 出版时间：March 2007
• 年：2007
• 卷：42
• 期：3
• 页码：384-391
• 全文大小：340 K

文摘

An extracellular lipase from Yarrowia lipolytica (YlLip2) has been purified by ion exchange chromatography on Q sepharose FF, followed by hydrophobic interaction chromatography on butyl sepharose FF. SDS-PAGE showed that the molecular weight of this lipase is about 38 kDa. N-terminal amino acid sequencing and MALDI-TOF mass spectral analysis showed that this lipase is encoded by gene LIP2 (GenBank accession no. AJ012632). Enzymatic deglycosylation showed that this lipase is a glycosylated protein which contains about 12 % sugar. The corresponding deglycosylated lipase remained 88 % specific activity of untreated lipase. There was a high amino acid sequence identity (91 % ) between YlLip2 and Candida deformans lipase CdLip1 (GenBank accession no. AJ428393). The optima temperature and pH for the purified lipase was 40 °C and 8.0, respectively. The lipase showed a preference for long chain fatty acid methyl esters (C12–C16), with the highest activity toward methyl myristate (C14). Lipase activity was stimulated by Ca²⁺ and Mg²⁺ and inhibited by Zn²⁺, Ni²⁺ and Cu²⁺, whereas EDTA had no effect on its activity. A 0.1 % of Tween 80 and Span 65 increased slightly the enzyme activity and SDS inhibited it.