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Characterization and heterologous expression of a novel lysophospholipase gene from Antrodia cinnamomea
Author(s) -
Hsu K.H.,
Wang S.Y.,
Chu F.H.,
Shaw J.F.
Publication year - 2010
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2009.04569.x
Subject(s) - library science , political science , humanities , art , computer science
Aims: A novel lysophospholipase (LysoPL) from the basidiomycetous fungi Antrodia cinnamomea named ACLysoPL was cloned, heteroexpressed in Escherichia coli and characterized. Methods and Results: The gene encoding ACLysoPL was obtained from expressed sequence tags from A. cinnamomea . The full length of this gene has a 945 ‐bp open reading frame encoding 314 amino acids with a molecular weight of 35·5 kDa. ACLysoPL contains a lipase consensus sequence (GXSXG) motif and a Ser–His–Asp catalytic triad. A putative peroxisomal targeting signal type 1 was found in the C‐terminal. Heterologous expression of ACLysoPL in E. coli showed that the enzyme preferentially hydrolyses long‐chain acyl esterases at pH 7 and 30°C. ACLysoPL is a psychrophilic enzyme about 40% of whose maximum activity remained at 4°C. The LysoPL activities with lysophospholipids as substrate were analysed by gas chromatography/mass spectrometry. Conclusion: We have identified and characterized a gene named ACLysoPL encoding a protein performing LysoPL and esterase activities. Significance and Impact of the Study: This is the first LysoPL of A. cinnamomea identified and characterized at the molecular level.