Characterization and heterologous expression of a novel lysophospholipase gene from Antrodia cinnamomea |
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| Authors: | K.‐H. Hsu S.‐Y. Wang F.‐H. Chu J.‐F. Shaw |
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| Affiliation: | 1. School of Forestry and Resource Conservation, Nation Taiwan University, Taipei, Taiwan;2. Department of Forestry, National Chung Hsing University, Taichung, Taiwan;3. Department of Forestry, National Chung Hsing University, Taichung, Taiwan;4. Department of Food Science and Biotechnology, National Chung Hsing University, Taichung, Taiwan |
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| Abstract: | 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. |
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| Keywords: | Antrodia cinnamomea esterase expression lysophospholipase |
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