Characterization and heterologous expression of a novel lysophospholipase gene from Antrodia cinnamomea.
OBJECTIVE
A novel lysophospholipase (LysoPL) from the basidiomycetous fungi Antrodia cinnamomea named ACLysoPL was cloned, heteroexpressed in Escherichia coli and characterized.


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 degrees C. ACLysoPL is a psychrophilic enzyme about 40% of whose maximum activity remained at 4 degrees C. The LysoPL activities with lysophospholipids as substrate were analysed by gas chromatography/mass spectrometry.


CONCLUSIONS
We have identified and characterized a gene named ACLysoPL encoding a protein performing LysoPL and esterase activities.


CONCLUSIONS
This is the first LysoPL of A. cinnamomea identified and characterized at the molecular level.
Hsu
K-H
KH
School of Forestry and Resource Conservation, Nation Taiwan University, Taipei, Taiwan.
Wang
S-Y
SY
Chu
F-H
FH
Shaw
J-F
JF
D000595:Amino Acid Sequence; D055491:Antrodia / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D008245:Lysophospholipase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D011994:Recombinant Proteins; D008245:Lysophospholipase