delta(6)-desaturase of Mucor rouxii with high similarity to plant delta(6)-desaturase and its heterologous expression in Saccharomyces cerevisiae

Gamma-linolenic acid (GLA, gamma-C18:3) is an essential fatty acid that plays a vital role in biological structures and cellular functions. Based on available sequence information and using polymerase chain reaction (PCR) technique, we cloned from the fungus Mucor rouxii the entire coding sequence o...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2000-12, Vol.279 (1), p.17-22
Main Authors: Laoteng, K, Mannontarat, R, Tanticharoen, M, Cheevadhanarak, S
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
Cloning, Molecular
desaturase
DNA Primers
DNA, Complementary
Fatty Acid Desaturases - chemistry
Fatty Acid Desaturases - genetics
Fatty Acid Desaturases - metabolism
Linoleoyl-CoA Desaturase
Molecular Sequence Data
Mucor - enzymology
Mucor rouxii
Phylogeny
Plants - enzymology
Saccharomyces cerevisiae
Saccharomyces cerevisiae - enzymology
Sequence Homology, Amino Acid
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Summary:Gamma-linolenic acid (GLA, gamma-C18:3) is an essential fatty acid that plays a vital role in biological structures and cellular functions. Based on available sequence information and using polymerase chain reaction (PCR) technique, we cloned from the fungus Mucor rouxii the entire coding sequence of a delta(6)-desaturase enzyme, which is responsible for the transformation of linoleic acid into GLA. The deduced amino acid sequence of M. rouxii gene showed the highest homology with the plant delta(6)-desaturase. It comprises the characteristics of membrane-bound desaturases, including histidine-rich motifs and hydrophobic regions. A cytochrome b(5)-like domain was observed at the N-terminus. In addition to three conserved histidine-rich motifs, we found an additional histidine-rich motif, HKHHSH, downstream of the cytochrome b(5)-like domain, which is not present in previously cloned delta(6)-desaturase genes. Heterologous expression of the M. rouxii cDNA in Saccharomyces cerevisiae resulted in the synthesis and accumulation of GLA.
ISSN:0006-291X
DOI:10.1006/bbrc.2000.3856