Molecular cloning and functional characterization of a Δ6-fatty acid desaturase gene from Rhizopus oryzae

The objective was to screen for and isolate a novel enzyme with the specific activity of a Δ6‐fatty acid desaturase from Rhizopus oryzae. In this study, R. oryzae was identified as a novel fungal species that produces large amounts of γ‐linolenic acid. A full‐length cDNA, designated here as RoD6D, w...

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Bibliographic Details
Published in:Journal of basic microbiology 2013-09, Vol.53 (9), p.773-777
Main Authors: Zhu, Yu, Zhang, Bi-Bo
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Cloning
Cloning, Molecular
Cluster Analysis
gamma-Linolenic Acid - metabolism
Gene Expression
Gene Library
Introns
Linoleoyl-CoA Desaturase - genetics
Linoleoyl-CoA Desaturase - metabolism
Membrane Proteins - genetics
Membrane Proteins - metabolism
Molecular Sequence Data
Open Reading Frames
Phylogeny
Protein Structure, Tertiary
Rhizopus - enzymology
Rhizopus - genetics
Rhizopus oryzae
Saccharomyces cerevisiae - genetics
Sequence Analysis, DNA
Sequence Homology
γ-Linolenic acid
Δ6-Fatty acid desaturase gene
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Summary:The objective was to screen for and isolate a novel enzyme with the specific activity of a Δ6‐fatty acid desaturase from Rhizopus oryzae. In this study, R. oryzae was identified as a novel fungal species that produces large amounts of γ‐linolenic acid. A full‐length cDNA, designated here as RoD6D, with high homology to fungal Δ6‐fatty acid desaturase genes was isolated from R. oryzae by using the rapid amplification of cDNA ends method. It had an open reading frame of 1176 bp encoding a deduced polypeptide of 391 amino acids. Bioinformatics analysis characterized the putative RoD6D protein as a typical membrane‐bound desaturase, including three conserved histidine‐rich motifs, a hydropathy profile, and a cytochrome b5‐like domain in the N terminus. When the coding sequence was expressed in the Saccharomyces cerevisiae strain INVScl, the encoded product of RoD6D exhibited Δ6‐fatty acid desaturase activity that led to the accumulation of γ‐linolenic acid. The corresponding genomic sequence of RoD6D was 1565 bp in length, with five introns. This is the first report on the characterization and gene cloning of a Δ6‐fatty acid desaturase of R. oryzae from Douchi.
ISSN:0233-111X
1521-4028
DOI:10.1002/jobm.201200189