Identification of delta5-desaturase from Mortierella alpina by heterologous expression in bakers' yeast and canola

A DNA fragment with homology to Delta6-desaturases from borage and cyanobacteria was isolated after polymerase chain reaction amplification of Mortierella alpina cDNA with oligonucleotide primers corresponding to the conserved regions of known Delta6-desaturase genes. This fragment was used as a pro...

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Bibliographic Details
Published in:The Journal of biological chemistry 1998-11, Vol.273 (45), p.29360-29366
Main Authors: Knutzon, D.S. (Abbott Laboratories, Columbus, OH.), Thurmond, J.M, Huang, Y.S, Chaudhary, S, Bobik, E.G. Jr, Chan, G.M, Kirchner, S.J, Mukerji, P
Format: Article
Language:English
Subjects:
ACIDE ARACHIDONIQUE
ACIDE GRAS
ACIDE LINOLEIQUE
ACIDE OLEIQUE
ACIDO ARAQUIDONICO
ACIDO LINOLEICO
ACIDO OLEICO
ACIDOS GRASOS
ACTIVIDAD ENZIMATICA
ACTIVITE ENZYMATIQUE
ADN
Amino Acid Sequence
AMINO ACID SEQUENCES
ARACHIDONIC ACID
Base Sequence
BRASSICA NAPUS
CHEMICAL COMPOSITION
Cloning, Molecular
COMPLEMENTARY DNA
COMPOSICION QUIMICA
COMPOSITION CHIMIQUE
DIHOMO-GAMMA-LINOLENIC ACID
DNA
DNA Primers
DNA, Complementary
ENZYMIC ACTIVITY
EXPRESION GENICA
EXPRESSION DES GENES
Fatty Acid Desaturases - genetics
Fatty Acid Desaturases - metabolism
FATTY ACIDS
Gas Chromatography-Mass Spectrometry
GENBANK/AF067654
GENE EXPRESSION
GENE TRANSFER
GENETIC TRANSFORMATION
GRAINE
LINOLEIC ACID
LONG CHAIN FATTY ACIDS
MOLECULAR SEQUENCE DATA
MORTIERELLA
Mortierella - enzymology
NUCLEOTIDE SEQUENCE
OLEIC ACID
OPEN READING FRAMES
OXIDOREDUCTASES
OXIDORREDUCTASAS
OXYDOREDUCTASE
OXYGENASES
PLANTAS TRANSGENICAS
PLANTE TRANSGENIQUE
Plants, Genetically Modified
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
SACCHAROMYCES CEREVISIAE
Saccharomyces cerevisiae - genetics
SECUENCIA NUCLEOTIDICA
SEEDS
Seeds - genetics
SEMILLA
Sequence Homology, Amino Acid
SEQUENCE NUCLEOTIDIQUE
TRANSFERENCIA DE GENES
TRANSFERT DE GENE
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
TRANSGENIC PLANTS
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Description
Summary:A DNA fragment with homology to Delta6-desaturases from borage and cyanobacteria was isolated after polymerase chain reaction amplification of Mortierella alpina cDNA with oligonucleotide primers corresponding to the conserved regions of known Delta6-desaturase genes. This fragment was used as a probe to isolate a cDNA clone with an open reading frame encoding 446 amino acids from a M. alpina library. Expression of this open reading frame from an inducible promoter in Saccharomyces cerevisiae in the presence of various substrates revealed that the recombinant product had Delta5-desaturase activity. The effects of growth and induction conditions as well as host strain on activity of the recombinant Delta5-desaturase in S. cerevisiae were evaluated. Expression of the M. alpina Delta5-desaturase cDNA in transgenic canola seeds resulted in the production of taxoleic acid (Delta5,9-18:2) and pinolenic acid (Delta5,9,12-18:3), which are the Delta5-desaturation products of oleic and linoleic acids, respectively.
ISSN:0021-9258
1083-351X