Identification of mutation sites on delta6 desaturase genes from Mortierella alpina 1S-4 mutants

Three Delta6 desaturase-defective mutants, designated YB214, HR95, and ST66, were newly isolated from Mortierella alpina 1S-4, after treating wild-type spores with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). These three mutants and Mut49, isolated previously, are capable of accumulating 5,11,1...

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Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2005-05, Vol.69 (5), p.1021-1024
Main Authors: Abe, T. (Kyoto Univ. (Japan)), Sakuradani, E, Asano, T, Kanamaru, H, Ioka, Y, Shimizu, S
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Arachidonic Acids - biosynthesis
Base Sequence
DNA
Fatty Acid Desaturases - chemistry
Fatty Acid Desaturases - genetics
ISOLATION
MOLECULAR CLONING
Molecular Sequence Data
MORTIERELLA
Mortierella - enzymology
Mortierella - genetics
MUTAGENS
MUTANTS
Mutation
NITROSOGUANIDINE
NUCLEOTIDE SEQUENCE
PCR
UNSATURATED FATTY ACIDS
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Summary:Three Delta6 desaturase-defective mutants, designated YB214, HR95, and ST66, were newly isolated from Mortierella alpina 1S-4, after treating wild-type spores with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). These three mutants and Mut49, isolated previously, are capable of accumulating 5,11,14-cis-eicosatrienoic acid (20:3Delta5). Two functional Delta6 desaturases (Delta6I and Delta6II) were found to exist in M. alpina 1S-4. The mutation sites on the Delta6I gene in the Delta6 desaturase-defective mutants were identified. The mutations each resulted in an amino acid replacement (W314Stop, T375K, and G390D) in Delta6I from ST66, HR95, and YB214 respectively, and uncorrected transcription of the Delta6I gene in Mut49 was caused by disappearance of the GT-terminal of the second intron, resulting in low Delta6 desaturation activity in these mutants. On the other hand, there was no mutation site on the Delta6II genes of the mutants.
ISSN:0916-8451
1347-6947