Identification and functional expression of a Δ9-fatty acid desaturase from Psychrobacter urativorans in Escherichia coli

Yan Li; Matthias Dietrich; Rolf D. Schmid; Bingfang He; Pingkai Ouyang; Vlada B. Urlacher

doi:10.1007/s11745-007-3150-5

Identification and functional expression of a Δ9-fatty acid desaturase from Psychrobacter urativorans in Escherichia coli

Yan Li, Matthias Dietrich, Rolf D. Schmid, Bingfang He, Pingkai Ouyang, Vlada B. Urlacher

School of Pharmaceutical Sciences

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

The Δ9-fatty acid desaturase is a key enzyme in the synthesis of unsaturated fatty acids. The fatty acid composition of membrane phospholipids in Psychrobacter urativorans is characterized by a high degree of desaturation at Δ9 position. Based on CODEHOP-mediated PCR strategy, a novel gene designated as PuFAD9, putatively encoding a Δ9-fatty acid desaturase (PuFAD9), was isolated from P. urativorans. The gene consists of 1,455 bp and codes for 484 amino acids. Analysis of the amino acid sequence reveals three histidine clusters and a hydropathy profile, typical for membrane-bound desaturases. Activity of the PuFAD9 protein, recombinantly expressed in Escherichia coli was confirmed by GC-MS analysis of the cellular fatty acid composition. It was found that the ratio between palmitoleic and palmitic acid in E. coli cells heterologously expressing the PuFAD9 gene was significantly affected by IPTG induction and the growth temperature.

Original language	English
Pages (from-to)	207-213
Number of pages	7
Journal	Lipids
Volume	43
Issue number	3
DOIs	https://doi.org/10.1007/s11745-007-3150-5
State	Published - Mar 2008

Keywords

Degenerate primer
Fatty acid composition
Gene expression
Δ9-fatty acid desaturase

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title = "Identification and functional expression of a Δ9-fatty acid desaturase from Psychrobacter urativorans in Escherichia coli",

abstract = "The Δ9-fatty acid desaturase is a key enzyme in the synthesis of unsaturated fatty acids. The fatty acid composition of membrane phospholipids in Psychrobacter urativorans is characterized by a high degree of desaturation at Δ9 position. Based on CODEHOP-mediated PCR strategy, a novel gene designated as PuFAD9, putatively encoding a Δ9-fatty acid desaturase (PuFAD9), was isolated from P. urativorans. The gene consists of 1,455 bp and codes for 484 amino acids. Analysis of the amino acid sequence reveals three histidine clusters and a hydropathy profile, typical for membrane-bound desaturases. Activity of the PuFAD9 protein, recombinantly expressed in Escherichia coli was confirmed by GC-MS analysis of the cellular fatty acid composition. It was found that the ratio between palmitoleic and palmitic acid in E. coli cells heterologously expressing the PuFAD9 gene was significantly affected by IPTG induction and the growth temperature.",

keywords = "Degenerate primer, Fatty acid composition, Gene expression, Δ9-fatty acid desaturase",

author = "Yan Li and Matthias Dietrich and Schmid, {Rolf D.} and Bingfang He and Pingkai Ouyang and Urlacher, {Vlada B.}",

year = "2008",

month = mar,

doi = "10.1007/s11745-007-3150-5",

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T1 - Identification and functional expression of a Δ9-fatty acid desaturase from Psychrobacter urativorans in Escherichia coli

AU - Li, Yan

AU - Dietrich, Matthias

AU - Schmid, Rolf D.

AU - He, Bingfang

AU - Ouyang, Pingkai

AU - Urlacher, Vlada B.

PY - 2008/3

Y1 - 2008/3

N2 - The Δ9-fatty acid desaturase is a key enzyme in the synthesis of unsaturated fatty acids. The fatty acid composition of membrane phospholipids in Psychrobacter urativorans is characterized by a high degree of desaturation at Δ9 position. Based on CODEHOP-mediated PCR strategy, a novel gene designated as PuFAD9, putatively encoding a Δ9-fatty acid desaturase (PuFAD9), was isolated from P. urativorans. The gene consists of 1,455 bp and codes for 484 amino acids. Analysis of the amino acid sequence reveals three histidine clusters and a hydropathy profile, typical for membrane-bound desaturases. Activity of the PuFAD9 protein, recombinantly expressed in Escherichia coli was confirmed by GC-MS analysis of the cellular fatty acid composition. It was found that the ratio between palmitoleic and palmitic acid in E. coli cells heterologously expressing the PuFAD9 gene was significantly affected by IPTG induction and the growth temperature.

AB - The Δ9-fatty acid desaturase is a key enzyme in the synthesis of unsaturated fatty acids. The fatty acid composition of membrane phospholipids in Psychrobacter urativorans is characterized by a high degree of desaturation at Δ9 position. Based on CODEHOP-mediated PCR strategy, a novel gene designated as PuFAD9, putatively encoding a Δ9-fatty acid desaturase (PuFAD9), was isolated from P. urativorans. The gene consists of 1,455 bp and codes for 484 amino acids. Analysis of the amino acid sequence reveals three histidine clusters and a hydropathy profile, typical for membrane-bound desaturases. Activity of the PuFAD9 protein, recombinantly expressed in Escherichia coli was confirmed by GC-MS analysis of the cellular fatty acid composition. It was found that the ratio between palmitoleic and palmitic acid in E. coli cells heterologously expressing the PuFAD9 gene was significantly affected by IPTG induction and the growth temperature.

KW - Degenerate primer

KW - Fatty acid composition

KW - Gene expression

KW - Δ9-fatty acid desaturase

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DO - 10.1007/s11745-007-3150-5

M3 - 文章

C2 - 18256868

AN - SCOPUS:41149128662

SN - 0024-4201

VL - 43

SP - 207

EP - 213

JO - Lipids

JF - Lipids

IS - 3

ER -

Identification and functional expression of a Δ9-fatty acid desaturase from Psychrobacter urativorans in Escherichia coli

Abstract

Keywords

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