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Identification and Functional Expression of a Δ9-Fatty Acid Desaturase from Psychrobacter urativorans in Escherichia coli

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Lipids

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.

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Abbreviations

IPTG:

Isopropyl-β-d-thiogalacto-pyranoside

MOPS:

3-Morpholinopropanesulfonic acid

PMSF:

Phenylmethylsulfonyl fluoride

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Acknowledgments

This work is part of the joint project between Nanjing University of Technology and the University of Stuttgart on Biosystems Engineering of Desaturation, which is in the framework of a Baden-Wuerttemberg—Shanghai/Jiangsu Exchange Program on Systems Biology and Biosystems Engineering.

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Correspondence to Vlada B. Urlacher.

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Li, Y., Dietrich, M., Schmid, R.D. et al. Identification and Functional Expression of a Δ9-Fatty Acid Desaturase from Psychrobacter urativorans in Escherichia coli . Lipids 43, 207–213 (2008). https://doi.org/10.1007/s11745-007-3150-5

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  • DOI: https://doi.org/10.1007/s11745-007-3150-5

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