The desaturase from Bacillus subtilis, a promising tool for the selective olefination of phospholipids

• Published in: Journal of biotechnology Vol. 121; no. 1; pp. 49 - 53
• Main Authors: Bonamore, Alessandra, Macone, Alberto, Colotti, Gianni, Matarese, Rosa Marina, Boffi, Alberto
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 02.01.2006; Amsterdam Elsevier; New York, NY
• Subjects: Bacillus subtilis; Bacillus subtilis - enzymology; Bacillus subtilis - genetics; Bacterial desaturases; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Biological and medical sciences; Biotechnology; Cyclopropane fatty acid synthase; dienoic fatty acids; enzyme activity; Escherichia coli; Escherichia coli - enzymology; Escherichia coli - genetics; Fatty Acid Desaturases - chemistry; Fatty Acid Desaturases - genetics; fatty acid esters; Fatty acid methyl esters; Fundamental and applied biological sciences. Psychology; Gas chromatography–mass spectrometry; monounsaturated fatty acids; Oxidation-Reduction; oxygenases; phospholipids; Phospholipids - chemistry; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Unsaturated fatty acids; Unsaturated fatty acids; Bacterial desaturases; Cyclopropane fatty acid synthase; Bacillus subtilis; Fatty acid methyl esters; Gas chromatography–mass spectrometry; Acyltransferases; Enzyme; Transferases; Phospholipid; Selectivity; Gas chromatography-mass spectrometry; Bacillaceae; Fatty-acid synthase; Gas chromatography; Bacillales; Cyclopropane; Bacteria; Unsaturated fatty acid; Fatty acid methyl ester; Mass spectrometry
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2005.07.008

The Δ5-desaturase from Bacillus subtilis has been cloned in Escherichia coli BL21 cells and its enzyme activity has been investigated as a function of temperature and oxygenation by analyzing methyl ester adducts from the total lipid extract in GC–MS measurements. The present data bring out that the activity of recombinant Δ5-desaturase, at 20–22 °C and 20% oxygen, is surprisingly high yielding 22% of C16:1,Δ5 (5- cis-palmitoleic acid) and 13% C18:2,Δ5Δ11 (efedrenic acid). Lower amounts of other mono- and doubly-Δ5-unsaturated fatty acids were also detected. These findings demonstrate that Δ5-desaturase can accept a multiplicity of substrates and is endowed with an unprecedented activity among other acyl-lipid desaturases thus representing a unique tool for the production of rare Δ5 unsaturated fatty acid derivatives.