Production of 5,8-dihydroxy-9,12(Z,Z)-octadecadienoic acid from linoleic acid by whole recombinant Escherichia coli cells expressing diol synthase from Aspergillus nidulans.
Diol synthase from Aspergillus nidulans was cloned and expressed in Escherichia coli. Recombinant E. coli cells expressing diol synthase from A. nidulans converted linoleic acid to a product that was identified as 5,8-dihydroxy-9,12(Z,Z)-octadecadienoic acid by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The recombinant cells and the purified enzyme showed the highest activity for linoleic acid among the fatty acids tested. The optimal reaction conditions for the production of 5,8-dihydroxy-9,12(Z,Z)-octadecadienoic acid from linoleic acid using whole recombinant E. coli cells expressing diol synthase were pH 7.5, 35°C, 250 rpm, 5 g l(-1) linoleic acid, 23 g l(-1) cells, and 20% (v/v) dimethyl sulfoxide in a 250-ml baffled flask. Under these optimized conditions, whole recombinant cells expressing diol synthase produced 4.98 g l(-1) 5,8-dihydroxy-9,12(Z,Z)-octadecadienoic acid for 150 min without detectable byproducts, with a conversion yield of 99% (w/w) and a productivity of 2.5 g l(-1) h(-1). This is the first report on the biotechnological production of dihydroxy fatty acid using whole recombinant cells expressing diol synthase.
Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, South Korea.
Min-Ju
MJ
Seo
Kyung-Chul
KC
Shin
Deok-Kun
DK
Oh
D001233:Aspergillus nidulans / Q000201:enzymology* / Q000235:genetics; D001711:Biotransformation; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D064429:Fatty Acid Synthases / Q000235:genetics / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D019787:Linoleic Acid / Q000378:metabolism*; D013058:Mass Spectrometry; D060847:Metabolic Engineering; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature
D005231:Fatty Acids, Unsaturated; D011994:Recombinant Proteins; C027369:octadecadienoic acid; D019787:Linoleic Acid; D064429:Fatty Acid Synthases