Abstract
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.
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Acknowledgments
This study was supported by grants from the Bio-industry Technology Development Program, Ministry for Food, Agriculture, Forestry and Fisheries (No. 112002-3) and the Korea Healthcare Technology R&D Project, Ministry for Health & Welfare, Republic of Korea (No. 2012-009).
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Seo, MJ., Shin, KC. & Oh, DK. 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 . Appl Microbiol Biotechnol 98, 7447–7456 (2014). https://doi.org/10.1007/s00253-014-5709-4
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DOI: https://doi.org/10.1007/s00253-014-5709-4