Production of 8-hydroxy-9,12(Z,Z)-octadecadienoic acid from linoleic acid by recombinant cells expressing H1004A-C1006S variant of Aspergillus nidulans diol synthase
2015
Jeong, Yeon-Ju | Seo, Min-Ju | Shin, Kyung-Chul | Oh, Deok-Kun
An H1004A-C1006S variant of Aspergillus nidulans diol synthase was identified as an 8-dioxygenase. Whole recombinant Escherichia coli cells expressing the double-site variant showed the highest activity for converting linoleic acid to 8-hydroperoxy-9,12(Z,Z)-octadecadienoic acid (8-HPODE), which in turn was reduced to 8-hydroxy-9,12(Z,Z)-octadecadienoic acid (8-HODE) by the addition of a reducing agent. The optimization of 8-HPODE production was performed by response surface methodology, and the optimal conditions were pH 8.3, 27.2°C, 22.7% dimethyl sulfoxide, 27.0gl⁻¹ cells, and 16.5gl⁻¹ linoleic acid in a 100ml baffled flask containing a 10ml reaction mixture with agitation at 236rpm. The reduction of 8-HPODE to 8-HODE was highest when Tris(2-carboxyethyl)phosphine hydrochloride (TECP-HCl) was added at a final concentration of 25mM. Under the optimized reaction and reduction conditions, whole cells expressing H1004A-C1006S variant of A. nidulans diol synthase produced 5.0gl⁻¹ 8-HODE for 1h without the formation of di-hydroxy fatty acids. This is the first report of the biotechnological production of 8-HODE.
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