5,8‐Dihydroxy‐9,12,15(Z,Z,Z)‐Octadecatrienoic Acid Production by Recombinant Cells Expressing Aspergillus nidulans Diol Synthase

Abstract

Whole cells of recombinant Escherichia coli expressing diol synthase from Aspergillus nidulans produced 5,8‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid from α‐linolenic acid via 8‐hydroperoxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid as an intermediate. The optimal conditions for 5,8‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid production using whole recombinant cells were exhibited at pH 7.0, 40 °C, and 250 rpm with 40 g/L cells, 12 g/L, α‐linolenic acid, and 5 % (v/v) dimethyl sulfoxide in a 250‐mL baffled flask containing 50 mL reaction solution. Under these conditions, whole recombinant cells produced 9.1 g/L 5,8‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid for 100 min, with a conversion yield of 75 % (w/w), a volumetric productivity of 5.5 g/L/h, and specific productivity of 137 mg/g‐cells/h. As an intermediate, 8‐hydroperoxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid was observed at approximately 1.4 g/L after 100 min. With regard to dihydroxy fatty acid production, this is the highest reported volumetric and specific productivities thus far. This is the first report on the biotechnological production of 5,8‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid.

Production of 8,11‐dihydroxy and 8‐hydroxy unsaturated fatty acids from unsaturated fatty acids by recombinant Escherichia coli expressing 8,11‐linoleate diol synthase from Penicillium chrysogenum

“…The volumetric and specific productivities of 8,11‐DiHODE by recombinant cells expressing 8,11‐LDS from P. chrysogenum were 3.5‐ and 3.0‐fold higher than those of 5,8‐DiHODE, as a positional isomer product, by recombinant cells expressing 5,8‐LDS from A. nidulans , respectively, whereas the conversion yield of 8,11‐DiHODE was 39% lower . The volumetric and specific productivities and conversion yield of 8,11‐DiHOTrE was 1.3‐ and 1.9‐fold and 3% lower than those of 5,8‐DiHOTrE, as a positional isomer product, by recombinant cells expressing 5,8‐LDS from A. nidulans . The biotechnological production of 8‐HHME was not compared because its positional isomer product was not identified.…”

“…The optimal pH and temperature for the conversion of unsaturated fatty acids to 5,8-dihydroxy fatty acids by recombinant cells expressing 5,8-LDS from A. nidulans were ranging from 7.0 and 358C to 7.5 and 408C, respectively. [9][10][11] The maximum conversion rate of linoleic acid to 7,8-DiHODE by 7,8-LDS from G. cingulate occurred at pH 7.0 and 408C. 17 The optimal pH values of LDSs were similar.…”

“…The maximal conversion rates of linioleic acid, α‐linolenic acid, palmitoleic acid, and oleic acid to 8,11‐DiHODE, 8,11‐DiHOTrE, 8‐HHME, and 8‐HOME, respectively, were determined at pH 7.0 and 25°C; pH 6.0 and 25°C; pH 7.0 and 25°C; and pH 8.5 and 30°C, respectively (Figure ). The optimal pH and temperature for the conversion of unsaturated fatty acids to 5,8‐dihydroxy fatty acids by recombinant cells expressing 5,8‐LDS from A. nidulans were ranging from 7.0 and 35°C to 7.5 and 40°C, respectively . The maximum conversion rate of linoleic acid to 7,8‐DiHODE by 7,8‐LDS from G. cingulate occurred at pH 7.0 and 40°C .…”

“…Although orthogonal experiments can be achieved more accurately optimal reaction conditions, the optimization for the production of hydroxy fatty acids has been mainly performed using single factor experiments , . The interactions between pH and temperature and between cell density and substrate concentration were a little (Supporting Information Figure S1).…”

“…Pseudomonas aeruginosa has been used to produce 7,10-dihydroxy-8(E)-hexadecenoic acid, 5 7,10-dihydroxy-8(E)-octadecenoic acid, 6 7,10-dihydroxy-8,12(E,Z)-octadecadienoic acid, 7 and 9,12-dihydroxy-10(E)-eicosenoic acid 8 from palmitoleic acid, oleic acid, linoleic acid, and eicosenoic acid, respectively. Recombinant Escherichia coli expressing 5,8-linoleate diol synthase (5, from Aspergillus nidulans has been used to produce 5,8-dihydroxy-9(Z)-octadecenoic acid (5,8-DiHOME), 9 5,8dihydroxy-9,12(Z,Z)-octadecadienoic acid (5,, 10 and 5,8-dihydroxy-9,12,15(Z,Z,Z)-octadecatrienoic acid (5, 11 from oleic acid, linoleic acid, and a-linolenic acid, respectively. 10S-Dioxygenase from P. aeruginosa has been used to produce 10S-hydroxy-8(E)-octadecenoic acid (10S-HOME) from oleic acid.…”