Metabolic Engineering of Mortierella alpina for Enhanced Arachidonic Acid Production through the NADPH-Supplying Strategy.

ABSTRACT:

Unlabelled

NADPH is known to be a key cofactor required for fatty acid synthesis and desaturation. Various enzymatic reactions can generate NADPH. To determine the effect of NADPH sources on lipogenesis, glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (PGD), isocitrate dehydrogenase (IDH), and malic enzyme (ME) were overexpressed in Mortierella alpina Our results showed that G6PD2 had the most significant effect on fatty acid synthesis, with a 1.7-fold increase in total fatty acid, whereas ME2 was more effective in desaturation, with a 1.5-fold increase in arachidonic acid (AA) content over control. Co-overexpression of G6PD2 and ME2 improved both fatty acid synthesis and desaturation. Within 96 h of fermentation using the fed-batch method, the co-overexpressing strain accumulated AA at a productivity of 1.9 ± 0.2 g/(liter · day), which was 7.2-fold higher than that in the M. alpina control that was cultured in a flask.

Importance

This study proved that the pentose phosphate pathway is the major NADPH contributor during fatty acid synthesis in M. alpina The NADPH sources may be differently responsible for fatty acid synthesis or desaturation. Co-overexpression of G6PD2 and ME2 significantly increases AA production.

SUBMITTER: Hao G 

PROVIDER: S-EPMC4959234 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature