A strategy for L-isoleucine dioxygenase screening and 4-hydroxyisoleucine production by resting cells.

ABSTRACT: L-Isoleucine dioxygenase (IDO) specifically converts L-isoleucine(L-Ile) to 4-hydroxyisoleucine(4-HIL). To obtain IDO with improved activity, a strategy was developed that is dependent on the restoration of succinate-minus E. coli cell growth by the coupling of L-Ile hydroxylation and the oxidation of α-ketoglutarate(α-KGA) to succinate. Five mutants were obtained with this strategy, and the characteristics of IDO^M3, which exhibited the highest activity, were studied. The catalytic efficiency, thermal stability and catalytic rate of IDO^M3 were significantly improved compared with those of wild-type IDO. Moreover, an efficient method for the biotransformation of 4-HIL by resting cells expressing IDO^M3 was developed, with which 151.9 mmol of 4-HIL/L (22.4 g/L

SUBMITTER: Zhang C 

PROVIDER: S-EPMC5972919 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature