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  • Title: A strategy for L-isoleucine dioxygenase screening and 4-hydroxyisoleucine production by resting cells.
    Author: Zhang C, Ma J, Li Z, Liang Y, Xu Q, Xie X, Chen N.
    Journal: Bioengineered; 2018 Jan 01; 9(1):72-79. PubMed ID: 28430004.
    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 IDOM3, which exhibited the highest activity, were studied. The catalytic efficiency, thermal stability and catalytic rate of IDOM3 were significantly improved compared with those of wild-type IDO. Moreover, an efficient method for the biotransformation of 4-HIL by resting cells expressing IDOM3 was developed, with which 151.9 mmol of 4-HIL/L (22.4 g/L) was synthesized in 12 h while the substrates seldom exhibited additional consumption.
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