1.14.14.40 | synthesis |
biosynthetic pathway for the production of phenylacetonitrile in Escherichia coli utilizing enzymes from the plant glucosinolate-biosynthetic and bacterial aldoxime-nitrile pathways, i.e. introducing the genes encoding cytochrome P450 (CYP) 79A2 and CYP reductase from Arabidopsis thaliana, yielding the E,Z-phenylacetaldoxime-producing transformant and further introducing the aldoxime dehydratase (Oxd) gene from Bacillus sp. strain OxB-1, yielding the phenylacetonitrile-producing transformant. Expression of active CYP79A2 and concentration of biomass are improved by the combination of the autoinduction method, coexpression of groE, encoding the heat shock protein GroEL/GroES, N-terminal truncation of CYP79A2, and optimization of the culture conditions, yielding a more than 60fold concentration of E,Z-phenylacetaldoxime up to 2.9 mM |
729046 |