Efficient production of (R)-(−)-mandelic acid with highly substrate/product tolerant and enantioselective nitrilase of recombinant Alcaligenes sp.

• DOI: 10.1016/j.procbio.2010.02.011
• Publication Date: 2010-02-17T09:39:50Z
• Authors: Zhi-Jun Zhang; Jian-He Xu; Yu-Cai He; Li-Ming Ouyang; You-Yan Liu; Tadayuki Imanaka
• Keywords: 01 natural sciences; 0104 chemical sciences

Abstract For efficient production of ( R )-(−)-mandelic acid, a nitrilase gene from Alcaligenes sp. ECU0401 was cloned and overexpressed in Escherichia coli . After simple optimization of the culture conditions, the biocatalyst production was greatly increased from 500 to 7000 U/l. The recombinant E. coli whole cells showed strong tolerance against a high substrate concentration of up to 200 mM, and the concentration of ( R )-(−)-mandelic acid after only 4 h of transformation reached 197 mM with an enantiomeric excess ( ee p ) of 99%. In a fed-batch reaction with 600 mM mandelonitrile as the substrate, the cumulative production of ( R )-(−)-mandelic acid after 17.5 h of conversion reached 520 mM. The recombinant E. coli cells could also be repeatedly used in the biotransformation, retaining 40% of the initial activity after 10 batches of reaction. The highly substrate/product tolerable and enantioselective nature of this recombinant nitrilase suggests that it is of great potential for the practical production of optically pure ( R )-(−)-mandelic acid.