Hydroxylation of L-proline to cis-3-hydroxy-L-proline by recombinant Escherichia coli expressing a synthetic L-proline-3-hydroxylase gene

Abstract A synthetic gene encoding a Streptomyces l -proline-3-hydroxylase was constructed and used to produce the hydroxylase protein in recombinant Escherichia coli . A fermentation process for growth of this recombinant E. coli for enzyme production was scaled-up to 250 L. A biotransformation process was developed using cell suspensions of the recombinant E. coli and subsequently scaled-up to 10 L for conversion of l -proline to cis -3-hydroxy- l -proline. A reaction yield of 85 M% and d.e. of 99.9% was obtained for cis -3-hydroxy- l -proline.