The invention discloses a
gene segment provided with coded and highly-active trans-4-hydroxyl-L-prolyl hydroxylase and an application thereof.
Gene codons of trans-4-hydroxyl-L-prolyl hydroxylase (
GenBank ID: BAA 20094.1) in Dactylosporangiumsp. RH1 are optimized to obtain a p4hyd
gene; the optimized p4hyd
gene is truncated to obtain a P4Hyd1-257 gene; recombinant plasmids including pET-M-3C-P4Hyd and pET-M-3C-P4Hyd1-257 are built respectively,
prokaryotic expression is performed through
escherichia coli, and trans-4-hydroxyl-L-
proline is produced in a conversion manner by utilizing thalli as an
enzyme source. An experimental result shows that L-
proline is taken as a substrate, compared with pET-M-3C-P4Hyd, the highest conversion rate of trans-4-hydroxyl-L-
proline generated by pET-M-3C-P4Hyd1-257 in the conversion manner is increased from 91.4% to 97.4%, the conversion time is shortened from 80 hours to 60 hours, so that the conversion rate is improved remarkably, and the conversion time is shortened remarkably. The gene segment provided with coded and highly-active trans-4-hydroxyl-L-prolyl hydroxylase and the application thereof can be used for producing L-proline by a
bioconversion method, and have better industrial application prospect.