Identification of reaction intermediates and optimization of enzymatic production of prostaglandin F2α

The cyclooxygenase from Agarophyton vermiculophyllum (GVCOX) is useful for prostaglandin F2α (PGF2α) production. In this study, 15-hydroperoxyPGF2α, 15-hydroperoxyPGE2, and 15-hydroperoxyPGD2 were identified as intermediates of PGF2α production from arachidonic acid (ARA) using recombinant Escherichia coli expressing the A. vermiculophyllum GVCOX gene. As a result of evaluation of the experimental conditions, the addition of 12.5 mM SnCl2 during the GVCOX catalytic reaction enabled efficient PGF2α production (6.6 mM PGF2α from 30 mM ARA). Furthermore, efficient PGF2α production without SnCl2 was achieved by a combination of prostaglandin F2α 9-dehydrogenase (PDH) which catalyzes PGE2 reduction to PGF2α.