Production of alkanedioic acids by cytochrome P450(BM-3) monooxygenase: Oxidation of 16-hydroxyhexadecanoic acid to hexadecane-1,16-dioic acid

Cytochrome P450(BM-3) monooxygenase [E.C.1.14.14.1] from Bacillus megateriu m ATCC 14581 was used to devise a production route towards long chain alkan edioic acids. Both cell lysates and intact cells of an Escherichia coil JM1 01 recombinant, expressing the cyp102 gene that encodes cytochrome P450(BM- 3) monooxygenase, carried out the biotransformation of 16-hydroxyhexadecano ic acid to 1,16-hexadecanedioic acid. The intermediates of 1 6-hydroxyhexad ecanoic acid oxidation by whole cells synthesizing cytochrome P450(BM-3) mo nooxygenase were identified by gas chromatography mass spectrometry. Oxidat ion of 16-hydroxyhexadecanoic acid occurred with concomitant accumulation o f small amounts of intermediates, one of which was tentatively identified a s 16-dihydroxyhexadecanoic acid. Products of side reactions were 13,16-dihy droxyhexadecanoic acid and 12,16-dihydroxyhexadecanoic acid. Thr oxidation reactions did not take place in E. coli JM101 carrying the vector-DNA witho ut the cyp102 gene. These data demonstrate for the first time that cytochrome P450(BM-3) monoox ygenase is able to oxidize omega-hydroxy long chain fatty acids to the corr esponding dicarboxylic acids.