caa a22 4500 397558368 CHVBK 20180308164809.0 cr unu---uuuuu 161202e199603 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021260 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021260 Cytochrome P450foxy, a Catalytically Self-Sufficient Fatty Acid Hydroxylase of the Fungus Fusarium oxysporum [Elektronische Daten] [Norikazu Nakayama, Asako Takemae, Hirofumi Shoun] We have purified membrane-bound fatty acid (ω-1-ω-3) hydroxylase of the fungus Fusarium oxysporum MT-811 and found that the activity depends on a single polypeptide with an apparent Mr value of 118,000. The purified hydroxylase exhibited spectral characteristics of cytochrome P450 (P450), and could catalyze the hydroxylation without the aid of any other proteinaceous components, such as NADPH-P450 reductase. These properties of the fungal hydroxylase are the same as those of bacterial P450BM3 of Bacillus megaterium, a catalytically self-sufficient fused protein of P450 and its reductase. Other properties of the two enzymes, such as molecular weight, high catalytic turnover, and the regiospecificity of the hydroxylating position, were also almost identical. Further, the fungal hydroxylase reacted with the antibody to P450BM3. It was thus shown that the fungal fatty acid hydroxylase structurally and functionally bears a close resemblance to P450BM3, although it is membrane-bound, unlike the bacterial counterpart.On the other hand,a unique phenomenon was found with the fungal hydroxylase: its NADPH-cyto-chrome c- or NADPH-menadione reductase activity was enhanced enormously upon binding of its substrate (fatty acid). This appears to be the first instance in which the reactivity of P450 reductase against an artificial electron acceptor was enhanced by the binding of the substrate (to be hydroxylated) to P450. These results raise interesting questions about the molecular evolution of P450. Here we term the fungal hydroxylase cytochrome P450foxy. © by the Journal of Biochemistry Regular Papers nationallicence cytochrome P450 nationallicence fatty acid hydroxylase nationallicence fungus nationallicence fused protein nationallicence NADPH-cytochrome P450 reductase nationallicence Nakayama Norikazu Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305 aut Takemae Asako Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305 aut Shoun Hirofumi Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305 aut The Journal of Biochemistry Oxford University Press 119/3(1996-03), 435-440 0021-924X 119:3<435 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021260 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021260 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nakayama Norikazu Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305 aut NATIONALLICENCE 700 1- Takemae Asako Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305 aut NATIONALLICENCE 700 1- Shoun Hirofumi Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/3(1996-03), 435-440 0021-924X 119:3<435 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford