%0 Journal Article
%T Exploring the Electron Transfer Pathway in the Oxidation of Avermectin by CYP107Z13 in Streptomyces ahygroscopicus ZB01
%A Mei Li
%A Yujie Zhang
%A Lin Zhang
%A Xiaoyan Yang
%A Xiliang Jiang
%J PLOS ONE
%D 2014
%I Public Library of Science (PLoS)
%R 10.1371/journal.pone.0098916
%X Streptomyces ahygroscopicus ZB01 can effectively oxidize 4¡å-OH of avermectin to form 4¡å-oxo-avermectin. CYP107Z13 is responsible for this site-specific oxidation in ZB01. In the present study, we explored the electron transfer pathway in oxidation of avermectin by CYP107Z13 in ZB01. A putative [3Fe-4S] ferredoxin gene fd68 and two possible NADH-dependent ferredoxin reductase genes fdr18 and fdr28 were cloned from the genomic DNA of ZB01. fd68 gene disruption mutants showed no catalytic activity in oxidation of avermectin to form 4¡å-oxo-avermectin. To clarify whether FdR18 and FdR28 participate in the electron transfer during avermectin oxidation by CYP107Z13, two whole-cell biocatalytic systems were designed in E. coli BL21 (DE3), with one co-expressing CYP107Z13, Fd68 and FdR18 and the other co-expressing CYP107Z13, Fd68 and FdR28. Both of the two biocatalytic systems were found to be able to mediate the oxidation of avermectin to form 4¡å-oxo-avermectin. Thus, we propose an electron transfer pathway NADH¡úFdR18/FdR28¡úFd68¡úCYP107Z13 for oxidation of avermectin to form 4¡å-oxo-avermectin in ZB01.
%U http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0098916