Cytochrome P450 Enzyme-based Biocatalytic Platform Construction And Application

Streptomyces species belong to the Actinomycete genus of Gram-positive bacteria and are found in soil and marine habitats. Streptomyces produce a vast array of antibiotics that play important role. Gene sequence of Streptomyces often contains a Cytochrome P450 gene. We hope to found some P450 genes in Streptomyces of our group utilize Genome mining approach. Genome sequence information enables direct cloning of these P450 genes using polymerase chain reaction (PCR), the cloned genes then be expressed in Escherichia coli, so that we obtain these new P450 Biocatalysts and establish a Cytochrome P450-based Biocatalysis platform. Then screen the function and activity of these enzymes, explore their applications in Drug Synthesis.Streptomyces TP-A2060 is the producing strain of antitumor antibiotic yatakemycin. We mining the P450 genes in this strain through Bioinformatics analysis. Analyzed these biocatalyst using online software, and they were divided into different CYP families. Cloned and expressed these genes by molecular biological methods. We purified 8 enzymes, and other 5 soluble proteins. So the Cytochrome P450 complement of Streptomyces TP-A2060 was established on the whole. We carried out some research on these enzymes, but no catalytic activity was detected at present. Further study of the fuction is in progress.