Construction of terminal oxidation systems for hydrocabon chain A use of a variety of yeast cytochrome P450 genes

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The aim of this research was to construct microbial systems which produce oxidized hydrocabons, such as long chain dicarboxylic acids. For this purpose, we cloned various genes which are encoding cytochromes P450 with different specificities, but of CYP52 family members. Those were 8 ALK genes from the yeast Candida maltosa and also 8 YlALK genes from Yarrowia lipolytica. The product of ALK1 gene that is prominently induced and expressed in C. maltosa oxidized very end of n-alkanes, whereas ALK5 gene product also oxidized omega position of fatty acids. ALK7 and ALK8 gene products only oxidize fatty acids. These gene were functionally expressed in heterologous yeast Saccharomyces cerevisiae. The eight cytochromes P450ALK from Y. lipolytica form a different CYP52 sub family. Among the products of YlALK genes, Ylalklp seemed principally to oxidize n-decane and Ylalk2p had some activity to n-hexadecane.These genes encoding different kind of cytochromes P450, properties of which have been elucidated by our current research, thus can be introduced into a new host and be utilized to construct a new production system of oxidized alkanes.