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

本研究的目的是建立微生物系统，产生氧化碳氢化合物，如长链二羧酸。为此，我们克隆了不同特异性的编码细胞色素P450的基因，但不包括CYP52家族成员。其中8个ALK基因来自麦芽糖念珠菌，8个YlALK基因来自解脂耶氏菌。在麦芽糖中显著诱导和表达的ALK1基因产物氧化了正构烷烃的极端，而ALK5基因产物也氧化了脂肪酸的ω位置。ALK7和ALK8基因产物只氧化脂肪酸。这些基因在异源酵母酿酒酵母（Saccharomyces cerevisiae）中功能表达。脂质体y的8个细胞色素P450ALK形成一个不同的CYP52亚家族。在YlALK基因的产物中，Ylalklp似乎主要氧化正癸烷，Ylalk2p对正十六烷有一定的活性。这些基因编码不同种类的细胞色素P450，我们目前的研究已经阐明了它们的性质，因此可以引入到新的宿主中，并利用它们构建新的氧化烷烃生产系统。

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.