A process for the hydroxylation of long-chain alkanes, fatty acids and other alkyl compounds

摘要

A microbial hydroxylation process is disclosed for selectively oxidising regions of long-chain alkanes, fatty acids and other alkyl compounds. The process should be easy to carry out and produce good yields of oxidation products, in particular hydroxylated fatty acids and long-chain dicarboxylic acids. The object of the invention is to modify yeast by genetic engineering so that when it is cultivated it expresses the required enzymes. The disclosed process is characterised in that the long-chain alkanes, fatty acids and other alkyl compounds are treated with monooxygenase systems that consist of cytochrom P450 and NADPOH cytochrom P450 reductase, and the hydroxylation products are then separated. The monooxygenase systems are produced in the reaction mixture by simultaneous expression of their components in yeast, preferably saccharomyces cerevisiae. The invention relates essentially to a vector for modifying saccharomyces by genetic engineering. On the basis of the structure Yep 51, the vector contains reductase cDNA between the restriction sites Sa1K and BamHI and a second expression cassette bound in the restriction site NruI. The expression cassette consists of the GAL10 promoter, the sequence that codes for cytochrom P450 and the ADH1 terminator.