Functional expression and characterisation of human cytochrome P45017α in Pichia pastoris

摘要: Human cytochrome P45017alpha (CYP17), present in mammalian adrenal and gonadal tissues, catalyses both steroid 17-hydroxylation C17,20 lyase reactions, producing intermediates for the glucocorticoid androgenic pathways, respectively. The characterisation of this complex enzyme was initially hampered due to low level vivo expression CYP17. Heterologous systems have contributed greatly our current knowledge CYP17's dual catalytic activity. However, hydrophobic nature membrane-bound protein, primarily truncated modified forms CYP17 are currently being expressed heterologously. Although N-terminally has been well characterised, protein structure function studies still necessitate unmodified, wild-type We report here a catalytically active, unmodified human industrial methylotrophic yeast, Pichia pastoris. A typical P450 carbon monoxide difference spectrum, with an absorption maximum at 448nm substrate-induced type I spectrum were recorded using detergent-solubilised cellular fraction containing catalysed conversion progesterone 17-hydroxyprogesterone as 16-hydroxyprogesterone, product unique chimpanzee This is first showing heterologous fully functional steroidogenic P.