Assessment of human drug metabolism by expression of cytochrome P-450 in yeasts.

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The purpose of this project is development of a new assessment method for human drug metabolism by using mixture of cytochrome P-450 expressed in yeast cells. We first cloned CYP3A4, a major human cytochrome P-450, and expressed it in yeast cells. On the other hand, we expressed human NADPH-cytochrome P-450 reductase and cytochrome b_5 in yeast cells. Then we improved the expression vectors to co-express different vectors in the same cells . We succeeded in co-expressing CYP3A4, NADPH-cytochrome P-450 reductase and cytochrome b_5 in the same cells and we observed high enzymic activities for several substrates. These results indicates further possible way to express sever al different cytochrome P-450 by this method.We, then, try to increase the yield of cytochrome P-450 in yeast cells changing by conditions of the incubation. Thus, we concluded that this approach is useful method for the assessment of human drug metabolism by using a mixture of human cytochrome P-450 expressed in yeast cells.