| 초록 |
To increase the efficiency of electron transfer for P450 reaction, PCNA(Proliferating cell nuclear antigen) is used as linker for three proteins which are putidaredoxin reductase (pdr), putidaredoxin (pdx) andP450cam.PCNA found in Sulfolobus solfataricus forms a heterotrimer. As fusing each domain to Pdr, Pdx and P450cam, stable and self-assembly heterotrimeric complex (PCNA-utilized protein complex of P450 and its two electron transfer-related proteins, PUPPET) was constructed.To use the enzyme as catalyst in industry, in vivo system should be developed because of the expensive cost for enzyme purification. Therefore, the PCNA effects for P450 activity using whole cell should be evaluated and the condition for expression of three proteins and reaction should be optimized. Here, we constructed two plasmid to express three PCNA fused proteins and transformed to E.coli. Finally, we confirm the PCNA effects for P450 reaction in vivo, too. |
