Yong-Chan Kwon, In-Seok Oh, Nahum Lee, Kyung-Ho Lee, Yeo Joon Yoon, Eun Yeol Lee, Byung-Gee Kim and Dong-Myung Kim Integrating cell-free biosyntheses of heme prosthetic group and apoenzyme for the synthesis of functional P450 monooxygenase Biotechnology and Bioengineering 110
Functional holoenzyme of P450 monooxygenase was produced in a cell-free synthesis system. Step reactions required for the syntheses of apoenzyme and prosthetic group have been designed and implemented into a cell-free synthesis system so that these two separate pathways take place in the same reaction mixture, while being insulated from each other. Combined pathways for the synthesis of functional P450 monooxygenase were then further integrated with in situ assay reactions to enable real-time measurement of enzymatic activity during its synthesis.
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