Integrating cell-free biosyntheses of heme prosthetic group and apoenzyme for the synthesis of functional P450 monooxygenase
- 주제(키워드) Cell-free protein synthesis , Cytochrome P450 monooxygenase , Heme , In vitro metabolic engineering , Prosthetic group
- 등재 SCIE, SCOPUS
- 발행기관 Wiley - V C H Verlag GmbbH & Co.
- 발행년도 2013
- 총서유형 Journal
- URI http://www.dcollection.net/handler/ewha/000000094726
- 본문언어 영어
- Published As http://dx.doi.org/10.1002/bit.24785
초록/요약
Harnessing the isolated protein synthesis machinery, cell-free protein synthesis reproduces the cellular process of decoding genetic information in artificially controlled environments. More often than not, however, generation of functional proteins requires more than simple translation of genetic sequences. For instance, many of the industrially important enzymes require non-protein prosthetic groups for biological activity. Herein, we report the complete cell-free biogenesis of a heme prosthetic group and its integration with concurrent apoenzyme synthesis for the production of functional P450 monooxygenase. Step reactions required for the syntheses of apoenzyme and the prosthetic group have been designed so that these two separate pathways take place in the same reaction mixture, 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. © 2012 Wiley Periodicals, Inc.
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