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Integrating cell-free biosyntheses of heme prosthetic group and apoenzyme for the synthesis of functional P450 monooxygenase

Title
Integrating cell-free biosyntheses of heme prosthetic group and apoenzyme for the synthesis of functional P450 monooxygenase
Authors
Kwon Y.-C.Oh I.-S.Lee N.Lee K.-H.Yoon Y.J.Lee E.Y.Kim B.-G.Kim D.-M.
Ewha Authors
윤여준
SCOPUS Author ID
윤여준scopus
Issue Date
2013
Journal Title
Biotechnology and Bioengineering
ISSN
0006-3592JCR Link
Citation
vol. 110, no. 4, pp. 1193 - 1200
Indexed
SCI; SCIE; SCOPUS WOS scopus
Abstract
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.
DOI
10.1002/bit.24785
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자연과학대학 > 화학·나노과학전공 > Journal papers
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