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KMID : 0545120100200030574
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Journal of Microbiology and Biotechnology
2010 Volume.20 No. 3 p.574 ~ p.578
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Regioselective Oxidation of Lauric Acid by CYP119, An Orphan Cytochrome P450 from Sulfolobus acidocaldarius
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Lim Young-Ran
Eun Chang-Yong
Park Hyoung-Goo
Han Song-Hee
Han Jung-Soo
Cho Kyoung-Sang
Chun Young-Jin
Kim Dong-Hak
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Abstract
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Archaebacteria Sulfolobus acidocaldarius contains the highly thermophilic cytochrome P450 enzyme (CYP119). CYP119 possesses the stable enzymatic activity at up to 85 ℃. However, this enzyme is still considered as an orphan P450 without known physiological function with endogenous or xenobiotic substrates. We characterized the regioselectivity of lauric acid by CYP119 using the auxiliary redox partner proteins, putidaredoxin (Pd) and putidaredoxin reductase (PdR). Purified CYP119 protein showed the tight binding affinity to lauric acid (Kd = 1.1 ± 0.1 μM) and dominantly hydroxylated (ω-1) position of lauric acid. We determined the steady-state kinetic parameters; kcat (10.8 min-1) and Km (12 μM). The increased ratio to ω-hydroxylated production of lauric acid catalyzed by CYP119 was observed with increase in the reaction temperature. These studies suggested that the regioselectivity of CYP119 provide the critical clue for the physiological enzyme function in this thermophilic archaebacteria. In addition, regiospecificity control of CYP119 without altering its thermostability can lead to the development of novel CYP119-based catalysts through protein engineering.
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KEYWORD
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P450, CYP119, Lauric acid, Oxidation, Regioselectivity
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