NDLI: Tuning the substrate specificity by engineering the active site of cytochrome P450cam: A rational approach

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Manna, Soumen Kanti Mazumdar, Shyamalava
Description	Author Affiliation: Manna SK ( Department of Chemical Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Colaba, Mumbai 400 005, India.)
Abstract	Rational design of the active site of cytochrome P450cam has been carried out to catalyse oxygenation of various potentially important chemical reactions. The modeling studies showed that the distal pocket of the heme consisting of the Y96, T101, F87 and L244 residues could be suitably mutated to change the substrate specificity of the enzyme . We found that the mutant enzymes could catalyse oxygenation of indole to produce indigo. While Y96F was found to be several times better as a catalyst for conversion of indole to indigo, the double mutant Y96F/L244A showed the highest NADH oxidation rate as well as yield of indigo. The oxidative catalysis using $H_{2}O_{2}$ as the oxygen source was found to produce a higher purity of indigo, and lesser or no formation of indirubin was detected. The enzymatic oxygenation of aromatic hydrocarbons such as coumarin and analogues was also found to be enhanced on mutation of Y96 and L244 residues in the enzyme . The studies also showed that mutation of suitable residues can alter the regio-selectivity of hydroxylation of the aromatic hydrocarbons.
ISSN	14779226
Issue Number	12
Volume Number	39
e-ISSN	13645447
Journal	Dalton Trans.
Language	English
Publisher	Royal Society of Chemistry
Publisher Date	2010-03-28
Publisher Place	Great Britain (UK)
Access Restriction	Subscribed
Subject Keyword	Camphor 5-Monooxygenase Catalytic Domain Protein Engineering Methods Chemistry Genetics Metabolism Coloring Agents Chemical Synthesis Ferredoxins Isolation & Purification Hydrogen Peroxide Indigo Carmine Indoles Molecular Structure Mutagenesis, Site-Directed NAD NADH, NADPH Oxidoreductases Oxidants Oxidation-Reduction Substrate Specificity Journal Article Research Support, Non-U.S. Gov't
Content Type	Text
Resource Type	Article
Subject	Inorganic Chemistry

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