Exploring the structure and activity of haloalkane dehalogenase from Sphingomonas paucimobilis UT26: evidence for product and water mediated inhibition

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Publikace nespadá pod Ekonomicko-správní fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.
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OAKLEY Aaron PROKOP Zbyněk BOHÁČ Michal KMUNÍČEK Jan JEDLIČKA Tomáš MONINCOVÁ Marta KUTÁ-SMATANOVÁ Ivana NAGATA Yuji DAMBORSKÝ Jiří WILCE Matthew

Rok publikování 2002
Druh Článek v odborném periodiku
Časopis / Zdroj Biochemistry
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
www http://ncbr.chemi.muni.cz/~jiri/ABSTRACTS/biochem02.html
Obor Biochemie
Klíčová slova X-RAY; DEHALOGENATION; ENZYME; PROTEIN ENGINEERING; INHIBITION
Popis The hydrolysis of haloalkanes to their corresponding alcohols and inorganic halides is catalysed by a/b-hydrolases called haloalkane dehalogenases. The study of haloalkane dehalogenases is vital for the development of these enzymes if they are to be utilized for bioremediation of organohalide-contaminated industrial waste. We report the kinetic and structural analysis of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26 (LinB) in complex with each of 1,2-dichloroethane and 1,2-dichloropropane and the reaction product of 1-chlorobutane turnover. Activity studies showed very weak, but detectable activity of LinB with 1,2-dichloroethane (0.012 nmol.s-1.mg-1 of enzyme) and 1,2-dichloropropane (0.027 nmol.s-1.mg-1 of enzyme). These activities are much weaker compared, for example, to activity of LinB with 1-chlorobutane (68.169 nmol.s-1.mg-1 of enzyme). Inhibition analysis reveals that both 1,2-dichloroethane and 1,2-dichloropropane act as simple competitive inhibitors of the substrate 1-chlorobutane and that 1,2-dichloroethane binds to LinB with lower affinity than 1,2-dichloropropane. Docking calculations on the enzyme in the absence of active site water molecules and halide ions confirms that these compounds could bind productively. However, when these moieties were included in the calculations, they bound in the manner similar to that observed in the crystal structure. These data provide an explanation for the low activity of LinB with small, chlorinated alkanes and show the importance of active site water molecules and reaction products in molecular docking.
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