Weak Activity of Haloalkane Dehalogenase LinB with 1,2,3-Trichloropropane Revealed by X-ray Crystallography and Microcalorimetry

Authors

Monincova, M., Prokop, Z., Vevodova, J., Nagata, Y., Damborsky, J.

Source

APPLIED AND ENVIRONMENTAL MICROBIOLOGY 73: 2005-2008 (2007)

Abstract

1,2,3-trichloropropane (TCP) is a highly toxic and recalcitrant compound. Haloalkane dehalogenases are bacterial enzymes catalysing the cleavage of a carbon-halogen bond in a wide range of organic halogenated compounds. Haloalkane dehalogenase LinB from Sphingobium japonicum UT26 has, for a long time, been considered inactive with TCP since the reaction can not be easily detected by conventional analytical methods. Here we demonstrate detection of the weak activity (kcat = 0.005 s-1) of LinB with TCP using X-ray crystallography and microcalorimetry. This observation makes LinB a useful starting material for development of new biocatalyst towards TCP by protein engineering. Microcalorimetry is proposed to be a universal method for detection of weak enzymatic activities. Detection of these activities is becoming increasingly important for engineering of novel biocatalysts using the scaffolds of proteins with promiscuous activities.

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Citation

Monincova, M., Prokop, Z., Vevodova, J., Nagata, Y., Damborsky, J., 2007: Weak Activity of Haloalkane Dehalogenase LinB with 1,2,3-Trichloropropane Revealed by X-ray Crystallography and Microcalorimetry. Applied and Environmental Microbiology 73: 2005-2008.

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