Abstract:
:1,2,3-Trichloropropane (TCP) is a highly toxic and recalcitrant compound. Haloalkane dehalogenases are bacterial enzymes that catalyze 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 cannot be easily detected by conventional analytical methods. Here we demonstrate detection of the weak activity (k(cat) = 0.005 s(-1)) of LinB with TCP using X-ray crystallography and microcalorimetry. This observation makes LinB a useful starting material for the development of a new biocatalyst toward TCP by protein engineering. Microcalorimetry is proposed to be a universal method for the detection of weak enzymatic activities. Detection of these activities is becoming increasingly important for engineering novel biocatalysts using the scaffolds of proteins with promiscuous activities.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Monincová M,Prokop Z,Vévodová J,Nagata Y,Damborsky Jdoi
10.1128/AEM.02416-06subject
Has Abstractpub_date
2007-03-01 00:00:00pages
2005-8issue
6eissn
0099-2240issn
1098-5336pii
AEM.02416-06journal_volume
73pub_type
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