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@article{404591,
author = {Bosma, Tjibbe and Damborský, Jiří and Stucki, Gerhard and Janssen, Dick},
article_number = {7},
keywords = {TRICHLOROPROPANE; PROTEIN ENGINEERING; COMPUTER MODELLING; DOCKING; MUTANT},
language = {eng},
issn = {1098-5336},
journal = {Applied and Environmental Microbiology},
title = {Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene},
url = {http://ncbr.chemi.muni.cz/~jiri/ABSTRACTS/aem02a.html},
volume = {68},
year = {2002}
}
TY - JOUR
ID - 404591
AU - Bosma, Tjibbe - Damborský, Jiří - Stucki, Gerhard - Janssen, Dick
PY - 2002
TI - Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene
JF - Applied and Environmental Microbiology
VL - 68
IS - 7
SP - 3582
EP - 3582
SN - 10985336
KW - TRICHLOROPROPANE
KW - PROTEIN ENGINEERING
KW - COMPUTER MODELLING
KW - DOCKING
KW - MUTANT
UR - http://ncbr.chemi.muni.cz/~jiri/ABSTRACTS/aem02a.html
N2 - Using a combined strategy of random mutagenesis of haloalkane dehalogenase and genetic engineering of a chloropropanol-utilizing bacterium, we obtained an organism that is capable of growth on 1,2,3-trichloropropane (TCP). The highly toxic and recalcitrant chemical TCP is a waste product generated from the manufacture of the industrial commodity chemical epichlorohydrin. Attempts to select and enrich bacterial cultures that can degrade TCP have been unsuccessful, prohibiting the development of a biological process for groundwater treatment. The key step in the aerobic degradation of TCP is the initial dehalogenation to 2,3-dichloro-1-propanol by a haloalkane dehalogenase. We used random mutagenesis and screening on eosine-methylene blue agar plates to improve the activity on TCP of the haloalkane dehalogenase from Rhodococcus sp. m15-3 (DhaA). A second-generation mutant containing two amino acid substitutions, Cys176Tyr and Tyr273Phe, was nearly eight times more efficient in dehalogenating TCP than wild type dehalogenase. The 2,3-dichloro-1-propanol utilizing bacterium Agrobacterium radiobacter AD1 expressing the evolved haloalkane dehalogenase under control of a constitutive promoter was able to utilize TCP as sole carbon- and energy source. These results demonstrated that directed evolution of a key catabolic enzyme and its subsequent recruitment by a suitable host organism can be used for the construction of bacteria for the degradation of toxic and environmentally recalcitrant chemicals.
ER -
BOSMA, Tjibbe, Jiří DAMBORSKÝ, Gerhard STUCKI a Dick JANSSEN. Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene. \textit{Applied and Environmental Microbiology}. 2002, roč.~68, č.~7, s.~3582-3587. ISSN~1098-5336.
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