Detailed Information on Publication Record
2002
Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene
BOSMA, Tjibbe, Jiří DAMBORSKÝ, Gerhard STUCKI and Dick JANSSENBasic information
Original name
Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene
Authors
BOSMA, Tjibbe (528 Netherlands), Jiří DAMBORSKÝ (203 Czech Republic, guarantor), Gerhard STUCKI (756 Switzerland) and Dick JANSSEN (528 Netherlands)
Edition
Applied and Environmental Microbiology, 2002, 1098-5336
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10600 1.6 Biological sciences
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 3.691
RIV identification code
RIV/00216224:14310/02:00006264
Organization unit
Faculty of Science
UT WoS
000176631600054
Keywords in English
TRICHLOROPROPANE; PROTEIN ENGINEERING; COMPUTER MODELLING; DOCKING; MUTANT
Změněno: 19/3/2010 10:54, prof. Mgr. Jiří Damborský, Dr.
Abstract
V originále
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.
Links
ME 276, research and development project |
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MSM 143100005, plan (intention) |
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