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 JANSSEN. Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene. Applied and Environmental Microbiology. 2002, vol. 68, No 7, p. 3582-3587. ISSN 1098-5336.

Basic 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

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: United States of America
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• 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
• Tags: COMPUTER MODELLING, DOCKING, MUTANT, Protein engineering, TRICHLOROPROPANE

Abstract

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: Name: Racionální re-design mikrobiálních enzymů podílejících se na degradaci toxických organických polutantů

Investor: Ministry of Education, Youth and Sports of the CR, Rational re-design of microbial enzymes involved in degradation of toxic organic pollutants.

• MSM 143100005, plan (intention): Name: Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Investor: Ministry of Education, Youth and Sports of the CR, Biomolecular Structure-function Relationships and their role in the Metabolism