Biodegradation of 1,2,3-trichloropropane through directed
evolution and heterologous expression of a haloalkane
dehalogenase gene

J 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 JANSSEN

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

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:

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

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

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

Citovat

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.

@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 and 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, vol.~68, No~7, p.~3582-3587. ISSN~1098-5336.

Detailed Information on Publication Record