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@article{1213426,
author = {Ghufrana, S. and Pavlová, Martina and Arif, M.I. and Postema, C.P. and Damborský, Jiří and Janssen, D.B.},
article_number = {17},
doi = {http://dx.doi.org/10.1128/AEM.01620-14},
keywords = {1; 2;3-Trichloropropane; biodegradation; Pseudomonas putida MC4},
language = {eng},
issn = {0099-2240},
journal = {Applied and Environmental Microbiology},
title = {A Pseudomonas putida Strain Genetically Engineered for 1,2,3-Trichloropropane Bioremediation.},
volume = {80},
year = {2014}
}
TY - JOUR
ID - 1213426
AU - Ghufrana, S. - Pavlová, Martina - Arif, M.I. - Postema, C.P. - Damborský, Jiří - Janssen, D.B.
PY - 2014
TI - A Pseudomonas putida Strain Genetically Engineered for 1,2,3-Trichloropropane Bioremediation.
JF - Applied and Environmental Microbiology
VL - 80
IS - 17
SP - 5467-5476
EP - 5467-5476
SN - 00992240
KW - 1
KW - 2;3-Trichloropropane
KW - biodegradation
KW - Pseudomonas putida MC4
N2 - 1,2,3-Trichloropropane (TCP) is a toxic compound that is recalcitrant to biodegradation in the environment and attempts to isolate TCP-degrading organisms using enrichment cultivation have failed. A potential biodegradation pathway starts with hydrolytic dehalogenation to 2,3 dichloro-1-propanol (DCP), followed by oxidative metabolism. To obtain a practically applicable TCP-degrading organism, we introduced an engineered haloalkane dehalogenase with improved TCP degradation activity into the DCP-degrading bacterium Pseudomonas putida MC4. For this, the dehalogenase gene (dhaA31) was cloned behind the constitutive dhlA promoter and introduced into the genome of strain MC4 using a transposon delivery system. The transposon-located antibiotic resistance marker was subsequently removed using a resolvase step. Growth of the resulting engineered bacterium P. putida MC4-5222 on TCP was indeed observed, and all organic chlorine was released as chloride. A packed-bed reactor with immobilized cells of strain MC4 5222 degraded >95% of influent TCP (0.33 mM) under continuous flow conditions, with stoichiometric release of inorganic chloride. The results show the use of a laboratory-evolved dehalogenase and genetic engineering for obtaining an effective plasmid-free and stable whole-cell biocatalyst for the aerobic bioremediation of a recalcitrant chlorinated hydrocarbon.
ER -
GHUFRANA, S., Martina PAVLOVÁ, M.I. ARIF, C.P. POSTEMA, Jiří DAMBORSKÝ a D.B. JANSSEN. A Pseudomonas putida Strain Genetically Engineered for 1,2,3-Trichloropropane Bioremediation. \textit{Applied and Environmental Microbiology}. 2014, roč.~80, č.~17, s.~5467-5476. ISSN~0099-2240. Dostupné z: https://dx.doi.org/10.1128/AEM.01620-14.
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