A Pseudomonas Putida Strain Genetically Engineered for 1,2,3-Trichloropropane Bioremediation

Authors

Ghufrana, S., Pavlova, M., Arif, M.I., Postema, C.P., Damborsky, J., Janssen, D.B.

Source

APPLIED AND ENVIRONMENTAL MICROBIOLOGY 80: 5467-5476 (2014)

Abstract

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.

Full text

Citation

Ghufrana, S., Pavlova, M., Arif, M.I., Postema, C.P., Damborsky, J., Janssen, D.B., 2014: A Pseudomonas Putida Strain Genetically Engineered for 1,2,3-Trichloropropane Bioremediation. Applied and Environmental Microbiology 80: 5467-5476.

Ondrej Vavra celebrating success with his presentation on CaverDock
Lecture by Dr. Stavrakis from ETH Zurich
High evaluation of the team’s research performance
Klaudia Chmelova and Veronika Liskova passed doctoral state exam
ELIXIR CZ annual conference 15 – 16 Nov, 2017
New video explaining the goals of Rafts4Biotech EU project
Invitation to a presentation by Synthetic Genomics