ArticleApplied and environmental microbiology2002
Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene.
Article in Applied and environmental microbiology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 128 citations in OpenAlex.
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- Dehalogenases: From Improved Performance to Potential Microbial Dehalogenation Applications.Molecules (Basel, Switzerland) · 2018Review
- Targeted mutagenesis: A sniper-like diversity generator in microbial engineering.Synthetic and systems biotechnology · 2017Review
- Biochemical characterization of two haloalkane dehalogenases: DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea.Protein science : a publication of the Protein Society · 2016Article
- Exacerbation of substrate toxicity by IPTG in Escherichia coli BL21(DE3) carrying a synthetic metabolic pathway.Microbial cell factories · 2015Article
- Biodegradation of hexachlorobenzene by a constructed microbial consortium.World journal of microbiology & biotechnology · 2015Article
- Selection and screening of microbial consortia for efficient and ecofriendly degradation of plastic garbage collected from urban and rural areas of Bangalore, India.Environmental monitoring and assessment · 2015Article
- A Pseudomonas putida strain genetically engineered for 1,2,3-trichloropropane bioremediation.Applied and environmental microbiology · 2014Article
- The predictability of molecular evolution during functional innovation.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Biochemical characterization of a haloalkane dehalogenase DadB from Alcanivorax dieselolei B-5.PloS one · 2014Article
- Improvement of biocatalysts for industrial and environmental purposes by saturation mutagenesis.Biomolecules · 2013Article
- Novel dehalogenase mechanism for 2,3-dichloro-1-propanol utilization in Pseudomonas putida strain MC4.Applied and environmental microbiology · 2012Article
- Transformation and biodegradation of 1,2,3-trichloropropane (TCP).Environmental science and pollution research international · 2012Review
- Development of a dehalogenase-based protein fusion tag capable of rapid, selective and covalent attachment to customizable ligands.Current chemical genomics · 2012Article
- CAVER 3.0: a tool for the analysis of transport pathways in dynamic protein structures.PLoS computational biology · 2012Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Using a combined strategy of random mutagenesis of haloalkane dehalogenase and genetic engineering of a chloropropanol-utilizing bacterium, we constructed an organism that is capable of growth on 1,2,3-trichloropropane (TCP). This highly toxic and recalcitrant compound is a waste product generated from the manufacture of the industrial chemical epichlorohydrin. Attempts to select and enrich bacterial cultures that can degrade TCP from environmental samples have repeatedly been unsuccessful, prohibiting the development of a biological process for groundwater treatment. The critical step in the aerobic degradation of TCP is the initial dehalogenation to 2,3-dichloro-1-propanol. We used random mutagenesis and screening on eosin-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. Molecular modeling of the mutant dehalogenase indicated that the Cys176Tyr mutation has a global effect on the active-site structure, allowing a more productive binding of TCP within the active site, which was further fine tuned by Tyr273Phe. The evolved haloalkane dehalogenase was expressed under control of a constitutive promoter in the 2,3-dichloro-1-propanol-utilizing bacterium Agrobacterium radiobacter AD1, and the resulting strain was able to utilize TCP as the 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 a toxic and environmentally recalcitrant chemical.
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