Construction of a synthetic metabolic pathway for degradation of important environmental pollutant using protein and metabolic engineering
Project goals
Anthropogenic compound 1,2,3-trichloropropane (TCP) is released into the environment as a result of its manufacture and use as a solvent. Since its half-life under groundwater conditions is estimated to be up to 100 years, TCP is likely to persist for long period of time in the groundwater environment. TCP is very resistant to biodegradation, which can be attributed to a single biochemical step, lacking dehalogenase efficiently converting TCP to dichloropropanol. We have recently constructed dehalogenase with improved catalysis of this biochemical step (Pavlová et al. 2009, Nat. Chem. Biol. 5: 727-733). The objective of this project is to assemble entire biochemical pathway by cloning the genes optimized by protein engineering to native bacteria, leading to construction of TCP-degrading bacterium. This bacterium will grow on TCP as a sole carbon and energy source. To best of our knowledge, this bacterium will be the first example of a microorganism constructed by protein and matabolic engineering, applicable for the cleanup of contaminated water in the closed flow systems.
Keywords
biodegradation environmental pollutant protein engineering synthetic biology 123-trichloropropane
Public support
Provider
Czech Science Foundation
Programme
Standard projects
Call for proposals
Standardní projekty 15 (SGA02012GA-ST)
Main participants
Masarykova univerzita / Přírodovědecká fakulta
Contest type
VS - Public tender
Contract ID
P503-12-0572
Alternative language
Project name in Czech
Konstrukce syntetické metabolické dráhy pro degradaci důležitého environmentálního polutantu proteinovým a metabolickým inženýrstvím
Annotation in Czech
Antropogenní sloučenina 1,2,3-trichlorpropan (TCP) je uvolňována do životního prostředí v důsledku výroby a použití jako rozpouštědlo a extrakční činidlo. Poločas rozpadu může v podzemních podmínkách dosahovat až 100 let a TCP přetrvává v prostředí podzemních vod dlouhou dobu. TCP je velmi odolný vůči biodegradaci v důsledku absence dehalogenázy efektivně přeměňující TCP na dichlorpropanol. Proteinovým inženýrstvím a řízenou evolucí jsme nedávno zkonstruovali dehalogenázu s významně lepší katalýzou tohoto biochemického kroku (Pavlová et al. 2009, Nat. Chem. Biol. 5: 727-733). Cílem tohoto projektu je sestavit kompletní biochemickou dráhu klonováním genů enzymů optimalizovaných proteinovým inženýrstvím do přírodních bakterií, vedoucí ke konstrukci bakterie degradující TCP. Tato bakterie bude růst na TCP jako jediném zdroji uhlíku a energie. Podle našich informací bude tato bakterie prvním mikroorganismem zkonstruovaným proteinovým a matabolickým inženýrstvím, použitelným k čištění vod kontaminovaných antropogenní sloučeninou v uzavřených průtočných systémech.
Scientific branches
R&D category
ZV - Basic research
CEP classification - main branch
DJ - Pollution and water control
CEP - secondary branch
CE - Biochemistry
CEP - another secondary branch
EI - Biotechnology and bionics
10511 - Environmental sciences (social aspects to be 5.7)
10608 - Biochemistry and molecular biology
10609 - Biochemical research methods
20801 - Environmental biotechnology
20802 - Bioremediation, diagnostic biotechnologies (DNA chips and biosensing devices) in environmental management
20803 - Environmental biotechnology related ethics
20901 - Industrial biotechnology
20902 - Bioprocessing technologies (industrial processes relying on biological agents to drive the process) biocatalysis, fermentation
20903 - Bioproducts (products that are manufactured using biological material as feedstock) biomaterials, bioplastics, biofuels, bioderived bulk and fine chemicals, bio-derived novel materials
30401 - Health-related biotechnology
30402 - Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction)
30403 - Technologies involving identifying the functioning of DNA, proteins and enzymes and how they influence the onset of disease and maintenance of well-being (gene-based diagnostics and therapeutic interventions [pharmacogenomics, gene-based therapeutics])
30404 - Biomaterials (as related to medical implants, devices, sensors)
30405 - Medical biotechnology related ethics
40401 - Agricultural biotechnology and food biotechnology
40402 - GM technology (crops and livestock), livestock cloning, marker assisted selection, diagnostics (DNA chips and biosensing devices for the early/accurate detection of diseases) biomass feedstock production technologies, biopharming
40403 - Agricultural biotechnology related ethics
Completed project evaluation
Provider evaluation
V - Vynikající výsledky projektu (s mezinárodním významem atd.)
Project results evaluation
The main objective of this project was construction and optimization of metabolic pathway for degradation of emerging environmental pollutant 1,2,3-trichloropropane (TCP) and its introduction into bacterial host cells. Apart from Escherichia coli host system, the pathway was also introduced into the soil bacterium Pseudomonas putida KT2440. The enzym haloalkane dehalogenase – was studied too.
Solution timeline
Realization period - beginning
Jan 1, 2012
Realization period - end
Dec 31, 2016
Project status
U - Finished project
Latest support payment
Apr 5, 2016
Data delivery to CEP
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data delivery code
CEP17-GA0-GA-U/03:1
Data delivery date
Jun 28, 2017
Finance
Total approved costs
8,980 thou. CZK
Public financial support
8,980 thou. CZK
Other public sources
0 thou. CZK
Non public and foreign sources
0 thou. CZK
Basic information
Recognised costs
8 980 CZK thou.
Public support
8 980 CZK thou.
100%
Provider
Czech Science Foundation
CEP
DJ - Pollution and water control
Solution period
01. 01. 2012 - 31. 12. 2016