Publication Details
Computer-Assisted Engineering of Synthetic Pathway for Biodegradation of Toxic Persistent Pollutant
Dvořák Pavel
Bendl Jaroslav, Ing., Ph.D.
Brezovský Jan
Prokop Zbyněk
Damborský Jiří, prof. Mgr., Dr. (UMEL)
activity, enantioselectivity, kinetic modeling, protein and metabolic engineering, synthetic pathway, toxicity
Anthropogenic halogenated compounds had been unknown to the nature until industrial revolution and microorganisms have not had sufficient time to evolve enzymes for their degradation. The lack of efficient enzymes and natural metabolic pathways can be addressed by combination of protein and metabolic engineering. We have assembled a synthetic pathway for conversion of highly toxic and recalcitrant 1,2,3-trichloropropane to glycerol in Escherichia coli and used it for systematic study of pathway bottlenecks. The optimal ratios of enzymes for maximal production of glycerol and minimal toxicity of metabolites were predicted using a mathematical model. The strains containing predicted ratios of enzymes were constructed and characterized for their viability and degradation efficiency. An excellent agreement between predicted and experimental data has been observed. The validated model has been used to quantitatively describe the kinetic limitations of currently available enzyme variants and predict the improvements required for further pathway optimization. Presented concept highlights the potential of rigorous rational engineering of microorganisms for degradation of toxic anthropogenic compounds.
@article{BUT133483,
author="Nagendra {Kurumbang} and Pavel {Dvořák} and Jaroslav {Bendl} and Jan {Brezovský} and Zbyněk {Prokop} and Jiří {Damborský}",
title="Computer-Assisted Engineering of Synthetic Pathway for Biodegradation of Toxic Persistent Pollutant",
journal="ACS Synthetic Biology",
year="2014",
volume="3",
number="3",
pages="172--181",
doi="10.1021/sb400147n",
issn="2161-5063",
url="http://pubs.acs.org/doi/pdf/10.1021/sb400147n"
}