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@article{1213471, author = {Kurumbang, Nagendra Prasad and Dvořák, Pavel and Bendl, Jaroslav and Brezovský, Jan and Prokop, Zbyněk and Damborský, Jiří}, article_number = {3}, doi = {http://dx.doi.org/10.1021/sb400147n}, keywords = {activity; enantioselectivity; kinetic modeling; protein and metabolic engineering;synthetic pathway; toxicity}, language = {eng}, issn = {2161-5063}, journal = {ACS Synthetic Biology}, title = {Computer-Assisted Engineering of Synthetic Pathway for Biodegradation of Toxic Persistent Pollutant.}, volume = {3}, year = {2014} }
TY - JOUR ID - 1213471 AU - Kurumbang, Nagendra Prasad - Dvořák, Pavel - Bendl, Jaroslav - Brezovský, Jan - Prokop, Zbyněk - Damborský, Jiří PY - 2014 TI - Computer-Assisted Engineering of Synthetic Pathway for Biodegradation of Toxic Persistent Pollutant. JF - ACS Synthetic Biology VL - 3 IS - 3 SP - 172-181 EP - 172-181 SN - 21615063 KW - activity KW - enantioselectivity KW - kinetic modeling KW - protein and metabolic engineering;synthetic pathway KW - toxicity N2 - Anthropogenic halogenated compounds were unknown to Nature until the industrial revolution, and microorganisms have not had sufficient time to evolve enzymes for their degradation. The lack of efficient enzymes and natural pathways can be addressed through a combination of protein and metabolic engineering. We have assembled a synthetic route for conversion of the highly toxic and recalcitrant 1,2,3-trichloropropane to glycerol in Escherichia coli, and used it for a systematic study of pathway bottlenecks. Optimal ratios of enzymes for the maximal production of glycerol, and minimal toxicity of metabolites were predicted using a mathematical model. The strains containing the expected optimal ratios of enzymes were constructed and characterized for their viability and degradation efficiency. Excellent agreement between predicted and experimental data was observed. The validated model was used to quantitatively describe the kinetic limitations of currently available enzyme variants, and predict improvements required for further pathway optimization. This highlights the potential of forward engineering of microorganisms for the degradation of toxic anthropogenic compounds. ER -
KURUMBANG, Nagendra Prasad, Pavel DVOŘÁK, Jaroslav BENDL, Jan BREZOVSKÝ, Zbyněk PROKOP a Jiří DAMBORSKÝ. Computer-Assisted Engineering of Synthetic Pathway for Biodegradation of Toxic Persistent Pollutant. \textit{ACS Synthetic Biology}. 2014, roč.~3, č.~3, s.~172-181. ISSN~2161-5063. Dostupné z: https://dx.doi.org/10.1021/sb400147n.
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