In vivo interpretation of model predicted inhibition in acrylate pathway engineered Lactococcus lactis
| Autor: | Priyadharshini Chandrasekran, Mark A. Eiteman, Ramalingam Subramanian, Padmapriya Ganapathyraman, Anitha Janet Roshni Yesudhas, Sowmiya Balasubramanian |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Bioengineering 01 natural sciences Applied Microbiology and Biotechnology Models Biological Enzyme catalysis Metabolic engineering 03 medical and health sciences chemistry.chemical_compound 010608 biotechnology Computer Simulation biology Lactococcus lactis Substrate (chemistry) biology.organism_classification Lactic acid Metabolic pathway 030104 developmental biology chemistry Biochemistry Acrylates Metabolic Engineering Dehydratase Steady state (chemistry) Biotechnology |
| Zdroj: | Biotechnology and bioengineeringREFERENCES. 117(12) |
| ISSN: | 1097-0290 |
| Popis: | To maximize the productivity of engineered metabolic pathway, in silico model is an established means to provide features of enzyme reaction dynamics. In our previous study, Escherichia coli engineered with acrylate pathway yielded low propionic acid titer. To understand the bottleneck behind this low productivity, a kinetic model was developed that incorporates the enzymatic reactions of the acrylate pathway. The resulting model was capable of simulating the fluxes reported under in vitro studies with good agreement, suggesting repression of propionyl-CoA transferase (Pct) by carboxylate metabolites as the main limiting factor for propionate production. Furthermore, the predicted flux control coefficients of the pathway enzymes under steady state conditions revealed that the control of flux is shared between Pct and lactoyl-CoA dehydratase. Increase in lactate concentration showed gradual decrease in flux control coefficients of Pct that in turn confirmed the control exerted by the carboxylate substrate. To interpret these in silico predictions under in vivo system, an organized study was conducted with a lactic acid bacteria strain engineered with acrylate pathway. Analysis reported a decreased product formation rate on attainment of inhibitory titer by suspected metabolites and supported the model. |
| Databáze: | OpenAIRE |
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