Synthetic design of microbial consortia with beneficial interactions for efficient production of value-added biochemicals from non-fermentable substrates

2

Carbon flux redistribution at malonyl-CoA node for efficient conversion of 3-hydroxypropionic acid from acetate in engineered Escherichia coli

2

Circuit-guided synthetic acclimation for improving biochemical production by microbial consortia

2

Circuit-guided Synthetic Acclimation of Microbial Consortia for Improving Biochemical Production

2

Bicontinuous Interfacially Jammed Emulsion Gels (bijels) as Media for Enabling Enzymatic Reactive Separation of a Highly Water Insoluble Substrate

Abstract Although enzymes are efficient catalysts capable of converting various substrates into desired products with high specificity under mild conditions, their effectiveness as catalysts is substantially reduced when substrates are poorly water-soluble. In this study, to expedite the enzymatic conversion of a hydrophobic substrate, we use a bicontinuous interfacially jammed emulsion gel (bijel) which provides large interfacial area between two immiscible liquids: oil and water. Using lipase-catalyzed hydrolysis of tributyrin as a model reaction in a batch mode, we show that bijels can be used as media to enable enzymatic reaction. The bijel system gives a four-fold increase in the initial reaction rate in comparison to a stirred biphasic medium. Our results demonstrate that bijels are powerful biphasic reaction media to accelerate enzymatic reactions with various hydrophobic reagents. This work also demonstrates that bijels can potentially be used as reaction media to enable continuous reactive separations

Design of mutualistic microbial consortia for stable conversion of carbon monoxide to value-added chemicals

Carbon monoxide (CO) is a promising carbon source for producing value-added biochemicals via microbial fermentation. However, its microbial conversion has been challenging because of difficulties in genetic engineering of CO-utilizing microorganisms and, more importantly, maintaining CO consumption which is negatively affected by the toxicity of CO and accumulated byproducts. To overcome these issues, we devised mutualistic microbial consortia, co-culturing Eubacterium limosum and genetically engineered Escherichia coli for the production of 3-hydroxypropionic acid (3-HP) and itaconic acid (ITA). During the co-culture, E. limosum assimilated CO and produced acetate, a toxic by-product, while E. coli utilized acetate as a sole carbon source. We found that this mutualistic interaction dramatically stabilized and improved CO consumption of E. limosum compared to monoculture. Consequently, the improved CO consumption allowed successful production of 3-HP and ITA from CO. This study is the first demonstration of value-added biochemical production from CO using a microbial consortium. Moreover, it suggests that synthetic mutualistic microbial consortium can serve as a powerful platform for the valorization of CO.11Nsciescopu

Plug -in repressor library for Precise metabolic flux regulation in E. coli

2

Plug-in repressor library for precise regulation of metabolic flux for efficient production of biochemicals in E. coli

2

Efficient Conversion of Acetate to 3-Hydroxypropionic Acid by Engineered <i>Escherichia coli</i>

Acetate, which is an abundant carbon source, is a potential feedstock for microbial processes that produce diverse value-added chemicals. In this study, we produced 3-hydroxypropionic acid (3-HP) from acetate with engineered Escherichia coli. For the efficient conversion of acetate to 3-HP, we initially introduced heterologous mcr (encoding malonyl-CoA reductase) from Chloroflexus aurantiacus. Then, the acetate assimilating pathway and glyoxylate shunt pathway were activated by overexpressing acs (encoding acetyl-CoA synthetase) and deleting iclR (encoding the glyoxylate shunt pathway repressor). Because a key precursor malonyl-CoA is also consumed for fatty acid synthesis, we decreased carbon flux to fatty acid synthesis by adding cerulenin. Subsequently, we found that inhibiting fatty acid synthesis dramatically improved 3-HP production (3.00 g/L of 3-HP from 8.98 g/L of acetate). The results indicated that acetate can be used as a promising carbon source for microbial processes and that 3-HP can be produced from acetate with a high yield (44.6% of the theoretical maximum yield)

Plug-in Repressor Library for Metabolic Flux Redistribution in Escherichia coli

2