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Malonyl-CoA is an important central metabolite for the production of diverse valuable chemicals including natural products, but its intracellular availability is often limited due to the competition with essential cellular metabolism. Several malonyl-CoA biosensors have been developed for high-throughput screening of targets increasing the malonyl-CoA pool. However, they are limited for use only in Escherichia coli and Saccharomyces cerevisiae and require multiple signal transduction steps. Here we report development of a colorimetric malonyl-CoA biosensor applicable in three industrially important bacteria: E. coli , Pseudomonas putida , and Corynebacterium glutamicum RppA, a type III polyketide synthase producing red-colored flaviolin, was repurposed as a malonyl-CoA biosensor in E. coli Strains with enhanced malonyl-CoA accumulation were identifiable by the colorimetric screening of cells showing increased red color. Other type III polyketide synthases could also be repurposed as malonyl-CoA biosensors. For target screening, a 1,858 synthetic small regulatory RNA library was constructed and applied to find 14 knockdown gene targets that generally enhanced malonyl-CoA level in E. coli These knockdown targets were applied to produce two polyketide (6-methylsalicylic acid and aloesone) and two phenylpropanoid (resveratrol and naringenin) compounds. Knocking down these genes alone or in combination, and also in multiple different E. coli strains for two polyketide cases, allowed rapid development of engineered strains capable of enhanced production of 6-methylsalicylic acid, aloesone, resveratrol, and naringenin to 440.3, 30.9, 51.8, and 103.8 mg/L, respectively. The malonyl-CoA biosensor developed here is a simple tool generally applicable to metabolic engineering of microorganisms to achieve enhanced production of malonyl-CoA-derived chemicals., Competing Interests: Conflict of interest statement: S.Y.L., D.Y., and S.M.Y. declare that the sRNA technology described here is patent filed including, but not limited to, KR 10-1575587, US 9388417, EP 13735942.8, CN 201380012767.X, KR 10-1690780, KR 10-1750855, US 15317939, CN 201480081132.X for potential commercialization. Additionally, S.Y.L. and D.Y. have conflict of interest as the RppA biosensor technology is of commercial interest and is patent filed including, but not limited to KR 10-2018-0066323.

Bio-based production of industrially important chemicals provides an eco-friendly alternative to current petrochemical-based processes. Because of the limited supply of fossil fuel reserves, various technologies utilizing microbial host strains for the sustainable production of platform chemicals from renewable biomass have been developed. Corynebacterium glutamicum is a non-pathogenic industrial microbial species traditionally used for L-glutamate and L-lysine production. It is a promising species for industrial production of bio-based chemicals because of its flexible metabolism that allows the utilization of a broad spectrum of carbon sources and the production of various amino acids. Classical breeding, systems, synthetic biology, and metabolic engineering approaches have been used to improve its applications, ranging from traditional amino-acid production to modern biorefinery systems for production of value-added platform chemicals. This review describes recent advances in the development of genetic engineering tools and techniques for the establishment and optimization of metabolic pathways for bio-based production of major C2-C6 platform chemicals using recombinant C. glutamicum.

Abstract: The fermentative production of platform chemicals in biorefineries is a sustainable alternative to current petroleum‐refining processes. Industrial microorganisms, such as Escherichia coli, Saccharomyces cerevisiae, and Corynebacterium glutamicum, have been engineered as microbial cell factories that are able to utilize biomass for the production of value‐added platform chemicals and polymers. Compared to E. coli and S. cerevisiae, C. glutamicum displays weak carbon catabolite repression and can co‐utilize mixed sugars as carbon sources, without any significant growth retardation. Pathways for the utilization of alternative carbon sources, such as d‐xylose and l‐arabinose from lignocellulosic biomass, lactose and galactose from whey, glycerol from biodiesel, and methanol from natural gas refineries, have been evaluated for chemical production. However, the application of C. glutamicum in biorefineries is limited because it does not secrete hydrolases for the efficient utilization of cellulose, xylan, and starch from lignocellulosic and starch biomass. To solve the limitation, C. glutamicum has been engineered for the consolidated bioprocessing of biomass by the heterologous expression of amylolytic and cellulolytic enzymes. Recently, C. glutamicum has been extensively engineered for polyamide monomer production owing to its ability to produce l‐lysine and l‐glutamate. This review summarizes recent advances in the development of C. glutamicum strains that can utilize renewable biomass resources for the production of industrially important chemicals. It highlights recent progress in metabolic engineering for the production of polyamide monomers. © 2018 Society of Chemical Industry and John Wiley & Sons, Ltd [ABSTRACT FROM AUTHOR]

Bio-based production of industrially important chemicals provides an eco-friendly alternative to current petrochemical-based processes. Because of the limited supply of fossil fuel reserves, various technologies utilizing microbial host strains for the sustainable production of platform chemicals from renewable biomass have been developed. Corynebacterium glutamicum is a non-pathogenic industrial microbial species traditionally used for l-glutamate and l-lysine production. It is a promising species for industrial production of bio-based chemicals because of its flexible metabolism that allows the utilization of a broad spectrum of carbon sources and the production of various amino acids. Classical breeding, systems, synthetic biology, and metabolic engineering approaches have been used to improve its applications, ranging from traditional amino-acid production to modern biorefinery systems for production of value-added platform chemicals. This review describes recent advances in the development of genetic engineering tools and techniques for the establishment and optimization of metabolic pathways for bio-based production of major C2-C6 platform chemicals using recombinant C. glutamicum. [ABSTRACT FROM AUTHOR]