Engineering co-culture system for production of apigetrin in Escherichia coli

Autor:	Amit Kumar Chaudhary, Duong Van Cuong, Nguyen Huy Thuan, Nguyen Xuan Cuong
Rok vydání:	2018
Předmět:	0301 basic medicine Chalcone synthase Spectrometry Mass Electrospray Ionization Coumaric Acids Chemistry Pharmaceutical DNA Recombinant Bioengineering medicine.disease_cause Applied Microbiology and Biotechnology Mixed Function Oxygenases Metabolic engineering Industrial Microbiology 03 medical and health sciences chemistry.chemical_compound Coenzyme A Ligases Escherichia coli medicine Apigenin Isomerases Chromatography High Pressure Liquid chemistry.chemical_classification DNA ligase biology Chemistry Temperature Protein engineering Industrial microbiology Coculture Techniques De novo synthesis 030104 developmental biology Metabolic Engineering Biochemistry biology.protein Propionates Acyltransferases Plasmids Biotechnology
Zdroj:	Journal of Industrial Microbiology and Biotechnology. 45:175-185
ISSN:	1476-5535 1367-5435
DOI:	10.1007/s10295-018-2012-x
Popis:	Microbial cells have extensively been utilized to produce value-added bioactive compounds. Based on advancement in protein engineering, DNA recombinant technology, genome engineering, and metabolic remodeling, the microbes can be re-engineered to produce industrially and medicinally important platform chemicals. The emergence of co-culture system which reduces the metabolic burden and allows parallel optimization of the engineered pathway in a modular fashion restricting the formation of undesired byproducts has become an alternative way to synthesize and produce bioactive compounds. In this study, we present genetically engineered E. coli-based co-culture system to the de novo synthesis of apigetrin (APG), an apigenin-7-O-β-d-glucopyranoside of apigenin. The culture system consists of an upstream module including 4-coumarate: CoA ligase (4CL), chalcone synthase, chalcone flavanone isomerase (CHS, CHI), and flavone synthase I (FNSI) to synthesize apigenin (API) from p-coumaric acid (PCA). Whereas, the downstream system contains a metabolizing module to enhance the production of UDP-glucose and expression of glycosyltransferase (PaGT3) to convert API into APG. To accomplish this improvement in titer, the initial inoculum ratio of strains for making the co-culture system, temperature, and media component was optimized. Following large-scale production, a yield of 38.5 µM (16.6 mg/L) of APG was achieved. In overall, this study provided an efficient tool to synthesize bioactive compounds in microbial cells.
Databáze:	OpenAIRE
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