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

Author:

Thuan Nguyen Huy1,Chaudhary Amit Kumar2,Van Cuong Duong3,Cuong Nguyen Xuan4

Affiliation:

1. grid.444918.4 Center for Molecular Biology Duy Tan University 03 Quang Trung Street, Haichau District Danang Vietnam

2. 0000 0001 0666 4105 grid.266813.8 Department of Pharmaceutical Sciences University of Nebraska Medical Center 68198 Omaha NE USA

3. 0000 0004 4911 9571 grid.472370.5 Faculty of Biotechnology and Food Technology Thainguyen University of Agriculture and Forestry Thainguyen Vietnam

4. 0000 0001 2105 6888 grid.267849.6 Lab of Marine Medicinal Materials, Institute of Marine Biochemistry (IMBC) Vietnam Academy of Science and Technology (VAST) 18 Hoang Quoc Viet Hanoi Vietnam

Abstract

Abstract 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.

Funder

National Foundation for Science and Technology Development of Vietnam

Publisher

Oxford University Press (OUP)

Subject

Applied Microbiology and Biotechnology,Biotechnology,Bioengineering

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