High-Yield Resveratrol Production in Engineered Escherichia coli

Abstract

Plant polyphenols have been the subject of several recent scientific investigations since many of the molecules in this class have been found to be highly active in the human body, with a plethora of healthpromoting activities against a variety of diseases, including heart disease, diabetes, and cancer, and with even the potential to slow aging. Further development of these potent natural therapeutics hinges on the formation of robust industrial production platforms designed using specifically selected as well as engineered protein sources along with the construction of optimal expression platforms. In this work, we first report the investigation of various stilbene synthases from an array of plant species considering structure-activity relationships, their expression efficiency in microorganisms, and their ability to synthesize resveratrol. Second, we looked into the construct environment of recombinantly expressed stilbene synthases, including different promoters, construct designs, and host strains, to create an Escherichia coli strain capable of producing superior resveratrol titers sufficient for commercial usage. Further improvement of metabolic capabilities of the recombinant strain aimed at improving the intracellular malonyl-coenzyme A pool, a resveratrol precursor, resulted in a final improved titer of 2.3 g/liter resveratrol.

A reliable protocol for the stable transformation of non-embryogenic cells cultures of grapevine (Vitis vinifera L.) and Taxus x media

“…Elicitation is an empirical approach done to stimulate cell cultures to produce secondary metabolites; for instance, grapevine cell cultures elicited with methylated cyclodextrins (MBCD) alone [21] or combined with methyl jasmonate (MeJA) [22,23] has proven an efficient system for the bioproduction of t-R, with 30% overall conversion of feeding sucrose carbon into resveratrol carbon [24]. Metabolic engineering has been currently applied to microorganisms as an alternative way for the bioproduction of t-R and analogues [25][26][27][28] due to the obvious advantages over plant cell cultures to develop transformed strains and to upscale the productive process. Yet the yield of elicited grapevine cell cultures is still unbeaten.…”

Metabolic Engineering of Microorganisms for the Production of Natural Compounds

“…Conversion of phenylalanine into cinnamic acid and tyrosine into p-coumaric acid are known to be rate-controlling steps because PAL/TAL have relatively low activities. [119] In a study of resveratrol production in S. cerevisiae, TAL from Rhodobacter sphaeroides did not show any activity toward the production of p-coumaric acid, whereas coexpression of 4CL from A. thaliana and STS from Vitis vinifera successfully produced resveratrol from the fed p-coumaric acid. [120] After analyzing the codon frequency of TAL, two regions with high percentages of low-frequency codons were found.…”

Development of bio-based fine chemical production through synthetic bioengineering

“…Recently, the stilbene resveratrol was biosynthesized at high yields (2.3 g/L) by an E. coli strain [114] that was genetically engineered to enhance the production of malonyl-CoA to increase the supply of malonyl-CoA, which is used to synthesize fatty acids (see [115] for a review). Such metabolic engineering may further improve production.…”