Synthetic Biology Approaches to Engineer Saccharomyces cerevisiae towards the Industrial Production of Valuable Polyphenolic Compounds

Abstract

Polyphenols are plant secondary metabolites with diverse biological and potential therapeutic activities such as antioxidant, anti-inflammatory and anticancer, among others. However, their extraction from the native plants is not enough to satisfy the increasing demand for this type of compounds. The development of microbial cell factories to effectively produce polyphenols may represent the most attractive solution to overcome this limitation and produce high amounts of these bioactive molecules. With the advances in the synthetic biology field, the development of efficient microbial cell factories has become easier, largely due to the development of the molecular biology techniques and by the identification of novel isoenzymes in plants or simpler organisms to construct the heterologous pathways. Furthermore, efforts have been made to make the process more profitable through improvements in the host chassis. In this review, advances in the production of polyphenols by genetically engineered Saccharomyces cerevisiae as well as by synthetic biology and metabolic engineering approaches to improve the production of these compounds at industrial settings are discussed.

Keywords: Saccharomyces cerevisiae; heterologous production; metabolic engineering; polyphenols biosynthesis; synthetic biology.

Figure 1

Chemical structure of some polyphenols.

Figure 2

Pathways involved in the polyphenol biosynthesis. 4CL—4-coumarate-CoA ligase; C4H—cinnamate-4-hydroxylase; CM—chorismate mutase; CS—chorismate synthase; DAHP—3-deoxy-D-arabino-heptulosonate-7-phosphate; DAHPS—3-deoxy-D-arabino-heptulosonate-7-phosphate synthase; DHQ—3-dehydroquinate dehydratase; DHQS—3-dehydroquinate synthase; E4P—D-erythrose-4-phosphate; EPSPS—5-enolpyruvylshikimate 3-phosphate synthase; HPP-AT—4-hydroxyphenylpyruvate aminotransferase; PAL—phenylalanine ammonia lyase; PDC—phenylpyruvate decarboxylase; PDH—prephenate dehydrogenase; PDT—prephenate dehydratase; PEP—Phosphoenolpyruvic acid; PPY-AT—phenylpyruvate aminotransferase; SDH—shikimate dehydrogenase; SK—shikimate kinase; TAL—tyrosine ammonia lyase.

Figure 3

Steps involved in the hydroxycinnamic acids, flavonoids, stilbenoids, polyphenolic amides and curcuminoids biosynthesis from p-coumaric acid. 3GT—anthocyanidin 3-O-glycosyltransferase; 4CL—4-coumarate-CoA ligase; AAT—anthocyanin acyltransferase; AMT—anthocyanin methyltransferase; ANS—anthocyanidin synthase; C3H—4-coumarate 3-hydroxylase; COMT—caffeic acid 3-O-methyltransferase; CCoAOMT—caffeoyl-CoA 3-O methyltransferase; CHI—chalcone isomerase; CHS—chalcone synthase; CS3′H—p-coumaroyl 5-O-shikimate 3′-hydroxylase; CST—p-coumaroyl shikimate transferase, CURS—curcumin synthase; CUS—curcuminoid synthase; DCS—diketide-CoA synthase; DFR—dihydroflavonol 4-reductase; F3H—flavanone 3-hydroxylase; F3′H—flavonoid 3′-hydroxylase; F3′5′H—flavonoid 3′5′-hydroxylase; FLS—flavonol synthase; FNS—flavone synthase; HCT—hydroxycinnamoyl-CoA: Shikimate/quinate hydroxycinnamoyl transferase; IFS—isoflavone synthase; LAR—leucoanthocyanidin 4-reductase; ROMT—resveratrol O-methyltransferase; STS—stilbene synthase.

Figure 4

Multi-copy integration of a heterologous pathway at delta sites (δ) mediated by the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated caspase 9 endonuclease (Cas9) [63]. DSB—Double strand break.