Engineering yeast for the production of breviscapine by genomic analysis and synthetic biology approaches

Abstract

The flavonoid extract from Erigeron breviscapus, breviscapine, has increasingly been used to treat cardio- and cerebrovascular diseases in China for more than 30 years, and plant supply of E. breviscapus is becoming insufficient to satisfy the growing market demand. Here we report an alternative strategy for the supply of breviscapine by building a yeast cell factory using synthetic biology. We identify two key enzymes in the biosynthetic pathway (flavonoid-7-O-glucuronosyltransferase and flavone-6-hydroxylase) from E. breviscapus genome and engineer yeast to produce breviscapine from glucose. After metabolic engineering and optimization of fed-batch fermentation, scutellarin and apigenin-7-O-glucuronide, two major active ingredients of breviscapine, reach to 108 and 185 mg l–1, respectively. Our study not only introduces an alternative source of these valuable compounds, but also provides an example of integrating genomics and synthetic biology knowledge for metabolic engineering of natural compounds.

Reconfigured Cyanogenic Glucoside Biosynthesis in Eucalyptus cladocalyx Involves a Cytochrome P450 CYP706C55

“…CYP706C55 was identified as the second enzyme in the prunasin biosynthetic pathway in E. cladocalyx. Three CYP706 members from other plant species have been functionally characterized previously and demonstrated to catalyze oxygenation reactions: CYP706B1 from cotton (Gossypium arboruem; Luo et al, 2001), CYP706M1 from Alaska cedar (Callitropsis nootkatensis), and a CYP706X from Erigeron breviscapus (Liu et al, 2018). In contrast, CYP706C55 catalyzes an unusual CYP reaction, which is the dehydration of phenylacetaldoxime to phenylacetonitrile.…”

Expanding the repertoire of aromatic chemicals by microbial production

“…trans-Cinnamic acid and p-hydroxycinnamic acid serve as starting compounds for the production of many commercially valuable chemicals, such as styrene, 4-hydroxystyrene, caffeic acid and flavonoids (Figs 3 and 4). [79][80][81] Liu et al 79 reported the production of breviscapine (one of the flavonoids) derived from an extended L-Phe pathway in engineered yeast in which the two major active ingredients of breviscapine reached 108 and 185 mg L −1 , respectively. 46 trans-Cinnamic acid also can be converted into p-hydroxycinnamic acid catalyzed by cinnamate 4-hydroxylase (C4H).…”

“…To solve these problems, the microbial production of these compounds has been studied extensively. [79][80][81] Liu et al 79 reported the production of breviscapine (one of the flavonoids) derived from an extended L-Phe pathway in engineered yeast in which the two major active ingredients of breviscapine reached 108 and 185 mg L −1 , respectively.…”

Comparative Genome Analysis ofScutellaria baicalensisandScutellaria barbataReveals the Evolution of Active Flavonoid Biosynthesis

“…In S. baicalensis , the biosynthetic genes of the root-specific compounds baicalein and norwogonin have been functionally identified, providing an important basis for studying the biosynthesis and regulation of the natural products that make up Huang Qin [14,15]. Recently, the in vitro production of baicalein and scutellarein in Escherichia coli and Saccharomyces cerevisiae has been carried out based on the guidance of synthetic biology [16,17], but the metabolic engineering of these compounds still faces considerable challenges, including the discovery and optimization of biological components. The Salvia miltiorrhiza genome from the Lamiaceae family provides useful information associated with secondary metabolism for the rapid functional identification of biosynthetic and regulatory genes [18−23].…”