Biosynthesis of plant-specific alkaloids tetrahydroprotoberberines in engineered Escherichia coli

Abstract

Tetrahydroprotoberberines (THPBs), mainly sourced from the medicinal plants of genus Stephania and Corydalis, exhibit a variety of pharmacological activities. Plant-sourced THPBs are limited by low yields, and their chemical synthesis...

Construction of ajmalicine and sanguinarine de novo biosynthetic pathways using stable integration sites in yeast

“…Moreover, when we diverted more carbon flux toward the branch point intermediate ( S )‐reticuline, the production of sanguinarine was only slightly increased (Figure 3b), indicating that reticuline oxidase (BBE) has become the bottleneck for sanguinarine biosynthesis in the engineered yeast strain (SAN006). Zhao et al found that BBE was mainly expressed as inclusion bodies in Escherichia coli and the functional expression level could be significantly increased by fusion expression with maltose binding protein and glutathione S‐transferase (W. Zhao et al, 2021). In addition to fusion expression with hydrophilic chaperons, we can also focus on searching for orthologous BBE candidates (X. Liu et al, 2017; Xu et al, 2021) or improving BBE activity via protein engineering to optimize the sanguinarine biosynthetic pathway.…”

Construction of biocatalytic cascades for the synthesis of benzylisoquinoline alkaloids from p-coumaric acid derivatives and dopamine

“…Moreover, the biocatalytic cascades could serve as interesting starting points for further improvement, such as adding methyltransferases, to synthesize methylated BIAs. 17…”

“…The conversions for (S)-14 and (S)-15 were lower than that of (S)-3 (90%). HPLC analysis showed that large amounts of styrenes (4-vinylguaiacol (16) and 4-vinylcatechol (17)) accumulated in the reaction mixtures (Fig. S4 †), and could be further converted to epoxides by StyAB.…”

“…Moreover, the biocatalytic cascades could serve as interesting starting points for further improvement, such as adding methyltransferases, to synthesize methylated BIAs. 17 Synthesis of (S)-norcoclaurine from dopamine (2) and lignocellulosic biomass hydrolysate Lignocellulosic biomass is the most abundant carbohydrate resource in the world, and its effective utilization is meaningful in terms of environmental and economic feasibility. [49][50][51] Aromatics derived from lignocellulosic biomass, such as 4, have been used as precursors for the synthesis of various value-added natural products, including naringenin, gastrodin, and salidroside.…”

“…11,12 The most-used in vivo method uses simple sugars as starting materials by constructing heterologous pathways in microbes, such as Saccharomyces cerevisiae or Escherichia coli. [13][14][15][16][17] This strategy was designed to simulate the natural biosynthetic pathway in plants, in which the synthesis of BIAs starts from the condensation of 4-hydroxyphenylacetaldehyde (1) and dopamine (2) to form (S)-norcoclaurine [(S)-3] catalyzed by norcoclaurine synthase (NCS) (Scheme 1A). However, this process contains multi-step enzyme reactions, which often leads to a low conversion rate.…”