Engineering Saccharomyces cerevisiae with the deletion of endogenous glucosidases for the production of flavonoid glucosides

Wang, Huimin; Yang, Yan; Lin, Lin; Zhou, Wenlong; Liu, Minzhi; Cheng, Kedi; Wang, Wei

doi:10.1186/s12934-016-0535-2

Cited by 47 publications

(31 citation statements)

References 36 publications

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“…Another future area of development for these enzymes includes metabolic engineering, which was not addressed herein but is an active area of research (Anasontzis, Kourtoglou, Villas-Boas, Hatzinikolaou, & Christakopoulos, 2016; K. S. Lee et al, 2011; Yanfeng Liu et al, 2014; Nic Lochlainn & Caffrey, 2009; Rodríguez-Díaz, Rubio-del-Campo, & Yebra, 2012; Wang et al, 2016; Zabala, Braña, Flórez, Salas, & Méndez, 2013). New biochemical and structural data summarized in this review have shed light on intriguing mechanistic features of the PHMs that were previously unappreciated.…”

Section: Discussionmentioning

confidence: 99%

Biology, Mechanism, and Structure of Enzymes in the α- d -Phosphohexomutase Superfamily

Stiers

Muenks

Beamer

2017

Advances in Protein Chemistry and Structural Biology

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Enzymes in the α-D-phosphohexomutases (PHM) superfamily catalyze the reversible conversion of phosphosugars, such as glucose 1-phosphate and glucose 6-phosphate. These reactions are fundamental to primary metabolism across the kingdoms of life, and are required for a myriad of cellular processes, ranging from exopolysaccharide production to protein glycosylation. The subject of extensive mechanistic characterization during the latter half of the twentieth century, these enzymes have recently benefitted from biophysical characterization, including X-ray crystallography, NMR, and hydrogen-deuterium exchange studies. This work has provided new insights into the unique catalytic mechanism of the superfamily, shed light on the molecular determinants of ligand recognition, and revealed the evolutionary conservation of conformational flexibility. Novel associations with inherited metabolic disease and the pathogenesis of bacterial infections have emerged, spurring renewed interest in the long-appreciated functional roles of these enzymes.

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Section: Discussionmentioning

confidence: 99%

Biology, Mechanism, and Structure of Enzymes in the α- d -Phosphohexomutase Superfamily

Stiers

Muenks

Beamer

2017

Advances in Protein Chemistry and Structural Biology

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“…In general, fewer glucosylation target sites were found per aglycone than those reported for MhGT1 (6). However, this comparison is made more complicated by the different assay formats (in vitro enzyme assays for MhGT1), the possible variability of the uptake of the aglycones by the cells expressing BbGT86-BbMT85, or the partial degradation of some glycosides in vivo (see the next section) (28).…”

Section: The Bbgt86-bbmt85 Pair Acts As a Broad Spectrum Methylglucosmentioning

confidence: 96%

“…In host tissues, small-molecule glycosides are frequently broken down to their constituent aglycones by glycoside hydrolases of the host or its associated microbiota. Some aglycones show reduced bioactivities (28), but conversely, glycoside hydrolysis may also be exploited in a prodrug strategy to release a highly active aglycone in situ. Modulation of the sensitivity of glycosides to host glycoside hydrolases is thus important to optimize the pharmacokinetic and pharmacodynamic (PK/PD) properties of drugs.…”

Section: Significancementioning

confidence: 99%

Methylglucosylation of aromatic amino and phenolic moieties of drug-like biosynthons by combinatorial biosynthesis

Xie

Zhang

Wang

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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Glycosylation is a prominent strategy to optimize the pharmacokinetic and pharmacodynamic properties of drug-like small-molecule scaffolds by modulating their solubility, stability, bioavailability, and bioactivity. Glycosyltransferases applicable for "sugarcoating" various small-molecule acceptors have been isolated and characterized from plants and bacteria, but remained cryptic from filamentous fungi until recently, despite the frequent use of some fungi for whole-cell biocatalytic glycosylations. Here, we use bioinformatic and genomic tools combined with heterologous expression to identify a glycosyltransferase-methyltransferase (GT-MT) gene pair that encodes a methylglucosylation functional module in the ascomycetous fungus The GT is the founding member of a family nonorthologous to characterized fungal enzymes. Using combinatorial biosynthetic and biocatalytic platforms, we reveal that this GT is a promiscuous enzyme that efficiently modifies a broad range of drug-like substrates, including polyketides, anthraquinones, flavonoids, and naphthalenes. It yields both- and -glucosides with remarkable regio- and stereospecificity, a spectrum not demonstrated for other characterized fungal enzymes. These glucosides are faithfully processed by the dedicated MT to afford 4--methylglucosides. The resulting "unnatural products" show increased solubility, while representative polyketide methylglucosides also display increased stability against glycoside hydrolysis. Upon methylglucosidation, specific polyketides were found to attain cancer cell line-specific antiproliferative or matrix attachment inhibitory activities. These findings will guide genome mining for fungal GTs with novel substrate and product specificities, and empower the efficient combinatorial biosynthesis of a broad range of natural and unnatural glycosides in total biosynthetic or biocatalytic formats.

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“…Lastly, the production levels in the 5-L bioreactor reached 1.7 g/L. Use of S. cerevisiae in synthesis of glycosylated natural products has been reported widely [16,28,29]. The findings from this study may be used to synthesize other glycosylated natural products using S. cerevisiae, and the cost of production can be reduced by using glycerin to synthesize natural products.…”

Section: Resultsmentioning

confidence: 64%

“…Wang et al overexpressed the gene PGM2 and UGP1 genes encoding phosphoglucomutase 2 and UTPglucose-1-phosphate uridylyltransferase 1, respectively, in S. cerevisiae that are involved in the glucose synthesis system of uridine diphosphate. The conversion rate of scutellarein into its glucoside increased from 75 to 92% [16].…”

Section: Open Accessmentioning

confidence: 98%

Promotion of compound K production in Saccharomyces cerevisiae by glycerol

et al. 2020

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Background: Ginsenoside compound K (CK), one of the primary active metabolites of protopanaxadiol-type ginsenosides, is produced by the intestinal flora that degrade ginseng saponins and exhibits diverse biological properties such as anticancer, anti-inflammatory, and anti-allergic properties. However, it is less abundant in plants. Therefore, enabling its commercialization by construction of a Saccharomyces cerevisiae cell factory is of considerable significance. Results:We induced overexpression of PGM2, UGP1, and UGT1 genes in WLT-MVA5, and obtained a strain that produces ginsenoside CK. The production of CK at 96 h was 263.94 ± 2.36 mg/L, and the conversion rate from protopanaxadiol (PPD) to ginsenoside CK was 64.23 ± 0.41%. Additionally, it was observed that the addition of glycerol was beneficial to the synthesis of CK. When 20% glucose (C mol) in the YPD medium was replaced by the same C mol glycerol, CK production increased to 384.52 ± 15.23 mg/L, which was 45.68% higher than that in YPD medium, and the PPD conversion rate increased to 77.37 ± 3.37% as well. As we previously observed that ethanol is beneficial to the production of PPD, ethanol and glycerol were fed simultaneously in the 5-L bioreactor fed fermentation, and the CK levels reached 1.70 ± 0.16 g/L. Conclusions:In this study, we constructed an S. cerevisiae cell factory that efficiently produced ginsenoside CK. Glycerol effectively increased the glycosylation efficiency of PPD to ginsenoside CK, guiding higher carbon flow to the synthesis of ginsenosides and effectively improving CK production. CK production attained in a 5-L bioreactor was 1.7 g/L after simultaneous feeding of glycerol and ethanol.

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Engineering Saccharomyces cerevisiae with the deletion of endogenous glucosidases for the production of flavonoid glucosides

Cited by 47 publications

References 36 publications

Biology, Mechanism, and Structure of Enzymes in the α- d -Phosphohexomutase Superfamily

Biology, Mechanism, and Structure of Enzymes in the α- d -Phosphohexomutase Superfamily

Methylglucosylation of aromatic amino and phenolic moieties of drug-like biosynthons by combinatorial biosynthesis

Promotion of compound K production in Saccharomyces cerevisiae by glycerol

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