One-Pot Synthesis of Ginsenoside Rh2 and Bioactive Unnatural Ginsenoside by Coupling Promiscuous Glycosyltransferase from <i>Bacillus subtilis</i> 168 to Sucrose Synthase

Dai, Longhai; Liu, Can; Li, Jiao; Dong, Caixia; Yang, Jiangang; Dai, Zhaoli; Zhang, Xueli; Sun, Yi

doi:10.1021/acs.jafc.8b00597

Cited by 72 publications

(61 citation statements)

References 36 publications

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“…UDP is involved in the formation of UDPG in the SuSy-catalyzed hydrolysis of sucrose. In addition, as a byproduct of the UGT-catalyzed reaction, a high concentration of UDP could inhibit UGT activity [29]. Only 1.30 mM Cy7G was formed in the presence of 0.1 mM UDP.…”

Section: Optimization Of Ugt88e18-susy Cascade Reaction For Cy7g Syntmentioning

confidence: 99%

Biocatalytic Synthesis of Calycosin-7-O-β-D-Glucoside with Uridine Diphosphate–Glucose Regeneration System

Min

et al. 2020

Catalysts

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Calycosin-7-O-β-D-glucoside (Cy7G) is one of the principal components of Radix astragali. This isoflavonoid glucoside is regarded as an indicator to assess the quality of R. astragali and exhibits diverse pharmacological activities. In this study, uridine diphosphate-dependent glucosyltransferase (UGT) UGT88E18 was isolated from Glycine max and expressed in Escherichia coli. Recombinant UGT88E18 could selectively and effectively glucosylate the C7 hydroxyl group of calycosin to synthesize Cy7G. A one-pot reaction by coupling UGT88E18 to sucrose synthase (SuSy) from G. max was developed. The UGT88E18–SuSy cascade reaction could recycle the costly uridine diphosphate glucose (UDPG) from cheap sucrose and catalytic amounts of uridine diphosphate (UDP). The important factors for UGT88E18–SuSy cascade reaction, including UGT88E18/SuSy ratios, different temperatures, and pH values, different concentrations of dimethyl sulfoxide (DMSO), UDP, sucrose, and calycosin, were optimized. We produced 10.5 g L−1 Cy7G in the optimal reaction conditions by the stepwise addition of calycosin. The molar conversion of calycosin was 97.5%, with a space–time yield of 747 mg L−1 h−1 and a UDPG recycle of 78 times. The present study provides a new avenue for the efficient and cost-effective semisynthesis of Cy7G and other valuable isoflavonoid glucosides by UGT–SuSy cascade reaction.

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Section: Optimization Of Ugt88e18-susy Cascade Reaction For Cy7g Syntmentioning

confidence: 99%

Biocatalytic Synthesis of Calycosin-7-O-β-D-Glucoside with Uridine Diphosphate–Glucose Regeneration System

Min

et al. 2020

Catalysts

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“…Antiproliferative activity of ginsenosides Rd12 and Rg3 against colon cancer cells Lovo, gastric cancer cell line SNU719, and lung cancer cell line DMS5 was performed as described previously [23]. The cancer cells were seeded into the 96-well plates with a density of 1 × 10 5 cells/mL and incubated for 24 h. Subsequently, the cancer cell lines were incubated with variable concentrations of ginsenosides Rd12 and Rg3 for another 48 h. MTT solution (15 µL, 5 mg/mL) was added to the wells and the cancer cell lines were incubated for 4 h. Finally, the medium was removed and DMSO (150 µL) was added into the plates.…”

Section: Cell Viability Assaymentioning

confidence: 99%

“…An Agilent 1260 HPLC system equipped with a C18 column was applied to analyze the samples as described previously [23]. The column was eluted with phase A (double-distilled water/0.1% formic acid, v/v) and phase B (chromatography-grade acetonitrile/0.1% formic acid, v/v).…”

Section: Hplc and Hplc-esi-ms Analysis Of The Glycosylated Productmentioning

confidence: 99%

“…A scale-up reaction (100 mL) was conducted for the preparation of unnatural ginsenoside Rd12. Ginsenoside Rd12 was purified by an Agilent 1260 preparative HPLC system as described previously [23]. The preparative column was eluted with double-distilled water (phase A) and chromatography-grade methanol (phase B) using a flow rate of 10 mL/min and a gradient of 25-100% phase B in 0-60 min.…”

Section: Structural Elucidation Of Ginsenoside Rd12 By Nmr Spectroscopymentioning

confidence: 99%

“…Bs-YjiC could glucosylate the C3, C6, and C12 hydroxyl groups of PPT to synthesize unnatural ginsenosides [10]. Bs-YjiC could also catalyze a continuous two-step glucosylation of the C3 and C12 hydroxyl groups of PPD to synthesize ginsenoside Rh2 and an unnatural ginsenoside F12 [23]. However, the regiospecificity of Bs-YjiC toward other ginsenosides have not been thoroughly elucidated.…”

Section: Introductionmentioning

confidence: 99%

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Biocatalytic Synthesis of a Novel Bioactive Ginsenoside Using UDP-Glycosyltransferase from Bacillus subtilis 168

et al. 2020

Catalysts

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Ginsenoside Rg3 is a bioactive compound from Panax ginseng and exhibits diverse notable biological properties. Glycosylation catalyzed by uridine diphosphate-dependent glycosyltransferase (UGT) is the final biosynthetic step of ginsenoside Rg3 and determines its diverse pharmacological activities. In the present study, promiscuous UGT Bs-YjiC from Bacillus subtilis 168 was expressed in Escherichia coli and purified via one-step nickel chelate affinity chromatography. The in vitro glycosylation reaction demonstrated Bs-Yjic could selectively glycosylate the C12 hydroxyl group of ginsenoside Rg3 to synthesize an unnatural ginsenoside Rd12. Ginsenoside Rd12 was about 40-fold more water-soluble than that of ginsenoside Rg3 (90 μM). Furthermore, in vitro cytotoxicity of ginsenoside Rd12 against diverse cancer cells was much stronger than that of ginsenoside Rg3. Our studies report the UGT-catalyzed synthesis of unnatural ginsenoside Rd12 for the first time. Ginsenoside Rd12 with antiproliferative activity might be further exploited as a potential anticancer drug.

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FlavonoidC‐Glycosyltransferases: Function, Evolutionary Relationship, Catalytic Mechanism and Protein Engineering

Dai

Chen

et al. 2021

ChemBioEng Reviews

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C‐glycosylflavonoids are polyphenolic phytochemicals that are ubiquitously distributed in the plant kingdom and confer a wealth of health‐promoting benefits to human body. These compounds harbor one or more sugar moieties that are attached to various regions of the flavonoid scaffold through rigid C–C glycosidic bonds. The diverse structures of C‐glycosylflavonoids are granted by the prominent reactions of flavonoid C‐glycosyltransferases (CGTs). This review gives an extensive overview of CGTs that are involved in the biosynthesis of C‐glycosylflavonoids, focusing on their evolutionary relationship and substrate specificity. Furthermore, structural insights into the catalytic mechanism and protein engineering of flavonoid CGTs are summarized. This review provides an important guidance for identification of novel flavonoid CGTs, biosynthesis of C‐glycosylflavonoids, and elucidation of the function‐structure relationship of flavonoid CGTs.

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One-Pot Synthesis of Ginsenoside Rh2 and Bioactive Unnatural Ginsenoside by Coupling Promiscuous Glycosyltransferase from Bacillus subtilis 168 to Sucrose Synthase

Cited by 72 publications

References 36 publications

Biocatalytic Synthesis of Calycosin-7-O-β-D-Glucoside with Uridine Diphosphate–Glucose Regeneration System

Biocatalytic Synthesis of Calycosin-7-O-β-D-Glucoside with Uridine Diphosphate–Glucose Regeneration System

Biocatalytic Synthesis of a Novel Bioactive Ginsenoside Using UDP-Glycosyltransferase from Bacillus subtilis 168

FlavonoidC‐Glycosyltransferases: Function, Evolutionary Relationship, Catalytic Mechanism and Protein Engineering

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