New Triterpenoid from Novel Triterpenoid 15-O-Glycosylation on Ganoderic Acid A by Intestinal Bacteria of Zebrafish

Chang, Te‐Sheng; Chiang, Chien-Min; Wang, Tzi‐Yuan; Lee, Chun-Hsien; Lee, Yu-Wen; Wu, Jiumn-Yih

doi:10.3390/molecules23092345

Cited by 13 publications

(14 citation statements)

References 39 publications

(48 reference statements)

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“…The mass data showed an [M − H] − ion peak at m / z : 677.67 in the ESI-MS spectrum, corresponding to C 36 H 53 O 12 that indicated the glucosylation of GAA. The structure of the metabolites was further confirmed as GAA-15- O -β-glucoside using NMR data, by comparison with that in our previously reported study [ 28 ]. The NMR spectrum of the two metabolites from either BsUGT398 or BsUGT489 are shown in Figure S3 -S16.The biotransformation process of GAA by either BsUGT398 or BsUGT489, where d -glucose was tentatively assumed, is shown in Figure 5 .…”

Section: Resultssupporting

confidence: 65%

Uridine Diphosphate-Dependent Glycosyltransferases from Bacillus subtilis ATCC 6633 Catalyze the 15-O-Glycosylation of Ganoderic Acid A

Chang

Wang

et al. 2018

IJMS

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Bacillus subtilis ATCC (American type culture collection) 6633 was found to biotransform ganoderic acid A (GAA), which is a major lanostane triterpenoid from the medicinal fungus Ganoderma lucidum. Five glycosyltransferase family 1 (GT1) genes of this bacterium, including two uridine diphosphate-dependent glycosyltransferase (UGT) genes, BsUGT398 and BsUGT489, were cloned and overexpressed in Escherichia coli. Ultra-performance liquid chromatography confirmed the two purified UGT proteins biotransform ganoderic acid A into a metabolite, while the other three purified GT1 proteins cannot biotransform GAA. The optimal enzyme activities of BsUGT398 and BsUGT489 were at pH 8.0 with 10 mM of magnesium or calcium ion. In addition, no candidates showed biotransformation activity toward antcin K, which is a major ergostane triterpenoid from the fruiting bodies of Antrodia cinnamomea. One biotransformed metabolite from each BsUGT enzyme was then isolated with preparative high-performance liquid chromatography. The isolated metabolite from each BsUGT was identified as ganoderic acid A-15-O-β-glucoside by mass and nuclear magnetic resonance spectroscopy. The two BsUGTs in the present study are the first identified enzymes that catalyze the 15-O-glycosylation of triterpenoids.

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Section: Resultssupporting

confidence: 65%

Uridine Diphosphate-Dependent Glycosyltransferases from Bacillus subtilis ATCC 6633 Catalyze the 15-O-Glycosylation of Ganoderic Acid A

Chang

Wang

et al. 2018

IJMS

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“…UGT73C11 can regiospecifically transfer a glucosyl moiety from UDP-glucose (UDPG) to the free C3 hydroxyl group of GA. YjiC1 was also demonstrated to glycosylate the free C3 hydroxyl group of GA with UDPG as a sugar donor. Microbial UGTs are generally recognized as effective tools for glycodiversification of natural products due to their broad acceptor tolerance [23]. Hence, we aim to seek microbial UGTs for biosynthesis of novel GA glycosides.…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Synthesis of Novel Glycyrrhizic Acid Glucosides Using a Promiscuous Bacillus Glycosyltransferase

Dai

Yang

et al. 2018

Catalysts

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Glycyrrhetinic acid (GA) and glycyrrhizin (GA-3-O-[β-d-glucuronopyranosyl-(1→2)-β-d-glucuronopyranoside], GL) are the major bioactive components of Glycyrrhiza uralensis and possess multifarious notable biological activities. UDP-glycosyltransferase (UGT)–catalyzed glycosylation remarkably extends the structural and functional diversification of GA-glycoside derivatives. In this study, six glucosides (1–6) of GA and GL were synthesized using a Bacillus subtilis 168–originated flexible UDP-glycosyltransferase Bs-YjiC. Bs-YjiC could transfer a glucosyl moiety from UDP-glucose to the free C3 hydroxyl and/or C30 carboxyl groups of GA and GL and further elongate the C30 glucosyl chain via a β-1-2-glycosidic bond. Glycosylation significantly increased the water solubility of these novel glucosides by 4–90 folds. In vitro assays showed that GA monoglucosides (1 and 2) showed stronger antiproliferative activity against human liver cancer cells HepG2 and breast cancer cells MCF-7 than that of GL and GL glucosides. These findings provide significant insights into the important role of promiscuous UGTs for the enzymatic synthesis of novel bioactive GA derivatives.

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“…Our previous study identified two Bacillus strains with the ability to biotransform GAA to GAA-15- O -β-glucoside, B. subtilis ATCC 6633 [4] and Bacillus sp. GA A07 [18]. To compare the biotransformation activity between the two strains, fermentation broths of the two strains fed with GAA were analyzed by ultra-performance liquid chromatography (UPLC) during cultivation.…”

Section: Resultsmentioning

confidence: 99%

“…Our previous study identified an intestinal bacterium of zebrafish, Bacillus sp. GA A07 strain, which could biotransform GAA into GAA-15- O -β-glucoside [18]. In order to identify the GTs of the GA A07 strain responsible for this triterpenoid biotransformation, the complete genome of the strain was resolved using both Nanopore long-read and BGI short-read sequencing technologies.…”

Section: Introductionmentioning

confidence: 99%

A Genome-Centric Approach Reveals a Novel Glycosyltransferase from the GA A07 Strain of Bacillus thuringiensis Responsible for Catalyzing 15-O-Glycosylation of Ganoderic Acid A

Chang

Wang

Hsueh

et al. 2019

IJMS

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Strain GA A07 was identified as an intestinal Bacillus bacterium of zebrafish, which has high efficiency to biotransform the triterpenoid, ganoderic acid A (GAA), into GAA-15-O-β-glucoside. To date, only two known enzymes (BsUGT398 and BsUGT489) of Bacillus subtilis ATCC 6633 strain can biotransform GAA. It is thus worthwhile to identify the responsible genes of strain GA A07 by whole genome sequencing. A complete genome of strain GA A07 was successfully assembled. A phylogenomic analysis revealed the species of the GA A07 strain to be Bacillus thuringiensis. Forty glycosyltransferase (GT) family genes were identified from the complete genome, among which three genes (FQZ25_16345, FQZ25_19840, and FQZ25_19010) were closely related to BsUGT398 and BsUGT489. Two of the three candidate genes, FQZ25_16345 and FQZ25_19010, were successfully cloned and expressed in a soluble form in Escherichia coli, and the corresponding proteins, BtGT_16345 and BtGT_19010, were purified for a biotransformation activity assay. An ultra-performance liquid chromatographic analysis further confirmed that only the purified BtGT_16345 had the key biotransformation activity of catalyzing GAA into GAA-15-O-β-glucoside. The suitable conditions for this enzyme activity were pH 7.5, 10 mM of magnesium ions, and 30 °C. In addition, BtGT_16345 showed glycosylation activity toward seven flavonoids (apigenein, quercetein, naringenein, resveratrol, genistein, daidzein, and 8-hydroxydaidzein) and two triterpenoids (GAA and antcin K). A kinetic study showed that the catalytic efficiency (kcat/KM) of BtGT_16345 was not significantly different compared with either BsUGT398 or BsUGT489. In short, this study identified BtGT_16345 from B. thuringiensis GA A07 is the catalytic enzyme responsible for the 15-O-glycosylation of GAA and it was also regioselective toward triterpenoid substrates.

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New Triterpenoid from Novel Triterpenoid 15-O-Glycosylation on Ganoderic Acid A by Intestinal Bacteria of Zebrafish

Cited by 13 publications

References 39 publications

Uridine Diphosphate-Dependent Glycosyltransferases from Bacillus subtilis ATCC 6633 Catalyze the 15-O-Glycosylation of Ganoderic Acid A

Uridine Diphosphate-Dependent Glycosyltransferases from Bacillus subtilis ATCC 6633 Catalyze the 15-O-Glycosylation of Ganoderic Acid A

Enzymatic Synthesis of Novel Glycyrrhizic Acid Glucosides Using a Promiscuous Bacillus Glycosyltransferase

A Genome-Centric Approach Reveals a Novel Glycosyltransferase from the GA A07 Strain of Bacillus thuringiensis Responsible for Catalyzing 15-O-Glycosylation of Ganoderic Acid A

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