scholarly journals Glycosylation of Ganoderic Acid G by Bacillus Glycosyltransferases

2021 ◽  
Vol 22 (18) ◽  
pp. 9744
Author(s):  
Jiumn-Yih Wu ◽  
Hsiou-Yu Ding ◽  
Tzi-Yuan Wang ◽  
Yun-Rong Zhang ◽  
Te-Sheng Chang
Keyword(s):  
Bacillus Subtilis ◽  
Ganoderma Lucidum ◽  
Aqueous Solubility ◽  
Triterpenoid Saponin ◽  
Bioactive Components ◽  
Ganoderic Acid ◽  
Triterpenoid Saponins ◽  
Medicinal Fungus ◽  
Bacillus Subtilis Atcc ◽  
New Compounds

Ganoderma lucidum is a medicinal fungus abundant in triterpenoids, its primary bioactive components. Although numerous Ganoderma triterpenoids have already been identified, rare Ganoderma triterpenoid saponins were recently discovered. To create novel Ganoderma saponins, ganoderic acid G (GAG) was selected for biotransformation using four Bacillus glycosyltransferases (GTs) including BtGT_16345 from the Bacillus thuringiensis GA A07 strain and three GTs (BsGT110, BsUGT398, and BsUGT489) from the Bacillus subtilis ATCC 6633 strain. The results showed that BsUGT489 catalyzed the glycosylation of GAG to GAG-3-o-β-glucoside, while BsGT110 catalyzed the glycosylation of GAG to GAG-26-o-β-glucoside, which showed 54-fold and 97-fold greater aqueous solubility than that of GAG, respectively. To our knowledge, these two GAG saponins are new compounds. The glycosylation specificity of the four Bacillus GTs highlights the possibility of novel Ganoderma triterpenoid saponin production in the future.

scholarly journals One-Pot Bi-Enzymatic Cascade Synthesis of Novel Ganoderma Triterpenoid Saponins

Catalysts ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 580
Author(s):  
Te-Sheng Chang ◽  
Chien-Min Chiang ◽  
Tzi-Yuan Wang ◽  
Yu-Li Tsai ◽  
Yu-Wei Wu ◽  
...  
Keyword(s):  
High Performance ◽  
Aqueous Solubility ◽  
Major Product ◽  
Cyclodextrin Glucanotransferase ◽  
Bioactive Constituents ◽  
Ganoderic Acid ◽  
One Pot ◽  
Triterpenoid Saponins ◽  
Medicinal Fungus ◽  
Triterpenoid Glycosides

Ganoderma lucidum is a medicinal fungus whose numerous triterpenoids are its main bioactive constituents. Although hundreds of Ganoderma triterpenoids have been identified, Ganoderma triterpenoid glycosides, also named triterpenoid saponins, have been rarely found. Ganoderic acid A (GAA), a major Ganoderma triterpenoid, was synthetically cascaded to form GAA-15-O-β-glucopyranoside (GAA-15-G) by glycosyltransferase (BtGT_16345) from Bacillus thuringiensis GA A07 and subsequently biotransformed into a series of GAA glucosides by cyclodextrin glucanotransferase (Toruzyme® 3.0 L) from Thermoanaerobacter sp. The optimal reaction conditions for the second-step biotransformation of GAA-15-G were found to be 20% of maltose; pH 5; 60 °C. A series of GAA glucosides (GAA-G2, GAA-G3, and GAA-G4) could be purified with preparative high-performance liquid chromatography (HPLC) and identified by mass and nucleic magnetic resonance (NMR) spectral analysis. The major product, GAA-15-O-[α-glucopyranosyl-(1→4)-β-glucopyranoside] (GAA-G2), showed over 4554-fold higher aqueous solubility than GAA. The present study demonstrated that multiple Ganoderma triterpenoid saponins could be produced by sequential actions of BtGT_16345 and Toruzyme®, and the synthetic strategy that we proposed might be applied to many other Ganoderma triterpenoids to produce numerous novel Ganoderma triterpenoid saponins in the future.

scholarly journals A New Triterpenoid Glucoside from a Novel Acidic Glycosylation of Ganoderic Acid A via Recombinant Glycosyltransferase of Bacillus subtilis

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3457 ◽  
Author(s):  
Te-Sheng Chang ◽  
Chien-Min Chiang ◽  
Yu-Han Kao ◽  
Jiumn-Yih Wu ◽  
Yu-Wei Wu ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
High Performance ◽  
Culture Collection ◽  
Subtilis Strain ◽  
Resonance Spectroscopy ◽  
Ganoderic Acid ◽  
Acidic Solutions ◽  
Medicinal Fungus ◽  
Bacillus Subtilis Atcc ◽  
Acidic Conditions

Ganoderic acid A (GAA) is a bioactive triterpenoid isolated from the medicinal fungus Ganoderma lucidum. Our previous study showed that the Bacillus subtilis ATCC (American type culture collection) 6633 strain could biotransform GAA into compound (1), GAA-15-O-β-glucoside, and compound (2). Even though we identified two glycosyltransferases (GT) to catalyze the synthesis of GAA-15-O-β-glucoside, the chemical structure of compound (2) and its corresponding enzyme remain elusive. In the present study, we identified BsGT110, a GT from the same B. subtilis strain, for the biotransformation of GAA into compound (2) through acidic glycosylation. BsGT110 showed an optimal glycosylation activity toward GAA at pH 6 but lost most of its activity at pH 8. Through a scaled-up production, compound (2) was successfully isolated using preparative high-performance liquid chromatography and identified to be a new triterpenoid glucoside (GAA-26-O-β-glucoside) by mass and nuclear magnetic resonance spectroscopy. The results of kinetic experiments showed that the turnover number (kcat) of BsGT110 toward GAA at pH 6 (kcat = 11.2 min−1) was 3-fold higher than that at pH 7 (kcat = 3.8 min−1), indicating that the glycosylation activity of BsGT110 toward GAA was more active at acidic pH 6. In short, we determined that BsGT110 is a unique GT that plays a role in the glycosylation of triterpenoid at the C-26 position under acidic conditions, but loses most of this activity under alkaline ones, suggesting that acidic solutions may enhance the catalytic activity of this and similar types of GTs toward triterpenoids.

Production of a new triterpenoid disaccharide saponin from sequential glycosylation of ganoderic acid A by 2 Bacillus glycosyltransferases

2021 ◽  
Vol 85 (3) ◽  
pp. 687-690
Author(s):  
Te-Sheng Chang ◽  
Chien-Min Chiang ◽  
Jiumn-Yih Wu ◽  
Yu-Li Tsai ◽  
Huei-Ju Ting
Keyword(s):  
Ganoderma Lucidum ◽  
Aqueous Solubility ◽  
Ganoderic Acid ◽  
Medicinal Fungus

ABSTRACT Ganoderic acid A (GAA) is a lanostane-type triterpenoid, isolated from medicinal fungus Ganoderma lucidum, and possesses multiple bioactivities. In the present study, GAA was sequentially biotransformed by 2 recently discovered Bacillus glycosyltransferases (GT), BtGT_16345 and BsGT110, and the final product was purified and identified as a new compound, GAA-15,26-O-β-diglucoside, which showed 1024-fold aqueous solubility than GAA.

Functions of reactive oxygen species in apoptosis and ganoderic acid biosynthesis in Ganoderma lucidum

2019 ◽  
Vol 366 (23) ◽  
Author(s):  
Jing Zhu ◽  
Fengli Wu ◽  
Sining Yue ◽  
Chen Chen ◽  
Shuqi Song ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Ganoderma Lucidum ◽  
Reactive Oxygen ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Activity Levels ◽  
Oxygen Species ◽  
Ganoderic Acid ◽  
Medicinal Fungus ◽  
Intracellular Ros ◽  
Ga Biosynthesis

ABSTRACT Ganoderma lucidum is a medicinal fungus that is widely used in traditional medicine. Fungal PacC is recognized as an important transcription factor that functions during adaptation to environmental pH, fungal development and secondary metabolism. Previous studies have revealed that GlPacC plays important roles in mycelial growth, fruiting body development and ganoderic acid (GA) biosynthesis. In this study, using a terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay, we found that the apoptosis level was increased when PacC was silenced. The transcript and activity levels of caspase-like proteins were significantly increased in the PacC-silenced (PacCi) strains compared with the control strains. Silencing PacC also resulted in an increased reactive oxygen species (ROS) levels (∼2-fold) and decreased activity levels of enzymes involved in the antioxidant system. Further, we found that the intracellular ROS levels contributed to apoptosis and GA biosynthesis. Adding N-acetyl-cysteine and vitamin C decreased intracellular ROS and resulted in the inhibition of apoptosis in the PacCi strains. Additionally, the GA biosynthesis was different between the control strains and the PacCi strains after intracellular ROS was eliminated. Taken together, the findings showed that silencing PacC can result in an intracellular ROS burst, which increases cell apoptosis and GA biosynthesis levels. Our study provides novel insight into the functions of PacC in filamentous fungi.

scholarly journals Discovery of Ganoderma lucidum triterpenoids as potential inhibitors against Dengue virus NS2B-NS3 protease

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Shiv Bharadwaj ◽  
Kyung Eun Lee ◽  
Vivek Dhar Dwivedi ◽  
Umesh Yadava ◽  
Aleksha Panwar ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Ganoderma Lucidum ◽  
Denv Infection ◽  
Ganoderic Acid ◽  
Medicinal Fungus ◽  
Infection Inhibition ◽  
Dynamics Simulations ◽  
Potential Inhibitors ◽  
Ns3 Protease

AbstractDengue virus (DENV) infection causes serious health problems in humans for which no drug is currently available. Recently, DENV NS2B-NS3 protease has been proposed as a primary target for anti-dengue drug discovery due to its important role in new virus particle formation by conducting DENV polyprotein cleavage. Triterpenoids from the medicinal fungus Ganoderma lucidum have been suggested as pharmacologically bioactive compounds and tested as anti-viral agents against various viral pathogens including human immunodeficiency virus. However, no reports are available concerning the anti-viral activity of triterpenoids from Ganoderma lucidum against DENV. Therefore, we employed a virtual screening approach to predict the functional triterpenoids from Ganoderma lucidum as potential inhibitors of DENV NS2B-NS3 protease, followed by an in vitro assay. From in silico analysis of twenty-two triterpenoids of Ganoderma lucidum, four triterpenoids, viz. Ganodermanontriol (−6.291 kcal/mol), Lucidumol A (−5.993 kcal/mol), Ganoderic acid C2 (−5.948 kcal/mol) and Ganosporeric acid A (−5.983 kcal/mol) were predicted to be viral protease inhibitors by comparison to reference inhibitor 1,8-Dihydroxy-4,5-dinitroanthraquinone (−5.377 kcal/mol). These results were further studied for binding affinity and stability using the molecular mechanics/generalized Born surface area method and Molecular Dynamics simulations, respectively. Also, in vitro viral infection inhibition suggested that Ganodermanontriol is a potent bioactive triterpenoid.

scholarly journals Effect of 26-Oxygenosterols from Ganoderma lucidum and Their Activity as Cholesterol Synthesis Inhibitors

2005 ◽  
Vol 71 (7) ◽  
pp. 3653-3658 ◽  
Author(s):  
Hassan Hajjaj ◽  
Catherine Macé ◽  
Matthew Roberts ◽  
Peter Niederberger ◽  
Laurent B. Fay
Keyword(s):  
Ganoderma Lucidum ◽  
Cholesterol Synthesis ◽  
Biological Effects ◽  
Cholesterol Biosynthesis ◽  
Blood Cholesterol ◽  
Ganoderic Acid ◽  
Isolation And Identification ◽  
Medicinal Fungus ◽  
Lower Blood Cholesterol

ABSTRACT Ganoderma lucidum is a medicinal fungus belonging to the Polyporaceae family which has long been known in Japan as Reishi and has been used extensively in traditional Chinese medicine. We report the isolation and identification of the 26-oxygenosterols ganoderol A, ganoderol B, ganoderal A, and ganoderic acid Y and their biological effects on cholesterol synthesis in a human hepatic cell line in vitro. We also investigated the site of inhibition in the cholesterol synthesis pathway. We found that these oxygenated sterols from G. lucidum inhibited cholesterol biosynthesis via conversion of acetate or mevalonate as a precursor of cholesterol. By incorporation of 24,25-dihydro-[24,25-3H2]lanosterol and [3-3H]lathosterol in the presence of ganoderol A, we determined that the point of inhibition of cholesterol synthesis is between lanosterol and lathosterol. These results demonstrate that the lanosterol 14α-demethylase, which converts 24,25-dihydrolanosterol to cholesterol, can be inhibited by the 26-oxygenosterols from G. lucidum. These 26-oxygenosterols could lead to novel therapeutic agents that lower blood cholesterol.

scholarly journals Enzyme-Catalyzed Glycosylation of Curcumin and Its Analogues by Glycosyltransferases from Bacillus subtilis ATCC 6633

Catalysts ◽  
2019 ◽  
Vol 9 (9) ◽  
pp. 734 ◽  
Author(s):  
Yatian Cheng ◽  
Jian Zhang ◽  
Yan Shao ◽  
Yixiang Xu ◽  
Haixia Ge ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Food Additives ◽  
Aqueous Solubility ◽  
Clinical Applications ◽  
Polyphenolic Compound ◽  
Subtilis Atcc ◽  
Curcumin Analogue ◽  
Naturally Occurring ◽  
Enhanced Solubility ◽  
Bacillus Subtilis Atcc

Curcumin is a naturally occurring polyphenolic compound that is commonly used in both medicine and food additives, but its low aqueous solubility and poor bioavailability hinder further clinical applications. For assessing the effect of the glycosylation of curcumin on its aqueous solubility, two glycosyltransferase genes (BsGT1 and BsGT2) were cloned from the genome of the strain Bacillus subtilis ATCC 6633 and over-expressed in Escherichia coli. Then, the two glycosyltransferases were purified, and their glycosylation capacity toward curcumin and its two analogues was verified. The results showed that both BsGT1 and BsGT2 could convert curcumin and its two analogues into their glucosidic derivatives. Then, the structures of the derivatives were characterized as curcumin 4′-O-β-D-glucoside and two new curcumin analogue monoglucosides namely, curcumoid-O-α-D-glucoside (2a) and 3-pentadienone-O-α-D-glucoside (3a) by nuclear magnetic resonance (NMR) spectroscopy. Subsequently, the dissolvability of curcumin 4′-O-β-D-glucoside was measured to be 18.78 mg/L, while its aglycone could not be determined. Furthermore, the optimal catalyzing conditions and kinetic parameters of BsGT1 and BsGT2 toward curcumin were determined, which showed that the Kcat value of BsGT1 was about 2.6-fold higher than that of BsGT2, indicating that curcumin is more favored for BsGT2. Our findings effectively apply the enzymatic approach to obtain glucoside derivatives with enhanced solubility.

scholarly journals Sequential Biotransformation of Antcin K by Bacillus subtilis ATCC 6633

Catalysts ◽  
2018 ◽  
Vol 8 (9) ◽  
pp. 349 ◽  
Author(s):  
Te-Sheng Chang ◽  
Chien-Min Chiang ◽  
Yi-Yun Siao ◽  
Jiumn-Yih Wu
Keyword(s):  
Nuclear Magnetic Resonance ◽  
High Performance Liquid Chromatography ◽  
Bacillus Subtilis ◽  
High Performance ◽  
Fruiting Bodies ◽  
Subtilis Atcc ◽  
Bacillus Subtilis Atcc ◽  
Triterpenoid Glycosides ◽  
New Compounds ◽  
First Time

The biotransformation of antcin K, a major ergostane triterpenoid from the fruiting bodies of Antrodia cinnamomea, by Bacillus subtilis (B. subtilis) ATCC 6633 was studied. Four metabolites from the biotransformation were isolated with preparative high-performance liquid chromatography and identified as 25S-antcin K 26-O-β-glucoside, 25R-antcin K 26-O-β-glucoside, 25S-antcin K 26-O-β-(6′-O-succinyl)-glucoside, and 25R-antcin K 26-O-β-(6′-O-succinyl)-glucoside with mass and nuclear magnetic resonance spectral analysis. By using either 25S-antcin K 26-O-β-glucoside or 25R-antcin K 26-O-β-glucoside as the biotransformation precursor, it was proven that 25S-antcin K 26-O-β-(6′-O-succinyl)-glucoside and 25R-antcin K 26-O-β-(6′-O-succinyl)-glucoside were biotransformed from 25S-antcin K 26-O-β-glucoside and 25R-antcin K 26-O-β-glucoside, respectively. To the best of our knowledge, this is the first study on the glycosylation of triterpenoids from A. cinnamomea, and the first time the succinylation of triterpenoid glycosides by microorganisms has been found. In addition, all four antcin K glucoside derivatives are new compounds.

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

2018 ◽  
Vol 19 (11) ◽  
pp. 3469 ◽  
Author(s):  
Te-Sheng Chang ◽  
Jiumn-Yih Wu ◽  
Tzi-Yuan Wang ◽  
Kun-Yuan Wu ◽  
Chien-Min Chiang
Keyword(s):  
Bacillus Subtilis ◽  
Liquid Chromatography ◽  
High Performance ◽  
Ultra Performance Liquid Chromatography ◽  
Culture Collection ◽  
Resonance Spectroscopy ◽  
Uridine Diphosphate ◽  
Ganoderic Acid ◽  
Subtilis Atcc ◽  
Bacillus Subtilis Atcc

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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