Synthesis of novel oligomeric anionic alkyl glycosides using laccase/TEMPO oxidation and cyclodextrin glucanotransferase (CGTase)‐catalyzed transglycosylation

Ngo, Ngoc T.N.; Linares-Pastén, Javier A.; Grey, Carl; Adlercreutz, Patrick

doi:10.1002/bit.27770

Cited by 8 publications

(3 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is of great interest to note that both enzymes efficiently catalyze the transglucosylation of baicalin. This confirms the recent findings of Ngo et al [17], namely, that glucuronide can be glucosylated by the CGTase from Thermoanaerobacter sp. In addition to the high yield of transglusylation afforded by Toruzyme ® , the enzyme was more tolerant to pH variations than CGTase Amano in the range from 4 to 6 (Supplementary Data S1).…”

Section: Glucosylation Of Baicalin Baicalein and Baicalein-7-o-glucosidesupporting

confidence: 93%

See 1 more Smart Citation

Enzymatic Synthesis of α-Glucosyl-Baicalin through Transglucosylation via Cyclodextrin Glucanotransferase in Water

et al. 2023

View full text Add to dashboard Cite

Baicalin is a biologically active flavone glucuronide with poor water solubility that can be enhanced via glucosylation. In this study, the transglucosylation of baicalin was successfully achieved with CGTases from Thermoanaerobacter sp. and Bacillus macerans using α-cyclodextrin as a glucosyl donor. The synthesis of baicalin glucosides was optimized with CGTase from Thermoanaerobacter sp. Enzymatically modified baicalin derivatives were α-glucosylated with 1 to 17 glucose moieties. The two main glucosides were identified as Baicalein-7-O-α-D-Glucuronidyl-(1→4′)-O-α-D-Glucopyranoside (BG1) and Baicalein-7-O-α-D-Glucuronidyl-(1→4′)-O-α-D-Maltoside (BG2), thereby confirming recent findings reporting that glucuronyl groups are acceptors of this CGTase. Optimized conditions allowed for the attainment of yields above 85% (with a total glucoside content higher than 30 mM). BG1 and BG2 were purified via centrifugal partition chromatography after an enrichment through deglucosylation with amyloglucosidase. Transglucosylation increased the water solubility of BG1 by a factor of 188 in comparison to that of baicalin (molar concentrations), while the same value for BG2 was increased by a factor of 320. Finally, BG1 and BG2 were evaluated using antioxidant and anti-glycation assays. Both glucosides presented antioxidant and anti-glycation properties in the same order of magnitude as that of baicalin, thereby indicating their potential biological activity.

show abstract

Section: Glucosylation Of Baicalin Baicalein and Baicalein-7-o-glucosidesupporting

confidence: 93%

“…Ngo et al recently demonstrated that CGTase from Thermoanaeroacter sp. is able to transfer a glucosyl unit from α-cyclodextrin to octyl β-D-glucuronic acid on the C4 position of glucuronide [17], suggesting that baicalin could be an acceptor of this enzyme.…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Synthesis of α-Glucosyl-Baicalin through Transglucosylation via Cyclodextrin Glucanotransferase in Water

et al. 2023

View full text Add to dashboard Cite

show abstract

“…Some examples with main specificities for α-glucosyl groups, include cyclomaltodextrin glucanotransferases (CGTases in family GH13) [5,6]. CGTases can also be used for the synthesis of alkyl glycosides [7,8].…”

Section: Introductionmentioning

confidence: 99%

Modeled 3D-Structures of Proteobacterial Transglycosylases from Glycoside Hydrolase Family 17 Give Insight in Ligand Interactions Explaining Differences in Transglycosylation Products

et al. 2021

Self Cite

View full text Add to dashboard Cite

The structures of glycoside hydrolase family 17 (GH17) catalytic modules from modular proteins in the ndvB loci in Pseudomonas aeruginosa (Glt1), P. putida (Glt3) and Bradyrhizobium diazoefficiens (previously B. japonicum) (Glt20) were modeled to shed light on reported differences between these homologous transglycosylases concerning substrate size, preferred cleavage site (from reducing end (Glt20: DP2 product) or non-reducing end (Glt1, Glt3: DP4 products)), branching (Glt20) and linkage formed (1,3-linkage in Glt1, Glt3 and 1,6-linkage in Glt20). Hybrid models were built and stability of the resulting TIM-barrel structures was supported by molecular dynamics simulations. Catalytic amino acids were identified by superimposition of GH17 structures, and function was verified by mutagenesis using Glt20 as template (i.e., E120 and E209). Ligand docking revealed six putative subsites (−4, −3, −2, −1, +1 and +2), and the conserved interacting residues suggest substrate binding in the same orientation in all three transglycosylases, despite release of the donor oligosaccharide product from either the reducing (Glt20) or non-reducing end (Glt1, Gl3). Subsites +1 and +2 are most conserved and the difference in release is likely due to changes in loop structures, leading to loss of hydrogen bonds in Glt20. Substrate docking in Glt20 indicate that presence of covalently bound donor in glycone subsites −4 to −1 creates space to accommodate acceptor oligosaccharide in alternative subsites in the catalytic cleft, promoting a branching point and formation of a 1,6-linkage. The minimum donor size of DP5, can be explained assuming preferred binding of DP4 substrates in subsite −4 to −1, preventing catalysis.

show abstract

Kinetics guided engineering of cyclodextrin glycosyltransferase with enhanced intermolecular transglycosylation activity

Chen,

Ju,

Dong

et al. 2024

AIChE Journal

View full text Add to dashboard Cite

Cyclodextrin glycosyltransferase (CGTase) catalyzes intermolecular transglycosylation through either disproportionation or cyclization‐coupling pathway. Kinetics analysis reveals that the hesperidin glycosylation process catalyzed by a CGTase variant (M1) is primarily accomplished through the disproportionation pathway. The cyclization‐coupling pathway exhibits a lower reaction rate and competitively consumes glycosyl donor and yield byproducts that impair disproportionation. Under the guidance of reaction kinetics, mutagenesis was targeted at residues in the −3, +1, and +2 subsites, known to control the selectivity between disproportionation and cyclization. A quadruple variant was identified with 2.9 times hesperidin glycosylation activity compared to M1, and 20.3 times compared to the wild‐type. Kinetic analysis reveals a fourfold improvement of kcat/KmA for disproportionation and an 85.5% reduction in kcat/Km for cyclization after mutagenesis. Binding free energy analysis further confirms that the mutagenesis favors the binding of hesperidin, and destabilizes the binding of cyclodextrin.

show abstract

Synthesis of novel oligomeric anionic alkyl glycosides using laccase/TEMPO oxidation and cyclodextrin glucanotransferase (CGTase)‐catalyzed transglycosylation

Cited by 8 publications

References 28 publications

Enzymatic Synthesis of α-Glucosyl-Baicalin through Transglucosylation via Cyclodextrin Glucanotransferase in Water

Enzymatic Synthesis of α-Glucosyl-Baicalin through Transglucosylation via Cyclodextrin Glucanotransferase in Water

Modeled 3D-Structures of Proteobacterial Transglycosylases from Glycoside Hydrolase Family 17 Give Insight in Ligand Interactions Explaining Differences in Transglycosylation Products

Kinetics guided engineering of cyclodextrin glycosyltransferase with enhanced intermolecular transglycosylation activity

Contact Info

Product

Resources

About