Controlled processivity in glycosyltransferases: A way to expand the enzymatic toolbox

Guidi, Chiara; Biarnés, Xevi; Planas, Antoni; Mey, Marjan De

doi:10.1016/j.biotechadv.2022.108081

Cited by 9 publications

(4 citation statements)

References 173 publications

(282 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The capacity of biocatalysts to process long polymers has also been related to enzyme oligomerization. For instance, processive glycosyltransferases can produce high‐molecular‐weight polysaccharides through the formation of transmembrane channels or extensive carbohydrate‐binding domains, or through enzyme multimerization, operating in an intersubunit mode (Guidi et al, 2023). An example of the latter case are also glycogenins, which form functional dimers consisting of two subunits (Bilyard et al, 2018), allowing both of the complexed enzymes to generate a maltosaccharide chain that may extend from one subunit to the other.…”

Section: Discussionmentioning

confidence: 99%

Structural and molecular insights into a bifunctional glycoside hydrolase 30 xylanase specific to glucuronoxylan

Pentari,

Kosinas,

Nikolaivits

et al. 2024

Biotech & Bioengineering

View full text Add to dashboard Cite

Glycoside hydrolase (GH) 30 family xylanases are enzymes of biotechnological interest due to their capacity to degrade recalcitrant hemicelluloses, such as glucuronoxylan (GX). This study focuses on a subfamily 7 GH30, TtXyn30A from Thermothelomyces thermophilus, which acts on GX in an “endo” and “exo” mode, releasing methyl‐glucuronic acid branched xylooligosaccharides (XOs) and xylobiose, respectively. The crystal structure of inactive TtXyn30A in complex with 23‐(4‐O‐methyl‐α‐D‐glucuronosyl)‐xylotriose (UXX), along with biochemical analyses, corroborate the implication of E233, previously identified as alternative catalytic residue, in the hydrolysis of decorated xylan. At the −1 subsite, the xylose adopts a distorted conformation, indicative of the Michaelis complex of TtXyn30AEE with UXX trapped in the semi‐functional active site. The most significant structural rearrangements upon substrate binding are observed at residues W127 and E233. The structures with neutral XOs, representing the “exo” function, clearly show the nonspecific binding at aglycon subsites, contrary to glycon sites, where the xylose molecules are accommodated via multiple interactions. Last, an unproductive ligand binding site is found at the interface between the catalytic and the secondary β‐domain which is present in all GH30 enzymes. These findings improve current understanding of the mechanism of bifunctional GH30s, with potential applications in the field of enzyme engineering.

show abstract

Section: Discussionmentioning

confidence: 99%

Structural and molecular insights into a bifunctional glycoside hydrolase 30 xylanase specific to glucuronoxylan

Pentari,

Kosinas,

Nikolaivits

et al. 2024

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…It is important to understand processive catalysis at the molecular level and in the context of the evolutionary history of processive enzymes and how they form reaction product trajectories after catalytic events are completed. In a broader context, it is remarkable to observe that the formation of exit routes for hydrolytic products in hydrolases [ 25 ] and polysaccharides in GT2 transferases [ 15 ] bear some common characteristics in how cavities and channels are formed and how C–H–π dispersive forces are involved in reactant binding.…”

Section: Discussionmentioning

confidence: 99%

Molecular mechanisms of processive glycoside hydrolases underline catalytic pragmatism

Hrmová

Schwerdt

2023

Biochemical Society Transactions

View full text Add to dashboard Cite

Processive and distributive catalysis defines the conversion continuum, thus underpinning the transformation of oligo- and polymeric substrates by enzymes. Distributive catalysis follows an association–transformation–dissociation pattern during the formation of enzyme–reactant complexes, whereas during processive catalysis, enzymes partner with substrates and complete multiple catalytic events before dissociation from an enzyme–substrate complex. Here, we focus on processive catalysis in glycoside hydrolases (GHs), which ensures efficient conversions of substrates with high precision, and has the advantage over distributive catalysis in efficiency. The work presented here examines a recent discovery of substrate-product-assisted processive catalysis in the GH3 family enzymes with enclosed pocket-shaped active sites. We detail how GH3 β-d-glucan glucohydrolases exploit a transiently formed lateral pocket for product displacement and reactants sliding (or translocation motion) through the catalytic site without dissociation, including movements during nanoscale binding/unbinding and sliding. The phylogenetic tree of putative 550 Archaean, bacterial, fungal, Viridiplantae, and Metazoan GH3 entries resolved seven lineages that corresponded to major substrate specificity groups. This analysis indicates that two tryptophan residues in plant β-d-glucan glucohydrolases that delineate the catalytic pocket, and infer broad specificity, high catalytic efficiency, and substrate-product-assisted processivity, have evolved through a complex evolutionary process, including horizontal transfer and neo-functionalisation. We conclude that the definition of thermodynamic and mechano-structural properties of processive enzymes is fundamentally important for theoretical and practical applications in bioengineering applicable in various biotechnologies.

show abstract

“…Therefore, further exploration of new ginsenoside-related UGTs with functions from original plants containing ginsenosides holds great value. Combined with protein engineering techniques such as rational enzyme design, 113 the catalytic function of UGTs can be further improved to provide genetic elements for the low-cost preparation of a wider variety of rare ginsenosides. Additionally, considering the substrate flexibility, UGTs from other species that can participate in the glycosylation of ginsenosides can serve as alternative choices.…”

Section: Journal Ofmentioning

confidence: 99%

Progress in Identification of UDP-Glycosyltransferases for Ginsenoside Biosynthesis

Yuan,

Li,

et al. 2024

J. Nat. Prod.

View full text Add to dashboard Cite

Ginsenosides, the primary pharmacologically active constituents of the Panax genus, have demonstrated a variety of medicinal properties, including anticardiovascular disease, cytotoxic, antiaging, and antidiabetes effects. However, the low concentration of ginsenosides in plants and the challenges associated with their extraction impede the advancement and application of ginsenosides. Heterologous biosynthesis represents a promising strategy for the targeted production of these natural active compounds. As representative triterpenoids, the biosynthetic pathway of the aglycone skeletons of ginsenosides has been successfully decoded. While the sugar moiety is vital for the structural diversity and pharmacological activity of ginsenosides, the mining of uridine diphosphate-dependent glycosyltransferases (UGTs) involved in ginsenoside biosynthesis has attracted a lot of attention and made great progress in recent years. In this paper, we summarize the identification and functional study of UGTs responsible for ginsenoside synthesis in both plants, such as Panax ginseng and Gynostemma pentaphyllum, and microorganisms including Bacillus subtilis and Saccharomyces cerevisiae. The UGT-related microbial cell factories for large-scale ginsenoside production are also mentioned. Additionally, we delve into strategies for UGT mining, particularly potential rapid screening or identification methods, providing insights and prospects. This review provides insights into the study of other unknown glycosyltransferases as candidate genetic elements for the heterologous biosynthesis of rare ginsenosides.

show abstract

Controlled processivity in glycosyltransferases: A way to expand the enzymatic toolbox

Cited by 9 publications

References 173 publications

Structural and molecular insights into a bifunctional glycoside hydrolase 30 xylanase specific to glucuronoxylan

Structural and molecular insights into a bifunctional glycoside hydrolase 30 xylanase specific to glucuronoxylan

Molecular mechanisms of processive glycoside hydrolases underline catalytic pragmatism

Progress in Identification of UDP-Glycosyltransferases for Ginsenoside Biosynthesis

Contact Info

Product

Resources

About