Transglycosylation and transfer reaction activities of endo-α-N-acetyl-D-galactosaminidase from Diplococcus (Streptococcus) pneumoniae

Bardales, R; Bhavanandan, V. P.

doi:10.1016/s0021-9258(19)47194-9

Cited by 45 publications

(4 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The first reports that endoglycosidases could be employed for synthetic purposes emerged over a decade ago. Endo-F was shown to synthesize trans -glycosylation products with high mannose N -glycans, and an endo- N -acetyl-galactosaminidase from Diplococcus pneumoniae was characterized for acceptor specificity . Until recently, the synthetic utility of endo-glycosidases has not been fully realized.…”

Section: B Transfer Of Oligosaccharides1 Endo-glycosidasesmentioning

confidence: 99%

Synthesis of Complex Carbohydrates and Glycoconjugates: Enzyme-Based and Programmable One-Pot Strategies

2000

View full text Add to dashboard Cite

Section: B Transfer Of Oligosaccharides1 Endo-glycosidasesmentioning

confidence: 99%

Synthesis of Complex Carbohydrates and Glycoconjugates: Enzyme-Based and Programmable One-Pot Strategies

2000

View full text Add to dashboard Cite

“…One of the enzymatic alternatives comprises the use of glycoside hydrolases. Besides hydrolysis of glycosidic bonds, many of these enzymes are able to catalyze transglycosylation reactions to form new glycosidic bonds between donor and acceptor saccharides. − In retaining glycoside hydrolases, the transglycosylation reaction occurs through a double-displacement mechanism . In the first step the glycosidic oxygen is protonated by a catalytic acid, and the anomeric carbon is a target for a nucleophilic attack from the catalytic base, leading to cleavage of the glycosidic bond and to formation of a glycosyl–enzyme intermediate.…”

mentioning

confidence: 99%

Mutational Effects on Transglycosylating Activity of Family 18 Chitinases and Construction of a Hypertransglycosylating Mutant

et al. 2011

View full text Add to dashboard Cite

Enzymatic features that determine transglycosylating activity have been investigated through site-directed mutagenesis studies on two family 18 chitinases, ChiA and ChiB from Serratia marcescens, with inherently little transglycosylation activity. The activity was monitored for the natural substrate (GlcNAc)(4) using mass spectrometry and HPLC. Mutation of the middle Asp in the diagnostic DxDxE motif, which interacts with the catalytic Glu during the catalytic cycle, yielded the strongly transglycosylating mutants ChiA-D313N and ChiB-D142N, respectively. Mutation of the same Asp(313/142) to Ala or the mutation of Asp(311/140) to either Asn or Ala had no or much smaller effects on transglycosylating activity. Mutation of Phe(396) in the +2 subsite of ChiA-D313N to Trp led to a severalfold increase in transglycosylation rate while replacement of aromatic residues with Ala in the aglycon (sugar acceptor-binding) subsites of ChiA-D313N and ChiB-D142N led to a clear reduction in transglycosylating activity. Taken together, these results show that the transglycosylation properties of family 18 chitinases may be manipulated by mutations that affect the configuration of the catalytic machinery and the affinity for sugar acceptors. The hypertransglycosylating mutant ChiA-D313N-F396W may find applications for synthetic purposes.

show abstract

“…Even though oligosaccharides have been prepared by chemical synthesis (54,(59)(60)(61), taking advantage of transglycosylation activities of glycosidases (62,63) and also using various bacterial chondroitinases (5,64), the current strategy of generating a library of oligosaccharides from a parent polysaccharide opens up the opportunity to generate more diverse structures that exist naturally. The sequencing of such CS/DS oligosaccharides by NMR (5,65,66) requires large amounts of materials or sequential digestions with various chondroitinases in conjunction with high-resolution HPLC (5) or capillary electrophoresis (64).…”

Section: Discussionmentioning

confidence: 99%

Novel Sulfated Octa- and Decasaccharides from Squid Cartilage Chondroitin Sulfate E: Sequencing and Application for Determination of the Epitope Structure of the Monoclonal Antibody MO-225

et al. 2007

View full text Add to dashboard Cite

A mixture of octa- and decasaccharides obtained by the digestion with the hyaluronidase of chondroitin sulfate E derived from squid cartilage was subfractionated into 20 and 23 different components, respectively, by anion-exchange HPLC. MALDI-TOF/MS was used to assign the sugar and sulfate composition of the putative octa- and decasaccharides, and a disaccharide composition analysis revealed the building blocks to be A- [GlcUAbeta1-3GalNAc(4S)], C- [GlcUAbeta1-3GalNAc(6S)], and E- [GlcUAbeta1-3GalNAc(4S,6S)] units, where 4S and 6S represent 4-O- and 6-O-sulfate, respectively. The sequences of these octa- and decasaccharides were determined at low picomole amounts by a combination of enzymatic digestions with chondroitinases in conjunction with anion-exchange HPLC. Sequencing revealed that each fraction is a mixture of a major component together with one to three minor components, reflecting the heterogeneity of the parent polysaccharide. Among the 11 different octasaccharide sequences reported here, 8 are novel, while all of the 6 decasaccharide sequences are novel, and this is the first report of the sequencing of CS oligosaccharides longer than octasaccharides. The reactivity of the monoclonal antibody MO-225 with octa- and decasaccharides tested with an oligosaccharide microarray revealed that a CS-E decasaccharide is the minimal requirement for antibody recognition. Among the 6 decasaccharides, only E-E-E-E-C was recognized by MO-225, suggesting the requirement of a C-unit at the reducing end and also the importance of chain length, which in turn may indicate the importance of the conformation acquired by this specific sequence for antibody recognition.

show abstract

Transglycosylation and transfer reaction activities of endo-α-N-acetyl-D-galactosaminidase from Diplococcus (Streptococcus) pneumoniae

Cited by 45 publications

References 33 publications

Synthesis of Complex Carbohydrates and Glycoconjugates: Enzyme-Based and Programmable One-Pot Strategies

Synthesis of Complex Carbohydrates and Glycoconjugates: Enzyme-Based and Programmable One-Pot Strategies

Mutational Effects on Transglycosylating Activity of Family 18 Chitinases and Construction of a Hypertransglycosylating Mutant

Novel Sulfated Octa- and Decasaccharides from Squid Cartilage Chondroitin Sulfate E: Sequencing and Application for Determination of the Epitope Structure of the Monoclonal Antibody MO-225

Contact Info

Product

Resources

About