scholarly journals A novel α-glucosidase from the acidophilic archaeon Ferroplasma acidiphilum strain Y with high transglycosylation activity and an unusual catalytic nucleophile

2005 ◽  
Vol 391 (2) ◽  
pp. 269-276 ◽  
Author(s):  
Manuel Ferrer ◽  
Olga V. Golyshina ◽  
Francisco J. Plou ◽  
Kenneth N. Timmis ◽  
Peter N. Golyshin
Keyword(s):  
Glycosyl Hydrolase ◽  
Soluble Starch ◽  
Glycoside Hydrolases ◽  
Site Directed Mutagenesis ◽  
Preference Order ◽  
Glycosyl Hydrolase Family ◽  
Significant Similarity ◽  
Transglycosylation Activity ◽  
Membrane Bound ◽  
Hydrolase Family

Ferroplasma acidiphilum strain Y (DSM 12658), a ferrous iron-oxidizing, acidophilic and mesophilic archaeon, was found to produce a membrane-bound α-glucosidase (αGluFa) showing no significant similarity to any of the known glycoside hydrolases classified in different families and having an unusual catalytic site consisting of a threonine and a histidine residue. The highest α-glucosidase activity was found at low pH, 2.4–3.5, and the substrate preference order was: sucrose>maltose>maltotriose ≫maltotetraose≫malto-oligosaccharides from maltopentaose to maltoheptaose⋙soluble starch (kcat/Km was 293.0, 197.0, 18.8, 0.3 and 0.02 s−1·mM−1 respectively). The enzyme was able to transfer glucosyl groups from maltose as donor, to produce exclusively maltotriose (up to 300 g/l). Chemical modification and electrospray ionization MS analysis of 5-fluoro-α-D-glucopyranosyl-enzyme derivatives, coupled with site-directed mutagenesis, strongly suggested that the putative catalytic nucleophile in this enzyme is Thr212. Iron was found to be essential for enzyme activity and integrity, and His390 was shown to be essential for iron binding. These results suggest that the metalloenzyme αGluFa is a new member of the glycosyl hydrolase family that uses a novel mechanism for sugar glycosylation and/or transglycosylation.

scholarly journals Characterization of Two Novel α-Glucosidases from Bifidobacterium breve UCC2003

2008 ◽  
Vol 75 (4) ◽  
pp. 1135-1143 ◽  
Author(s):  
Karina Pokusaeva ◽  
Mary O'Connell-Motherway ◽  
Aldert Zomer ◽  
Gerald F. Fitzgerald ◽  
Douwe van Sinderen
Keyword(s):  
Glycosyl Hydrolase ◽  
Hydrolytic Activity ◽  
Glycosyl Hydrolase Family ◽  
Bifidobacterium Breve ◽  
Transglycosylation Activity ◽  
Encoding Genes ◽  
Temperature Optima ◽  
Hydrolase Family

ABSTRACT Two α-glucosidase-encoding genes (agl1 and agl2) from Bifidobacterium breve UCC2003 were identified and characterized. Based on their similarity to characterized carbohydrate hydrolases, the Agl1 and Agl2 enzymes are both assigned to a subgroup of the glycosyl hydrolase family 13, the α-1,6-glucosidases (EC 3.2.1.10). Recombinant Agl1 and Agl2 into which a His12 sequence was incorporated (Agl1His and Agl2His, respectively) exhibited hydrolytic activity towards panose, isomaltose, isomaltotriose, and four sucrose isomers—palatinose, trehalulose, turanose, and maltulose—while also degrading trehalose and, to a lesser extent, nigerose. The preferred substrates for both enzymes were panose, isomaltose, and trehalulose. Furthermore, the pH and temperature optima for both enzymes were determined, showing that Agl1His exhibits higher thermo and pH optima than Agl2His. The two purified α-1,6-glucosidases were also shown to have transglycosylation activity, synthesizing oligosaccharides from palatinose, trehalulose, trehalose, panose, and isomaltotriose.

scholarly journals Novel α-Glucosidase from Aspergillus nidulans with Strong Transglycosylation Activity

2002 ◽  
Vol 68 (3) ◽  
pp. 1250-1256 ◽  
Author(s):  
Naoki Kato ◽  
Sachie Suyama ◽  
Masao Shirokane ◽  
Masashi Kato ◽  
Tetsuo Kobayashi ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Aspergillus Nidulans ◽  
Glycosyl Hydrolase ◽  
Amino Acid Sequences ◽  
Glycosyl Hydrolase Family ◽  
Transglycosylation Activity ◽  
Single Polypeptide ◽  
Hydrolysis Of ◽  
Hydrolase Family

ABSTRACT Aspergillus nidulans possessed an α-glucosidase with strong transglycosylation activity. The enzyme, designated α-glucosidase B (AgdB), was purified and characterized. AgdB was a heterodimeric protein comprising 74- and 55-kDa subunits and catalyzed hydrolysis of maltose along with formation of isomaltose and panose. Approximately 50% of maltose was converted to isomaltose, panose, and other minor transglycosylation products by AgdB, even at low maltose concentrations. The agdB gene was cloned and sequenced. The gene comprised 3,055 bp, interrupted by three short introns, and encoded a polypeptide of 955 amino acids. The deduced amino acid sequence contained the chemically determined N-terminal and internal amino acid sequences of the 74- and 55-kDa subunits. This implies that AgdB is synthesized as a single polypeptide precursor. AgdB showed low but overall sequence homology to α-glucosidases of glycosyl hydrolase family 31. However, AgdB was phylogenetically distinct from any other α-glucosidases. We propose here that AgdB is a novel α-glucosidase with unusually strong transglycosylation activity.

scholarly journals New Chitosan-Degrading Strains That Produce Chitosanases Similar to ChoA of Mitsuaria chitosanitabida

2005 ◽  
Vol 71 (9) ◽  
pp. 5138-5144 ◽  
Author(s):  
ChoongSoo Yun ◽  
Daiki Amakata ◽  
Yasuhiro Matsuo ◽  
Hideyuki Matsuda ◽  
Makoto Kawamukai
Keyword(s):  
Southern Hybridization ◽  
Glycosyl Hydrolase ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
16S Rrna Gene Sequences ◽  
Glycosyl Hydrolase Family ◽  
Broad Variety ◽  
Hydrolase Family ◽  
The 16S Rrna Gene ◽  
Sequence Similarities

ABSTRACT The betaproteobacterium Mitsuaria chitosanitabida (formerly Matsuebacter chitosanotabidus) 3001 produces a chitosanase (ChoA) that is classified in glycosyl hydrolase family 80. While many chitosanase genes have been isolated from various bacteria to date, they show limited homology to the M. chitosanitabida 3001 chitosanase gene (choA). To investigate the phylogenetic distribution of chitosanases analogous to ChoA in nature, we identified 67 chitosan-degrading strains by screening and investigated their physiological and biological characteristics. We then searched for similarities to ChoA by Western blotting and Southern hybridization and selected 11 strains whose chitosanases showed the most similarity to ChoA. PCR amplification and sequencing of the chitosanase genes from these strains revealed high deduced amino acid sequence similarities to ChoA ranging from 77% to 99%. Analysis of the 16S rRNA gene sequences of the 11 selected strains indicated that they are widely distributed in the β and γ subclasses of Proteobacteria and the Flavobacterium group. These observations suggest that the ChoA-like chitosanases that belong to family 80 occur widely in a broad variety of bacteria.

Variants of glycosyl hydrolase family 2 β-glucuronidases have increased activity on recalcitrant substrates

2021 ◽  
Vol 145 ◽  
pp. 109742
Author(s):  
Caleb R. Schlachter ◽  
Amanda C. McGee ◽  
Pongkwan N. Sitasuwan ◽  
Gary C. Horvath ◽  
Nanda G. Karri ◽  
...  
Keyword(s):  
Glycosyl Hydrolase ◽  
Glycosyl Hydrolase Family ◽  
Increased Activity ◽  
Hydrolase Family

scholarly journals A constitutive expression system for glycosyl hydrolase family 7 cellobiohydrolases in Hypocrea jecorina

2015 ◽  
Vol 8 (1) ◽  
Author(s):  
Jeffrey G Linger ◽  
Larry E Taylor ◽  
John O Baker ◽  
Todd Vander Wall ◽  
Sarah E Hobdey ◽  
...  
Keyword(s):  
Glycosyl Hydrolase ◽  
Expression System ◽  
Constitutive Expression ◽  
Hypocrea Jecorina ◽  
Glycosyl Hydrolase Family ◽  
Hydrolase Family

scholarly journals Detecting the structural determinants of glycosyl hydrolase family 11 xylanase inhibition

2005 ◽  
Vol 61 (a1) ◽  
pp. c197-c197
Author(s):  
C. De Ranter ◽  
S. Sansen ◽  
K. Gebruers ◽  
K. Brijs ◽  
C. M. Courtin ◽  
...  
Keyword(s):  
Glycosyl Hydrolase ◽  
Glycosyl Hydrolase Family ◽  
Structural Determinants ◽  
Hydrolase Family
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