scholarly journals Crystal Structure of Cel44A, a Glycoside Hydrolase Family 44 Endoglucanase from Clostridium thermocellum

2007 ◽  
Vol 282 (49) ◽  
pp. 35703-35711 ◽  
Author(s):  
Yu Kitago ◽  
Shuichi Karita ◽  
Nobuhisa Watanabe ◽  
Masakatsu Kamiya ◽  
Tomoyasu Aizawa ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Glutamic Acid ◽  
Glycoside Hydrolase ◽  
Clostridium Thermocellum ◽  
Structural Features ◽  
Glycoside Hydrolase Family ◽  
Wild Type ◽  
H Nmr ◽  
Hydrolase Family

The crystal structure of Cel44A, which is one of the enzymatic components of the cellulosome of Clostridium thermocellum, was solved at a resolution of 0.96Å. This enzyme belongs to glycoside hydrolase family (GH family) 44. The structure reveals that Cel44A consists of a TIM-like barrel domain and a β-sandwich domain. The wild-type and the E186Q mutant structures complexed with substrates suggest that two glutamic acid residues, Glu186 and Glu359, are the active residues of the enzyme. Biochemical experiments were performed to confirm this idea. The structural features indicate that GH family 44 belongs to clan GH-A and that the reaction catalyzed by Cel44A is retaining type hydrolysis. The stereochemical course of hydrolysis was confirmed by a 1H NMR experiment using the reduced cellooligosaccharide as a substrate.

Structural and functional characterization of a glycoside hydrolase family 3 β-N-acetylglucosaminidase from Paenibacillus sp. str. FPU-7

2019 ◽  
Vol 166 (6) ◽  
pp. 503-515
Author(s):  
Takafumi Itoh ◽  
Tomomitsu Araki ◽  
Tomohiro Nishiyama ◽  
Takao Hibi ◽  
Hisashi Kimoto
Keyword(s):  
Crystal Structure ◽  
Glycoside Hydrolase ◽  
Catalytic Mechanism ◽  
Functional Characterization ◽  
Structural Features ◽  
Expression Cloning ◽  
Glycoside Hydrolase Family ◽  
Paenibacillus Sp ◽  
Hydrolase Family

Abstract Chitin, a β-1,4-linked homopolysaccharide of N-acetyl-d-glucosamine (GlcNAc), is one of the most abundant biopolymers on Earth. Paenibacillus sp. str. FPU-7 produces several different chitinases and converts chitin into N,N′-diacetylchitobiose ((GlcNAc)2) in the culture medium. However, the mechanism by which the Paenibacillus species imports (GlcNAc)2 into the cytoplasm and divides it into the monomer GlcNAc remains unclear. The gene encoding Paenibacillus β-N-acetyl-d-glucosaminidase (PsNagA) was identified in the Paenibacillus sp. str. FPU-7 genome using an expression cloning system. The deduced amino acid sequence of PsNagA suggests that the enzyme is a part of the glycoside hydrolase family 3 (GH3). Recombinant PsNagA was successfully overexpressed in Escherichia coli and purified to homogeneity. As assessed by gel permeation chromatography, the enzyme exists as a 57-kDa monomer. PsNagA specifically hydrolyses chitin oligosaccharides, (GlcNAc)2–4, 4-nitrophenyl N-acetyl β-d-glucosamine (pNP-GlcNAc) and pNP-(GlcNAc)2–6, but has no detectable activity against 4-nitrophenyl β-d-glucose, 4-nitrophenyl β-d-galactosamine and colloidal chitin. In this study, we present a 1.9 Å crystal structure of PsNagA bound to GlcNAc. The crystal structure reveals structural features related to substrate recognition and the catalytic mechanism of PsNagA. This is the first study on the structural and functional characterization of a GH3 β-N-acetyl-d-glucosaminidase from Paenibacillus sp.

scholarly journals Crystal structure of the N-terminal domain of a glycoside hydrolase family 131 protein from Coprinopsis cinerea

FEBS Letters ◽  
2013 ◽  
Vol 587 (14) ◽  
pp. 2193-2198 ◽  
Author(s):  
Takatsugu Miyazaki ◽  
Makoto Yoshida ◽  
Mizuki Tamura ◽  
Yutaro Tanaka ◽  
Kiwamu Umezawa ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Glycoside Hydrolase ◽  
Glycoside Hydrolase Family ◽  
Coprinopsis Cinerea ◽  
Terminal Domain ◽  
Hydrolase Family

Pseudomonas cellulosa expresses a single membrane-bound glycoside hydrolase family 51 arabinofuranosidase

2001 ◽  
Vol 358 (3) ◽  
pp. 599-605 ◽  
Author(s):  
Marie-Helene BEYLOT ◽  
Kaveh EMAMI ◽  
Vincent A. McKIE ◽  
Harry J. GILBERT ◽  
Gavin PELL
Keyword(s):  
Sugar Beet ◽  
Glycoside Hydrolase ◽  
Northern Blot Analysis ◽  
Glycoside Hydrolase Family ◽  
Wild Type ◽  
Mutant Bacterium ◽  
Membrane Bound ◽  
Hydrolase Family ◽  
Wild Type Bacterium ◽  
Glycoside Hydrolase Family 51

In the accompanying paper [Beylot, McKie, Voragen, Doeswijk-Voragen and Gilbert (2001) Biochem. J. 358, 607–614] the chromosome of Pseudomonas cellulosa was shown to contain two genes, abf51A and abf62A, that encode arabinofuranosidases belonging to glycoside hydrolase families 51 and 62, respectively. In this report we show that expression of Abf51A is induced by arabinose and arabinose-containing polysaccharides. Northern-blot analysis showed that abf51A was efficiently transcribed, whereas no transcript derived from abf62A was detected in the presence of arabinose-containing polysaccharides. Zymogram and Western-blot analyses revealed that Abf51A was located on the outer membrane of P. cellulosa. To investigate the importance of Abf51A in the release of arabinose from poly- and oligosaccharides, transposon mutagenesis was used to construct an abf51A-inactive mutant of P. cellulosa (Δabf51A). The mutant did not grow on linear arabinan or sugar beet arabinan, and utilized arabinoxylan much more slowly than the wild-type bacterium. Arabinofuranosidase activity in Δabf51A against aryl-α-arabinofuranosides, arabinan and α1,5-linked arabino-oligosaccharides was approx. 1% of the wild-type bacterium. The mutant bacterium did not exhibit arabinofuranosidase activity against arabinoxylan, supporting the view that abf62A is not expressed in P. cellulosa. These data indicate that P. cellulosa expresses a membrane-bound glycoside hydrolase family 51 arabinofuranosidase that plays a pivotal role in releasing arabinose from polysaccharides and arabino-oligosaccharides.

scholarly journals Crystal Structure and Mutational Analysis of Isomalto-dextranase, a Member of Glycoside Hydrolase Family 27

2015 ◽  
Vol 290 (43) ◽  
pp. 26339-26349 ◽  
Author(s):  
Yuka Okazawa ◽  
Takatsugu Miyazaki ◽  
Gaku Yokoi ◽  
Yuichi Ishizaki ◽  
Atsushi Nishikawa ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Glycoside Hydrolase ◽  
Mutational Analysis ◽  
Glycoside Hydrolase Family ◽  
Hydrolase Family

scholarly journals The structure of PfGH50B, an agarase from the marine bacterium Pseudoalteromonas fuliginea PS47

2020 ◽  
Vol 76 (9) ◽  
pp. 422-427
Author(s):  
Benjamin Pluvinage ◽  
Craig S. Robb ◽  
Roderick Jeffries ◽  
Alisdair B. Boraston
Keyword(s):  
Glycoside Hydrolase ◽  
Marine Bacterium ◽  
Structural Features ◽  
Degree Of Polymerization ◽  
Glycoside Hydrolase Family ◽  
Carbohydrate Binding ◽  
X Ray ◽  
Terminal Domain ◽  
Polysaccharide Utilization Locus ◽  
Hydrolase Family

The recently identified marine bacterium Pseudoalteromonas fuliginea sp. PS47 possesses a polysaccharide-utilization locus dedicated to agarose degradation. In particular, it contains a gene (locus tag EU509_06755) encoding a β-agarase that belongs to glycoside hydrolase family 50 (GH50), PfGH50B. The 2.0 Å resolution X-ray crystal structure of PfGH50B reveals a rare complex multidomain fold that was found in two of the three previously determined GH50 structures. The structure comprises an N-terminal domain with a carbohydrate-binding module (CBM)-like fold fused to a C-terminal domain by a rigid linker. The CBM-like domain appears to function by extending the catalytic groove of the enzyme. Furthermore, the PfGH50B structure highlights key structural features in the mobile loops that may function to restrict the degree of polymerization of the neoagaro-oligosaccharide products and the enzyme processivity.

scholarly journals Crystal structure of β-L-arabinobiosidase belonging to glycoside hydrolase family 121

PLoS ONE ◽  
2020 ◽  
Vol 15 (6) ◽  
pp. e0231513 ◽  
Author(s):  
Keita Saito ◽  
Alexander Holm Viborg ◽  
Shiho Sakamoto ◽  
Takatoshi Arakawa ◽  
Chihaya Yamada ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Glycoside Hydrolase ◽  
Glycoside Hydrolase Family ◽  
Hydrolase Family
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