Biochemical characterization of a novel ulvan lyase from Pseudoalteromonas sp. strain PLSV

Previous studies on this cortexless mutant of Bacillus cereus var. alesti indicated that the forespore membrane was the site of the biochemical lesion. This hypothesis is supported by the results presented here: fatty acid composition of sporulating cells of the mutant is altered, while in vegetative cells it is comparable to the parent; soluble precursors of peptidoglycan synthesis are accumulated in the mutant, at the time of cortex formation; homogenates of the mutant prepared at the time of cortex formation are unable to incorporate tritiated diaminopimelic acid into peptidoglycan, while homogenates of cells forming germ cell wall do so to an extent comparable to that of the parent; lipid-linked intermediates are formed by the mutant as in the parent. Apparently the mutant is unable either to transfer disaccharide penta-peptide units from the carrier lipid to the growing peptidoglycan acceptor, or to transport lipid-linked intermediates across the forespore membrane.

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Biochemical characterization of a 27kDa 1,3-β-d-glucanase from Trichoderma asperellum induced by cell wall of Rhizoctonia solani

Carbohydrate Polymers ◽

10.1016/j.carbpol.2011.09.001 ◽

2012 ◽

Vol 87 (2) ◽

pp. 1219-1223 ◽

Cited By ~ 15

Author(s):

Raquel da Silva Aires ◽

Andrei Stecca Steindorff ◽

Marcelo Henrique Soller Ramada ◽

Saulo José Linhares de Siqueira ◽

Cirano José Ulhoa

Keyword(s):

Cell Wall ◽

Rhizoctonia Solani ◽

Biochemical Characterization ◽

Trichoderma Asperellum

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Purification and characterization of iduronic acid-rich and glucuronic acid-rich proteoglycans implicated in human post-burn keloid scar

Carbohydrate Research ◽

10.1016/0008-6215(90)84056-z ◽

1990 ◽

Vol 207 (2) ◽

pp. 295-305 ◽

Cited By ~ 12

Author(s):

Hari G. Garg ◽

Eric W. Lippay ◽

D.Andrew R. Burd ◽

Peter J. Neame

Keyword(s):

Glucuronic Acid ◽

Purification And Characterization ◽

Keloid Scar ◽

Iduronic Acid

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Biochemical characterization of wheat straw cell wall with special reference to bioactive profile

International Journal of Food Properties ◽

10.1080/10942912.2018.1484759 ◽

2018 ◽

Vol 21 (1) ◽

pp. 1303-1310 ◽

Cited By ~ 6

Author(s):

Tabussam Tufail ◽

Farhan Saeed ◽

Muhammad Imran ◽

Muhammad Umair Arshad ◽

Faqir Muhammad Anjum ◽

...

Keyword(s):

Cell Wall ◽

Special Reference ◽

Wheat Straw ◽

Biochemical Characterization

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Identification and biochemical characterization of two novel UDP-2,3-diacetamido-2,3-dideoxy-α-D-glucuronic acid 2-epimerases from respiratory pathogens

Biochemical Journal ◽

10.1042/bj20070017 ◽

2007 ◽

Vol 405 (1) ◽

pp. 123-130 ◽

Cited By ~ 14

Author(s):

Erin L. Westman ◽

David J. Mcnally ◽

Martin Rejzek ◽

Wayne L. Miller ◽

Vellupillai Sri Kannathasan ◽

...

Keyword(s):

Capillary Electrophoresis ◽

Glucuronic Acid ◽

Biochemical Characterization ◽

Intermediate Compound ◽

Proton Spectrum ◽

Respiratory Pathogens ◽

Sodium Phosphate Buffer ◽

Mannuronic Acid ◽

Enzyme Substrate

The heteropolymeric O-antigen of the lipopolysaccharide from Pseudomonas aeruginosa serogroup O5 as well as the band-A trisaccharide from Bordetella pertussis contain the di-N-acetylated mannosaminuronic acid derivative, β-D-ManNAc3NAcA (2,3-diacetamido-2,3-dideoxy-β-D-mannuronic acid). The biosynthesis of the precursor for this sugar is proposed to require five steps, through which UDP-α-D-GlcNAc (UDP-N-acetyl-α-D-glucosamine) is converted via four steps into UDP-α-D-GlcNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-α-D-glucuronic acid), and this intermediate compound is then epimerized by WbpI (P. aeruginosa), or by its orthologue, WlbD (B. pertussis), to form UDP-α-D-ManNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-α-D-mannuronic acid). UDP-α-D-GlcNAc3NAcA, the proposed substrate for WbpI and WlbD, was obtained through chemical synthesis. His6–WbpI and His6–WlbD were overexpressed and then purified by affinity chromatography using FPLC. Capillary electrophoresis was used to analyse reactions with each enzyme, and revealed that both enzymes used UDP-α-D-GlcNAc3NAcA as a substrate, and reacted optimally in sodium phosphate buffer (pH 6.0). Neither enzyme utilized UDP-α-D-GlcNAc, UDP-α-D-GlcNAcA (UDP-2-acetamido-2,3-dideoxy-α-D-glucuronic acid) or UDP-α-D-GlcNAc3NAc (UDP-2,3-diacetamido-2,3-dideoxy-α-D-glucose) as substrates. His6–WbpI or His6–WlbD reactions with UDP-α-D-GlcNAc3NAcA produce a novel peak with an identical retention time, as shown by capillary electrophoresis. To unambiguously characterize the reaction product, enzyme–substrate reactions were allowed to proceed directly in the NMR tube and conversion of substrate into product was monitored over time through the acquisition of a proton spectrum at regular intervals. Data collected from one- and two-dimensional NMR experiments showed that His6–WbpI catalysed the 2-epimerization of UDP-α-D-GlcNAc3NAcA, converting it into UDP-α-D-ManNAc3NAcA. Collectively, these results provide evidence that WbpI and WlbD are UDP-2,3-diacetamido-2,3-dideoxy-α-D-glucuronic acid 2-epimerases.

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Genetic and Biochemical Characterization of the Cell Wall Hydrolase Activity of the Major Secreted Protein of Lactobacillus rhamnosus GG

PLoS ONE ◽

10.1371/journal.pone.0031588 ◽

2012 ◽

Vol 7 (2) ◽

pp. e31588 ◽

Cited By ~ 46

Author(s):

Ingmar J. J. Claes ◽

Geert Schoofs ◽

Krzysztof Regulski ◽

Pascal Courtin ◽

Marie-Pierre Chapot-Chartier ◽

...

Keyword(s):

Cell Wall ◽

Biochemical Characterization ◽

Lactobacillus Rhamnosus ◽

Hydrolase Activity ◽

Secreted Protein ◽

Lactobacillus Rhamnosus Gg ◽

Cell Wall Hydrolase

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The copolymeric structure of pig skin dermatan sulphate. Characterization of d-glucuronic acid-containing oligosaccharides isolated after controlled degradation of oxydermatan sulphate

Biochemical Journal ◽

10.1042/bj1430369 ◽

1974 ◽

Vol 143 (2) ◽

pp. 369-378 ◽

Cited By ~ 15

Author(s):

Lars-Åke Fransson ◽

Lars Cöster ◽

Anders Malmström ◽

Ingrid Sjöberg

Keyword(s):

General Formula ◽

Glucuronic Acid ◽

Periodate Oxidation ◽

Dermatan Sulphate ◽

Chondroitinase Abc ◽

Pig Skin ◽

Iduronic Acid ◽

Unsaturated Disaccharide ◽

Selective Periodate Oxidation

Selective periodate oxidation of unsubstituted l-iduronic acid residues in copolymeric dermatan sulphate chains was followed by reduction-hydrolysis or alkaline elimination. By this procedure the glucuronic acid-containing periods were isolated in oligosaccharide form; general formula: [Formula: see text] Further degradation of these oligosaccharides with chondroitinase-AC yielded three types of products: (a) sulphated trisaccharide containing an unsaturated uronosyl moiety in the non-reducing terminal and a C4 fragment in the reducing terminal, ΔUA-GalNAc-(-SO4)-R; (b) monosulphated, unsaturated disaccharide, ΔUA-GalNAc-SO4 when n is greater than or equal to 2; and (c) N-acetylgalactosamine with or without sulphate. Oligosaccharides containing a single glucuronic acid residue (n=1) comprised more than half of the glucuronic acid-containing oligosaccharides. The terminal N-acetylgalactosamine moiety of the shortest oligosaccharide was largely 4-sulphated, whereas higher oligosaccharides primarily contained 6-sulphated or unsulphated hexosamine moieties in the same position. Moreover, IdUA-SO4-containing oligosaccharides were encountered. These oligosaccharides were resistant to the action of chondroitinase-ABC.

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Cloning and Characterization of a Novel Invertase from the Obligate Biotroph Uromyces fabae and Analysis of Expression Patterns of Host and Pathogen Invertases in the Course of Infection

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-19-0625 ◽

2006 ◽

Vol 19 (6) ◽

pp. 625-634 ◽

Cited By ~ 67

Author(s):

Ralf T. Voegele ◽

Stefan Wirsel ◽

Ulla Möll ◽

Melanie Lechner ◽

Kurt Mendgen

Keyword(s):

Cell Wall ◽

Expression Analysis ◽

Biochemical Characterization ◽

Higher Plants ◽

Rust Fungus ◽

Expression Patterns ◽

Invertase Activity ◽

Pathogen Infection ◽

Uromyces Fabae

Invertases are key enzymes in carbon partitioning in higher plants. They gain additional importance in the distribution of carbohydrates in the event of wounding or pathogen attack. Although many researchers have found an increase in invertase activity upon infection, only a few studies were able to determine whether the source of this activity was host or parasite. This article analyzes the role of invertases involved in the biotrophic interaction of the rust fungus Uromyces fabae and its host plant, Vicia faba. We have identified a fungal gene, Uf-INV1, with homology to invertases and assessed its contribution to pathogenesis. Expression analysis indicated that transcription began upon penetration of the fungus into the leaf, with high expression levels in haustoria. Heterologous expression of Uf-INV1 in Saccharomyces cerevisiae and Pichia pastoris allowed a biochemical characterization of the enzymatic activity associated with the secreted gene product INV1p. Expression analysis of the known vacuolar and cell-wall-bound invertase isoforms of V. faba indicated a decrease in the expression of a vacuolar invertase, whereas one cell-wall-associated invertase exhibited increased expression. These changes were not confined to the infected tissue, and effects also were observed in remote plant organs, such as roots. These findings hint at systemic effects of pathogen infection. Our results support the hypothesis that pathogen infection establishes new sinks which compete with physiological sink organs.

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Extraction, quantification and biochemical characterization of cereal industry by‐product cell wall

Journal of Food Processing and Preservation ◽

10.1111/jfpp.15023 ◽

2020 ◽

Author(s):

Tabussam Tufail ◽

Farhan Saeed ◽

Imran Pasha ◽

Muhammad Umair Arshad ◽

Muhammad Afzaal ◽

...

Keyword(s):

Cell Wall ◽

Biochemical Characterization ◽

Cereal Industry

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Characterization of FcXTH2, a Novel Xyloglucan Endotransglycosylase/Hydrolase Enzyme of Chilean Strawberry with Hydrolase Activity

International Journal of Molecular Sciences ◽

10.3390/ijms21093380 ◽

2020 ◽

Vol 21 (9) ◽

pp. 3380 ◽

Cited By ~ 1

Author(s):

Luis Morales-Quintana ◽

Dina Beltrán ◽

Ángela Mendez-Yañez ◽

Felipe Valenzuela-Riffo ◽

Raúl Herrera ◽

...

Keyword(s):

Cell Wall ◽

Structural Model ◽

Biochemical Characterization ◽

Comparative Modeling ◽

Xyloglucan Endotransglycosylase ◽

Protein Ligand Interactions ◽

Modeling Methodology ◽

Ligand Interactions ◽

Km Value

Xyloglucan endotransglycosylase/hydrolases (XTHs) are cell wall enzymes with hydrolase (XEH) and/or endotransglycosylase (XET) activities. As they are involved in the modification of the xyloglucans, a type of hemicellulose present in the cell wall, they are believed to be very important in different processes, including growth, development, and fruit ripening. Previous studies suggest that XTHs might play a key role in development and ripening of Fragaria chiloensis fruit, and its characterization is pending. Therefore, in order to provide a biochemical characterization of the FcXTH2 enzyme to explain its possible role in strawberry development, the molecular cloning and the heterologous expression of FcXTH2 were performed. The recombinant FcXTH2 was active and displayed mainly XEH activity. The optimal pH and temperature are 5.5 and 37 °C, respectively. A KM value of 0.029 mg mL−1 was determined. Additionally, its protein structural model was built through comparative modeling methodology. The model showed a typically β-jelly-roll type folding in which the catalytic motif was oriented towards the FcXTH2 central cavity. Using molecular docking, protein-ligand interactions were explored, finding better interaction with xyloglucan than with cellulose. The data provided groundwork for understanding, at a molecular level, the enzymatic mechanism of FcXTH2, an important enzyme acting during the development of the Chilean strawberry.

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