Biomedical subjects

S Armand

Publications and source records attributed to S Armand.

17 recordsLinked to original sources

PubMed1996

Plant chitinases use two different hydrolytic mechanisms.

Bacterial, fungal, animal, and some plant chitinases form family 18 of glycosyl hydrolases. Most plant chitinases form the family 19. While some chitinases also have lysozyme activity, animal lysozymes belong to different families. For glycosyl hydrolases, two reaction mechanisms are possible, leading to either retention or inversion of the anomeric configuration. We analyzed by HPLC the stereochemical outcome of the hydrolysis catalyzed by cucumber and bean chitinases, belonging to families 18 and 19, respectively. Cucumber chitinase used the retaining mechanism as known for bacterial chitinases and hen egg white lysozyme for which the mechanism has been determined. In contrast, bean chitinase catalyzed the hydrolysis of chitooligosaccharides with overall inversion of anomeric configuration.

Catalysis↗

PubMed1996

Stereochemical course and reaction products of the action of beta-xylosidase from Thermoanaerobacterium saccharolyticum strain B6A-RI.

Beta-Xylosidases are grouped in families 39 and 43 of a general classification of glycosyl hydrolases based on amino acid sequence similarities. The Beta-xylosidase from Butyrivibrio fibrisolvens, which belongs to family 43, has been shown to operate by a molecular mechanism which results in the inversion of the anomeric configuration. Thermoanaerobacterium saccharolyticum B6A-RI Beta-xylosidase which belongs to family 39 was purified as a recombinant enzyme from Escherichia coli. The stereochemistry of the hydrolysis of p-nitrophenyl Beta-D-xylopyranoside was followed by 1H NMR. The spectrum recorded after 2 h hydrolysis showed a large signal centred at 4.47 ppm (J approximately 10 Hz) assignable to H1 of free Beta-xylose with a small amount of alpha-xylose (5.05 ppm, J approximately 3 Hz) attributable to mutarotation. This result indicates that T. saccharolyticum Beta-xylosidase operates with overall retention of the anomeric configuration. This result, with the lack of sequence similarity between the two families of Beta-xylosidases, suggests that these two families have major differences in their active-site geometries. Consistent with its retaining mechanism, Beta-xylosidase of T. saccharolyticum B6A-RI also displayed transglycosylating activity:reverse-phase HPLC showed approximately 30% conversion of p-nitrophenyl Beta-D-xylopyranoside into a number of higher nitrophenyl oligosaccharides after 5 min incubation with the enzyme. The structure of the most abundant oligosaccharides could be determined by total correlation spectroscopy NMR and showed that the enzyme can build Beta-1,4, Beta-1,3- and Beta-1,2-linked xylo-oligosaccharides.

Bacteria↗

PubMed1996

A modular family 19 chitinase found in the prokaryotic organism Streptomyces griseus HUT 6037.

The specificity of chitinase C-1 of Streptomyces griseus HUT 6037 for the hydrolysis of the beta-1,4-glycosidic linkages in partially acetylated chitosan is different from that of other microbial chitinases. In order to study the primary structure of this unique chitinase, the chiC gene specifying chitinase C-1 was cloned and its nucleotide sequence was determined. The gene encodes a polypeptide of 294 amino acids with a calculated size of 31.4 kDa. Comparison of the amino acid sequence of the deduced polypeptide with that of other proteins revealed a C-terminal catalytic domain displaying considerable sequence similarity to the catalytic domain of plant class I, II, and IV chitinases which form glycosyl hydrolase family 19. The N-terminal domain of the deduced polypeptide exhibits sequence similarity to substrate-binding domains of several microbial chitinases and cellulases but not to the chitin-binding domains of plant chitinases. The previously purified chitinase C-1 from S. griseus is suggested to be generated by proteolytic removal of the N-terminal chitin-binding domain and corresponds to the catalytic domain of the chitinase encoded by the chiC gene. High-performance liquid chromatography analysis of the hydrolysis products from N-acetyl chitotetraose revealed that chitinase C-1 catalyzes hydrolysis of the glycosidic bond with inversion of the anomeric configuration, in agreement with the previously reported inverting mechanism of plant class I chitinases. This is the first report of a family 19 chitinase found in an organism other than higher plants.

Amino Acid Sequence↗

PubMed1995

Stereochemistry of chitin hydrolysis by a plant chitinase/lysozyme and X-ray structure of a complex with allosamidin: evidence for substrate assisted catalysis.

The plant enzyme hevamine has both chitinase and lysozyme activity. HPLC analysis of the products of the hydrolysis of chitopentaose shows that hevamine acts with retention of the configuration, despite the absence of a nucleophilic or stabilizing carboxylate. To analyze the stabilization of a putative oxocarbonium ion intermediate, the X-ray structure of hevamine complexed with the inhibitor allosamidin was determined at 1.85 A resolution. This structure supports the role of Glu127 as a proton donor. The allosamizoline group binds in the center of the active site, mimicking a reaction intermediate in which a positive charge at C1 is stabilized intramolecularly by the carbonyl oxygen of the N-acetyl group at C2.

Acetylglucosamine↗

PubMed1994

Stereochemical course of the hydrolysis reaction catalyzed by chitinases A1 and D from Bacillus circulans WL-12.

Chitinases A1 and D were purified from the periplasmic proteins produced by Escherichia coli HB101 harbouring recombinant plasmids carrying respectively the chiA and chiD genes of Bacillus circulans WL-12. HPLC analysis indicated that during the hydrolysis of chitotriose, both chitinases initially produce N-acetylglucosamine and only one anomer of chitobiose. 1H NMR spectroscopy of the hydrolysis of chitotetraitol showed that this anomer corresponds to beta-chitobiose, demonstrating that chitinases A1 and D act by a molecular mechanism that retains the anomeric configuration. This mechanism is similar to that of lysozymes although both chitinases belong to a family of proteins sharing no demonstrable amino acid sequence similarity with lysozymes.

Bacillus↗

PubMed1993

The effect of sibling relationship on the periodontal condition.

The purpose of this investigation was to study clinically as well as microbiologically the effect of sibling relationship on the periodontal condition in a young population with a relatively high prevalence of periodontal disease and deprived from regular dental care. In this study, 23 family units consisting of 3 more siblings were evaluated. In all, 78 subjects aged 15 to 25 years were included in the study. The mean interproximal amount of loss of attachment in this population was 0.29 mm. The individual mean ranged from 0 to 1.27 mm. In 33% of the subjects, > or = 1 sites with a probing depth of 5 mm or more in conjunction with 2 mm of attachment loss were present. The results show a significant sibship effect for: plaque, calculus, loss of attachment, spirochetes on the tongue and in the pocket, Porphyromonas gingivalis on the gingiva and in the saliva and Prevotella intermedia in the saliva. These results support the hypothesis that periodontitis aggregates in families.

Adolescent↗

PubMed1991