Carbohydrate esterase family 4 enzymes: substrate specificity

Caufrier, Frederic; Martinou, Aggeliki; Dupont, Claude; Bouriotis, Vassilis

doi:10.1016/s0008-6215(03)00002-8

Cited by 120 publications

(125 citation statements)

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“…SpPgdA is a member of the family 4 carbohydrate esterases (CE-4) as defined in the CAZy database (http:͞͞afmb.cnrsmrs.fr͞CAZy), which has been reviewed recently (6). NodB, the first enzyme of this family to be described in detail, deacetylates a GlcNAc residue in the synthesis of Nod factors, bacterial signaling molecules that regulate the symbiotic relationship with leguminous plants (7).…”

mentioning

confidence: 99%

Structure and metal-dependent mechanism of peptidoglycan deacetylase, a streptococcal virulence factor

Blair

Schüttelkopf

MacRae

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

198

389

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Streptococcus pneumoniae peptidoglycan GlcNAc deacetylase (SpPgdA) protects the Gram-positive bacterial cell wall from host lysozymes by deacetylating peptidoglycan GlcNAc residues. Deletion of the pgda gene has been shown to result in hypersensitivity to lysozyme and reduction of infectivity in a mouse model. SpPgdA is a member of the family 4 carbohydrate esterases, for which little structural information exists, and no catalytic mechanism has yet been defined. Here we describe the native crystal structure and product complexes of SpPgdA biochemical characterization and mutagenesis. The structural data show that SpPgdA is an elongated three-domain protein in the crystal. The structure, in combination with mutagenesis, shows that SpPgdA is a metalloenzyme using a His-His-Asp zinc-binding triad with a nearby aspartic acid and histidine acting as the catalytic base and acid, respectively, somewhat similar to other zinc deacetylases such as LpxC. The enzyme is able to accept GlcNAc 3 as a substrate (Km ‫؍‬ 3.8 mM, kcat ‫؍‬ 0.55 s ؊1 ), with the N-acetyl of the middle sugar being removed by the enzyme. The data described here show that SpPgdA and the other family 4 carbohydrate esterases are metalloenzymes and present a step toward identification of mechanismbased inhibitors for this important class of enzymes.crystal structure ͉ metalloenzyme

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mentioning

confidence: 99%

Structure and metal-dependent mechanism of peptidoglycan deacetylase, a streptococcal virulence factor

Blair

Schüttelkopf

MacRae

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

198

389

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“…ClCDA is a representative member of the CE-4 family and shares a number of sequence motifs, covering several conserved histidine and aspartic acid residues ( Figure 1B). It appears from several kinetic studies that divalent cations may play a role in catalysis (12,19,28,29). Previous studies have shown that ClCDA is an endo-chitin deacetylase with four significant subsites, -2,-1, 0, and +1 (where 0 is the catalytic site), and that subsites -2 and 0 strongly recognize the N-acetyl groups on the GlcNAc sugars whereas subsite +1 recognizes a glucosamine moiety (30)(31)(32).…”

mentioning

confidence: 99%

Structure and Mechanism of Chitin Deacetylase from the Fungal Pathogen Colletotrichum lindemuthianum^,

et al. 2006

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The fungal pathogen Colletotrichum lindemuthianum secretes an endo-chitin de-N-acetylase (ClCDA) to modify exposed hyphal chitin during penetration and infection of plants. Although a significant amount of biochemical data is available on fungal chitin de-N-acetylases, no structural data exist. Here we describe the 1.8 Å crystal structure of a ClCDA product complex and the analysis of the reaction mechanism using Hammett linear free energy relationships, subsite probing, and atomic absorption spectroscopy studies. The structural data in combination with biochemical data reveal that ClCDA consists of a single domain encompassing a mononuclear metalloenzyme which employs a conserved His-HisAsp zinc-binding triad closely associated with the conserved catalytic base (aspartic acid) and acid (histidine) to carry out acid/base catalysis. The data presented here indicate that ClCDA possesses a highly conserved substrate-binding groove, with subtle alterations that influence substrate specificity and subsite affinity. Strikingly, the structure also shows that the hexahistidine purification tag appears to form a tight interaction with the active site groove. The enzyme requires occupancy of at least the 0 and +1 subsites by (GlcNAc) 2 for activity and proceeds through a tetrahedral oxyanion intermediate.Colletotrichum lindemuthianum is a plant fungal pathogen found extensively in tropical and subtropical regions. Colletotrichum species are the causative agent of anthracnose that affects economically important crop species (1). Furthermore, Colletotrichum sp. have recently been reported to cause subcutaneous and systemic infections among immunosuppressed patients (2). Colletotrichum sp. are facultative biotrophs. Before the fungal hyphae can successfully penetrate and gain access to host tissue, the fungus has to first evade plant antimicrobial hydrolases such as chitinases and -(1,3)glucanases (3, 4). The chitinases degrade fungal chitin, an insoluble linear polymer of -(1-4)-linked N-acetylglucosamine (GlcNAc). 1 The breakdown products may act as elicitors of active defense responses within the plant (5-7). Studies of cell wall composition of invasive fungal hyphae suggest that exposed fungal chitin polymers are partially de-N-acetylated during infection and initial growth within the host (8). Chitosan, the de-N-acetylated product, is a poor substrate for chitinases, which require the presence of N-acetyl moieties for recognition and catalysis (9). Thus, conversion of chitin to chitosan during plant extracellular colonization may protect pathogenic fungal hyphae from being lysed by secreted plant chitinases. The enzyme responsible for chitin modification is a developmentally regulated, secreted, chitin deacetylase (CDA) (10). C. lindemuthianum chitin deacetylase (ClCDA) is a member of the family 4 carbohydrate esterases (CE-4s) as defined by the CAZY database [http://afmb.cnrs-mrs.fr/∼cazy/CAZY (11)], which include several members that share the primary structure assigned as the "NodB homology domain" (12). Rh...

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“…Unlike other class-1, 2 and 3 CE class enzymes, CE class-4 enzymes are highly specific towards the acetyl xylan substrates and these AcXEs are not active against acetylgalactoglucomannan or acetylated manno-residues. However, the classification of chitin and peptidoglycan deacetylating AcXEs under CE class-4 explains the characteristic ability of CE class-4 AcXEs activity on chitin (Biely et al 1996b; Caufrier et al 2003; Blair et al 2005; Taylor et al 2006). The CE class-5 AcXEs show their specificity towards acetyl xylan residues, acetylated xylo-oligosaccharide, mannosides and cellulose acetate residues by deacetylating them on position 2.…”

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 99%

“…However, these structures can be majorly differentiated based on their protein structures; N-terminus of SpPgdA consists two domains and in SlCE4 C-terminus is involved in the dimerisation of the AcXE. It is reported that SlCE4 also exhibits a strong activity against chitin by acting as chitooligosaccharide de-N-acetylase (Caufrier et al 2003). …”

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 99%

Understanding the structural and functional properties of carbohydrate esterases with a special focus on hemicellulose deacetylating acetyl xylan esterases

Kameshwar

Qin

2018

Mycology

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Acetyl and methyl esterifications are two major naturally found substitutions in the plant cell-wall polysaccharides. The non-cellulosic plant cell-wall polysaccharides such as pectin and hemicellulose are differentially esterified by the O-acetyl and methyl groups to cease the action of various hydrolytic enzymes secreted by different fungi and bacterial species. Thus, microorganisms have emerged with a special class of enzymes known as carbohydrate esterases (CE). The CE catalyse O-de, N-deacetylation of acetylated saccharide residues (esters or amides, where sugars play the role of alcohol/amine/acid). Carbohydrate active enzyme (CAZy) database has classified CE into 16 classes, of which hemicellulose deacetylating CE were grouped into eight classes (CE-1 to CE-7 and CE-16). Various plant biomass degrading fungi and bacteria secretes acetyl xylan esterases (AcXE); however, these enzymes exhibit varied substrate specificities. AcXE and xylanases-coupled pretreatment methods exhibit significant applications, such as enhancing animal feedstock, baking industry, production of food additives, paper and pulp, xylitol production and biorefinery-based industries, respectively. Thus, understanding the structural and functional properties of acetyl xylan esterase will significantly aid in developing the efficient AcXE with wide range of industrial applications.

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Carbohydrate esterase family 4 enzymes: substrate specificity

Cited by 120 publications

References 15 publications

Structure and metal-dependent mechanism of peptidoglycan deacetylase, a streptococcal virulence factor

Structure and metal-dependent mechanism of peptidoglycan deacetylase, a streptococcal virulence factor

Structure and Mechanism of Chitin Deacetylase from the Fungal Pathogen Colletotrichum lindemuthianum^,

Understanding the structural and functional properties of carbohydrate esterases with a special focus on hemicellulose deacetylating acetyl xylan esterases

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Carbohydrate esterase family 4 enzymes: substrate specificity

Cited by 120 publications

References 15 publications

Structure and metal-dependent mechanism of peptidoglycan deacetylase, a streptococcal virulence factor

Structure and metal-dependent mechanism of peptidoglycan deacetylase, a streptococcal virulence factor

Structure and Mechanism of Chitin Deacetylase from the Fungal Pathogen Colletotrichum lindemuthianum,

Understanding the structural and functional properties of carbohydrate esterases with a special focus on hemicellulose deacetylating acetyl xylan esterases

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Structure and Mechanism of Chitin Deacetylase from the Fungal Pathogen Colletotrichum lindemuthianum^,