Multifunctional Xylooligosaccharide/Cephalosporin C Deacetylase Revealed by the Hexameric Structure of the Bacillus subtilis Enzyme at 1.9Å Resolution

Vincent, Florence; Charnock, Simon J.; Verschueren, K.H.G.; Turkenburg, J.P.; Scott, David J.; Offen, W.A.; Roberts, S.M.; Pell, G.; Gilbert, Harry J.; Davies, G.J.; Brannigan, J.A.

doi:10.1016/s0022-2836(03)00632-6

Cited by 67 publications

(89 citation statements)

References 68 publications

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“…Both BpAcXE-apo and BpAcXE-DEP are homo-hexamers, which can be further classified as dimer of two trimers with overall structure resembling a doughnut. The 3-D structures of BpAcXE-apo and BpAcXE-DEP proteins closely resemble the structure of BsAcXE ( Bacillus subtilis acetyl xylan esterase PDB ID: 1ODS (Vincent et al 2003)) with rmsd values of 0.38 Å and 0.41Å from the superimposition of 317 and 311 Cα atoms, respectively. When compared to the well-known α/β hydrolases fold, the subunit folds of BpAcXE-apo and BpAcXE-DEP differs significantly in the presence of extra three-helix bundle, also the presence of second insertion containing two helices and one β-strand near N-terminus (Vincent et al 2003).…”

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 59%

“…The 3-D structures of BpAcXE-apo and BpAcXE-DEP proteins closely resemble the structure of BsAcXE ( Bacillus subtilis acetyl xylan esterase PDB ID: 1ODS (Vincent et al 2003)) with rmsd values of 0.38 Å and 0.41Å from the superimposition of 317 and 311 Cα atoms, respectively. When compared to the well-known α/β hydrolases fold, the subunit folds of BpAcXE-apo and BpAcXE-DEP differs significantly in the presence of extra three-helix bundle, also the presence of second insertion containing two helices and one β-strand near N-terminus (Vincent et al 2003). According to Montoro-García et al (2011), three helix bundle insertion might play a crucial role in hexamer formation as this three-helix bundle interacts with one subunit of adjacent trimer and with another subunit from the same trimer, thus contacting the previous loop of α-helix (α11) on the C-terminal which also resides important His 298 residue of the catalytic triad.…”

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 59%

“…This might propose that these enzymes take part in the degradation of the acetylxylan in its final stages. The CE class-7 AXE’s exhibit a very low positional specificity, these enzymes also exhibit a cephalosporin C deacetylase activity, respectively (Vincent et al 2003; Kremnický et al 2004; Krastanova et al 2005; Biely et al 2007; Biely 2012; Levisson et al 2012). The activity of AcXe is affected by the substrate and positional specificity, respectively.…”

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 99%

“…The X-ray crystallographic structures were determined at resolution 1.9Å (BpAcXE-apo) and 2.7Å (BpAcXE-DEP), respectively (Montoro-García et al 2011). Previous studies have functionally characterised BPAcXE based on the sequence comparison (Vincent et al 2003); enzyme activity (Martínez-Martínez et al 2007) and molecular mass studies have proved that BPAcXE belongs to the AcXE/cephalosporin C deacetylases class enzymes. BPAcXE gene sequence showed sequence identity of 41%, 76% and 89% for AcXE encoding gene sequences of Thermotoga maritima, B. subtilis and B. pumilus strain P213, respectively (Vincent et al 2003; Montoro-García et al 2011).…”

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 99%

“…The AcXE from the above-mentioned organisms contain the CE class-7 conserved sequence motifs RGQ, GxSQG containing the serine residue and HE histidine residues for the catalytic triad formation (Vincent et al 2003; Montoro-García et al 2011). Both BpAcXE-apo and BpAcXE-DEP are homo-hexamers, which can be further classified as dimer of two trimers with overall structure resembling a doughnut.…”

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 99%

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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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Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 59%

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 59%

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 99%

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 99%

Section: Microbial Enzymes Depolymerisation Of Hemicellulosementioning

confidence: 99%

See 3 more Smart Citations

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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Endogenous carbohydrate esterases of Clostridium thermocellum are identified and disrupted for enhanced isobutyl acetate production from cellulose

Seo

Nicely

Trinh

2020

Biotech & Bioengineering

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Medium‐chain esters are versatile chemicals with broad applications as flavors, fragrances, solvents, and potential drop‐in biofuels. Currently, these esters are largely produced by the conventional chemical process that uses harsh operating conditions and requires high energy input. Alternatively, the microbial conversion route has recently emerged as a promising platform for sustainable and renewable ester production. The ester biosynthesis pathways can utilize either lipases or alcohol acyltransferase (AAT), but the AAT‐dependent pathway is more thermodynamically favorable in an aqueous fermentation environment. Even though a cellulolytic thermophile Clostridium thermocellum harboring an AAT‐dependent pathway has recently been engineered for direct conversion of lignocellulosic biomass into esters, the production is not efficient. One potential bottleneck is the ester degradation caused by the endogenous carbohydrate esterases (CEs) whose functional roles are poorly understood. The challenge is to identify and disrupt CEs that can alleviate ester degradation while not negatively affecting the efficient and robust capability of C. thermocellum for lignocellulosic biomass deconstruction. In this study, by using bioinformatics, comparative genomics, and enzymatic analysis to screen a library of CEs, we identified and disrupted the two most critical CEs, Clo1313_0613 and Clo1313_0693, that significantly contribute to isobutyl acetate degradation in C. thermocellum. We demonstrated that an engineered esterase‐deficient C. thermocellum strain not only reduced ester hydrolysis but also improved isobutyl acetate production while maintaining effective cellulose assimilation.

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Regioselective Deprotection of Peracetylated Disaccharides at the Primary Position Catalyzed by Immobilized Acetyl Xylan Esterase from Bacillus pumilus

et al. 2011

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In this work peracetylated lactose and related compounds have been tested as advanced intermediate substrates in enzymatic deprotections catalyzed by acetyl xylan esterase (AXE) from Bacillus pumilus. In particular, we present a simple and efficient enzymatic procedure to obtain products bearing only one free primary hydroxy group in the C6′ position of the galactoside moiety of “lacto” carbohydrates. In particular, good yields (60–70 %) have been achieved in the hydrolysis of 1‐O‐methyl lactosamine and hexaacetyl lactal. The products obtained are of great interest as advanced building blocks for the synthesis of very important oligosaccharides of biological relevance.

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Multifunctional Xylooligosaccharide/Cephalosporin C Deacetylase Revealed by the Hexameric Structure of the Bacillus subtilis Enzyme at 1.9Å Resolution

Cited by 67 publications

References 68 publications

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

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

Endogenous carbohydrate esterases of Clostridium thermocellum are identified and disrupted for enhanced isobutyl acetate production from cellulose

Regioselective Deprotection of Peracetylated Disaccharides at the Primary Position Catalyzed by Immobilized Acetyl Xylan Esterase from Bacillus pumilus

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