A Novel Family 8 Xylanase, Functional and Physicochemical Characterization

Collins, Tony; Meuwis, Marie‐Alice; Stals, Ingeborg; Claeyssens, Marc; Feller, Georges; Gerday, Charles

doi:10.1074/jbc.m204517200

Cited by 174 publications

(131 citation statements)

References 49 publications

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“…In the retention mechanism, hydrolyses are generally achieved through a double-displacement mechanism in which a glycosyl-enzyme intermediate is initially formed followed by its hydrolysis through oxocarbenium-ion transition states [70]. On the other hand, the GH-8 family includes enzymes that catalyze the glycosidic hydrolysis through a single-displacement inverting mechanism with α-configuration products [71,72]. In the inverting mechanism, a carboxyl group (acting as the general acid) and a carboxylate group (acting as the general base) are considered as the two main functional groups.…”

Section: Discussionmentioning

confidence: 99%

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Ibrahim¹,

Jones²,

Hossenya³

2013

J Pet Environ Biotechnol

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Lignocellulosic biomass has potential for bioethanol, a renewable fuel. A limitation is that bioconversion of the complex lignocellulosic material to simple sugars and then to bioethanol is a challenging process. Recent work has focused on the genetic engineering of a biocatalyst that may play a critical role in biofuel production. Escherichia coli have been considered a convenient host for biocatalysts in biofuel production for its fermentation of glucose into a wide range of short-chain alcohols and production of highly deoxygenated hydrocarbon. The bacterium Pectobacterium carotovorum subsp. carotovorum (P. carotovorum) is notorious for its maceration of the plant cell wall causing soft rot. The ability to destroy plants is due to the expression and secretion of a wide range of hydrolytic enzymes that include cellulases and polygalacturonases. P. carotovorum ATCC™ no. 15359 was used as a source of DNA for the amplification of celB, celC and peh. These genes encode 2 cellulases and a polygalacturonase, respectively. Primers were designed based on published gene sequences and used to amplify the open reading frames from the genomic DNA of P. carotovorum. The individual PCR products were cloned into the pTAC-MAT-2 expression vector and transformed into Escherichia coli. The deduced amino acid sequences of the cloned genes have been analyzed for their catalytically active domains. Estimation of the molecular weights of the expressed proteins was performed using SDS-PAGE analysis and celB, celC and peh products were approximately 29.5 kDa, 40 kDa 41.5 kDa, respectively. Qualitative determination of the cellulase and polygalacturonase activities of the cloned genes was carried out using agar diffusion assays.

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Section: Discussionmentioning

confidence: 99%

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Ibrahim¹,

Jones²,

Hossenya³

2013

J Pet Environ Biotechnol

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“…In addition, endo b-1,4 xylanase (pXyl), belonging to this family, was found in a culture supernatant of Pseudoalteromonas haloplanktis and characterized (72). The three dimensional structures of two GH8 enzymes were found to have an (a/a)6 barrel structure (clan GH-M) (73,74).…”

Section: A Model Of Inverting Gh Reducing End Xylose-releasing Exomentioning

confidence: 98%

“…The three dimensional structures of two GH8 enzymes were found to have an (a/a)6 barrel structure (clan GH-M) (73,74). The GH8 xylanase, pXyl, has the highest amino acid identity (32.6 z) with the protein encoded by the BH2105 gene (GenBank accession number: BAB05824) of Bacillus halodurans C-125, an alkalophilic bacterium whose genomic sequence is available (72,75).…”

Section: A Model Of Inverting Gh Reducing End Xylose-releasing Exomentioning

confidence: 99%

“…Collins et al (72,79) found that a mutation at a conserved tyrosine residue (Y203) in pXyl, which shares 32.6z amino acid identity with Rex, caused a marked decrease in the hydrolytic activity of the enzyme toward xylan. The residue formed a hydrogen bond with a nucleophilic water molecule that formed another hydrogen bond with the general base residue (D281).…”

Section: E Conversion Of Rex Into Glycosynthase E-1 Hehre Resynthesmentioning

confidence: 99%

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Conversion of inverting glycoside hydrolases into catalysts for synthesizing glycosides employing a glycosynthase strategy

Kitaoka

Honda

Fushinobu

et al. 2009

TIGG

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23Trends in Glycoscience and Glycotechnology Vol. 21 No. 117 (2009) AbstractReducing-end xylose-releasing exo-oligoxylanase (Rex, EC. 3.2.1.156) is an inverting xylanolytic enzyme, belonging to the glycoside hydrolase (GH) family 8, which hydrolyzes xylooligosaccharides to release xylose (X1) from its reducing end. Rex hydrolyzes a-xylobiosyl ‰uo-ride (a-X2F) to yield xylobiose (X2) only in the presence of X1, conˆrming the Hehre resynthesis-hydrolysis mechanism. A library of mutant Rex at the catalytic base (D263) was constructed by saturation mutagenesis, in which D263C accumulated the highest level of xylotriose (X 3 ) from a-X 2 F and X 1 . However, F -releasing activities of the mutants were much less than that of the wild type. Next, Y198 residue of Rex that forms a hydrogen bond with nucleophilic water was substituted with phenylalanine, causing a marked decrease in hydrolytic activity and a small increase in the F -releasing activity from a-X2F in the presence of X1. Y198F of Rex accumulated more product during the glycosynthase reaction than D263C. Recently, an inverting a-1,2-fucosidase belonging to GH95 was converted into glycosynthase by mutating a catalytic base residue. In both cases, the catalytic base should be intact.

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“…Xylan, a major component of the plant cell wall, consists of a backbone of β-(1,4)-linked d-xylosyl residues with substitutions by arabinosyl, acetyl, and glucuronosyl residues (Thomson 1993;Cazemier et al 1999;Collins et al 2002). The hydrolysis of the xylan backbone involves several enzymes, and the most important one is endo-β-(1,4)-xylanase (EC 3.2.1.8).…”

mentioning

confidence: 99%

Optimisation of solid-state fermentation of Aspergillus niger JL-15 for xylanase production and xylooligosaccharides preparation

Dai¹,

Liu²,

HongXiao³

et al. 2011

Czech J. Food Sci.

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The production of xylanase (XylA) by Aspergillus niger JL-15 in solid-state fermentation (SSF) on orange peel was optimised by the response surface methodology (RSM). The results revealed that four factors had significant effects on the XylA production (P < 0.05), that is the concentrations of the added glycerin and ammonium sulfate, the moisture content, and fermentation time. Exploying orange peel as the solid substrate, maximum xylanase activity (917.7 U/g dry fermentation product) was obtained at 4.2% glycerin, 3.1% (NH 4 ) 2 SO 4 , 61% moisture content, and 73.4-h fermentation, this activity being close to the predicted one and 3.2 times higher than that of the basic medium (218.5 U/g). Optimum temperature and pH for XylA activity were 55°C and pH 5.0, respectively. SDS-PAGE analysis showed that the relative molecular mass of XylA was about 30.0 kDa. XylA exhibited K m and V max values of 9.24 mg/ml and 54.05 μmol/min/ml, respectively. XylA liberated mainly xylotriose from birchwood xylan and wheat bran, respectively. XylA was an endo-acting xylanase with transglycosylation activity, with the ability to hydrolyse, xylobiose, xylotriose, xylotetraose, xylopentaose, and xylohexaose.

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A Novel Family 8 Xylanase, Functional and Physicochemical Characterization

Cited by 174 publications

References 49 publications

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Molecular Cloning and Expression of Cellulase and Polygalacturonase Genes in E. coli as a Promising Application for Biofuel Production

Conversion of inverting glycoside hydrolases into catalysts for synthesizing glycosides employing a glycosynthase strategy

Optimisation of solid-state fermentation of Aspergillus niger JL-15 for xylanase production and xylooligosaccharides preparation

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