Cloning and expression of a Paecilomyces thermophila xylanase gene in E. coli and characterization of the recombinant xylanase

Zhang, Min; Jiang, Zhengqiang; Yang, Shaoqing; Hua, Chengwei; Li-te, LI

doi:10.1016/j.biortech.2009.08.055

Cited by 43 publications

(11 citation statements)

References 35 publications

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“…The K m value of the enzyme is similar to that of xylanases reported from other bacteria and the same genus bacteria [18,27,30,31]. The specific activities of several commercial thermostable xylanases reported from Sigma-Aldrich (http://www.sigmaaldrich.com) are much lower than Xyn10A.…”

Section: Discussionsupporting

confidence: 74%

“…thermarum DSM 5069 exhibited the highest activity at temperature 95°C (Figure 1b). The optimal temperature was higher than the xylanases from Chaetomium sp ., Nonomuraea flexuosa , Paecilomyces thermophila , Talaromyces thermophilus and Cellulomonas flavigena , whose optimal temperature were ranged from 50°C to 80°C [3,7,27,28]. However, all of these thermophilic or hyperthermophilic xylanases share the similar optimal pH near 7.0.…”

Section: Discussionmentioning

confidence: 99%

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A novel highly thermostable xylanase stimulated by Ca2+ from Thermotoga thermarum: cloning, expression and characterization

Shi

Zhang

et al. 2013

Biotechnol Biofuels

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BackgroundXylanase is an important component of hemicellulase enzyme system. Since it plays an important role in the hydrolysis of hemicellulose into xylooligosaccharides (XOs), high thermostable xylanase has been the focus of much recent attention as powerful enzyme as well as in the field of biomass utilization.ResultsA xylanase gene (xyn10A) with 3,474 bp was cloned from the extremely thermophilic bacterium Thermotoga thermarum that encodes a protein containing 1,158 amino acid residues. Based on amino acid sequence homology, hydrophobic cluster and three dimensional structure analyses, it was attested that the xylanase belongs to the glycoside hydrolase (GH) families 10 with five carbohydrate binding domains. When the xylanase gene was cloned and expressed in Escherichia coli BL21 (DE3), the specific enzyme activity of xylanase produced by the recombinant strain was up to 145.8 U mg-1. The xylanase was optimally active at 95°C, pH 7.0. In addition, it exhibited high thermostability over broad range of pH 4.0-8.5 and temperature 55-90°C upon the addition of 5 mM Ca2+. Confirmed by Ion Chromatography System (ICS) analysis, the end products of the hydrolysis of beechwood xylan were xylose, xylobiose, xylotriose, xylotetraose, xylopentaose and xylohexaose.ConclusionsThe xylanase from T. thermarum is one of the hyperthermophilic xylanases that exhibits high thermostability, and thus, is a suitable candidate for generating XOs from cellulosic materials such as agricultural and forestry residues for the uses as prebiotics and precursors for further preparation of furfural and other chemicals.

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Section: Discussionsupporting

confidence: 74%

Section: Discussionmentioning

confidence: 99%

A novel highly thermostable xylanase stimulated by Ca2+ from Thermotoga thermarum: cloning, expression and characterization

Shi

Zhang

et al. 2013

Biotechnol Biofuels

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“…Transglycosylation by xylanases has been reported with another family 10 xylanase, XynB of Thermotoga maritima [30,31], and several family 11 xylanases, Xyn1 of Paenibacillus sp. strain W-61 [34], TfxA of Thermomonospora fusca, AnxA of Aspergillus niger, and their hybrid xylanase ATX [35], and XynA of Paecilomyces thermophilia [36]. The transglycosylation activity can be used for the synthesis of alkyl oligoglycosides [33,34].…”

Section: Analysis Of End Productsmentioning

confidence: 99%

Characterization of Xyn10J, a Novel Family 10 Xylanase from a Compost Metagenomic Library

Jeong

Kim

et al. 2012

Appl Biochem Biotechnol

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A gene encoding an extracellular xylanase was cloned from a compost metagenomic library. The xylanase gene, xyn10J, was 1,137 bp in length and was predicted to encode a protein of 378 amino acid residues with a putative signal peptide of 27 amino acid residues. The molecular mass of the mature Xyn10J was calculated to be 39,882 Da with a pI of 6.09. Xyn10J had a motif GVKVHFTEMDI characteristic of most members of glycosyl hydrolase family 10. The amino acid sequence of Xyn10J showed 60.0% identity to that of XynH, a xylanase from an uncultured soil bacterium and 55% identity to XylC of Cellvibrio mixtus. Site-directed mutagenesis of the expected active site based on the sequence analysis indicated that an aspartic acid residue (Asp207), in addition to the identified catalytic residues Glu165 and Glu270, plays a crucial role for the catalytic activity. The purified Xyn10J had a mass of about 40 kDa and was optimally active at pH 7.0 and 40 °C. Xyn10J hydrolyzed beechwood xylan > birchwood xylan > oat spelt xylan > arabinoxylan. Xyn10J hydrolyzed xylotetraose and xylohexaose exclusively to xylobiose, xylopentaose, and xylotriose mainly to xylobiose with transglycosylation activity. The saccharification of reed (Phragmites communis) powder by commercial enzymes was significantly increased by the addition of a small amount of Xyn10J to the commercial preparation. Xyn10J is the first xylanase screened directly from a compost metagenomic library, and the enzyme has the potential to be used in the conversion of biomass to fermentable sugars for biofuel production.

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“…Probably, not all GH10 xylanases form xylose as the main hydrolysis product of xylan, and some members of the GH10 family are structurally similar to GH11 xylanases. GH10 xylanases that did not form xylose were described previously (Waeonukul et al 2009; Zhang et al 2010). …”

Section: Discussionmentioning

confidence: 99%

Xylanases of Cellulomonas flavigena: expression, biochemical characterization, and biotechnological potential

et al. 2017

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Four xylanases of Cellulomonas flavigena were cloned, expressed in Escherichia coli and purified. Three enzymes (CFXyl1, CFXyl2, and CFXyl4) were from the GH10 family, while CFXyl3 was from the GH11 family. The enzymes possessed moderate temperature stability and a neutral pH optimum. The enzymes were more stable at alkaline pH values. CFXyl1 and CFXyl2 hydrolyzed xylan to form xylobiose, xylotriose, xylohexaose, xylopentaose, and xylose, which is typical for GH10. CFXyl3 (GH11) and CFXyl4 (GH10) formed the same xylooligosaccharides, but xylose was formed in small amounts. The xylanases made efficient saccharification of rye, wheat and oat, common components of animal feed, which indicates their high biotechnological potential.

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Cloning and expression of a Paecilomyces thermophila xylanase gene in E. coli and characterization of the recombinant xylanase

Cited by 43 publications

References 35 publications

A novel highly thermostable xylanase stimulated by Ca2+ from Thermotoga thermarum: cloning, expression and characterization

A novel highly thermostable xylanase stimulated by Ca2+ from Thermotoga thermarum: cloning, expression and characterization

Characterization of Xyn10J, a Novel Family 10 Xylanase from a Compost Metagenomic Library

Xylanases of Cellulomonas flavigena: expression, biochemical characterization, and biotechnological potential

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