Cloning, expression and characterization of glycoside hydrolase family 11 endoxylanase from Bacillus pumilus ARA

Qu, Wei; Shao, Weilan

doi:10.1007/s10529-011-0568-x

Cited by 20 publications

(6 citation statements)

References 23 publications

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“…Bioinformatic analysis of the spectra on the Mascot platform identified peptides belonging to the endo-1,4-β-xylanase of Bacillus sonorensis or uncultured bacteria with scores 304 and 101, respectively ( Table 2 ). The endo-1,4-β-xylanase is affiliated with glycosyl hydrolases [ 45 ] and is often found in bacteria of the genus Bacillus [ 12 , 14 – 16 , 18 – 21 , 23 , 26 , 46 , 47 ].…”

Section: Resultsmentioning

confidence: 99%

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Cloning, expression, and characterization of a recombinant xylanase from Bacillus sonorensis T6

et al. 2022

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Xylanase is one of industrial enzymes with diverse applications including the paper-bleaching industry and feed additives. Here, a strain having xylanolytic activity and identified as Bacillus sonorensis T6 was isolated from soil. A secretory enzyme was identified by mass-spectrometry as a xylanase of glycosyl hydrolase family 11, with a molecular weight of 23.3 kDa. The xylanase gene of Bacillus sonorensis T6 was cloned and expressed in Escherichia coli (yielding an enzyme designated as rXynT6-E) and in Pichia pastoris (yielding rXynT6-P). The recombinant xylanases were found to have optimal activity at 47–55°C and pH 6.0–7.0. The recombinant xylanase expressed in P. pastoris has 40% higher thermal stability than that expressed in E. coli. The recombinant xylanases retained 100% of activity after 10 h incubation in the pH range 3–11 and 68% of activity after 1 h at pH 2.0. The xylanase activities of rXynT6-E and rXynT6-P under optimal conditions were 1030.2 and 873.8 U/mg, respectively. The good stability in a wide range of pH and moderate temperatures may make the xylanase from Bacillus sonorensis T6 useful for various biotechnological applications, e.g., as an enzyme additive in the feed industry.

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Section: Resultsmentioning

confidence: 99%

“…Glycosylation did not influence pH stability of the XynT6 enzyme. There are data in the literature on pH stability of xylanases; the enzymes that have been studied to date retain no more than 80% of activity after incubation for 1 h in the pH range 5.0–9.6 [ 20 , 21 , 26 ].…”

Section: Resultsmentioning

confidence: 99%

Cloning, expression, and characterization of a recombinant xylanase from Bacillus sonorensis T6

et al. 2022

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“…Moreover, the pH stability of Pc Xyn11A in this study was much broader than those reported from A. awamori VTCC-F312 (pH 4.0–8.0) [42], Achaetomium sp. Xz-8 (pH 5.0–10.0) [9], Aspergillus fumigatus MKU1 (pH 4.0–8.0) [45], Bacillus pumilus ARA (pH 5.8–8.2) [46], Bacillus sp. GA1(6) (pH 4.0–7.0) [47] and Penicillium oxalicum (pH 3.0–5.0) [48].…”

Section: Resultsmentioning

confidence: 99%

High-Level Heterologous Expression of Endo-1,4-β-Xylanase from Penicillium citrinum in Pichia pastoris X-33 Directed through Codon Optimization and Optimized Expression

et al. 2019

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Most common industrial xylanases are produced from filamentous fungi. In this study, the codon-optimized xynA gene encoding xylanase A from the fungus Penicilium citrinum was successfully synthesized and expressed in the yeast Pichia pastoris. The levels of secreted enzyme activity under the control of glyceraldehyde-3-phosphate dehydrogenase (PGAP) and alcohol oxidase 1 (PAOX1) promoters were compared. The Pc Xyn11A was produced as a soluble protein and the total xylanase activity under the control of PGAP and PAOX1 was 34- and 193-fold, respectively, higher than that produced by the native strain of P. citrinum. The Pc Xyn11A produced under the control of the PAOX1 reached a maximum activity of 676 U/mL when induced with 1% (v/v) methanol every 24 h for 5 days. The xylanase was purified by ion exchange chromatography and then characterized. The enzyme was optimally active at 55 °C and pH 5.0 but stable over a broad pH range (3.0–9.0), retaining more than 80% of the original activity after 24 h or after pre-incubation at 40 °C for 1 h. With birchwood xylan as a substrate, Pc Xyn11A showed a Km(app) of 2.8 mg/mL, and a kcat of 243 s−1. The high level of secretion of Pc Xyn11A and its stability over a wide range of pH and moderate temperatures could make it useful for a variety of biotechnological applications.

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“…The SDS–PAGE analysis indicated that the recombinant xylanase was expressed at high levels (Fig. A), and the expression level and purification strategy was appropriate for the industrial production of the enzyme . The protein content was 6.47 mg ml −1 , and the specific activity of purified xynB xylanase was 1916 U mg −1 , which was approximately 1.5‐fold higher than xylanase B in A. niger (1250 U mg −1 ) .…”

Section: Discussionmentioning

confidence: 99%

Cloning and expression of a xylanase xynB from Aspergillus nigerIA‐001 in Pichia pastoris

Fang

Gao

Cao

et al. 2013

J. Basic Microbiol.

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The high-level expression of the xylanase GH11 gene from Aspergillus niger IA-001 called xynB was successfully completed in Pichia pastoris. The xynB gene encoding a mature xylanase of 225 amino acid was subcloned into the pPICZαA vector and was transformed into P. pastoris X-33 under the control of the alcohol oxidase I (AOX1) promoter. The xynB gene was ligated with a sequence encoding modified α-factor signal peptide (pPICZαmA) and the recombinant xylanase activity, which was measured 1280 U ml(-1), was 1.5-fold higher than when it was inserted into pPICZαA and was 19.39-fold greater than the native xylanase in the original strain. In a 10 L fermenter, the recombinant xylanase activity measured 10,035 U ml(-1) after 114 h. The SDS-PAGE analysis revealed that the purified xynB protein migrated as a single band with an apparent molecular weight of 24 kDa. The specific activity, using beechwood xylan as a substrate, was 1916 U mg(-1). The xylanase activity was optimal at pH 5.0 and at 50 °C. In addition, the xynB was active over a pH range of 2.2 to 10.0. The apparent Km and Vmax values were 4.429 mg ml(-1) and 1429 U mg(-1), respectively.

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Cloning, expression and characterization of glycoside hydrolase family 11 endoxylanase from Bacillus pumilus ARA

Cited by 20 publications

References 23 publications

Cloning, expression, and characterization of a recombinant xylanase from Bacillus sonorensis T6

Cloning, expression, and characterization of a recombinant xylanase from Bacillus sonorensis T6

High-Level Heterologous Expression of Endo-1,4-β-Xylanase from Penicillium citrinum in Pichia pastoris X-33 Directed through Codon Optimization and Optimized Expression

Cloning and expression of a xylanase xynB from Aspergillus nigerIA‐001 in Pichia pastoris

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