Cloning of β-1,3-1,4-glucanase gene from Bacillus licheniformis EGW039 (CGMCC 0635) and its expression in Escherichia coli BL21 (DE3)

Teng, Da; Wang, Jianhua; Fan, Ying; Yang, Yalin; Tian, Zhigang; Luo, Jianxun; Yang, Guanpin; Fan, Zhang

doi:10.1007/s00253-006-0329-2

Cited by 62 publications

(49 citation statements)

References 32 publications

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“…There was no significant differences between the native and the recombinant proteins. The recombinant protein exhibited maximal enzyme activity at 65°C with oat β-glucan as substrate, and showed a higher thermoresistance than previously reported for β-1,3-1,4-glucanases, such as from B. subtilis MA139 (40°C) (Qiao et al, 2009), B. licheniformis EGW039 (40°C) (Teng et al, 2006), B. amyloliquefaciens ATCC 23350 (50°C) (Sun et al, 2012), and thermophilic Rhizomucor miehei (60°C) (Tang et al, 2012). For the thermostability assay, the native and recombinant proteins were stable at 65°C for 1.5 h and had residual activity of 82.7 and 90.77%, respectively.…”

Section: Discussionmentioning

confidence: 76%

“…For the thermostability assay, the native and recombinant proteins were stable at 65°C for 1.5 h and had residual activity of 82.7 and 90.77%, respectively. For the native protein, it retained 67.83 and 33% of its activity after incubation at 75° and 85°C for 10 min, respectively, while for the recombinant protein the result was 72.88 and 42.36%, respectively, which is higher than that of β-1,3-1,4-glucanases obtained from B. amyloliquefaciens (50%, 4 min at 70°C) (Hofemeisrer et al, 2003), B. subtilis MA139 (10%, 5 min at 70°C) (Qiao et al, 2009), and B. licheniformis EGW039 (50%, 10 min at 70°C) (Teng et al, 2006). The high thermal stability of the protein may be related to its structure.…”

Section: Discussionmentioning

confidence: 92%

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Purification, characterization, and heterologous expression of an antifungal protein from the endophytic Bacillus subtilis strain Em7 and its activity against Sclerotinia sclerotiorum

Wang¹,

Gao²,

Yan³

et al. 2015

Genet. Mol. Res.

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ABSTRACT. An antifungal protein exhibiting a high activity against Sclerotinia sclerotiorum in vivo was purified by ammonium sulfate precipitation, hydrophobic chromatography, and gel filtration chromatography from the culture filtrate of the endophytic Bacillus subtilis strain Em7. The protein was characterized as a β-1,3-1,4-glucanase according to amino acid analysis, and showed excellent properties in thermal stability and acid resistance. At the same time, the antifungal protein was cloned and heterologously expressed in Escherichia coli BL21. The recombinant protein was purified and showed similar enzymatic properties to the native protein, exhibiting strong inhibitory activity against S. sclerotiorum. This shows that the β-1,3-1,4-glucanase may play a very important role in B. subtilis Em7 biocontrol function. In addition, many physiochemical properties of the native and purified recombinant protein were compared, including the effect of pH, temperature, metal cations, substrate specificity, and kinetic parameters. All parameters were similar between the native and recombinant pu- rified protein, indicating that the purified recombinant protein has potential for industrial applications.

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Section: Discussionmentioning

confidence: 76%

Section: Discussionmentioning

confidence: 92%

Purification, characterization, and heterologous expression of an antifungal protein from the endophytic Bacillus subtilis strain Em7 and its activity against Sclerotinia sclerotiorum

Wang¹,

Gao²,

Yan³

et al. 2015

Genet. Mol. Res.

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“…During the malting and saccharification process, β-1,3-1,4-glucanase is heat-inactivated, leaving a high content of β-glucan in wort (Teng et al, 2006). Immobilization can increase the stability of enzymes (Sakai et al, 1991).…”

Section: Discussionmentioning

confidence: 99%

“…Traditionally, β-1,3-1,4-glucanase is made by separation and purification from natural sources, such as Penicillium emersonii, Aspergillus niger, Bacillus subtilis, and Trichoderma reesei (Harman and Kubicek, 1998).microorganisms, including Lactococcus lactis , Escherichia coli (Teng et al, 2006;Qiao et al, 2009), Pichia pastoris (Teng et al, 2007), and Saccharomyces cerevisiae (Hinchliffe and Box, 1984;van Rensburg et al, 1997;Zhang Q. et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability

Guo

Zhang

et al. 2010

J. Zhejiang Univ. Sci. B

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Abstract:The aim of this work was to construct a novel food-grade industrial arming yeast displaying β-1,3-1,4-glucanase and to evaluate the thermal stability of the glucanase for practical application. For this purpose, a bi-directional vector containing galactokinase (GAL1) and phosphoglycerate kinase 1 (PGK1) promoters in different orientations was constructed. The β-1,3-1,4-glucanase gene from Bacillus subtilis was fused to α-agglutinin and expressed under the control of the GAL1 promoter. α-galactosidase induced by the constitutive PGK1 promoter was used as a food-grade selection marker. The feasibility of the α-galactosidase marker was confirmed by the growth of transformants harboring the constructed vector on a medium containing melibiose as a sole carbon source, and by the clear halo around the transformants in Congo-red plates owing to the expression of β-1,3-1,4-glucanase. The analysis of β-1,3-1,4-glucanase activity in cell pellets and in the supernatant of the recombinant yeast strain revealed that β-1,3-1,4-glucanase was successfully displayed on the cell surface of the yeast. The displayed β-1,3-1,4-glucanase activity in the recombinant yeast cells increased immediately after the addition of galactose and reached 45.1 U/ml after 32-h induction. The thermal stability of β-1,3-1,4-glucanase displayed in the recombinant yeast cells was enhanced compared with the free enzyme. These results suggest that the constructed food-grade yeast has the potential to improve the brewing properties of beer.

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“…β-1,3-1,4-Glucanase gene from different sources has been cloned and expressed in Escherichia coli (Hinchliffe 1984;Teng et al 2006), Bacillus (Kim 2003), Lactobacillus (Heng et al 1997), Escherichia faecalis (Ekinci et al 1997), and Saccharomyces cerevisiae (Cantwell et al 1986;Chen et al 1992). However, due to the low activity and stability, the application of these enzymes is not popularized in manufacture of commercial products.…”

mentioning

confidence: 99%

Codon optimization, expression and characterization of Bacillus subtilis MA139 β-1,3-1,4-glucanase in Pichia pastoris

et al. 2010

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β-1,3-1,4-Glucanase has been broadly used in feed and brewing industries. According to the codon bias of Pichia pastoris, the Bacillus subtilis MA139 β-1,3-1,4-glucanase gene was de novo synthesized and expressed in P. pastoris X-33 strain under the control of the alcohol oxidase 1 promoter. In a 10-L fermentor, the β-1,3-1,4-glucanase was overexpressed with a yield of 15,000 U/mL by methanol induction for 96 h. The recombinant β-1,3-1,4-glucanase exhibited optimal activity at 40• C and pH 6.4. The activity of the recombinant β-1,3-1,4-glucanase was not significantly affected by various metal ions and chemical reagents. To our knowledge, the expression of this β-1,3-1,4-glucanase from Bacillus sp. in P. pastoris is in relatively high level compared to previous reports. These biochemical characteristics suggest that the recombinant β-1,3-1,4-glucanase has a prospective application in feed and brewing industries.

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Cloning of β-1,3-1,4-glucanase gene from Bacillus licheniformis EGW039 (CGMCC 0635) and its expression in Escherichia coli BL21 (DE3)

Cited by 62 publications

References 32 publications

Purification, characterization, and heterologous expression of an antifungal protein from the endophytic Bacillus subtilis strain Em7 and its activity against Sclerotinia sclerotiorum

Purification, characterization, and heterologous expression of an antifungal protein from the endophytic Bacillus subtilis strain Em7 and its activity against Sclerotinia sclerotiorum

A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability

Codon optimization, expression and characterization of Bacillus subtilis MA139 β-1,3-1,4-glucanase in Pichia pastoris

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