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Purification and characterization of a new endo-β-1,3-glucanase exhibiting a high specificity for curdlan for production of β-1,3-glucan oligosaccharides

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Abstract

Endo-β-1,3-glucanase (Endo23) was purified from a Trichoderma reesei GIMCC 3.498 fermentation broth using anion exchange and 2-stage size exclusion chromatography. Purification of 44.5× and a 12% recovery yield of enzyme activity were achieved. The Mw and isoelectric point were estimated to be 24 kDa and 3.85 using SDS-PAGE and IEF, respectively. The highest substrate specificity was observed for water-insoluble curdlan. The optimal conditions for hydrolyzing curdlan were pH 5.0 and 50°C. The main hydrolytic products were glucobiose and glucotriose. Minor amounts of glucose and glucotetraose were detected. Hg2+, Fe2+, Fe3+, and Sn2+ inhibited the hydrolysis activity of Endo23 at 5 and 50 mM. K+ slightly promoted Endo23 activity. Endo23 belongs to the category EC3.2.1.39. The peptide sequences of Endo23 showed identity with conserved sequences that typically exist in β-1,3-glucanases of the glycoside hydrolase family. The Endo23 sequence was partially similar to a hypothetical lignocellulase from Penicillium oxalicum 114-2.

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Authors and Affiliations

  1. Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, 214122, China

    Jing Li, Xiao-Bei Zhan, Min Xu, Chi-Chung Lin & Zhi-Yong Zheng

  2. Jiangsu Rayguang Biotech Co. Ltd., Wuxi, Jiangsu, 214125, China

    Li Zhu & Xiao-Bei Zhan

  3. Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, Jiangsu, 214122, China

    Wei-Jiang Li

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  1. Jing Li
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Correspondence to Xiao-Bei Zhan.

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Li, J., Zhu, L., Zhan, XB. et al. Purification and characterization of a new endo-β-1,3-glucanase exhibiting a high specificity for curdlan for production of β-1,3-glucan oligosaccharides. Food Sci Biotechnol 23, 799–806 (2014). https://doi.org/10.1007/s10068-014-0108-2

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  • DOI: https://doi.org/10.1007/s10068-014-0108-2

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