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Purification and functional characterization of endo-β-mannanase MAN5 and its application in oligosaccharide production from konjac flour

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Abstract

MAN5, the main extracellular saccharide hydrolase from Bacillus sp. MSJ-5, is an endo-β-mannanase with a demand of at least five sugar moieties for effective cleavage. It has a pH optimum of 5.5 and a temperature optimum of 50°C and is stable at pH 5–9 or below 65°C. MAN5 has a very high ability to hydrolyze konjac flour, 10 U/mg of which could completely liquefy konjac flour gum in 10 min at 50°C. HPLC analysis showed that most glucomannan in the konjac flour was hydrolyzed into a large amount of oligosaccharides with DP of 2–6 and a very small amount of monosaccharide. With the culture supernatant as enzyme source, the optimum condition to prepare oligosaccharides from konjac flour was obtained as 10 mg/ml konjac flour incubated with 10 U/mg enzyme at 50°C for 24 h. With this condition, more than 90% polysaccharides in the konjac flour solution were hydrolyzed into oligosaccharides and a little monosaccharide (2.98% of the oligosaccharides). Konjac flour is an underutilized agricultural material with low commercial value in China. With MAN5, konjac flour can be utilized to generate high value-added oligosaccharides. The high effectiveness and cheapness of this technique indicates its potential in industry.

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Acknowledgments

The work was supported by the Program for New Century Excellent Talents in University (NCET-04-0637), Foundation for Young Excellent Scientists in Shandong Province (2004BS06001), and COMRA Program (DYXM-115-02-2-6).

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Authors and Affiliations

  1. The State Key Lab of Microbial Technology, Shandong University, Jinan, 250100, People’s Republic of China

    Min Zhang, Xiu-Lan Chen, Zhi-Hua Zhang, Cai-Yun Sun, Lei-Lei Chen, Hai-Lun He, Bai-Cheng Zhou & Yu-Zhong Zhang

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  1. Min Zhang
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  2. Xiu-Lan Chen
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  3. Zhi-Hua Zhang
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  4. Cai-Yun Sun
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Correspondence to Yu-Zhong Zhang.

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Min Zhang and Xiu-Lan Chen contributed equally to this work.

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Zhang, M., Chen, XL., Zhang, ZH. et al. Purification and functional characterization of endo-β-mannanase MAN5 and its application in oligosaccharide production from konjac flour. Appl Microbiol Biotechnol 83, 865–873 (2009). https://doi.org/10.1007/s00253-009-1920-0

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  • DOI: https://doi.org/10.1007/s00253-009-1920-0

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