Purification, characterization, and overexpression of an endo-1,4-β-mannanase from thermotolerant Bacillus sp. SWU60

  • Weeranuch Seesom
  • Polphet Thongket
  • Tomohiro Yamamoto
  • Shigeo Takenaka
  • Tatsuji Sakamoto
  • Wasana Sukhumsirichart
Original Paper


Endo-β-1,4-mannanases are important catalytic agents in several industries. The enzymes randomly cleave the β-1,4-linkage in the mannan backbone and release short β-1,4-mannooligosaccharides and mannose. In the present study, mannanase (ManS2) from thermotolerant Bacillus sp. SWU60 was purified, characterized, and its gene was cloned and overexpressed in Escherichia coli. ManS2 was purified from culture filtrate (300 ml) by using hydrophobic, ion-exchange, and size-exclusive liquid chromatography. The apparent molecular mass was 38 kDa. Optimal pH and temperature for enzyme activity were 6.0 and 60 °C, respectively. The enzyme was stable up to 60 °C for 1 h and at pH 5–9 at 4 °C for 16 h. Its enzyme activity was inhibited by Hg2+. The full-length mans2 gene was 1,008 bp, encoding a protein of 336 amino acids. Amino acid sequence analysis revealed that it belonged to glycoside hydrolase family 26. Konjac glucomannan was a favorable substrate for recombinant ManS2 (rManS2). rManS2 also degraded galactomannan from locust bean gum, indicating its potential for production of glucomanno- and galactomanno-oligosaccharides. Both native and recombinant ManS2 from Bacillus sp. SWU60 can be applied in several industries especially food and feed.


Endo-β-1,4-mannanases Bacillus sp. Recombinant enzyme Mannan Glucomannan Galactomannan 



This work was supported by grant from Srinakhariwirot university (017/2557) and performed under the Core-to-Core Program, which was financially supported by Japan Society for the Promotion of Science (JSPS), National Research Council of Thailand (NRCT), Vietnam Ministry of Science and Technology (MOST), the National University of Laos, Beuth University of Applied Sciences and Brawijaya University. The authors thank Dr. Daisuke Fujiwara of Osaka Prefecture University for assistance with the MALDI-TOF MS experiment. We would like to thank Editage ( for English language editing.


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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Weeranuch Seesom
    • 1
  • Polphet Thongket
    • 1
  • Tomohiro Yamamoto
    • 2
  • Shigeo Takenaka
    • 3
  • Tatsuji Sakamoto
    • 2
  • Wasana Sukhumsirichart
    • 1
  1. 1.Department of Biochemistry, Faculty of MedicineSrinakharinwirot UniversityBangkokThailand
  2. 2.Division of Applied Life Sciences, Graduate School of Life and Environmental SciencesOsaka Prefecture UniversityOsakaJapan
  3. 3.Division of Veterinary Science, Graduate School of Life and Environmental SciencesOsaka Prefecture UniversityOsakaJapan

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