Characterization of two β-mannanases from Cellulosimicrobium sp. YB-43

Cellulosimicrobium sp. YB-43에 의해 생산되는 2종류 β-mannanase의 특성분석

  • Yoon, Ki-Hong (Food Science and Biotechnology Major, Woosong University)
  • 윤기홍 (우송대학교 바이오식품과학전공)
  • Received : 2015.08.05
  • Accepted : 2015.09.04
  • Published : 2015.09.30


A bacterial strain producing extracellular mannanases was isolated from soil of chestnut tree farm located in Gongju city of Korea by enrichment culture using Avicel as a carbon source. 16S rDNA sequence of the isolate YB-43 was highly homologous to those of genus Cellulosimicrobium strains with sequence similarities of above 99.6%. Mannanase productivity was significantly increased when the Cellulosimicrobium sp. YB-43 was grown in the presence of locust bean gum (LBG) or konjac. The mannanases were partially purified to be mannanase A (ManA) and mannanase C (ManC) by DEAE-Sepharose column and Q-Sepharose column chromatography from the culture filtrate of Cellulosimicrobium sp. YB-43 grown in LB medium supplemented with 0.7% LBG for 24 h. The partially purified ManA showed the highest activity at $55^{\circ}C$ and pH 6.5, while ManC activity was optimal at $65^{\circ}C$ and pH 7.5. ManA was stable up to $40^{\circ}C$ for 1 h, but ManC activity decreased significantly even after 1 h at $20^{\circ}C$. ManA and ManC showed difference from each other according to their substrate specificities and predominant products resulting from the mannanase hydrolysis for mannooligosaccharides. As a result, Cellulosimicrobium sp. YB-43 was found to produce two different kinds of mannanases.


Cellulosimicrobium sp. YB-43;mannanases;productivity;property


  1. Dhawan, S. and Kaur, J. 2007. Microbial mannanases: an overview of production and applications. Crit. Res. Biotechnol. 27, 197-216.
  2. Fleuri, L.F., Kawaguti, H.Y., and Sato, H.H. 2009. Production, purification and application of extracellular chitinase from Cellulosimicrobium cellulans 191. Braz. J. Microbiol. 40, 623-630.
  3. Huang, J.L., Bao, L.X., Zou, H.Y., Che, S.G., and Wang, G.X. 2012. High-level production of a cold-active ${\beta}$-mannanase from Bacillus subtilis BS5 and its molecular cloning and expression. Mol. Gen. Mikrobiol. Virusol. 4, 14-17.
  4. Jiang, Z., Wei, Y., Li, D., Li, L., Chai, P., and Kusakabe, I. 2006. High-level production, purification and characterization of a thermostable ${\beta}$-mannanase from the newly isolated Bacillus subtilis WY34. Carbohydr. Polym. 66, 68-96.
  5. Kansoh, A.L. and Nagieb, Z.A. 2004. Xylanase and mannanase enzymes from Streptomyces galbus NR and their use in biobleaching of softwood kraft pulp. Antonie van Leeuwenhoek 85, 103-114.
  6. Kataoka, N. and Tokiwa, Y. 1998. Isolation and characterization of an active mannanase-producing anaerobic bacerium, Clostridium tertium KT-5A, from lotus soil. J. Appl. Microbiol. 84, 357-367.
  7. Kim, D.Y., Ham, S.J., Lee, H.J., Cho, H.Y., Kim, J.H., Kim, Y.J., Shin, D.H., Rhee, Y.H., Son, K.H., and Park, H.Y. 2011a. Cloning and characterization of a modular GH5 ${\beta}$-1,4-mannanase with high specific activity from the fibrolytic bacterium Cellulosimicrobium sp. strain HY-13. Bioresour. Technol. 102, 9185-9192.
  8. Kim, D.Y., Ham, S.J., Lee, H.J., Kim, Y.J., Shin, D.H., Rhee, Y.H., Son, K.H., and Park, H.Y. 2011b. A highly active endo-${\beta}$-1,4-mannanase produced by Cellulosimicrobium sp. strain HY-13, a hemicellulolytic bacterium in the gut of Eisenia fetida. Enzyme Microb. Technol. 48, 365-370.
  9. Kim, D.Y., Han, M.K., Lee, J.S., Oh, H.W., Park, D.S., Shin, D.H., Bae, K.S., Son, K.H., and Park, H.Y. 2009. Isolation and characterization of a cellulase-free endo-${\beta}$-1,4-xylanase produced by an invertebrate-symbiotic bacterium, Cellulosimicrobium sp. HY-13. Proc. Biochem. 44, 1055-1059.
  10. Kweun, M.A., Lee, M.S., Choi, J.H., Cho, K.H., and Yoon, K.H. 2004. Cloning of a Bacillus subtilis WL-7 mannanase gene and characterization of the gene product. J. Microbiol. Biotechnol. 14, 1295-1302.
  11. Lu, H., Zhang, H., Shi, P., Luo, H., Wang, Y., Yang, P., and Yao, B. 2013. A family 5 ${\beta}$-mannanase from the thermophilic fungus Thielavia arenaria XZ7 with typical thermophilic enzyme features. Appl. Microbiol. Biotechnol. 97, 8121-8128.
  12. Mok, C.H., Lee, J.H., and Kim, B.G. 2013. Effects of exogenous phytase and ${\beta}$-mannanase on ileal and total tract digestibility of energy and nutrient in palm kernel expeller-containing diets fed to growing pigs. Anim. Feed Sci. Technol. 186, 209-213.
  13. Nabti, E., Bensidhoum, L., Tabli, N., Dahel, D., Weiss, A., Rothballer, M., Schmid, M., and Hartmann, A. 2014. Growth stimulation of barley and biocontrol effect on plant pathogenic fungi by a Cellulosimicrobium sp. strain isolated from salt-affected rhizosphere soil in northwestern Algeria. Eur. J. Soil Biol. 61, 20-26.
  14. Oh, H.W., Heo, S.Y., Kim, D.Y., Park, D.S., Bae, K.S., and Park, H.Y. 2008. Biochemical characterization and sequence analysis of a xylanase produced by an exo-symbiotic bacterium of Gryllotalpa orientalis, Cellulosimicrobium sp. HY-12. Antonie van Leeuwenhoek 93, 437-442.
  15. Shi, P., Yuan, T., Zhao, J., Huang, H., Luo, H., Meng, K., Wang, Y., and Yao, B. 2010. Genetic and biochemical characterization of a protease-resistant mesophilic bmannanase from Streptomyces sp. S27. J. Ind. Microbiol. Biotechnol. 38, 451-458.
  16. Song, J.M. and Wei, D.Z. 2010. Production and characterization of cellulases and xylanases of Cellulosimicrobium cellulans grown in pretreated and extracted bagasse and minimal nutrient medium M9. Biomass Bioenerg. 34, 1930-1934.
  17. Srivastava, P.K. and Kapoor, M. 2014. Cost-effective endo-mannanase from Bacillus sp. CFR1601 and its application in generation of oligosaccharides from guar gum and as detergent additive. Prep. Biochem. Biotechnol. 44, 392-417.
  18. Tanabe, Y. and Oda, M. 2011. Molecular characterization of endo-1,3-${\beta}$-glucanase from Cellulosimicrobium cellulans: Effects of carbohydrate-binding module on enzymatic function and stability. Biochim. Biophy. Acta 1814, 1713-1719.
  19. Vijayalaxmi, S., Prakash, P., Jayalakshmi, S.K., Mulimani, V.H., and Sreeramulu, K. 2013. Production of extremely alkaliphilic, halotolerent, detergent, and thermostable mannanase by the free and immobilized cells of Bacillus halodurans PPKS-2. Purification and characterization. Appl. Biochem. Biotechnol. 171, 382-395.
  20. Yoon, K.H. 2011. Production and properties of hemicellulases by a Cellulosimicrobium sp. isolate. Kor. J. Microbiol. Biotechnol. 39, 252-258.
  21. Yoon, K.H. and Lim, B.L. 2007. Cloning and strong expression of a Bacillus subtilis WL-3 mannanase gene in B. subtilis. J. Microbiol. Biotechnol. 17, 1688-1694.
  22. You, J., Liu, J.F., Yang, S.Z., and Mu, B.Z. 2015. Low-temperatureactive and salt-tolerant ${\beta}$-mannanase from a newly isolated Enterobacter sp. strain N18. J. Biosci. Bioeng. (Article in press).
  23. Yuan, Y., Hu, Y., Hu, C., Leng, J., Chen, H., Zhao, X., Gao, J., and Zhou, Y. 2015. Overexpression and characterization of a glycoside hydrolase family 1 enzyme from Cellulosimicrobium cellulans sp. 21 and its application for minor ginsenosides production. J. Mol. Cataly. B: Enzym. 120, 60-67.
  24. Zhang, C., Chen, J.D., and Yang, F.Q. 2014. Konjac glucomannan, a promising polysaccharide for OCDDS. Carbohydr. Polym. 104, 175-181.
  25. Zhang, Y., Ju, J., Peng, H., Gao, F., Zhou, C., Zeng, Y., Xue, Y., Li, Y., Henrissat, B., Gao, G.F., et al. 2008. Biochemical and structural characterization of the intracellular mannanase AaManA of Alicyclobacillus acidocaldarius reveals a novel glycoside hydrolase family belonging to clan GH-A. J. Biol. Chem. 283, 31551-31558.
  26. Zhou, J., Zhang, R., Gao, Y., Li, J., Tang, X., Mu, Y., Wang, F., Li, C., Dong, Y., and Huang, Z. 2012. Novel low-temperature-active, salt-tolerant and proteases-resistant endo-1,4-${\beta}$-mannanase from a new Sphingomonas strain. J. Biosci. Bioeng. 113, 568-574.

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