Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

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Indian Journal of Animal Research, volume 51 issue 4 (august 2017) : 785-790

Isolation of endo-1,4-b-D-glucanase producing Bacillus subtilis sp. from fermented foods and enhanced enzyme production by developing the mutant strain

Eunhye Park, Sung Sill Lee, Byeongwoo Kim, Teaksoon Shin, Byungwook Cho, Seongkeun, Cho, K.J. Cho, Jong K. Ha, Jakyeom Seo
1<p>Department of Agricultural Biotechnology, College of Agriculture and Life Science,&nbsp;Seoul National University, Seoul 151-921, Republic of Korea.</p>
Cite article:- Park Eunhye, Lee Sill Sung, Kim Byeongwoo, Shin Teaksoon, Cho Byungwook, Seongkeun, Cho, Cho K.J., Ha K. Jong, Seo Jakyeom (2017). Isolation of endo-1,4-b-D-glucanase producing Bacillus subtilis sp. from fermented foods and enhanced enzyme production bydeveloping the mutant strain . Indian Journal of Animal Research. 51(4): 785-790. doi: 10.18805/ijar.v0iOF.7253.

Cellulolytic bacteria living in food can be applied to microbial feed additives to improve fiber digestion in animal feeds. In this study, a cellulase-producing bacteria was isolated from salted clam and treated with physical or chemical agents to enhance their enzyme production. The bacteria was identified as a strain of Bacillus subtilis on the basis of 16S rRNA analysis. Endo-1,4-b-D-glucanase (endoglucanase) was produced by the wild type using 0.4% carboxy-methyl-cellulose as a carbon source with maximal activity (0.04 U/mL) after 24 h incubation. Insoluble cellulose and oat spelt xylan were also used as carbon sources for investigation of exoglucanase and xylanase, however, these enzymes were not found in the culture supernatant. Maximum endoglucanase activity of Bacillus subtilis sp. was measured at 50°C and pH 5, respectively. Then, the strain was subjected to classical mutagenesis (UV-irradiation and chemical treatment) to improve endoglucanase production. A mutant strain, P11 treated with ethyl methyl sulfonate was finally selected. Mutant P11 was sub-cultured and tested for endoglucanase production, which was 0.05 U/mL after 24 h growth. The significant difference of endoglucanase production between wild type and mutant P11 was prolonged to 10th generation. Thus, the mutant strain was found to have enhanced endoglucanase production.


  1. Afzal, S., Saleem, M., Yasmin, R., Naz, M. and Imran, M. (2010). Pre and post cloning characterization of a ²-1,4- endoglucanase from Bacillus sp. Mol. Biol. Rep., 37: 1717-1723.

  2. Ayyachamy, M. and Vatsala, T.M. (2007). Production and partial characterization of cellulase free xylanase by Bacillus subtilis C01 using agri-residues and its application in bio-bleaching of non-woody plant pulps. Lett. Appl. Microbiol.,  45: 467-472. 

  3. Beukes, N. and Pletschke, B.I. (2006). Effect of sulfur containing compounds on Bacillus cellulosome–associated ‘CMCase’ and ‘Avicelase’ activities. FEMS Microbiol. Lett., 264: 226–231.

  4. Bischoff, K.M., Liu, S. and Hughes, S.R. (2007). Cloning and characterization of a recombinant family 5 endoglucanase from Bacillus licheniformis strain B-41361. Process Biochem., 42: 1150-1154.

  5. Chand, P., Aruna, A., Maqsood, A.M. and Rao, L.V. (2005). Novel mutation method for increased cellulase production. J. Appl. Microbiol., 98: 318-323.

  6. Doi, R.H. (2008). Cellulases of mesophilic microorganisms. Ann. N. Y. Acad. Sci., 1125: 267-279.

  7. Fan, L.T., Gharpuray, M.M. and Lee, Y.H. (1987). Cellulose Hydrolysis. Berlin: Springer-Verlag. pp. 1–68.

  8. Ghose, T.K. (1987). Measurement of cellulase activities. Pure Appl. Chem., 59: 257-268.

  9. Gopinath, K.P., Murugesan, S., Abraham, J. and Muthukumar, K. (2009). Bacillus sp. mutant for improved biodegradation of Congo red: Random mutagenesis approach. Bioresour. Technol., 100: 6295-6300.

  10. Hong, J., Tamaki, H., Akiba, S., Yamamoto, K. and Kumagai, H. (2001). Cloning of a gene encoding a highly stable endo-² -1,4-glucanase from Aspergillus niger and its expression in yeast. J. Biosci. Bioeng., 92: 434-441.

  11. Kotchoni, O.S., Shonukan, O.O. and Gachomo, W.E. (2003). Bacillus pumilus BpCRI6, a promising candidate for cellulase production under conditions of catabolite repression. Afr. J. Biotechnol., 2: 140–146.

  12. Li, W., Zhang, W.-W., Yang, M.-M. and Chen, Y.-L. (2008). Cloning of the thermostable cellulase gene from newly isolated Bacillus subtilis and its expression in Escherichia coli. Mol. Biotechnol., 40: 195-201.

  13. Li, W., Huan, X., Zhou, Y., Ma, Q. and Chen, Y. (2009). Simultaneous cloning and expression of two cellulase genes from Bacillus subtilis newly isolated from Golden Takin (Budorcas taxicolor Bedfordi). Biochem. Biophys. Res. Commun., 383: 397-400.

  14. Manimaran, A. and Vatsals, T.M. (2007). Biobleaching of banana fibre pulp using Bacillus subtilis C O1 xylanase produced from wheat bran under solid-state cultivation. J. Ind. Microbiol. Biotechnol., 34: 745-749.

  15. Mawadza, C., Hatti-Kaul, R., Zvauya, R. and Mattiasson, B. (2000). Purification and characterization of cellulases produced by two Bacillus strains. J. Biotechnol., 83: 177-187.

  16. Miller, G.L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem., 31: 426-428.

  17. Rastogi, G., Bhalla, A., Adhikari, A., Bischoff, K.M., Hughes, S.R., Christopher, L.P. and Sani, R.K. (2010). Characterization         of thermostable cellulases produced by Bacillus and Geobacillus strains. Bioresour. Technol., 101: 8798-8806.

  18. Stackebrandt, E. and Goodfellow, M. (1991). Nucleic acid techniques in bacterial systematics. Chichester, UK. 

  19. Seo, J.K., Park, T.S., Kwon, I.H., Piao, M.Y., Lee, C.H. and Ha, J.K. (2013). Characterization of cellulolytic and xylanolytic enzymes of Bacillus licheniformis JK7 isolated from the rumen of a native Korean goat. Asian-australas. J. Anim. Sci., 26: 50-58.

  20. Yang, D., Weng, H., Wang, M., Xu, W., Li, Y. and Yang, H. (2009). Cloning and expression of a novel thermostable cellulase from newly isolated Bacillus subtilis strain I15. Mol. Biol. Rep., 37: 1923-1929.

     

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