Characterization of Novel Alkaliphilic Isolate of Bacillus mannanilyticus, Strain IB-OR17, Displaying Chitinolytic and Antifungal Activities

A. I. Melentiev, N. F. Galimzianova, E. A. Gilvanova, E. A. Shchelchkova, L. Yu. Kuzmina, T. F. Boyko, N. G. Usanov, G. E. Aktuganov

The Institute of Biology, Ufa Research Center of Russian Academy of Sciences, Ufa, Russia.

DOI: 10.4236/aim.2014.48050                

Abstract

The novel alkaliphilic strain, designated as Bacillus sp. IB-OR17 and isolated from soda lake sediments, was identified and characterized. Isolated strain demonstrated slight antifungal activity against some plant pathogen fungi that are capable to grow under alkaline conditions. Based on its morphological, physiological and biochemical characteristics as well as on 16S rRNA gene analysis data, Bacillus sp. IB-OR17 were related to alkaliphilic species B. mannanilyticus. Such as type species, Bacillus sp. IB-OR17 produced extracellular β-mannanase but additionally it displayed also chitinolytic activity which is a new property reported for this species. Bacillus sp. IB-OR17 grew in pH range 8.0 - 11.0 with maximal intensity under 9.0 - 10.0 but not showed halophilic properties (growth limit under NaCl concentrations < 5%). Maximal production of chitinase is observed at the same pH interval after 96 h of submerged cultivation of the strain. Bacillus sp. IB-OR17 produced chitinase(s) in presence of colloidal chitin as main carbon source and sodium carbonate (0.25% - 1.0%) demonstrating high enzyme yield under enough low concentrations of the substrate (0.20%). Unlike chitinase, β-mannanase was constitutively produced by Bacillus sp. IB- OR17 in presence of various substrates including crab shell chitin. Probable involvement of the enzymes in antifungal activity of Bacillus sp. IB-OR17 is discussed shortly in terms of further researches and application of this strain.

Keywords

Bacillus mannanilyticus, Alkaliphilic Bacteria, Bacilli, 16S rRNA, Chitinase, β-Mannanase, Antifungal Activity, Microbial Antagonism

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Rossi, M., Ciaramella, M., Cannio, R., Pisani, F.M., Moracci, M. and Bartolucci, S. (2003) Extremophiles 2002. Journal of Bacteriology, 185, 3683-3689. http://dx.doi.org/10.1128/JB.185.13.3683-3689.2003
[2]	Horikoshi, K. (1999) Alkaliphiles: Some Applications of Their Products for Biotechnology. Microbiology and Molecular Biology Reviews, 63, 735-750.
[3]	Rajan, L.A., Dharini, J., Singh, K.H.P., Sivvaswaamy, S.N., Sheela, J.S. and Sundarm, N. (2010) Identification, Cloning and Sequence Analysis of Chitinase Gene in Bacillus halodurans Isolated from Salted Fish. Biotechnology, 9, 229-233. http://dx.doi.org/10.3923/biotech.2010.229.233
[4]	Sarethy, I.P., Saxena, Y., Kapoor, A., Sharma, M., Sharma, S.K., Gupta, V. and Gupta, S. (2011) Alkaliphilic Bacteria: Applications in Industrial Biotechnology. Journal of Industrial Microbiology and Biotechnology, 38, 769-790. http://dx.doi.org/10.1007/s10295-011-0968-x
[5]	Sakai, K., Yokota, A., Kurokawa, H., Wakayama, M. and Moriguchi, M. (1998) Purification and Characterization of Three Thermostable Endochitinases of a Noble Bacillus Strain, MH-1, Isolated from Chitin Containing Compost. Applied and Environmental Microbiology, 64, 3397-3402.
[6]	Bansode, V.B. and Bajekal, S.S. (2006) Characterization of Chitinases from Microorganisms Isolated from Lonar Lake. Indian Journal of Biotechnology, 5, 357-363.
[7]	Loni, P.P. and Bajekal, S.S. (2011) Alkaline Chitinase from Bacillus firmus SBPL-05 Isolated from Alkaline-Saline Environment of Lonar Lake. Indian Journal of Fundamental and Applied Life Sciences, 1, 161-165.
[8]	Bhushan, B. and Hoondal, G.S. (1998) Isolation, Purification and Properties of a Thermostable Chitinase from an Alkalophilic Bacillus sp. BG-11. Biotechnology Letters, 20, 157-159. http://dx.doi.org/10.1023/A:1005328508227
[9]	Bhushan, B. (2000) Production and Characterization of a Thermostable Chitinase from a New Alkalophilic Bacillus sp. BG-11. Journal of Applied Microbiology, 88, 800-808. http://dx.doi.org/10.1046/j.1365-2672.2000.01016.x
[10]	Horikoshi, K. and Akiba, T. (1982) Alkalophilic Microorganisms, a New Microbial World. Japan Scientific Societies Press und Springer Verlag, Tokyo und Berlin-Heidelberg-New York.
[11]	The Firmicutes (2009) In: Vos, P., Garrity, G., Jones, D., Krieg, N.R., Ludwig, W., Rainey, F.A., Schleife, K.-H. and Whitman, W.B., Eds., Bergey’s Manual of Systematic Bacteriology, 2nd Edition, Vol. 3, Springer, Berlin.
[12]	Higgins, D.G., Thompson, J.D. and Gibson, T.J. (1996) Using CLUSTAL for Multiple Sequence Alignments. Methods in Enzymology, 266, 383-402.
[13]	Hall, T. (2007) BioEdit. Biological Sequence Alignment Editor for Win95/98/NT/2K/XP. Carlsbad, CA: Ibis Biosciences. http://www.mbio.ncsu.edu/BioEdit/bioedit.html
[14]	Van de Peer, Y. and De Wachter, R. (1994) TREECON for Windows: A Software Package for the Construction and Drawing of Evolutionary Trees for the Microsoft Windows Environment. Computer Applications in the Biosciences, 10, 569-570.
[15]	Rodriguez-Kabana, R., Godoy, G., Morgan-Jones, G. and Shelby, R. (1983) The Determination of Soil Chitinase Activity: Conditions for Assay and Ecological Studies. Plant and Soil, 75, 95-106. http://dx.doi.org/10.1007/BF02178617
[16]	Imoto, T. and Yagishita, K. (1971) A Simple Activity Measurement of Lyzozyme. Agricultural and Biological Chemistry, 35, 1154-1156. http://dx.doi.org/10.1271/bbb1961.35.1154
[17]	Nogi, Y., Takami, H. and Horikoshi, K. (2005) Characterization of Alkaliphilic Bacillus Strains Used in Industry: Proposal of Five Novel Species. International Journal of Systematic and Evolutionary Microbiology, 55, 2309-2315. http://dx.doi.org/10.1099/ijs.0.63649-0
[18]	Akino, T., Nakamura, N. and Horikoshi, K. (1987) Production of β-Mannosidase and β-Mannanase by an Alkalophilic Bacillus sp. Applied Microbiology and Biotechnology, 26, 323-327. http://dx.doi.org/10.1007/BF00256662
[19]	Paavilainen, S., Helsito, P. and Korpela, T. (1994) Conversion of Carbohydrates to Organic Acids by Alkaliphilic Bacilli. Journal of Fermentation and Bioengineering, 78, 217-222. http://dx.doi.org/10.1016/0922-338X(94)90293-3
[20]	Patil, U. and Chaudhari, A. (2013) Production of Alkaline Protease by Solvent-Tolerant Alkaliphilic Bacillus circulans MTCC 7942 Isolated from Hydrocarbon Contaminated Habitat: Process Parameters Optimization. ISRN Biochemistry, 2013, Article ID: 942590.
[21]	Chauhan, P.S., Puri, N., Sharma, P. and Gupta, N. (2012) Mannanases: Microbial Sources, Production, Properties and Potential Biotechnological Application. Applied Microbiology and Biotechnology, 93, 1817-1830. http://dx.doi.org/10.1007/s00253-012-3887-5
[22]	Tsujibo, H., Kubota, T., Yamamoto, M., Myiamoto, K. and Inamori, Y. (2003) Characterization of Chitinase Genes from Alkaliphilic Actinomycete, Nocardiopsis prasina OPC-131. Applied and Environmental Microbiology, 69, 894-900. http://dx.doi.org/10.1128/AEM.69.2.894-900.2003
[23]	Lawton, E.M., Cotter, P.D., Hill, C. and Ross, R.P. (2007) Identification of a Novel Two-Peptide Lantibiotic, Haloduracin, Produced by the Alkaliphile Bacillus halodurans C-125. FEMS Microbiology Letters, 267, 64-71. http://dx.doi.org/10.1111/j.1574-6968.2006.00539.x