Cloning of Partial  β-Mannanase Gene from Indonesia Marine Bacteria Bacillus Subtilis LBF-005

Yopi yopi; Nanik Rahmani; Maghfirotul Amaniyah; Apridah Cameliawati Djohan

doi:10.15578/squalen.252

Cloning of Partial β-Mannanase Gene from Indonesia Marine Bacteria Bacillus Subtilis LBF-005

Yopi yopi, Nanik Rahmani, Maghfirotul Amaniyah, Apridah Cameliawati Djohan

Abstract

The strain LBF-005 from marine bacteria have already isolated and screened for mannanase degrading enzyme in submerged fermentation process. This strain was further identified by using 16S rRNA showed that bacterium is belong to Bacillus subtilis that could produce mannanase with activity around 9.5 U/mL. The optimum pH and temperature for the activity of crude enzyme for mannanase was 6.0 and 50 ^oC. Cloning of mannanase gene from B. subtilis was conducted using six primers set designed based on the homology analysis conserve region several mannanase from bacteria (Bacillus sp.) glycosyl hydrolase (GH) family 26. Optimization of PCR conditions was performed by gradient PCR to obtained PCR product of b-mannanase gene. The PCR product was obtained by third primer combination and was estimated to be around 972-bp. Analysis of the nucleotide sequence showed that sequences has similarity with mananase gene from other Bacillus sp., such as the B. subtilis strain WLY-12, B. subtilis strain WL-8, B. subtilis strain CICC 10260, B.subtilis strain CD-25, B.subtilis strain G1, and Bacillus sp. SWU60 were about 98%, 98%, 98%, 98%, 97% and 95%, respectively. The next research will to obtain a whole gene encoding β-mannanase by using in vitro cloning method, characterization of recombinant mannanase and application this enzyme for mannooligosaccharides production.

Keywords

marine bacteria, Bacillus subtilis, β-mannanase, partial gene

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References

Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W., and Lipman, D.J. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research. 25, 3389-3402. doi:10.1093/nar/25.17.3389

Benech, R.O., Xiaoming Li, Patton, D., Powlowski, J., Storms, R., Bourbonnais, R., Paice, M., and Tsang, A. (2007). Recombinant expression, characterization, and pulp prebleaching property of a Phanerochaete chrysosporium endo-β-1,4-mannanase. Enzyme and Microbial Technology, 41, 740–747.

doi:10.1016/j.enzmictec.2007.06.012

Blibech, M., Chaari, F., Bhiri, F., Dammak, I., Ghorbel, R.E., and Chaabouni, S.E. (2011). Production of manno-oligosaccharides from locust bean gum using immobilized Penicillium occitanis mannanase. Journal of Molecular Catalysis B: Enzymatic, 73, 111– 115. doi:10.1016/j.molcatb.2011.08.007

Chauhan, P.S., Puri, N., Sharma, P., and Gupta, N. (2012). Mannanases: microbial sources, production, properties and potential biotechnological applications. Applied Microbiology and Biotechnology, 93, 1817–1830. doi:10.1007/s00253-012-3887-5

Dhawan S. and Kaur J. (2007). Microbial mannanases: an overview of production and applications. Critical Reviews in Biotechnology, 27, 197–216. doi:10.1080/07388550701775919

Guo, B., Li P.Y., Yue Y.S., Zhao H.L., Song S.D., Sun, C.Y., Zhang, W.X., Chen, X.L., Zhang, X.Y., Zhou, B.C., and Zhang, Y.Z. (2013). Gene Cloning, Expression and Characterization of a Novel Xylanase from the Marine Bacterium, Glaciecola mesophila KMM241. Marine Drugs, 11, 1173-1187. doi:10.3390/md11041173

Hall, B.G. (2013). Building phylogenetic trees from molecular data with MEGA. Molecular Biology and Evolution, 1-7. doi:10.1093/molbev/mst012

Henrissat, B. (1991). A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochemical Journal, 280, 309-316. doi:10.1042/bj2800309

Jian, H.L., Zhu L.W., Zhang W.M., Sun D.F., and Jiang J.X. (2013). Enzymatic production and characterization of manno-oligosaccharides from Gleditsiasinensis galactomannan gum. International Journal of Biological Macromolecules, 55, 282– 288. http://dx.doi. org/10.1016/j.ijbiomac.2013.01.025

Lee, J.T., Bailey C.A., and Cartwright A.L. (2003). Beta mannanase ameliorates viscosity-associated depression of growth in broiler chickens fed guar germ and hull fractions. Poultry Science, 82, 1925-1931. doi:10.1093/ps/82.12.1925

Liepman, A.H., Nairn, C.J., Willats, W.G.T., Sørensen, I., Roberts, A.W., and Keegstra, K. 2007. Functional genomic analysis supports conservationof function among cellulose synthase-like A gene familymembers and suggest diverse roles of mannans in plants. Plant Physiology, 143, 1881–1893. http://dx.doi.org/10.1104/pp.106.093989

Liu, H.X., Gong, J.S., Li, H., Lu, Z.M., Li, H., Qian, J.Y., Xu, Z.H., and Shi, J.S.. (2015). Biochemical characterization and cloning of an endo-1,4- β-mannanase from Bacillus subtilis YH12 with unusually broad substrate profile. Process Biochemistry, 50, 712–721. http://dx.doi.org/10.1016/j.procbio.2015.02.011

Ma, Y., Xue, Y., Dou, Y., Xu, Z., Tao, W., and Zhou, P. (2004). Characterization and gene cloning of a novel β-mannanase from alkaliphilic Bacillus sp. N16-5. Extremophiles, 8, 447–454. doi:10.1007/s00792-004-0405-4.

Mendoza, N.S., Arai, M., Kawaguchi, T., Yoshida, T., and Joson, L.M. (1994). Purification and properties of mannanase from Bacillus subtilis. World Journal of Microbiology and Biotechnology, 10, 551–555. doi:10.1007/BF00367665

Moreira, L.R. and Filho. E.X. (2008). An overview of mannan structure and mannan degrading enzyme systems. Applied Microbiology and Biotechnology. 79, 165-178. doi:10.1007/s00253-008-1423-4

Politz, O., Krah, M.K., Thomsen, K., and Borriss. R. (2000). A highly thermostableendo-(1,4)-b-mannanase from the marine bacterium Rhodothermusmarinus. AppliedMicrobiology andBiotechnology, 53, 715-721. doi:10.1007/s002530000351

Schroder, R., Atkinson, R.G., Redgwell, R.J. (2009). Re-interpreting the role of endo-β-mannanases as mannanendotransglycosylase/hydrolases in the plant cell wall. Annals of Botany, 104, 197–204. doi:10.1093/aob/mcp120

Sievers, F., Andreas, W., David, D., Toby, J.G., Kevin, K., Weizhong, L., Rodrigo, L., Hamish, M., Michael, R., Johannes, S., Julie, D.T. and Desmond, G.H. (2011). Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Molecular Systems Biology, 7, 1-6.doi:10.1038/msb.2011.75

Summpunn, P., Chaijan, S., Isarangkul, D., Wiyakrutta, S., and Meevootisom, V. (2011). Characterization, gene cloning, andheterologous expression ofβ-mannanase from a thermophilic Bacillus subtilis. The Journal of Microbiology, 49, 86-93. doi: 10.1007/s12275-011-0357-1

Tamaru, Y., Araki, T., Morishita, T., Kimura, T., Sakka, K., and Ohmiya. K. (1997). Cloning, DNA sequencing, and expression of the P-1,4-mannanase gene from a marine bacterium, Vibrio sp. strain MA-138. Journal of Fermentation and Bioengering, 83, 201-205.doi: 10.1016/S0922-338X(97)83584-2

Wang, M., You, S., Zhang, S., Qi, W., Liu, Z., Wu, W., Su, R., and He, Z. (2013). Purification, characterization, and production of b-mannanase from Bacillus subtilis TJ-102 and its application in gluco-mannooligosaccharides preparation. European Food Research and Technology, 237, 399–408. doi:10.1007/s00217-013-2002-1

Zou, X.T., Qiao, X.J., and Xu, Z.R., (2006). Effect of β-mannanase (hemicell) on growth performance and immunity of broilers. Poultry Science, 85, 2176–2179. doi:10.1093/ps/85.12.

DOI: https://doi.org/10.15578/squalen.252