Characterization of mannanase-producing bacteria from sago hump

Sri Wahyuni, Andi Khaeruni R, Lianto Lianto, Sidarmin Sidarmin, Holilah Holilah, Wahyu Prasetyo Utomo, Asranudin Asranudin



Sago is a hardwood that can be used as source of fiber hemicellulose mannan. The abundance of mannanase waste is potential for the production of functional foods such as Manno oligosaccharides. Sago hump which is sago processing waste is very potential for growth of microorganisms because it contains lignin, cellulose, starch, minerals and vitamins that can be used as sources of carbon and energy for the growth. In this study, BLS.11-01 and BLS.11-02 isolates have been isolated and characterized from waste of sago hump. Locust bean gum was used as  substrate for measuring the activity of mannanase. Both isolates had optimum temperatures at 70°C and 60°C while they remain stable at temperature range of 30°C - 90°C. Mannanase from BLS.11-01 and BLS.11-02 had optimum pH at 6 and 7, respectively, and were also stable in wide pH range of 2-8. Co2+ was a mannanase activator for both isolates. Cu2+, Zn2+, Ni2+, Ca2+ and Fe2+ were  mannanase inhibitors for two isolates. Tween 20 (0.5% and 1.0%) and NaCl (0.5% and 1.0%) were chemical reagents that could enhance the activity of BLS.11-01 and BLS.11-02 mannanase isolates.  While tween 80 (0.5% and 1.0%), urea (0.5% and 1.0%) and SDS (0.5% and 1.0%) were chemical reagents that were not effective for both isolates mannanase activity

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Alsarrani, A. Q. 2011. Production of Mannan-degrading enzyme by Aspergillus niger. Journal of Thaibah University for Science, 5:1-6.

Abe, J., Hossain, Z. M., and Hizukuri, S. 1994. Isolation of β-mannanase producing microorganisms. Journal of Fermentation and Bioengeering, 3:259–261.

Adiguzel, G., Sonmez, Z., Adiguzel, A., and Nadaroglu, H. 2015. Purification and characterization of a thermostable endo-β-1,4 mannanase from Weissella viridescens LB37 and its application in fruit juice clarification. European Food Research and Technology, 1-8.

Bettiol, J.L. P., Boutique, J. P., Gualco. L.M. P., and Johnston, J.P. 2000. Nonaqueous liquid detergent compositions comprising a borate releasing compound and a mannanase. Patent EP1059351.

Blibech, M., Ghorbel, R. E., Fakhfakh, I., Ntarima, P., Piens, K., Bacha, A. B., and Chaabouni, S.E. 2010. Purification and characterization of a low molecular weight of β-mannanases from Penicillium occitanis Pol6. Applied Biochemistry Biotechnology, 160:1227–1240.

Blibech, M., Ghorbel, R. E., Chaari, F., Dammak, I., Bhiri, F., Neifar, M., and Chaabouni, S.E. 2011. Improved mannanase production from Penicillium occitanis by fed-batch fermentation using acacia seeds. ISRN Microbiology.

Bradford, M.M. 1976. Analytical Biochemistry. 72, pp 248–254.

Carr, E. L., Kampfer, P., Patel, B.K. C., Gurtler, V., and Seviour, R.J. 2003. Seven novel species of Acinetobacter isolated from activated sludge. International Journal of Systematic and Evolutionary Microbiology, 53:953–963.

Chandra, M.R. S., Lee, Y. S., Park, I. H., Zhou, Y., Kim, K. K., and Choi, Y. L. 2011. Isolation, purification and characterization of a thermostable β-mannanase from Paenibacillus sp. DZ3. journal of The Korean Society for Applied Biological Chemistry, 54(3):325–331.

Chauhan, P. S., Puri, N., Sharma, P., and Gupta, N. 2012. Mannanases: microbial sources, production, properties and potential biotechnological applications. Applied Microbiology and Biotechnology, 3(5):1817–1830.

Chen, X., Cao, Y., Ding, Y., Lu, W., and Li, D. 2007. Cloning, functional expression and characterization of Aspergillus sulphureus beta mannanase in Pichia pastoris. Journal of Biotechnology, 128(3):452–461.

Comfort, D.A., Swapnil, R., Chhabra, S.R., Conners, S.B., Chou, C.J., and Epting, K.L. 2004. Strategic biocatalysis with hyperthermophilic enzymes. Green Chemistry, 6:459–465.

de O. Petkowicz, C. L., Reicher, F., Chanzy, H., Taravel, F., and Vuong, R. 2001. Linear mannan in the endosperm of Schizolobium amazonicum. Carbohydrate Polymers, 44:107–112.

Dhawan, S., and Kaur, J. 2007. Microbial mannanases: an overview of production and applications. Critical Reviews in Biotechnology, 27(4):197–216.

Duffaud, G. D., C. M. McCutchen, P. Leduc, K. N. Parker, and R. M. Kelly. 1997. Purification and characterization of extremely thermostable beta-mannanase, beta-mannosidase, and alpha-galactosidase from the hyperthermophilic eubacterium Thermotoga neapolitana 5068. Applied Environmental and Microbiology. 63:169-177.

Gilbert, H.J. 2009. Understanding how diverse β-mannanases recognize heterogenous substrates. Biochemistry, 48:7009–7018.

He, X., Liu, N., Zhang, Z., Zhang, B., and Ma, Y. 2008. Inducible and constitutive expression of a novel thermostable alkaline β-mannanase from alkalophilic Bacillus sp. N16-5 in Pichia pastoris and characterization of the recombinant enzyme. Enzyme and Microbial Technology, 43:13–18.

Hossain, M. Z., Abe, J., and Hizukuri, S. 1996. Multiple forms of β-mannanase from Bacillus sp. KK01. Enzyme and Microbial Technology, 18:95–98.

Holt, J. G., Krieg, N. R., Sneath, P.H. A., Staley, J. T., and Williams, S.T. 1994. Bergey’s manual of determinative bacteriology. 9th ed. Williams & Wilkins USA.

Hsiao, Y. M., Liu, Y. F., Fang, M. C., Tseng, Y.H. 2010. Transcriptional regulation and molecular characterization of the manA gene encoding the biofilm dispersing enzyme mannan endo-1,4-β-mannosidase in Xanthomonas campestris. Journal of Agricultural and Food Chemistry, 58:1653–1663.

Huang, J. L., Bao, L. X., Zou, H. Y., Che, S. G., and Wang, G.X. 2012. High-level production of a cold-active β-mannanase from Bacillus subtilis Bs5 and its molecular cloning and expression. Molecular Genetics Microbiology and Virology, 27:147-153.

Jiang, Z. Y., Wei, D., Li, L., Li, P., Chai., and Kusakabe, I. 2006. High-level production, purification and characterization of a thermostable β-mannanase from the newly isolated Bacillus subtilis WY34. Carbohydrate Polymers, 66:88-96.

Johnvesly, B., and Naik, G.R. 2001. Studies on production of thermostable alkaline protease from thermophilic and alkaliphilic Bacillus sp. JB-99 in a chemically defined medium. Process Biochemistry, 37:139–144.

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. 2011. Cloning and characterization of a modular GH5 β-1,4-mannanase with high specific activity from the fibrolytic bacterium Cellulosimicrobium sp. strain HY-13. Bioresource Technology, 102:9185–9192.

Lehninger, A.L. 1982. Principles of Biochemistry. Worth Publishers, New York.

Li, Y., Yang, P., Meng, K., Wang, Y., Luo, H., Wu, N., Fan, Y., and Yao, B. 2008. Gene cloning, expression, and characterization of a novel beta-mannanase from Bacillus circulans CGMCC 1416. Journal of Microbiology and Biotechnology, 18(1):160–166.

Lu, H., Zhang, H., Shi, P., Luo, H., Wang, Y., Yang, P., and Yao, B. 2013. A family 5 β-mannanase from the thermophilic fungus Thielavia arenaria XZ7 with typical thermophilic enzyme. Applied Microbiology and Biotechnology, 97:8121–8128.

Mabrouk, M.E. M., and Ahwany, A.M.D.E.I. 2008. Production of β-mannanase by Bacillus amyloliquefaciens 10A1 cultured on potato peels. African Journal of Biotechnology, 7:1123–1128.

McCleary, B. V. 1978. A Simple Assay Procedure for β-D-Mannanase. Carbohydrate Research, 67,213-221.

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.

Miller, G. L. 1959. Use of Dinitrosalicylic acid reagent for determination of reducing sugar. Analitycal Chemistry, 31:426-428.

Moreira, L.R. S., Filho, E.X.F. 2008. An overview of mannan structure and mannan degrading enzyme systems. Applied Microbiology and Biotechnology, 79(2):165–178.

Mou, H., Zhou, F., Jiang, X., and Liu, Z. 2011. Production, purification and properties of β-mannanase from soil bacterium Bacillus circulans M-21. Journal of Food Biochemistry, 35:1451–1460.

Naughton, P. J,, Mikkelsen, L. L., and Jensen, B.B. 2001. Effects of nondigestible oligosaccharides on Salmonella enterica Serovar Typhimurium and nonpathogenic Escherichia coli in the pig small intestine in vitro. Applied and Environmental Microbiology, 67:3391–3395.

Pan, X., Zhou, J., Tian, A., Le, K., Yuan, H., Xue, Y., Ma, Y., and Lu, H. 2011. High level expression of a truncated β-mannanase from alkaliphilic Bacillus sp. N16-5 in Kluyveromyces cicerisporus. Biotechnology Letters, 33(3):565–570.

Parisi, G. C., Zilli, M., Miani, M. P., Carrara, M., Bottona, E., Verdianelli, G., Battaglia, G., Desideri, S., Faedo, A., Marzolino, C., Tono, A., Ermani, M., and Leandro, G. 2002. High-fibre diet supplementation in patients with irritable bowel syndrome (IBS): a multicenter, randomized, open trial comparison between wheat bran diet and partially hydrolyzed guar gum (PHGG). Digestive Diseases and Sciences, 47(8):1697–1704.

Pee, V., Ignatius, K. L., Speybroeck, V., Michel, M. P., and Jozef, V.P. 2002. Use of mannanases as a slime control agents. Patent EP0871596 Application Number: EP19960916095.

Politz, O., Krah, M., Thomsen, K. K., and Borriss, R. 2000. A highly thermostable endo-(1,4)-β-mannanase from the marine bacterium Rhodothermus marinus. Applied Microbiology and Biotechnology, 53(6):715–721.

Zhang, R., Zhou1, U., Yajie G., Yaping, G., Junjun, Li., Xianghua T., Xu, B., Junmei, D., and Zunxi, H. 2015. Molecular and biochemical characterizations of a new low-temperature active mannanase. Folia Microbiologica, 60:483–492.

Schröder, R., Atkinson, R. G., and Redgwell, R.J. 2009. Re-interpreting the role of endo-β-mannanases as mannan endotransglycosylase/hydrolases in the plant cell wall, Annals of Botany, 104:197–204.

Sachslehner, A., Nidetzky, B., Kulbe, K. D., and Haltrich, D. 1998. Induction of mannanase, xylanase, and endoglucanase activities in Sclerotium rolfsii. Applied Environmental and Microbiology, 64:594–600.

Shi, P., Yuan, T., Zhao, J., Huang, H., Luo, H., Meng, K., Wang, Y., and Yao, B. 2011. Genetic and biochemical characterization of a protease resistant mesophilic β-mannanase from Streptomycessp.S27. Journal of Industrial Microbiology and Biotechnology, 38:451–458.

Spencer, D. B., Chen, C. P., and Hulett, F. M. 1981. Effect of cobalt on synthesis and activity of Bacillus licheniformis alkaline phosphatase. Journal of Bacteriology, 145:926–933.

Sumardi, S., Antonius, T. S., Maggy and Tresnawati, P. 2006. Purification and characterization Of extracellular β-mannanase from a thermophilic bacterium, Geobacillus stearothermophilus L-07. Journal of Microbiology Indonesia, 57-62.

Syakir, M., Karmawati, E. 2013. Potency of sago palm (Metroxylon spp.) as an alternative for raw material of biofuel. Indonesian Center For Estate Crops Research and Development. Perpekstif, 12(2):57-64.

Titapoka, S., Keawsompong, S., Haltrich, D., and Nitisinprasert, S. 2008. Selection and characterization of mannanase producing bacteria useful for the formation of pre biotic manno oligosaccharides from copra meal. World Journal od Microbiology and Biotechnology, 24:1425–1433.

Varnai, A., Huikko, L., Pere, J., Siikaaho, M., and Viikari, L. 2011. Synergistic action of xylanase and mannanase improves the total hydrolysis of softwood. Bioresource Technology, 102:9096–9104.

Wahyuni, S., Lianto, Khaeruni, A. 2014. Isolation and characterization of mannolitic bacteria from sago farm. Journal of Agrotechnology, 3:174-179.

Wang, C., Zhang, J., Wang, Y., Niu, C., Ma, R., Wang, Y., Bai, Y., Luo, H., and Yao, B. 2016. Biochemical characterization of an acidophilic β-mannanase from Gloeophyllum trabeum CBS900.73 with significant transglycosylation activity and feed digesting ability. Food Chemistry, 197:474–481.

Wang, Y., Azhar, S., Gandini, R., Divne, C., Ezcurraa, I., and Aspeborg, H. 2015. Biochemical characterization of the novel endo-β-mannanase AtMan5-2 from Arabidopsis thaliana. Plant Science, 241:151–163.

Yang, H., Shi, P., Lu, H., Wang, H., Luo, H., Huang, H., Yang, P., and Yao, B. 2015. A thermophilic b-mannanase from Neosartorya fischeri P1 with broad pH stability and significant hydrolysis ability of various mannan. Polymers Food Chemistry, 173:283–289.

Yoo, H. Y., Pradeep, G. C., Kim, S. W., Park, D. H., Choi, Y. H., Suh, J. W., and Yoo, J. C. 2015. A novel low-molecular weight alkaline mannanase from Streptomyces tendae. Biotechnology and Bioprocess Engineering, 20:453-461.

Zhao, H. 2005. Effect of ions and other compatible solutes on enzyme activity, and its implication for biocatalysis using ionic liquids. Journal of Molecular Catalysis B: Enzymatic, 37:16–25.

Oda, Y., Komaki, T., and Tonomura, K. 1993. Purification and properties of extracellular β-mannanases produced by Enterococcus casseliflavus FL2121 isolated from decayed Konjac. Journal of Fermentation and Bioengineering, 76: 14-18.

Van Zyl, W.H., Rose, S.H., Trollope, K., and Gorgens, J.F. 2010. Fungal β-mannanase: mannan hydrolysis, heterologous production and biotechnology applications. Process Biochemistry, 45: 203-1213.

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