Isolation, Cellulase Activity Test and Molecular Identification of Selected Cellulolytic Bacteria Indigenous Rice Bran

https://doi.org/10.22146/ijc.26783

Akyunul Jannah(1*), Aulanni`am Aulanni`am(2), Tri Ardyati(3), Suharjono Suharjono(4)

(1) Department of Chemistry, Maulana Malik Ibrahim State Islamic University Malang, Jl. Gajayana 50 Dinoyo, Malang 65144, East Java, Indonesia
(2) Faculty of Veterinary Medicine, Brawijaya University, Jl. Puncak Dieng Dau, Malang 65151, East Java, Indonesia
(3) Department of Biology, Brawijaya University, Jl. Veteran 6, Malang 65144, East Java, Indonesia
(4) Department of Biology, Brawijaya University, Jl. Veteran 6, Malang 65144, East Java, Indonesia
(*) Corresponding Author

Abstract


Rice bran is the waste product of rice milling which is abundant in Indonesia, it can be used as a raw material for the manufacture of bioethanol by fermentation. Before being fermented, rice bran must be hydrolyzed into glucose by biomass degrading. This study was aimed to isolate indigenous cellulolytic bacteria from rice bran as producer of cellulolytic enzymes and resulted in 22 bacterial isolates that demonstrated cellulolytic activity being identified. Among them, BE 8 and BE 14 isolates showed the highest endoglucanase activity at 2.16 and 1.31 U/mL respectively. Identification of the 16S rDNA showed that BE 8 belongs to Bacillus subtilis and BE 14 in Bacillus cereus.

Keywords


cellulase enzyme; rice bran; 16S rDNA method; Bacillus subtillis; Bacillus cereus

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References

[1] Balat, M., and Balat, H., 2009, Recent trends in global production and utilization of bio-ethanol fuel, Appl. Energy, 86, 2273–2282.

[2] Nejadkoorki, F., Nicholson, K., Lake, I., and Davis, T., 2008, An approach for modelling CO2 emissions from road traffic in urban areas, Sci. Total Environ., 406 (1-2), 269–278.

[3] Saini, J.K., Saini, R., and Tewari, L., 2015, Lignocellulosic agriculture wastes as biomass feedstocks for second-generation bioethanol production: Concepts and recent developments, 3Biotech, 5 (4), 337–353.

[4] Toharisman, A., 2007, Etanol dari Tebu, P3GI, http://p3gi.net, accessed on 4 May 2017.

[5] Badan Ketahanan Pangan, 2016, Produksi Beras 2017 Ditarget 77 Juta Ton, http://www.republika.co.id/berita/ekonomi/makro/16/08/17/oc2agl368, accessed on 2 April 2017.

[6] Moongngarm, I., Daomukda, N., and Khumpika, S., 2012, Chemical compositions, phytochemicals, and antioxidant, APCBEE Procedia, 2, 73–79.

[7] Faria, S.A.S.C., Bassinello, P.Z., and Penteado, M.V.C., 2012, Nutritional composition of rice bran submitted to different stabilization procedures, Braz. J. Pharm. Sci., 48 (4), 651–657.

[8] Tiwari, S., Jadhav, S.K., and Tiwari, K.L., 2015, Bioethanol production from rice bran with optimization of parameters by Bacillus cereus strain McR-3, Int. J. Environ. Sci. Technol., 12 (12), 3819–3826.

[9] Nakazawa, H., Okada, K., Kobayashi, R., Kubota, T., Onodera, T., Ochiai, N., Omata, N., Ogasawara, W., Okada, H., and Morikawa, Y., 2008, Characterization of the catalytic domains of Trichoderma reesei endoglucanase I, II, and III expressed in Escherichia coli, Appl. Microbiol. Biotechnol., 81 (4), 681–689.

[10] Maki, M., Leung, K.T., and Qin, W., 2009, The prospects of cellulase-production bacteria for the bioconversion of lignocellulosic biomass, Int. J. Biol. Sci., 5 (5), 500–516.

[11] Li, X., and Gao, P., 1997, CMC-liquefying enzyme a low molecular mass initial cellulose decomposing cellulose responsible for fragmentation from Streptomyces sp. LX, J. Appl. Microbiol., 83 (1), 59–66.

[12] Ariffin, H., Abdullah, N., Kalsom, M.S.U., Shiari, Y., and Hassan, M.A., 2006, Production and characterization of cellulase by Bacillus pumilus EB3, Int. J. Eng. Technol., 3 (1), 47–53.

[13] Ahmed, A., and Vermette, P., 2008, Culture-based strategies to enhance cellulase enzyme production from Trichoderma reesei RUT-C30 in bioreactor culture conditions, Biochem. Eng. J., 40 (3), 399–407.

[14] Juhasz, T., Kozma, K., Szengyel, Z., and Réczey, K., 2003, Production of β-glucosidase in mixed culture of Aspergillus niger BKMF 1305 and Trichoderma reesei RUT C30, Food Technol. Biotechnol., 41 (1), 49–53.

[15] Kim, C.H., and Kim, D.S., 1993, Extracellular cellulolytic enzymes of Bacillus circulans are present as two multipleprotein complex, Appl. Biochem. Biotechnol., 42 (1), 83–94.

[16] Lin, L., Meng, X., Liu, P., Hong, Y., Wu, G., Huang, X., Li, C., Dong, J., Xiao, L., and Liu, Z., 2009, Improved catalytic efficiency of endo-beta-1,4-glucanase from Bacillus subtilis BME-15 by directed evolution, Appl. Microbiol Bitechnol., 82 (4), 671–479.

[17] Zhang, Y.H.P., Himmel, M.E., and Mielenz, J.R., 2006, Outlook for cellulase improvement: Screening and selection strategies, Biotechnol. Adv., 24 (5), 452–481.

[18] Nirmala, P., and Sindhu, A., 2011, Production of endoglucanase by optimizing the environmental conditions of Bacillus circulans on submerged fermentation, IJAER, 2 (2), 472–481.

[19] Liang, Y.L., Zhang, Z., Wu, M., Wu, Y., and Feng, J.X., 2014, Isolation, screening, and identification of cellulolytic bacteria from natural reserves in the subtropical region of China and optimization of cellulase production by Paenibacillus terrae ME27-1, BioMed Res. Int., 2014, 512497.

[20] 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, AJAS, 26 (1), 50–58.

[21] Yang, W., Meng, F., Peng, J., Han, P., Fang, F., Ma, L., and Cao, B., 2014, Isolation and identification of a cellulolytic bacterium from the Tibetan pig’s intestine and investigation of its cellulose production, Electron. J. Biotechnol., 17 (6), 262–267.

[22] Meng, F., Ma, L., Yang, W., and Cao, B., 2014, Isolation and characterization of Bacillus subtilis strain BY-3, a thermophilic and efficient cellulase-producing bacterium on untreated plant biomass, Lett. Appl. Microbiol., 59 (3), 306–312.

[23] Gundllapalli, S.B., Pretorius, I.S., and Cordero Otero, R.R., 2007, Effect of the cellulose-binding domain on the catalytic activity of a beta-glucosidase from Saccharomycopsis fibuligera, J. Ind. Microbiol. Biotechnol., 34 (6), 413–421.

[24] Singh, A.K., Maharana, A.K., Masih, H., Kumar, Y., and Mishra, S.K. 2012, Production, optimization and purification of bacterial cellulase by solid state bioprocessing of agro biomass, RJPBCS, 3 (2), 977–989.

[25] Bajaj, B.K., Pangotra, H., Wani, M.A., Sharma, P., and Sharma, A., 2009, Partial purification and characterization of a highly thermostable and pH stable endoglucanase from a newly isolated Bacillus strain M-9, Indian J. Chem. Technol., 16. 382–387.

[26] Yin, L.J., Lin, H.H., and Xiao, Z.R., 2010, Purification and characterization of a cellulase from Bacillus subtilis YJ1, J. Mar. Sci. Technol., 18 (3), 466–471.

[27] Kim, Y.K., Lee, S.C., Cho, Y.Y., Oh, H.J., and Ko, Y.H., 2012, Isolation of cellulolytic Bacillus subtilis strains from agricultural environments, ISRN Microbiol., 2012, 650563.

[28] Yan, H., Dai, Y., Zhang, Y., Yang, L., and Liu. D., 2011. Purification and characterization of an endo-1,4-β-glucanase from Bacillus cereus, Afr. J. Biotechnol., 10 (72), 16277–16285.

[29] Fatema, K., and Manchur, M.A., 2015, Isolation, identification and cellulase production by Bacillus brevis from the Acacia forest soil, IJRAF, 2 (9), 14–22.

[30] Zainudin, M.H., Hassan, M.A., Tokura, M., and Shirai, Y., 2013, Indigenous cellulolytic and hemicellulolytic bacteria enhanced rapid co-composting of lignocellulose oil palm empty fruit bunch with palm oil mill effluent anaerobic sludge, Bioresour. Technol., 147, 632–635.

[31] 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 (22), 8798–8806.



DOI: https://doi.org/10.22146/ijc.26783

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