Isolation and identification of cellulase producing Bacillus subtilis EG7 and its potential in composting of agricultural wastes
Paper Details
Isolation and identification of cellulase producing Bacillus subtilis EG7 and its potential in composting of agricultural wastes
Abstract
In the current study, some bacterial isolates capable of utilizing Rice straw as an agricultural waste were obtained. Among them, the cellulolytic activity of the isolate EG7, detected both qualitatively and quantitatively, was promising. This isolate degraded the different cellulosic substrates used; Carboxymethylcellulose, Avicel, Cotton seed linters and Rice straw. The isolate was identified by classical bacteriological examination, metabolic fingerprinting using Biolog MicroPlate and phylogenetic analysis of 16S ribosomal Ribo-Nucleic Acid (16SrRNA) gene nucleotide sequence as Bacillus subtilis, GenBank accession number (KX817281). Cellulase enzyme of the isolate EG7 was produced under the determined optimum conditions. Among these conditions, static incubation for three days at initial pH 7.00 and incubation temperature 40oC with cellulose concentration 10 % (w/v) were favorable. Under these conditions, the enzyme activity recorded 45.00 UmL-1. Purification of the produced enzyme was performed and the results revealed that the enzyme activity recorded 64.50UmL-1. Molecular weight determination revealed the presence of two distinct bands of about 43 and 37 KDa. The use of the isolate EG7 in composting of Rice straw and Banana peels waste resulted in enhancing the process and reducing time of composting into five to seven days. Our results indicated the potential of using the isolate EG7 in enhancing composting of agricultural wastes, besides, the use of its cellulase in the different industrial applications.
Abdullah R, Zafar W, Nadeem M, Iqtedar M, Kaleem A, Naz S. 2016. Partial Purification and Characterization of Cellulases Produced by bacillus Strain. Romanian Biotechnological Letters 21(1).
Annamalai N, Rajeswari MV, Sivakumar N. 2016. Cellobiohydrolases: Role, Mechanism, and Recent Developments. In book: Microbial Enzymes in Bioconversions of Biomass, Gupta VK. ed. Biofuel and Biorefinery Technologies 3, 29-35.
Chandna P, Nain L, Singh S, Kuhad RC. 2013. Assessment of bacterial diversity during composting of agricultural byproducts. Biomed Central Microbiology 13,99. http://doi.org/10.1186/1471-2180-13-99
Fariq A. 2016. Microbial Cellulases: Production and Applications. Journal of Biotechnology Science Research 3(1), 122-127.
Irfan M, Mushtaq Q, Tabssum F, Shakir HA, Qazi JI. 2017. Carboxymethyl cellulase production optimization from newly isolated thermophilic Bacillus subtilis K-18 for scarification using response surface methodology. AMB Express 7, 29. http://dx.doi.org/10.1186/s13568-017-0331-3
Kasana RC, Salwan R, Dhar H, Dutt S, Gulati A. 2008. A rapid and easy method for the detection of microbial cellulases on agar plates using gram’s iodine. Current Microbiology57(5), 503-507. http://dx.doi.org/10.1007/s00284-008-9276-8
Khatiwada P, Ahmed J, Sohag MH, Islam K, Azad AK. 2016. Isolation, Screening and Characterization of Cellulase Producing Bacterial Isolates from Municipal Solid Wastes and Rice Straw Wastes. Journal of Bioprocessing and Biotechniques 6,280. http://dx.doi.org/10.4172/2155-9821.1000280
Khianngam S, Pootaeng-on Y, Techakriengkrai T, Tanasupawat S. 2014. Screening and identification of cellulase producing bacteria isolated from oil palm meal. Journal of Applied Pharmaceutical Science 4(4), 090-096. http://dx.doi.org/10.7324/JAPS.2014.40416
Kulkarni AG, Vedamurthy AB. 2015. Isolation and Characterization of Cellulolytic Bacteria from Soil. International Journal of Environmental Biology 5(3), 57-65.
Kumar S, Stecher G, Tamura K. 2016. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33, 1870-1874. http://dx.doi.org/10.1093/molbev/msw054
Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophageT4. Nature 277, 680-685. http://dx.doi.org/10.1038/227680a0
Logan NA, De Vos P. 2009. Bacillus; In Bergey’s Manual of Systematic Bacteriology, 2nd ed. De Vos P, Garrity GM, Jones D, Krieg NR, Ludwig W, Rainey FR, Schleifer KH, Whitman WB. New York: Springer 3, 21–128.
Lokhande S, Musaddiq M. 2015. Isolation of cellulolytic bacterial strains for bioconversion of municipal solid waste. International Journal of Applied Research 1(11), 902-905.
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with Folin-phenol reagent. Journal of Biological Chemistry 193, 265-275.
Lugani Y, Singla R, Sooch BS. 2015. Optimization of Cellulase Production from Newly Isolated Bacillus sp. Y3. Journal of Bioprocessing and Biotechniques 5(11), 264-269. http://dx.doi.org/10.4172/2155-9821.1000264
Menendez E, Garcia-Fraile P, Rivas R. 2015. Biotechnological applications of bacterial cellulases. AIMS Bioengineering 2(3), 163-182. http://dx.doi.org/10.3934/bioeng.2015.3.163
Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry 31, 426-428. http://dx.doi.org/10.1021/ac60147a030
Nandimath AP, Kharat KR, Gupta SG, Kharat AS. 2016. Optimization of cellulase production for Bacillus sp. and Pseudomonas sp. soil isolates. African Journal of Microbiology Research 10(13), 410-419. http://dx.doi.org/10.5897/AJMR2016.7954
Pokhrel B, Bashyal B, Magar RT. 2014. Production, Purification and Characterization of Cellulase from Bacillus subtilis Isolated from Soil. European Journal of Biotechnology and Bioscience 2(5), 31-37.
Venkateswar Reddy K, Vijaya Lakshmi T, Vamshi Krishna Reddy A, Hima bindu V, Lakshmi Narasu M. 2016. Isolation, Screening, Identification and Optimized Production of Extracellular Cellulase from Bacillus subtilis Sub.sps using Cellulosic Waste as Carbon Source. International Journal of Current Microbiology and Applied Sciences 5(4), 442-451. http://dx.doi.org/10.20546/ijcmas.2016.504.052
Essam Hussein, Abdel-Shakour, 2017. Isolation and identification of cellulase producing Bacillus subtilis EG7 and its potential in composting of agricultural wastes. Int. J. Biosci., 11(4), 150-163.
Copyright © 2017 by the Authors. This article is an open access article and distributed under the terms and conditions of the Creative Commons Attribution 4.0 (CC BY 4.0) license.