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http://dx.doi.org/10.3839/jabc.2018.009

Isolation and characterization of cellulolytic bacteria, Bacillus sp. EFL1, EFL2, and EFP3 from the mixed forest  

Park, Hwa Rang (Department of Forest Resources, Gyeongnam National University of Science and Technology)
Oh, Ki-Cheol (Nakdong River Basin Environmental Office)
Kim, Bong-Gyu (Department of Forest Resources, Gyeongnam National University of Science and Technology)
Publication Information
Journal of Applied Biological Chemistry / v.61, no.1, 2018 , pp. 59-67 More about this Journal
Abstract
This study was conducted to isolate the cellulolytic bacteria able to grow on LB- Carboxymethyl cellulose (CMC) agar trypan blue medium from the mixed forest and Larix leptolepis stands. Three bacterial strains with high activity against both CMC and xylan were isolated. Both API kit test and 16S rRNA gene sequence analysis revealed that the three different isolates belong to the gene Bacillus. Therefore, the isolates named as Bacillus sp. EFL1, Bacillus sp. EFL2, and Bacillus sp. EFP3. The optimum growth temperature of Bacillus sp. EFL1, EFL2, and EFP3 were $37^{\circ}C$. The optimum temperature for CMCase and xylanase from Bacillus sp. EFL1 were $50^{\circ}C$. The optimum pH of Bacillus sp. EFL1 xylanase was pH 5.0 but the optimum pH of CMCase from Bacillus sp. EFL1 was pH 6.0. The optimum temperature of CMCase and xylanase from Bacillus sp. EFL2 was $60^{\circ}C$, respectively. The optimum pH of CMCase of Bacillus sp. EFL2 was 5.0, whereas xylanase showed high activity at pH 3.0-9.0. The optimum temperature for CMCase and xylanase of Bacillus sp. EFP3 was $50^{\circ}C$. The optimum pH for CMCase and xylanse was 5.0 and 4.0, respectively. CMCases from Bacillus sp. EFL1, EFL2, and EFP3 were thermally unstable. Although xylanase from Bacillus sp. EFL1 and EFP3 showed to be thermally unstable, xylanase from Bacillus sp. EFL2 showed to be thermally stable. Therefore, Bacillus sp. EFL2 has great potential for animal feed, biofuels, and food industry applications.
Keywords
Bacillus sp.; Carboxymethyl cellulase; Lignocellulose; Xylanase;
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1 Oh SH, Kim MS, So S, Suj HJ (2003) Studies on the Production of cellulase by Trichoderma sp. SO-571 and the Enzyme Treatment for Cellulosic Fabrics. J. Microbiol. Biotechnol 31(3): 42-45
2 Robson LM, Chambliss GH (1989) Cellulases of bacterial origin. Enzyme and Microbial Technology, 11(10): 626-644   DOI
3 Saito K, Kawamura Y, Oda Y (2003) Role of the pectinolytic enzyme in the lactic acid fermentation of potato pulp by Rhizopus oryzae. Journal of industrial microbiology & biotechnology 30(7): 440-444   DOI
4 Singh S, Madlala AM, Prior BA (2003) Thermomyces lanuginosus: properties of strains and their hemicellulases. FEMS Microbiology Reviews 2003; 27: 3-16
5 Sohail M, Siddiqi R, Ahmad A, Khan SA (2009) Cellulase production from Aspergillus niger MS82: effect of temperature and pH. New Biotechnol 25(6): 437-41   DOI
6 Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 30: 2725-2729   DOI
7 Tjalsma H, Antelmann H, Jongbloed JD, Braun PG, Darmon E, Dorenbos R, Kuipers OP (2004) Proteomics of protein secretion by Bacillus subtilis: separating the "secrets" of the secretion. Microbiology and Molecular Biology Reviews 68(2): 207-233   DOI
8 Whitaker JR (1990) Cellulase production and application. Food Biotechnol 4: 669-697   DOI
9 Wilson DB (2011) Microbial diversity of cellulose hydrolysis. Current opinion in microbiology 14(3): 259-263   DOI
10 Yang JK, Zhang JJ, Yu HY, Cheng JW, Miao LH (2014) Community composition and cellulase activity of cellulolytic bacteria from forest soils planted broad-leaved deciduous and evergreen trees. Appl Microbiol Biotechnol 98: 1449-1458   DOI
11 Kim YK, Lee SC, Cho YY, Oh HJ (2012) Isolation cellulolytic Bacillus subtilis strains from agricultural environments. ISRN Microbiol Article ID 650563: 9
12 Zang XZ, Zhang YHP (2011) Simple, fast and high-efficiency transformation system for directed evolution of cellulase in Bacillus subtilis. Microbial Biotechnology 4: 98-105   DOI
13 Agnelli A, Ascher J, Corti G, Ceccherini MT, Nanniieri P, Pietramellra G (2004) Distribution of microbial communities in a forest soil profile investigated by microbial biomass, soil respiration and DGGE of total and extracellular DNA. Soil Biol Biochem 36: 859-868   DOI
14 Bayer EA, Lamed R, Himmel ME (2007) The potential of cellulases and cellulosomes for cellulosic waste management. Current opinion in Biotechnology 18(3): 237-245   DOI
15 Cai YJ, Chapman SJ, Buswell JA, Chang ST (1999) Production and distribution of endoglucanase, cellobiohydrolase, and beta-glucosidase components of the cellulolytic system of Volvariella volvacea, the edible straw mushroom. Appl Environ Microbiol 65: 553-559
16 Cantarel BL, Coutinho PM, Rancurel C, Bernard T, Lombard V, Henrissat B (2014) The carbohydrate-active enzymes database (CAZy) in 2013. Nucleic Acids Res 42: D490-495   DOI
17 Jang MY, Park HR, Lee CG, Choo GC, Cho HS, Park SB, Oh KC, Kim BG (2017) Isolation and biochemical characterization of acid tolerance xylanase producing Bacteria, Bacillus sp. GJY from city park soil. J Appl Biol Chem 60(1): 79-86   DOI
18 Khandeparker R, Verma P, Deobagkar D (2011) A novel halotolerant xylanase from marine isolate Bacillus subtilis cho40: gene cloning and sequencing. N Biotechnol 28: 814-821   DOI
19 Kulkarni N, Shendye A, Rao M (1999) Molecular and biotechnological aspects of xylanases. FEMS Microbiology Reviews 1999; 23: 411-456   DOI
20 Kumar R, Singh S, Singh OV (2008) Bioconversion of lignocellulosic biomass: biochemical and molecular perspectives. J Ind Microbiol Bioechnol 35: 377-391   DOI
21 Lee CK, Jang MY, Park HR, Choo GC, Cho HS, Park SB, Oh KC, An JB, Kim BG (2016) Cloning and characterization of xylanase in cellulolytic Bacillus sp. strain JMY1 isolated from forest soil. App Biol Chem 59(3): 415-423   DOI
22 Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar, Analytical Chemistry, 31(3): 426-428   DOI
23 Lee YJ, Kim BK, Lee BH, Jo KI, Lee NK, Chung CH, Lee JW (2008) Purification and characterization of cellulase produced by Bacillus amyoliquefaciens DL-3 utilizing rice hull. Bioresource technology 99(2): 378-386   DOI
24 Li XH, Yang HJ, Roy B, Wang D, Yue WF, Jiang LJ, Miao YG (2009) The most stirring technology in future: Cellulase enzyme and biomass utilization. African Journal of Biotechnology 8(11)
25 Lynd LR, Weimer PJ, Van Zyl WH, Pretorius IS (2002) Microbial cellulose utilization: fundamentals and biotechnology. Microbiology and molecular biology reviews 66(3): 506-577   DOI
26 Mosolova TP, Kalyuzhnyi SV, Varfolomeyev SD, Velikodvorskaya GA (1993) Purification and properties of Clostridium thermocellum endoglucanase 5 produced in Escherichia coli. Applied biochemistry and biotechnology 42(1): 9-18   DOI
27 Murashima K, Nishimura T, Nakamura Y, Koga J, Moriya T, Sumida N, Kono T (2002) Purification and characterization of new endo-1, 4-$\beta$-Dglucanases from Rhizopus oryzae. Enzyme and Microbial Technology 30(3): 319-326   DOI
28 Nikaus PA, Wardle DA, Tate KR (2006) Effects of plant species diversity and composion on nitrogen cycling and the trace gas balance of soils, Plant Soil 282: 83-98   DOI