Toxicology International (Formerly Indian Journal of Toxicology)

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Microbial Production and Purification of Cellulase Enzyme on Waste as a Substrate


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1 Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, Lucknow – 227105, Uttar Pradesh, India
     

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The cellulase is of industrial importance and there is a high demand to produce low cost cellulase. The cellulase production was carried out by Aspergillus niger on wastes like saw dust and wheat straw, as it is a need to reduce and reuse waste from the environment. The production was optimized at 72 hrs for saw dust and wheat straw with enzyme activity of 249.10 U/ ml/min and 338.02 U/ml/min respectively. The produced enzyme was purified, the specific enzyme activity of Cellulase after precipitation at 80% (NH4)2SO4 was observed as 4.71 U/min/mg for saw dust and 6.91 U/min/mg for wheat straw respectively. After ion-exchange chromatography best specific enzyme activity was observed in saw dust at 0.6 M NaCl with 3498.04 U/min/mg and for wheat straw at 0.4 M NaCl with 1692.6 U/min/mg. The industrial importance is to produce cellulase at low cost which can be achieved using waste as substrate as well as reduces toxicity of waste.

Keywords

Aspergillus niger, Cellulase, Fermentation, Purification.
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  • Kubicek CP, Messner R, Guber F, Mach RL, Kubicek-Pranz EM. The Trichoderma cellulase regulatory puzzle: From the interior life of a secretory fungus. Enzyme and Microbial Technology. 1993; 15: 90–5. https://doi.org/10.1016/01410229(93)90030-6

  • Lee SM, Koo YM. Pilot-scale production of cellulase using Trichoderma reesei Rut C-30 in fed-batch mode. Journal of Microbiology and Biotechnology. 2001 April; 11(2): 229– 33.

  • Singh J, Kaur P. Optimization of process parameters for cellulase production from Bacillus sp. JS14 in solid substrate fermentation using response surface methodology. Brazilian Archives of Biology and Technology. 2012; 55(4). https://doi.org/10.1590/S1516-89132012000400004

  • Walsh G. Industrial enzymes: Proteases and carbohydrases. Proteins; Biochemistry and Biotechnology, John Wiley and Sons. Ltd; 2002.

  • Himmel ME, Baker JO, Overend RP. Approaches to cellulase purification. Enzymatic Conversion of Biomass for Fuel Production, ACS symposium series 566; 1994.

  • Jahangeer S, Khan N, Jahangeer S, Sohail M, Shahzad S, Ahmad A, Khan SA, Screening and characterization of fungal cellulases isolated from the native environmental source. Pakistan Journal of Botany. 2005; 37(3): 739–48.

  • Gong CS, Cao NJ, Tsao GT. Ethanol production from renewable resources. Advances in Biochemical Engineering/ Biotechnology. Recent Progress in Bioconversion of Lignocellulosics, Berlin: Springer- Verlag. 1999; 65: 207–41. https://doi.org/10.1007/3-540-49194-5_9

  • Himmel ME, Ruth MF, Wyman CE. Cellulase for commodity products from cellulosic biomass. Current Opinion in Biotechnology. 1999; 10: 358–64. https://doi.org/10.1016/S0958-1669(99)80065-2

  • Bai S, Kumar MR, Kumar DJM, Balashanmugam P, Kumaran MDB, Kalaichelvan PT. Cellulase production by Bacillus subtilis isolated from cow dung. Archive of Applied Science Research. 2012; 4(1): 269–79.

  • Bakri YP, Jacques P, Thonart. Xylanase production by Penicillum canescens 10-10c in solid state fermentation. Applied Biochemistry and Biotechnology 2003; 108(1-3): 737–48. https://doi.org/10.1385/ABAB:108:1-3:737

  • Peij N, Gielkens MMC, Verles RP, Visser K, Graff LH. The transcriptional activator X in R regulates both xylanolytic endoglucanase gene expression in Aspergillus niger. Applied and Environmental Microbiology. 1998; 64: 3615– 17. PMid:9758775 PMCid:PMC106473

  • Abdullah JJ, Greetham D, Pensupa N, Tucker GA, Du C. Optimizing cellulase production from Municipal Solid Waste (MSW) using Solid State Fermentation (SSF). Journal of Fundamentals of Renewable Energy and Applications. 2016; 6: 3. https://doi.org/10.4172/2090-4541.1000206

  • Bairagi S. Optimization of cellulase enzyme from vegetable waste by using Trichoderma atroviride in solid state fermentation. IOSR Journal of Environmental Science, Toxicology and Food Technology. 2016 May; 10(5): 68–73.

  • Jadhav AR, Girde AV, More SM, More SB, Khan S. Cellulase Production by utilizing agricultural wastes. Research Journal of Agriculture and Forestry Sciences. 2013 Aug; 1(7): 6–9.

  • Singhania RR, Sukumaran RK, Patel AK, Larroche C, Pandey A. Advancement and comparative profiles in the production technologies using solid-state and submerged fermentation for microbial cellulases. Enzyme and Microbial Technology. 2010; 46: 541–9. https://doi.org/10.1016/j.enzmictec.2010.03.010

  • Goyal M, Kalra KL, Sareen VK, Soni G. Xylanase production with xylan rich lignocellulasic waste by a local soil isolatee of Trichoderma viride. Brazilian Journal of Microbiology. 2008; 39: 535–41. https://doi.org/10.1590/S1517-83822008000300025. PMid:24031262. PMCid:PMC3768426

  • Khan JA, Singh SK. Production of cellulase using cheap substrates by solid state fermentation, International Journal of Plant Animal and Environmental Sciences. 2011; 1(3): 179–87.

  • UdoHölker, JürgenLenz, Solid-state fermentation - are there any biotechnological advantages? Current Opinion in Microbiology. 2005; 8(3): 301–6. https://doi.org/10.1016/j.mib.2005.04.006. PMid:15939353

  • Atlas RM, Parks LC. Handbook of Microbiological Media. Florida CRC Press; 1996.

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin-Phenol reagents. Journal of Biological Chemistry. 1951 May 28; 193 :265–75. PMid:14907713

  • Dubois M, Gilles KA, Hamilton JK, Rebers PK, Smith F. Colorimetric method for determination of sugar and related substrate. Analytical Chemistry. 1956; 28: 350–6. https://doi.org/10.1021/ac60111a017

  • Mandels M, Weber J. The production of cellulases. American Chemical Society, Advances in Chemistry, ACS publishers; 1969; 95: 391–414.

  • Dunn MJ. Protein determination of total protein concentration. Harris ELV, Angal S, editors, Protein Purification Methods, Oxford: IRL Press. 1992; 59: 209–12.

  • Gaur R, Tiwari S. Isolation, production, purification and characterization of an organic-solvent-thermostable alkalophilic cellulase from Bacillus vallismortis RG-07. BMC Biotechnology. 2015 Mar 19; 15(19): 1–12. https://doi.org/10.1186/s12896-015-0129-9

  • Garg N, Ashfaque M. Mango peel as substate for production of extracellular polygalacturonase from Aspergillus fumigatus. Indian Journal of Horticulture. 2010 Mar; 67(1): 140–3.

  • Sriariyanun M, Tantayotai P, Yasurin P, Pornwongthong P, Cheenkachorn K. Production, purification and characterization of an ionic liquid tolerant cellulase from Bacillus sp. isolated from rice paddy field soil. Electronic Journal of Biotechnology. 2016; 19: 23–8. https://doi.org/10.1016/j.ejbt.2015.11.002


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  • Microbial Production and Purification of Cellulase Enzyme on Waste as a Substrate

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Authors

Mohit Nigam
Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, Lucknow – 227105, Uttar Pradesh, India
Shoaa Muzaffar Husain
Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, Lucknow – 227105, Uttar Pradesh, India
Garima Awasthi
Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, Lucknow – 227105, Uttar Pradesh, India

Abstract


The cellulase is of industrial importance and there is a high demand to produce low cost cellulase. The cellulase production was carried out by Aspergillus niger on wastes like saw dust and wheat straw, as it is a need to reduce and reuse waste from the environment. The production was optimized at 72 hrs for saw dust and wheat straw with enzyme activity of 249.10 U/ ml/min and 338.02 U/ml/min respectively. The produced enzyme was purified, the specific enzyme activity of Cellulase after precipitation at 80% (NH4)2SO4 was observed as 4.71 U/min/mg for saw dust and 6.91 U/min/mg for wheat straw respectively. After ion-exchange chromatography best specific enzyme activity was observed in saw dust at 0.6 M NaCl with 3498.04 U/min/mg and for wheat straw at 0.4 M NaCl with 1692.6 U/min/mg. The industrial importance is to produce cellulase at low cost which can be achieved using waste as substrate as well as reduces toxicity of waste.

Keywords


Aspergillus niger, Cellulase, Fermentation, Purification.

References





DOI: https://doi.org/10.18311/ti%2F2018%2Fv25i2%2F22222