ISOLATION AND IDENTIFICATION OF A CELLULOLYTIC FUNGUS FROM ANIMAL MANURE
Abstract
Successful conversion of lignocellulose waste to fermentable sugars should open the door for the production of many desired metabolites. Fungi are the best cellulose decomposers in nature. The present study describes the isolation of cellulose-degrading fungal strain, AGE-1, from cow manure. AGE-1 fungal strain was tested for the cellulases production under various pHs, incubation temperatures, incubation time, and carbon sources. The AGE-1 isolate showed the maximum activity for CMCase of 1.44 U/ml ± 0.02 on day 7 when the fungal isolate was incubated at 30 °C. The maximum activity for FPase was on day 7 and it reaches 0.88 U/ml ± 0.03 when the isolate incubated at 30ºC. The strain was molecularly identified using the 18S rRNA gene and ITS as Aspergillus terreus. The strain was tested for cellulases production using solidstate fermentation and showed maximum expression of FPase and CMCase at 30ºC and pH7. Various agricultural wastes were tested as substrates for cellulase production by solid-state fermentation, and sugarcane bagasse (SCB) was the best sole source of carbon for cellulase production followed by wheat straw. The isolated strain can be used as a starting step for degrading agricultural waste that can be used for the production of biofuel and the production of desired metabolites from cellulose.
References
Acourene S., and Ammouche A., (2012). Optimization of Ethanol, Citric Acid, and α-Amylase Production from Date Wastes by Strains of Saccharomyces cerevisiae, Aspergillus niger, and Candida guilliermondii. Journal of Industrial Microbiology and Biotechnology 39, no. 5 (May 2012): 759-66. https://doi.org/10.1007/s10295-011-1070-0
Adney B., and Nrel J. B., (2008). Measurement of Cellulase Activities Laboratory Analytical Procedure.” Renewable Energy, no. January: 8.
Aggarwal Kumar Neeraj, Goyal Varsha, Saini Anita, Yadav Anita, and Gupta R., (2017). Enzymatic Saccharification of Pretreated Rice Straw by Cellulases from Aspergillus Niger BK01. 3 Biotech 7, no. 3 (July 1, 2017): 1-10. https://doi.org/10.1007/s13205-017-0755-0
Aliyah Andi, Alamsyah Gandhi, Ramadhani Rizky, and Hermansyah H., (2017). Production of α-Amylase and β- Glucosidase from Aspergillus Niger by Solid State Fermentation Method on Biomass Waste Substrates from Rice Husk, Bagasse and Corn Cob. In Energy Procedia, 136:418-423. Elsevier Ltd, 2017. https://doi.org/10.1016/j.egypro.2017.10.269
Altschul Stephen F., Alejandro Madden Thomas L., Schäffer A., Zhang Jinghui, Zhang Zheng, Miller Webb, and Lipman D. J., (1997). Gapped BLAST and PSI-BLAST: A New Generation of Protein Database Search Programs. Nucleic Acids Research. Oxford University Press, September 1, 1997. https://doi.org/10.1093/nar/25.17.3389
Baldrian Petr, and Valášková V., (2008). Degradation of Cellulose by Basidiomycetous Fungi. FEMS Microbiology Reviews. Oxford Academic, May 1. https://doi.org//10.1111/j.1574-6976.2008.00106.x
Bansal Namita, Tewari Rupinder, Soni Raman, and Soni S. K., (2012). Production of Cellulases from Aspergillus Niger NS-2 in Solid State Fermentation on Agricultural and Kitchen Waste Residues. Waste Management 32, no. 7 (July 1, 2012): 1341-46. https://doi.org/10.1016/j.wasman.2012.03.006
Devi Mc., and Kumar M., (2012). Production, Optimization and Partial Purification of Cellulase by Aspergillus Niger Fermented with Paper and Timber Sawmill Industrial Wastes. Journal of Microbiology and Biotechnology Research 2, no. 1: 120-28.
Ellilä Simo, Lucas Fonseca, Uchima Cristiane, Cota Junio, Goldman Gustavo Henrique, Saloheimo Markku, Sacon Vera, and SiikaAho M., (2017). Development of a Low-Cost Cellulase Production Process Using Trichoderma Reesei for Brazilian Biorefineries. Biotechnology for Biofuels 10, no. 1: 30. https://doi.org/10.1186/s13068-017-0717-0
Florencio Camila, Couri Sonia, and Farinas C. S., (2012). Correlation between Agar Plate Screening and Solid-State Fermentation for the Prediction of Cellulase Production by Trichoderma Strains. Enzyme Research 2012. https://doi.org/10.1155/2012/793708
Gao Jianmin, Weng Haibo, Daheng Zhu, Yuan Mingxue, Guan Fangxia, and Xi Y., (2008). Production and Characterization of Cellulolytic Enzymes from the Thermoaci-dophilic Fungal Aspergillus Terreus M11 Under Solid-State Cultivation of Corn Stover. Bioresource Technology 99, no. 16: 7623-29. https://doi.org/10.1016/j.biortech.2008.02.005
Ghose T. K., (1987). Measurement of Cellulase Activities. Pure and Applied Chemistry 59, no. 2: 257-68. https://doi.org/10.1351/pac198759020257
Gleason F. H., Marano A. V., Digby A. L., Al-Shugairan N., Lilje O., Steciow M. M., Barrera M. D., Inaba S., and Nakagiri A., (2011). Patterns of Utilization of Different Carbon Sources by Chytridiomycota. Hydrobiologia 659, no. 1:55-64. https://doi.org/10.1007/s10750-010-0461-y
Gusakov Alexander V., (2011). Alternatives to Trichoderma Reesei in Biofuel Production. Trends in Biotechnology. Elsevier. https://doi.org/10.1016/j.tibtech.2011.04.004
Gutierrez-Correa Marcel, and Tengerdy R. P., (1997). Production of Cellulase on Sugar Cane Bagasse by Fungal Mixed Culture Solid Substrate Fermentation. Biotechnology Letters 19, no.7:665-67. https://doi.org/10.1023/A:1018342916095
Henry Travis, Iwen Peter C., and Hinrichs S. H., (2000). Identification of Aspergillus Species Using Internal Transcribed Spacer Regions 1 and 2. Journal of Clinical Microbiology 38, no.4:1510-15. https://doi.org/10.1128/jcm.38.4.1510-1515.2000
Hernández Christian, Milagres Adriane M. F., Vázquez-Marrufo Gerardo, Muñoz-Páez Karla María, José Antonio García-Pérez, and E. Alarcón (2018). An Ascomycota Coculture in Batch Bioreactor Is Better than Polycultures for Cellulase Production. Folia Microbiologica 63, no. 4: 467-78. https://doi.org/10.1007/s12223-018-0588-1
Ismail Shaymaa A., and Hassan A., (2020). Optimizing the Production of Rice Straw Hydrolytic Cellulase under Solid-State Fermentation Using Aspergillus Terreus RS2. Egyptian Pharmaceutical Journal 19, no. 1 (2020):7. https://doi.org/10.4103/epj.epj_44_19
Juhász T., Szengyel Z., Réczey K., Siika- Aho M., and Viikari L., (2005). Characterization of Cellulases and Hemicellulases Produced by Trichoderma Reesei on Various Carbon Sources. Process Biochemistry 40, no. 11: 3519-25. https://doi.org/10.1016/j.procbio.2005.03.057
Kamm B., and Kamm M., (2004). Principles of Biorefineries. Appl Microbiol Biotechnol, April 2004. https://doi.org/10.1007/s00253-003-1537-7
Kasana Ramesh Chand, Salwan Richa, Dhar Hena, Dutt Som, and Gulati A., (2008). A Rapid and Easy Method for the Detection of Microbial Cellulases on Agar Plates Using Gram’s Iodine. Current Microbiology 57, no. 5: 503-7. https://doi.org/10.1007/s00284-008-9276-8
Kumar Sudhir, Stecher Glen, and Tamura K., (2016). MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular Biology and Evolution 33, no. 7: 1870-74. https://doi.org/10.1093/molbev/msw054
Liang Yan Ling, Zhang Zheng, Wu Min, Wu Yuan, 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 Research International 2014. https://doi.org/10.1155/2014/512497
Lynd Lee R., Weimer Paul J., Van Zyl Willem H., and Isak S., (2002). Microbial Cellulose Utilization: Fundamentals and Biotechnology Microbial Cellulose Utilization: Fundamentals and Biotechnology. Microbiology and Molecular Biology Reviews 66, no. 3: 506-577. https://doi.org/10.1128/MMBR.66.3.506-577.2002
Mrudula Soma, and Murugammal R., (2011). Production of Cellulase by Aspergillus Niger under Submerged and Solid State Fermentation Using Coir Waste as a Substrate. Brazilian Journal of Microbiology 42, no. 3 (2011): 1119-27. https://doi.org/10.1590/S1517-83822011000300033
Panzer Katrin, Yilmaz Pelin, Weiß Michael, Reich Lothar, Richter Michael, Wiese Jutta, Schmaljohann Rolf, et al., (2015). Identification of Habitat-Specific Biomes of Aquatic Fungal Communities Using a Comprehensive Nearly Full-Length 18S RRNA Dataset Enriched with Contextual Data. PLoS ONE 10, no. 7 (2015): 1-20. https://doi.org/10.1371/journal.pone.0134377
Prasanna H. N., Ramanjaneyulu G., and Reddy B. R., (2016). Optimization of Cellulase Production by Penicillium Sp. 3 Biotech 6, no. 2. https://doi.org/10.1007/s13205-016-0483-x
Sari, Wenny Novita, Safika Darmawi, and Fahrimal Y., (2017). Isolation and Identification of a Cellulolytic Enterobacter from Rumen of Aceh Cattle. Veterinary World 10, no. 12: 1515-20. https://doi.org/10.14202/vetworld.2017.1515-1520
Shahriarinour Mahdi, Abdul Wahab Mohd Noor, Mohamad Rosfarizan, Mustafa Shuhaimi, and Ariff A. B., (2011). Effect of Medium Composition and Cultural Condition on Cellulase Production by Aspergillus Terreus. African Journal of Biotechnology 10, no. 38: 7459-67. https://doi.org/10.5897/AJB11.199
Sohail Muhammad, Siddiqi Roquya, Ahmad Aqeel, and Khan S. A., (2009). Cellulase Production from Aspergillus Niger MS82: Effect of Temperature and PH. New Biotechnology 25, no. 6: 437-41. https://doi.org/10.1016/j.nbt.2009.02.002
Sugita Takashi, Nakajima Masamitsu, Ikeda Reiko, Matsushima Toshiharu, and Shinoda T., (2002). Sequence Analysis of the Ribosomal DNA Intergenic Spacer 1 Regions of Trichosporon Species. Journal of Clinical Microbiology 40, no. 5: 1826-30. https://doi.org/10.1128/JCM.40.5.1826-1830.2002
Sukumaran Rajeev K., Rani Reeta Singhania, Mathew Gincy Marina, and Pandey A., (2009). Cellulase Production Using Biomass Feed Stock and Its Application in Lignocellulose Saccharification for Bio-Ethanol Production. Renewable Energy 34, no. 2: 421-24. https://doi.org/10.1016/j.renene.2008.05.008
Szijártó Nóra, Szengyel Zsolt, Lidén Gunnar, and Réczey K., (2004). Dynamics of Cellulase Production by Glucose Grown Cultures of Trichoderma Reesei Rut-C30 as a Response to Addition of Cellulose. In Proceedings of the Twenty-Fifth Symposium on Biotechnology for Fuels and Chemicals Held May 4- 7, 2003, in Breckenridge, CO, 115-24. Humana Press, 2004. https://doi.org/10.1007/978-1-59259-837-3_10
Teather R. M., and Wood P. J., (1982). Use of Congo Red-Polysaccharide Interactions in Enumeration and Characterization of Cellulolytic Bacteria from the Bovine Rumen. Applied and Environmental Microbiology 43, no. 4: 777-80. https://doi.org/10.1128/aem.43.4.777-780.1982
Toor Yasmin, and Ilyas U., (2014). Optimization of Cellulase Production by Aspergillus ornatus by the Solid State Fermentation of Cicer Arietinum. Vol. 2. http://www.usa-journals.com
Vega Karin, Gretty Villena K., Victor Sarmiento H., Ludeña Yvette, Vera Nadia, and Gutiérrez-Correa M., (2012). Production of Alkaline Cellulase by Fungi Isolated from an Undisturbed Rain Forest of Peru. Biotechnology Research International 2012: 1-7. https://doi.org/10.1155/2012/934325
Vries Ronald de P., and Visser J., (2001). Aspergillus Enzymes Involved in Degradation of Plant Cell Wall Polysaccharides. Microbiology and Molecular Biology Reviews 65, no. 4: 497-522. https://doi.org/10.1128/mmbr.65.4.497-522.2001
Woo Patrick C. Y., Shui Yee Leung, Kelvin K. W. To, Jasper F. W., Chan, Ngan Antonio H. Y., Vincent Cheng C. C., Lau Susanna K. P., and Yuen K. Y., (2010). Internal Transcribed Spacer Region Sequence Heterogeneity in Rhizopus Microsporus: Implications for Molecular Diagnosis in Clinical Microbiology Laboratories. Journal of Clinical Microbiology 48, no. 1: 208-14. https://doi.org/10.1128/JCM.01750-09
Yoon Li Wan, Nam Ang Teck, Ngoh Gek Cheng, and Chua A. S. M., (2014). Fungal Solid-State Fermentation and Various Methods of Enhancement in Cellulase Production. Biomass and Bioenergy 67: 319-38. https://doi.org/10.1016/j.biombioe.2014.05.013
Zheng Linyong, Jia Dinghong, Fei Xiaofan, Luo Xia, and Yang Z., (2009). An Assessment of the Genetic Diversity within Ganoderma Strains with AFLP and ITS PCR-RFLP. Microbiological Research 164, no. 3: 312-21. https://doi.org/10.1016/j.micres.2007.02.002