Hyper production of carboxy methyl cellulase by Thermomyces dupontii utilizing physical and chemical mutagenesis

  • K. Nisar
  • R. Abdullah
  • A. Kaleem
  • M. Iqtedar
  • F. Saleem
Keywords: Molecular identification, Fungi, CMCase, Mutation


Utilization of cellulases as substitute of chemical process gained huge momentum in the field of biotechnology  Now there is dire need to find out un explore reveres of fungi possessing greater potential for efficient cellulase production.  This boosted isolation of novel thermo tolerant fungal strains capable of producing the targeted product. In this investigation 70 thermophilic cellulolytic fungal strains were isolated. All the strains were screened via submerged fermentation. The strain showing highest CMCase activity was identified by conventional method i.e.  based on morphology and microscopic features and confirmed by 18S rDNA gene sequencing, using specific ITS primers. The modified CTAB method was used for rapid extraction of DNA from thermo tolerant strain.  The selected strain subsequently subjected to sequencing and phylogenetic analysis. The result indicates the selected strain was found to be T. dupontii. For strain improvement the T. dupontii was subjected to random mutagenesis by using (UV) irradiation and EMS treatment. Out of 40 screened mutant T. dupontii EMS 15 had 2.4 fold more yield (21.8 U/ml/min) as compared to parental strain. The Five fermentation media were also screened. The Medium3 gave higher titer of cellulase activity (29.3 U/ml/min) and found to be the best medium.


Ashwani, K., Saida, L. and Reddy, K.V. (2014). Isolation, screening, and characterization of cellulolytic bacteria from forest soil sample. International Journal of Current Microbiology and Applied Sciences 3, 679-685.

Aslam, S., Tahir, A., Aslam, M.F., Alam, M.W., Shedayi, A.A. and Sadia, S. (2017). Recent advances in molecular techniques for the identification of phytopathogenic fungi–a mini review. Journal of Plant Interactions 12, 493-504.

Brito, C. C., Gama, A.R., Jesuino, R., Faria, F.P. and Bataus, L. (2015). Production of cellulases from a novel thermophilic Streptomyces thermocerradoensis I3 using agricultural waste residue as substrate. Journal of Agriculture and Environmental Sciences 4, 90-99.

Burlacu, A., Israel-Roming, F. and Cornea, C.P. (2017). Fungal strains improvement for xylanase over production through physical and chemical mutagenesis. Agro Life Scientific Journal 6, 40-7.

Coral, G., Arikan, B., Unaldi, M.N. and Güvenmez, H. (2002). Some properties of crude carboxymethyl cellulase of Aspergillus niger Z10 wild-type strain. Turkish Journal of Biology 26, 209-213.

Das, A., Paul, T., Halder, S.K., Maity, C., Mohapatra, P.D., Pati, B.R. and Mondal, K.C. (2013). Study on regulation of growth and biosynthesis of cellulolytic enzymes from newly isolated Aspergillus fumigatus ABK9. Polish Journal of Microbiology 62, 31-43.

Gao, J., Weng, H., Zhu, D., Yuan, M., Guan, F. and Xi, Y. (2008). Production and characterization of cellulolytic enzymes from the thermoacidophilic fungal Aspergillus terreus M11 under solid-state cultivation of corn stover. Bioresource Technology 99, 7623-7629.

Ho, H. and Ho, K. (2015). Fungal strain improvement of Aspergillus brasiliensis for overproduction of xylanase in submerged fermentation through UV irradiation and chemicals mutagenesis. Journal of Advances in Biology and Biotechnology 3, 117 -131.

Jafari, N., Jafarizadeh, M.H., Hamzeh, M.M. and Adibpour, M. (2017). Optimization of UV irradiation mutation conditions for cellulase production by mutant fungal strains of Aspergillus niger through solid state fermentation. Green Processing and Synthesis 6, 333-40.

Keller, N.P., Turner, G. and Bennett, J.W. (2005). Fungal secondary metabolism—from biochemistry to genomics. Nature Reviews Microbiology 3, 937-947.

Kocher, G., Kalra, K. and Banta, G. (2008). Optimization of cellulase production by submerged fermentation of rice straw by Trichoderma harzianum Rut-C 8230. The Internet Journal of Microbiology 5, 21.

Kumar, S., Stecher, G. and Tamura, K. (2016). Mega7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33, 1870-1874.

Lynd, L.R., Weimer, P.J., Van Zyl, W.H. and Pretorius, I.S. (2002). Microbial cellulose utilization: Fundamentals and Biotechnology. Microbiology and Molecular Biology Reviews. 66, 506-577.

Mandel, M. (1969). Exoglucanase activity by microorganisms. Advances in Chemistry 95, 391-414.

Mejad, A. N. (2013). Response of some fungal species to the effect of copper, magnesium and zinc under the laboratory condition. European Journal of Experimental Biology 3, 535-40.

Oliveira, P., De Brito, A., Pimentel, A., Soares, G., Pacheco, C., Santana, N., da Silva, E., Fernandes, A.d.A., Ferreira, M. and Oliveira, J. (2019). Cocoa shell for the production of endoglucanase by Penicillium roqueforti ATCC 10110 in solid state fermentation and biochemical properties. Revista Mexicana de Ingeniería Química 18, 777-87.

Paw, H.M. and Izwikow, M. (2016). Cellulolytic activity of Trichoderma viride with regard to selected lignocellulosic waste materials. Journal of Ecological Engineering 17. 119-122.

Porter, T.M. and Brian Golding, G. (2011). Are similarity‐or phylogeny‐based methods more appropriate for classifying internal transcribed spacer (ITS) metagenomic amplicons? New Phytologist 192, 775-782.

Prabha, T., Revathi, K., Vinod, M., Shanthakumar, S. and Bernard, P. (2013). A simple method for total genomic DNA extraction from water moulds. Current Science 104, 345-347.

Prasanna, H., Ramanjaneyulu, G. and Reddy, B.R. (2016). Optimization of cellulase production by Penicillium sp. 3 Biotech 6, 162.

Radha, S., Babu, R.H., Sridevi, A., Prasad, N. and Narasimha, G. (2012). Development of mutant fungal strains of Aspergillus niger for enhanced production of acid protease in submerged and solid state fermentation. European Journal of Experimental Biology 2, 1517-28

Rathnan, R., Nair, P. and Balasaravanan, T. (2012). Isolation, identification and characterization of efficient cellulolytic fungi from natural resources. International Journal of Microbial Resource Technology 1, 379-387.
Salar, R.K. and Aneja, K. (2007). Thermophilic fungi: Taxonomy and Biogeography. Journal of Agricultural Technology 3, 77-107.

Shafique, S., Bajwa, R. and Shafique, S. (2011). Strain improvement in Trichoderma viride through mutation for overexpression of cellulase and characterization of mutants using random amplified polymorphic DNA (RAPD). African Journal of Biotechnology 10, 19590-19597.

Shahriarinour, M., Wahab, M.N.A., Ariff, A. and Mohamad, R. (2011). Screening, isolation and selection of cellulolytic fungi from oil palm empty fruit bunch fibre. Biotechnology 10, 108-113.

Velmurugan, P., Hur, H., Balachandar, V., Kamala-Kannan, S., Lee, K.J., Lee, S.M., Chae, J.C., Shea, P.J. and Oh, B.T. (2011). Monascus pigment production by solid-state fermentation with corn cob substrate. Journal of Bioscience and Bioengineering 112, 590-594.

Wati, L., Kumari, S. and Kundu, B. (2007). Paddy straw as substrate for ethanol production. Indian Journal of Microbiology 47, 26-29.
How to Cite
Nisar, K., Abdullah, R., Kaleem, A., Iqtedar, M., & Saleem, F. (2019). Hyper production of carboxy methyl cellulase by Thermomyces dupontii utilizing physical and chemical mutagenesis. Revista Mexicana De Ingeniería Química, 19(2), 617-625. https://doi.org/10.24275/rmiq/Bio823