Cellulase Production by Myceliophthora thermophila in Solid State Fermentation and Its Utility in Saccharification of Rice Straw


Singh, B., Anu, A., Singh, D., Kumar, V., Kumar, V., & Malik, V. (2022). Cellulase Production by Myceliophthora thermophila in Solid State Fermentation and Its Utility in Saccharification of Rice Straw. New Energy Exploitation and Application, 1(2), 10–17.


  • Bijender Singh
    Department of Biotechnology, Central University of Haryana, Jant-Pali, Mahendergarh-123031, Haryana, India
  • Anu Anu Laboratory of Bioprocess Technology, Department of Microbiology, Maharshi Dayanand University, Rohtak- 124001, Haryana, India.
  • Davender Singh Department of Physics, RPS Degree College, Balana, Mahendergarh-123031, Haryana, India
  • Vinod Kumar Department of Chemistry, Central University of Haryana, Jant-Pali, Mahendergarh-123031, Haryana, India
  • Vijay Kumar Department of Botany, Shivaji College (University of Delhi), Ring Road Raja Garden, New Delhi 110027, India
  • Vinay Malik Department of Zoology, Maharshi Dayanand University, Rohtak- 124001, Haryana, India.
Optimization of cellulase production by thermophilic mould Myceliophthora thermophila BJTLRMDU3 was studied in solid state fermentation. Myceliophthora thermophila produced maximum cellulase (45.81 U/g DMR) at substrate to moisture ratio of 1:3 with 5-d old inoculum at water activity 0.95, ammonium sulfate (0.5%) and PEG 20000 (0.5%) at 45 °C using “one variable at a time” approach. Further supplementation of Tween-20 (0.5%) and K2HPO4 (0.25%) enhanced the cellulase production (56.06 U/g DMR) by M. thermophila in SSF. Optimization of saccharification by partially purified cellulase of M. thermophila (20 U), liberated maximum reducing sugars at pH 5.0 (185.56 mg/g substrate) and 60 °C (190.83 mg/g substrate) after 24 h (203.91 mg/g substrate) from sodium carbonate pretreated rice straw as compared to untreated biomass. Liberated reducing sugars were higher in sodium carbonate pretreated rice straw than untreated rice straw.


Rice straw M. thermophila BJTLRMDU3 Optimization Cellulase Sodium carbonate Saccharification


  1. Anu, Kumar, A., Rapoport, A., et al., 2020a. Multifarious pretreatment strategies for the lignocellulosic substrates for the generation of sustainable biofuels. Renewable Energy. 160, 1228-1252.
  2. Anu, Singh, B., Kumar, A., 2020b. Process development for sodium carbonate pretreatment and enzymatic saccharification of rice straw for bioethanol production. Biomass Bioenergy. 138, 105574.
  3. Anu, Kumar, A., Jain, KK., et al., 2020c. Process Optimization for Chemical Pretreatment of Rice Straw for Bioethanol Production. Renewable Energy. 156, 1233-1243.
  4. Dahiya, S., Kumar, A., Singh, B., 2020. Enhanced endoxylanase production by Myceliophthora thermophila using rice straw and its synergism with phytase in improving nutrition. Process Biochemistry. 94, 235-242.
  5. Bala, A., Singh, B., 2017. Concomitant production of cellulase and xylanase by thermophilic mould Sporotrichum thermophile in solid state fermentation and
  6. their applicability in bread making. World Journal of Microbiology & Biotechnology. 33, 109.
  7. Bala, A., Singh, B., 2016. Cost-effective production of biotechnologically important hydrolytic enzymes by Sporotrichum thermophile. Bioprocess and Biosystems Engineering. 39, 181-191.
  8. Mehboob, N., Asad, M.J., Asgher, M., et al., 2014. Exploring thermophilic cellulolytic enzyme production potential of Aspergillus fumigatus by the solid-state fermentation of wheat straw. Applied Biochemistry & Biotechnology. 172, 3646-3655.
  9. Singh, B., 2016. Myceliophthora thermophile syn. Sporotrichum thermophile: a thermophilic mould of biotechnological potential. Critical Reviews in Biotechnology. 36, 59-69.
  10. Kuhad, R.C., Gupta, R., Singh, A., 2011. Microbial cellulases and their industrial applications. Enzyme Research.
  11. Alokika, Singh, B., 2020. Enhanced production of bacterial xylanase and its utility in saccharification of sugarcane bagasse. Bioprocess and Biosystems Engineering. pp. 1-11.
  12. Madadi, M., Tu, Y., Abbas, A., 2017. Recent status on enzymatic saccharification of lignocellulosic biomass for bioethanol production. Electronic Journal of Biology. 13, 135-143.
  13. Emerson, R., 1941. An experimental study on the life cycles and taxonomy of Allomyces. Lloydia. 4, 77-144.
  14. Miller, G.L., 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry. 31, 426-428.
  15. El-Bakry, M., Abraham, J., Cerda, A., et al., 2015. From wastes to high value added products: novel aspects of SSF in the production of enzymes. Critical Reviews in Environmental Science and Technology.45, 1999-2042.
  16. Sapna, Singh, B., 2014. Phytase production by Aspergillus oryzae in solid-state fermentation and its applicability in dephytinization of wheat bran. Applied Biochemistry & Biotechnology. 173, 1885-1895.
  17. Kar, S., Gauri, S.S., Das, A., et al., 2013. Process optimization of xylanase production using cheap solid substrate by Trichoderma reesei SAF3 and study on the alteration of behavioral properties of enzyme obtained from SSF and SmF. Bioprocess and Biosystems Engineering. 36, 57-68.
  18. Maan, P., Bharti, A.K., Gautam, S., et al., 2016. Screening of important factors for xylanase and cellulase production from the fungus C. cinerea RM-1 NFCCI-3086 through Plackett-Burman experimental design. BioResources. 11, 8269-8276.
  19. Jain, K.K., Dey, T.B., Kumar, S., et al., 2014. Production of thermostable hydrolases (cellulases and xylanase) from Thermoascus aurantiacus RCKK: a potential fungus. Bioprocess and Biosystems Engineering. 38, 787-796.
  20. Dahiya, S., Singh, B., 2019. Enhanced endoxylanase production by Myceliophthora thermophila with applicability in saccharification of agricultural substrates. 3 Biotech. 9, 214.
  21. Hemansi, Gupta, R., Kuhad, R.C., Saini, J.K., 2018. Cost effective production of complete cellulase system by newly isolated Aspergillus niger RCKH3 for efficient enzymatic saccharification: medium engineering by overall evaluation criteria approach (OEC). Biochemical Engineering Journal. 132, 182-190.
  22. Matsakas, L., Antonopoulou, I., Christakopoulos, P., 2015a. Evaluation of Myceliopthora thermophila as an enzyme factory for the production of thermophilic cellulolytic enzymes. BioResources. 10, 5140-5158.
  23. Matsakas, L., Christakopoulos, P., 2015b. Ethanol production from enzymatically treated dried food waste using enzymes produced on-site. Sustainability. 7, 1446-1458.
  24. Moretti, M.M.D.S., Bonfa, E.C., Chierotti, M.C.M., et al., 2014. Fibrolytic enzyme production of Myceliophthora thermophila M. 7.7. using inexpensive carbon sources and mineral nutrients. African Journal of Microbiology Research. 8, 4013-4019.
  25. Sipos, B., Szilágyi, M., Sebestyén, Z., et al., 2011. Mechanism of the positive effect of poly (ethylene glycol) addition in enzymatic hydrolysis of steam pretreated lignocelluloses. Comptes Rendus Biologies. 334, 812-823.
  26. Ramanjaneyulu, H.N.P.G., Reddy, B.R., 2016. Optimization of cellulase production by Penicillium sp. 3 Biotech. 6, 162.
  27. Chugh, P., Soni, R., Soni, S.K., 2016. Deoiled rice bran: a substrate for coproduction of a consortium of hydrolytic enzymes by Aspergillus niger P-19. Waste and Biomass Valorization. 7, 513-525.
  28. Jain, P., Jain, R.K., 2016. Enhanced cellulase production from isolated fungus Aspergillus niger RKJP and its application in lignocellulosic saccharification for bioethanol production. Biotechnological Research. 2, 61-68.
  29. Singh, V., Haque, S., Niwas, R., et al., 2017. Strategies for fermentation medium optimization: an indepth review. Frontiers in Microbiology. 7, 2087.
  30. Bibi, Z., Ansari, A., Zohra, R.R., et al., 2014. Production of xylan degrading endo-1, 4-β-xylanase from thermophilic Geobacillus stearothermophilus KIBGE-IB29. Journal of Radiation Research and Applied Sciences. 7, 478-485.
  31. Salihu, A., Bala, S.M., Olagunju, A., 2015. A statistical design approach for xylanase production by Aspergillus niger using soybean hulls: optimization and determining the synergistic effects of medium components on the enzyme production. Jordan Journal of Biological Sciences. 147, 3388.
  32. Alrumman, S.A., 2016. Enzymatic saccharification and fermentation of cellulosic date palm wastes to glucose and lactic acid. Brazilian Journal of Microbiology. 47, 110-119.
  33. Saratale, G., Jung, M., Oh, M., 2016 Reutilization of green liquor chemicals for pretreatment of whole rice waste biomass and its application to 2, 3-butanediol production. Bioresource Technology. 205, 90-96.
  34. Jin, Y., Huang, T., Geng, W., et al., 2013. Comparison of sodium carbonate pretreatment for enzymatic hydrolysis of wheat straw stem and leaf to produce fermentable sugars. Bioresource Technology. 137, 294-301.
  35. Kahar, P., 2013. Synergistic effects of pretreatment process on enzymatic digestion of rice straw for efficient ethanol fermentation. Environmental Biotechnology-New Approaches and Prospective Applications. pp. 73-77.
  36. Khaleghian, H., Karimi, K., Behzad, T., 2015. Ethanol production from rice straw by sodium carbonate pretreatment and Mucor hiemalis fermentation. Industrial Crops and Products. 76, 1079-1085.
  37. Salehi, S., Karimi, K., Behzad, T., et al., 2012. Efficient conversion of rice straw to bioethanol using sodium carbonate pretreatment. Energy Fuels. 26, 7354-7361.
  38. Ashoor, S., Sukumaran, R.K., 2020. Mild alkaline pretreatment can achieve high hydrolytic and fermentation efficiencies for rice straw conversion to bioethanol. Preparative Biochemistry & Biotechnology. pp. 1-6.
  39. Prasad, S., Kumar, S., Yadav, K.K., et al., 2020. Screening and evaluation of cellulytic fungal strains for saccharification and bioethanol production from rice residue. Energy. 190, 116422.
  40. Shen, Z., Zhang, K., Si, M., et al., 2018. Synergy of lignocelluloses pretreatment by sodium carbonate and bacterium to enhance enzymatic hydrolysis of rice straw. Bioresource Technology. 249, 154-160.
  41. Łukajtis, R., Kucharska, K., Hołowacz, I., et al., 2018. Comparison and optimization of saccharification conditions of alkaline pre-treated triticale straw for acid and enzymatic hydrolysis followed by ethanol fermentation. Energies. 11, 639.
  42. Prajapati, B.P., Kango, N., 2021. Rice straw saccharification using cellulolytic cocktail from Aspergillus tubingensis and structure alterations studies of the wall polymer. Biomass Conversion and Biorefinery. pp. 1-15.
  43. Bera, S., Banerjee, T., Samanta, A., 2021. Evaluation of enzymatic delignification of rice straw residues for bioethanol production. International Journal of Renewable Energy Technology. 12, 99-117.
  44. Singh, B., Bala, A., Anu, et al., 2021. Biochemical properties of cellulolytic and xylanolytic enzymes from Sporotrichum thermophile and their utility in bioethanol production using rice straw. Preparative Biochemistry & Biotechnology. pp. 1-13.
  45. Guo, J.M., Wang, Y.T., Cheng, J.R., et al., 2020. Enhancing enzymatic hydrolysis and fermentation efficiency of rice straw by pretreatment of sodium perborate. Biomass Conversion and Biorefinery. pp. 1-10.