Skip to main content
SpringerLink
Log in

Production of cellulolytic enzymes by Myceliophthora thermophila and their applicability in saccharification of rice straw

  • Original Article
  • Published:
Biomass Conversion and Biorefinery Aims and scope Submit manuscript

Abstract

Optimization of cellulase production by Myceliophthora thermophila BJTLRMDU3 was studied in solid state fermentation. Sequential use of Plackett-Burman and response surface methodology (RSM) resulted in the production of 98.81, 243.19, and 316.48 U/g dry moldy residue (DMR) of FPase, CMCase, and β-glucosidase, respectively. Statistical optimization has resulted in more than 4.0-fold increase in the production of cellulases. Optimization of saccharification of untreated and pretreated rice straw was carried out by cellulolytic enzymes of thermophilic mold M. thermophila. The liberation of reducing sugars was higher using fungal cellulases in ammonia-pretreated rice straw as compared with untreated biomass. Maximum liberation of reducing sugars was attained at 60 °C (224.24 mg/g substrate) and pH 5.0 (246.33 mg/g substrate) after an incubation time of 24 h (345.61 mg/g substrate) using enzyme dose of 20 U/g in ammonia-pretreated rice straw. Supplementation of xylanase further enhanced the saccharification (488.78 mg/g substrate) of pretreated rice straw. Analysis using high-performance liquid chromatography (HPLC) indicated the presence of various monomeric and oligomeric sugars in the enzymatic hydrolysate of pretreated rice straw. Both, Fourier-transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) revealed the structural changes in rice straw after ammonia pretreatment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Baeyens J, Kang Q, Appels L, Dewil R, Lv Y, Tan T (2015) Challenges and opportunities in improving the production of bio-ethanol. Prog Energy Combust Sci 47:60–88

    Article  Google Scholar 

  2. U.S. Department of Energy, available from: http://bioenergywiki.net/.

  3. Manivannan A, Narendhirakannan RT (2014) Response surface optimization for co-production of cellulase and xylanase enzymes by Trichoderma reesei NRRL–3652. Int J ChemTech Res 6:3883–3888

    Google Scholar 

  4. Singh B, Poças-Fonseca MJ, Johri BN, Satyanarayana T (2016) Thermophilic molds: biology and applications. Crit Rev Microbiol 42:985–1006

    Article  Google Scholar 

  5. Kaur P, Bhardwaj S, Bhardwaj NK, Sharma J (2018) Lignocellulosic waste as a sole substrate for production of crude cellulase from Bacillus subtilis PJK6 under solid state fermentation using statistical approach. J Carbohydr 1(1):1–14

    Google Scholar 

  6. Bala A, Singh B (2016) Cost-effective production of biotechnologically important hydrolytic enzymes by Sporotrichum thermophile. Bioprocess Biosyst Eng 39:181–191

    Article  Google Scholar 

  7. Behera SS, Ray RC (2016) Solid state fermentation for production of microbial cellulases: recent advances and improvement strategies. Int J Biol Macromol 86:656–669

    Article  Google Scholar 

  8. Bhat MK (2000) Cellulases and related enzymes in biotechnology. Biotechnol Adv 18:355–383

    Article  Google Scholar 

  9. Mehboob N, Asad MJ, Asgher M, Gulfraz M, Mukhtar T, Mahmood RT (2014) Exploring thermophilic cellulolytic enzyme production potential of Aspergillus fumigatus by the solid-state fermentation of wheat straw. Appl Biochem Biotechnol 172:3646–3655

    Article  Google Scholar 

  10. Kalogeris E, Christakopoulos P, Katapodis P, Alexiou A, Vlachou S, Kekos D, Macris BJ (2003) Production and characterization of cellulolytic enzyme from the thermophilic fungus Thermoascus aurantiacus under solid state cultivation of agricultural wastes. Process Biochem 38:1099–1104

    Article  Google Scholar 

  11. Kamra P, Satyanarayana T (2004) Xylanase production by the thermophilic mold Humicola lanuginosa in solid-state fermentation. Appl Biochem Biotechnol 119:145–157

    Article  Google Scholar 

  12. Emerson R (1941) An experimental study of the life cycles and taxonomy of Allomyces. Lloydia 4:77–144

    Google Scholar 

  13. Dahiya S, Singh B (2019) Enhanced endoxylanase production by Myceliophthora thermophila with applicability in saccharification of agricultural substrates. 3 Biotech 9:214

    Article  Google Scholar 

  14. Plackett RL, Burman JP (1946) The design of optimum multifactorial experiments. Biometrika 33:305–325

    Article  MathSciNet  MATH  Google Scholar 

  15. Kumar V, Satyanarayana T (2014) Biochemical and thermodynamic characteristics of thermo-alkali-stable xylanase from a novel polyextremophilic Bacillus halodurans TSEV1. Extremophiles 17:797–808

    Article  Google Scholar 

  16. Ghose TK (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268

    Article  Google Scholar 

  17. Kyu KL, Ratanakhanokchai K, Uttapap D, Tanticharoen M (1994) Induction of xylanase in Bacillus circulans B6. Bioresour Technol 48:163–167

    Article  Google Scholar 

  18. Miller GL (1959) Use of dinitrosalicyclic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  Google Scholar 

  19. Xin F, Geng A (2010) Horticultural waste as the substrate for cellulase and hemicellulase production by Trichoderma reesei under solid-state fermentation. Appl Biochem Biotechnol 162:295–306

    Article  Google Scholar 

  20. Bala A, Singh B (2019) Development of an environmental-benign process for efficient pretreatment and saccharification of Saccharum biomasses for bioethanol production. Renew Energy 130:12–24

    Article  Google Scholar 

  21. Sandhu SK, Oberoi HS, Babbar N, Miglani K, Chadha BS, Nanda DK (2013) Two-stage statistical medium optimization for augmented cellulase production via solid-state fermentation by newly isolated Aspergillus niger HN-1 and application of crude cellulase consortium in hydrolysis of rice straw. J Agric Food Chem 61:12653–12661

    Article  Google Scholar 

  22. Chugh P, Soni R, Soni SK (2016) Deoiled rice bran: a substrate for coproduction of a consortium of hydrolytic enzymes by Aspergillus niger P-19. Waste Biomass Valor 7:513–525

    Article  Google Scholar 

  23. Gao W, Kim YJ, Chung CH, Li J, Lee JW (2010) Optimization of mineral salts in medium for enhanced production of pullulan by Aureobasidium pullulans HP-2001 using an orthogonal array method. Biotechnol Bioprocess Eng 15:837–845

    Article  Google Scholar 

  24. Sharma R, Kocher GS, Rao SS, Oberoi HS (2019) Improved production of multi-component cellulolytic enzymes using sweet sorghum bagasse and thermophilic Aspergillus terreus rwy through statistical process optimization. Waste Biomass Valoriz. https://doi.org/10.1007/s12649-019-00670-5.

  25. Narra M, Dixit G, Divecha J, Madamwar D, Shah AR (2012) Production of cellulases by solid state fermentation with Aspergillus terreus and enzymatic hydrolysis of mild alkali-treated rice straw. Bioresour Technol 121:355–361

    Article  Google Scholar 

  26. de Cassia PJ, Paganini Marques N, Rodrigues A, Brito de Oliveira T, Boscolo M, Da Silva R, Gomes E, Bocchini Martins DA (2015) Thermophilic fungi as new sources for production of cellulases and xylanases with potential use in sugarcane bagasse saccharification. J Appl Microbiol 118:928–939

    Article  Google Scholar 

  27. Kilikian BV, Afonso LC, Souza TFC, Ferreira RG, Pinheiro IR (2014) Filamentous fungi and media for cellulase production in solid state cultures. Braz J Microbiol 45:279–286

    Article  Google Scholar 

  28. Moretti M, Bocchini-Martins DA, Silva RD, Rodrigues A, Sette LD, Gomes E (2012) Selection of thermophilic and thermotolerant fungi for the production of cellulases and xylanases under solid-state fermentation. Braz J Microbiol 43:1062–1071

    Article  Google Scholar 

  29. Zanelato AI, Shiota VM, Gomes E, Silva RD, Thoméo JC (2012) Endoglucanase production with the newly isolated Myceliophthora sp. i-1d3b in a packed bed solid state fermentor. Braz J Microbiol 43(4):1536–1544

    Article  Google Scholar 

  30. Soni R, Chadha B, Saini HS (2008) Novel sources of fungal cellulases of thermophilic/thermotolerant for efficient deinking of composite paper waste. Bioresources 3:234–246

    Google Scholar 

  31. Badhan AK, Chadha BS, Kaur J, Saini HS, Bhat MK (2007) Production of multiple xylanolytic and cellulolytic enzymes by thermophilic fungus Myceliophthora sp. IMI 387099. Bioresour Technol 98:504–510

    Article  Google Scholar 

  32. da Silva T, de Cássia V, de Souza Coto AL, de Carvalho SR, Bertoldi Sanchez Neves M, Gomes E, Bonilla-Rodriguez GO (2016) Effect of pH, temperature, and chemicals on the endoglucanases and β-glucosidases from the thermophilic fungus Myceliophthora heterothallica F.2.1. 4. Obtained by solid-state and submerged cultivation. Biochem Res Int 2016:1–9

    Article  Google Scholar 

  33. Kadowaki MA, Higasi P, de Godoy MO, Prade RA, Polikarpov I (2018) Biochemical and structural insights into a thermostable cellobiohydrolase from Myceliophthora thermophila. FEBS J 285:559–579

    Article  Google Scholar 

  34. Bonfá EC, de Souza Moretti MM, Gomes E, Bonilla-Rodriguez GO (2018) Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M. 7.7. Biocatal Agric Biotechnol 13:311–318

    Article  Google Scholar 

  35. Irshad MN, Anwar Z, But HI, Afroz A, Ikram N, Rashid U (2013) The industrial applicability of purified cellulase complex indigenously produced by Trichoderma viride through solid-state bio-processing of agro-industrial and municipal paper wastes. BioResources 8:145–157

    Google Scholar 

  36. Kumar B, Bhardwaj N, Alam A, Agrawal K, Prasad H, Verma P (2018) Production, purification and characterization of an acid/alkali and thermo tolerant cellulase from Schizophyllum commune NAIMCC-F-03379 and its application in hydrolysis of lignocellulosic wastes. AMB Express 8(1):173

    Article  Google Scholar 

  37. Li J, Yei X, Ruan S, Cai L, Chang Z (2010) Enzymatic hydrolysis of rice straw pretreated with ammonia. Conference on Environmental Pollution and Public Health, Scientific Research Publishing 482–485.

  38. Kaur K, Phutela UG (2018) Morphological and structural changes in paddy straw influenced by alkali and microbial pretreatment. Detritus 3:30–36

    Google Scholar 

  39. Park YC, Kim JS (2012) Comparison of various alkaline pretreatment methods of lignocellulosic biomass. Energy 47:31–35

    Article  Google Scholar 

  40. Chang KL, Thitikorn-amorn J, Hsieh JF, Ou BM, Chen SH, Ratanakhanokchai K, Huang PJ, Chen ST (2011) Enhanced enzymatic conversion with freeze pretreatment of rice straw. Biomass Bioenergy 35:90–95

    Article  Google Scholar 

  41. Alrumman SA (2016) Enzymatic saccharification and fermentation of cellulosic date palm wastes to glucose and lactic acid. Braz J Microbiol 47:110–119

    Article  Google Scholar 

  42. Rahnama N, Foo HL, Rahman NAA, Ariff A, Shah UKM (2014) Saccharification of rice straw by cellulase from a local Trichoderma harzianum SNRS3 for biobutanol production. BMC Biotechnol 14:103

    Article  Google Scholar 

  43. Jung YH, Kim IJ, Han JI, Choi IG, Kim KH (2011) Aqueous ammonia pretreatment of oil palm empty fruit bunches for ethanol production. Bioresour Technol 102:9806–9809

    Article  Google Scholar 

  44. Jung YH, Kim S, Yang TH, Lee HJ, Seung D, Park YC, Seo JH, Choi IG, Kim KH (2012) Aqueous ammonia pretreatment, saccharification, and fermentation evaluation of oil palm fronds for ethanol production. Bioprocess Biosyst Eng 35:1497–1503

    Article  Google Scholar 

  45. Sajja HK, Chowdary GV (2000) Optimization of simultaneous saccharification and fermentation for the production of ethanol from lignocellulosic biomass. J Agric Food Chem 48:1971–1976

    Article  Google Scholar 

  46. Zhu Y, Wang W, Wang Y, Jin Y (2015) Effects of pH and sulfonated lignin on the enzymatic saccharification of acid bisulfite-and green liquor-pretreated poplar wood. BioRes 10:7361–7371

    Google Scholar 

  47. Takano M, Hoshino K (2018) Bioethanol production from rice straw by simultaneous saccharification and fermentation with a statistical optimized cellulase cocktail and fermenting fungus. Bioresour Bioprocess 5:16

    Article  Google Scholar 

  48. Kahar P (2013) Synergistic effects of pretreatment process on enzymatic digestion of rice straw for efficient ethanol fermentation. In: Environmental Biotechnology-New Approaches and Prospective Applications, pp 65–87

    Google Scholar 

  49. Motamedi H, Hedayatkhah A, Varzi HN (2015) Improvement of the hydrolysis and fermentation of rice straw by Saccharomyces cerevisiae by ammonia-based pretreatments. BioResources 10:4360–4374

    Article  Google Scholar 

  50. Shi T, Lin J, Li J, Zhang Y, Jiang C, Lv X, Fan Z, Xiao W, Xu Y, Liu Z (2019) Pre-treatment of sugarcane bagasse with aqueous ammonia–glycerol mixtures to enhance enzymatic saccharification and recovery of ammonia. Bioresour Technol 289:121628

    Article  Google Scholar 

  51. Song HT, Gao Y, Yang YM, Xiao WJ, Liu SH, Xia WC, Liu ZL, Yi L, Jiang ZB (2016) Synergistic effect of cellulase and xylanase during hydrolysis of natural lignocellulosic substrates. Bioresour Technol 219:710–715

    Article  Google Scholar 

  52. Arantes V, Saddler JN (2011) Cellulose accessibility limits the effectiveness of minimum cellulase loading on the efficient hydrolysis of pre-treated lignocellulosic substrates. Biotechnol Biofuels 4:3

    Article  Google Scholar 

  53. Hu J, Arantes V, Saddler JN (2011) The enhancement of enzymatic hydrolysis of lignocellulosic substrates by the addition of accessory enzymes such as xylanase: is it an additive or synergistic effect? Biotechnol Biofuels 4(1):36

    Article  Google Scholar 

  54. Phitsuwan P, Permsriburasuk C, Baramee S, Teeravivattanakit T, Ratanakhanokchai K (2017) Structural analysis of alkaline pretreated rice straw for ethanol production. Int J Polym Sci 2017:1–9

    Article  Google Scholar 

  55. Binod P, Satyanagalakshmi K, Sindhu R, Janu KU, Sukumaran RK, Pandey A (2012) Short duration microwave assisted pretreatment enhances the enzymatic saccharification and fermentable sugar yield from sugarcane bagasse. Renew Energy 37:109–116

    Article  Google Scholar 

  56. Cha YL, Yang J, Ahn JW, Moon YH, Yoon YM, Yu GD, An GH, Choi IH (2014) The optimized CO2-added ammonia explosion pretreatment for bioethanol production from rice straw. Bioprocess Biosyst Eng 37:1907–1915

    Article  Google Scholar 

  57. Birhade S, Pednekar M, Sagwal S, Odaneth A, Lali A (2017) Preparation of cellulase concoction using differential adsorption phenomenon. Prep Biochem Biotechnol 47:520–529

    Article  Google Scholar 

  58. Ghaffar A, Yameen M, Aslam N, Jalal F, Noreen R, Munir B, Mahmood Z, Saleem S, Rafiq N, Falak S, Tahir IM, Noman M, Farooq MU, Qasim S, Latif F (2017) Acidic and enzymatic saccharification of waste agricultural biomass for biotechnological production of xylitol. Chem Cent J 11:97

    Article  Google Scholar 

Download references

Acknowledgments

The Department of Science and Technology [No. 1196 SR/FST/LS-I/2017/4] is highly acknowledged for providing basic infrastructure facility at Maharshi Dayanand University, Rohtak for carrying out this research work.

Funding

The authors acknowledge the Haryana State Council for Science and Technology (HSCST/R&D/2017/62 and No. 1743, dated 12/04/2017) for financial assistance in the form of a research project and fellowship, respectively during the tenure of this research work.

Author information

Authors and Affiliations

  1. Laboratory of Bioprocess Technology, Department of Microbiology, Maharshi Dayanand University, Rohtak, Haryana, 124001, India

    Anu & Bijender Singh

  2. Department of Botany, Pt. N.R.S. Govt. College, Rohtak, Haryana, 124001, India

    Anil Kumar

  3. Department of Physics, RPS Degree College, Balana, Mahendergarh, Haryana, 123031, India

    Davender Singh

  4. Department of Chemistry, Central University of Haryana, Jant-Pali, Mahendergarh, Haryana, 123031, India

    Vinod Kumar

  5. Department of Biotechnology, Central University of Haryana, Jant-Pali, Mahendergarh, Haryana, 123031, India

    Bijender Singh

Authors
  1. Anu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anil Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Davender Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vinod Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bijender Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bijender Singh.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(PDF 187 kb).

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Anu, Kumar, A., Singh, D. et al. Production of cellulolytic enzymes by Myceliophthora thermophila and their applicability in saccharification of rice straw. Biomass Conv. Bioref. 12, 2649–2662 (2022). https://doi.org/10.1007/s13399-020-00783-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13399-020-00783-1

Keywords

Search

Navigation