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Biosynthesis and extraction of cellulase produced by Trichoderma on agro-wastes

  • R. KalsoomEmail author
  • S. Ahmed
  • M. Nadeem
  • S. Chohan
  • M. Abid
Original Paper
  • 52 Downloads

Abstract

Cellulase is the most important enzyme in the world enzyme market, which can be synthesized by growing fungi on lignocellulosic substrates. In this study, cellulase was produced by using Trichoderma species. Twenty-three Trichoderma species were isolated and screened out for their cellulase-producing ability. Selected species, Trichoderma reesei, was further optimized on Leptochloa fusca, a perennial grass. For the higher production of enzyme, different culture conditions were optimized in flask fermenters. Our study points out that overall cellulase production was highest as 1.165 IU/ml/min at 70% moisture level, 120-h incubation period, 30 °C incubation temperature, 6 initial pH, 20% inoculum size, 0.3% NH4NO3 concentration and 0.3% concentration of surfactant (Tween 80), respectively. Under solid-state fermentation, the recovery of the cellulase from fermented substrate was optimized which yielded 1.785 IU/ml/min. Among different solvents tested, optimal extraction was attained by using citrate buffer. The optimal conditions for extraction were 90 min soaking time, 1:5 solid-to-solvent ratio, 140 revolution per minute agitation. It was detected that two washes were enough for maximum leaching of the enzyme. Results specify the admirable scope of utilizing kallar grass for biosynthesis of cellulase in solid-state fermentation employing Trichoderma reesei commercially.

Keywords

Trichoderma reesei Kallar grass (Leptochloa fusca L.Kunth) Cellulase NH4NO3 Surfactants Extraction 

Notes

Acknowledgements

Authors want to thank Dr. Shahjahan Baig for providing the laboratory facilities for enzyme analysis at Pakistan Council of Scientific and Industrial Research (PCSIR) Lahore, Pakistan.

Supplementary material

13762_2018_1717_MOESM1_ESM.xlsx (82 kb)
Supplementary material 1 (XLSX 81 kb)

References

  1. Acharya PB, Acharya DK, Modi HA (2008) Optimization for cellulose production by Aspergillus niger using saw dust as substrate. Afr J Biotechnol 22:4147–4152Google Scholar
  2. Alam Z, Muhammad N, Mahmat ME (2005) Production of cellulase from oil palm biomass as substrate by solid state bioconversion. J Appl Sci 2:569–572CrossRefGoogle Scholar
  3. Ariffin H, Abdullah N, Umikalsom MS, Shirai Y, Hassan MA (2008) Production of bacterial endoglucanase from pretreated oil palm empty fruit bunch by Bacillus pumilus EB3. J Biosci Bioeng 106:231–236CrossRefGoogle Scholar
  4. Bilal T, Malik B, Reiaz RU, Kumar M (2015) Influence of various parameters on cellulase and xylanase production by different strains of trichoderma species. Aust J Anal Pharm Chem 2(1):1034Google Scholar
  5. Bisen PS, Sharma A (2012) Introduction to instrumentation in life sciences. CRC Press, Boca RatonCrossRefGoogle Scholar
  6. Brijwani KH, Oberoi S, Vadlani PV (2010) Production of a cellulolytic enzyme system in mixed-culture solid state fermentation of soybean hulls supplemented with wheat bran. Process Biochem 45:120–128CrossRefGoogle Scholar
  7. Chandra MA, Kalra PK, Sharma H, Kumar, Sangwan RS (2010) Optimization of cellulases production by Trichoderma citrinoviride on marc of Artemisia annua and its application for bioconversion process. Biomass Bioenerg Article in pressGoogle Scholar
  8. Chinedu SN, Okochi IV (2003) Cellulase production by wild-type Aspergillus niger, Penicillium chrysogenum and Trichoderma harzianum using waste cellulosic materials. Idi-Araba, Lagos, NigeriaGoogle Scholar
  9. Dedavid e Silva LA, Lopes FC, Silveira ST, Brandelli A (2009) Production of cellulolytic enzymes by Aspergillus phoenicis in grape waste using response surface methodology. Appl Biochem Biotechnol 152:295–305CrossRefGoogle Scholar
  10. Fadel M (2000) Production physiology of cellulases and β-glucosidase enzymes of Aspergillus niger grown under solid state fermentation conditions. J Biol Sci 1:401–411Google Scholar
  11. Fatokun EN, Nwodo UU, Okoh AI (2016) Classical optimization of cellulase and xylanase production by a marine streptomyces species. Appl Sci 6:286.  https://doi.org/10.3390/app6100286 CrossRefGoogle Scholar
  12. Fernandes P, Marques MP, Carvalho F, Cabral J (2009) A simple method for biocatalyst immobilization using PVA-based hydrogel particles. J Chem Technol Biotechnol 84:561–564CrossRefGoogle Scholar
  13. Gashe BA (1992) Cellulase production and activity by Trichoderma sp. A-001. J Appl Microbiol 73(1):79–82.  https://doi.org/10.1111/j.1365-2672.1992.tb04973.x Google Scholar
  14. Guoweia S, Man H, Shikai W, He C (2011) Effect of some factors on Production of cellulase by Trichoderma reesei HY07. Procedia Environ Sci 8:357–361CrossRefGoogle Scholar
  15. Ilyas U, Ahmed S, Majeed A, Nadeem M (2012) Biohydrolysis of Saccharum spontaneum for cellulase production by Aspergillus terreus. Afr J Biotechnol 11(21):4914–4920.  https://doi.org/10.5897/AJB11.1194 Google Scholar
  16. Ilyas U, Gohar F, Saeed S, Bukhari Z, Ilyas H (2013) Screening of locally isolated Aspergillus species for their cellulolytic potential and their optimization on Vigna mungo in solid state fermentation. Br Biotechnol J 3(3):350–358CrossRefGoogle Scholar
  17. Kretschmer B, Allen B, Hart K (2012) Mobilising Cereal Straw in the EU to feed Advanced Biofuel Production. Report produced for Novozymes. Institute for European Environmental Policy (IEEP), LondonGoogle Scholar
  18. Latifian M, Hamidi-Esfahani Z, Barzegar M (2007) Evaluation of culture conditions for cellulase production by two Trichoderma reesei mutants under solid-state fermentation conditions. Bioresource Technol 98:3634–3637CrossRefGoogle Scholar
  19. Liu J, Yang J (2007) Cellulase Production by Trichoderma koningii AS3.4262 in Solid-State Fermentation Using Lignocellulosic Waste from the Vinegar Industry. Food Technol Biotechnol 45:420–425Google Scholar
  20. Murphy RA, Horgan KA (2005) Antibiotics, enzymes and chemical commodities from Fungi. Biol Appl, Fungi, pp 113–143Google Scholar
  21. Pal A, Khanum F (2010) Production and extraction optimization of xylanase from Aspergillus niger DFR-5 through solid-state-fermentation. Bioresource Technol 101:7563–7569CrossRefGoogle Scholar
  22. Pardo AG (1996) Effect of surfactants on cellulase production by Nectria catalinensis. Curr Microbiol 33:275.  https://doi.org/10.1007/s002849900113 CrossRefGoogle Scholar
  23. Pirota RDPB, Miotto LS, Delabona PS, Farinas CS (2013) Improving the extraction conditions of endoglucanase produced by Aspergillus niger under solid-state fermentation. Braz J Chem Eng 30:117CrossRefGoogle Scholar
  24. Pirzadah T, Garg S, Singh J, Vyas A, Kumar M, Gaur N (2014) Characterization of Actinomycetes and Trichoderma spp. for cellulase production utilizing crude substrates by response surface methodology. BMC Springer Plus 3: 622Google Scholar
  25. Pothiraj C, Balaji P, Eyini M (2006) Enhanced production of Cellulases by various fungal cultures in solid state fermentation of cassava waste. Afr J Biotechnol 5:1882–1885Google Scholar
  26. Rajoka MI (2004) Influence of various fermentation variables on exo-glucanase production in Cellulomonas flavigena. Electr J Biotechnol 7(3):0717–3458CrossRefGoogle Scholar
  27. Samuels JG, Hebbar KP (2015) Trichoderma identification and agricultural applications. The American phytopathological society St. Paul, Minnesota, USAGoogle Scholar
  28. Singhania RR, Sukurmaran RK, Pillai A, Prema P, Szakacs G, Panday A (2006) Solid state fermentation of lignocellulosic substrates for cellulase production by Trichoderma reesei NRRL 11460. Indian J Biotechnol 5:332–336Google Scholar
  29. Steel RGD, Torrie JH, Dickey DA (1997) Principles and procedures of statistics: A biometrical approach, 3rd edn. McGraw Hill Book Co. Inc, New York, pp 400–438Google Scholar
  30. Toor Y, Ilyas U (2014) Optimization of Cellulase Production by Aspergillus ornatus by the Solid State Fermentation of Cicer arietinum. Am J Res Commun 2(1):125–141Google Scholar
  31. Zhang Z, Lohr L, Escalante C, Wetzstein M (2010) Food versus fuel: what do prices tell us? Energy Policy 38:445–451CrossRefGoogle Scholar

Copyright information

© Islamic Azad University (IAU) 2018

Authors and Affiliations

  1. 1.Institute of Agricultural SciencesUniversity of the PunjabLahorePakistan
  2. 2.Pakistan Council of Scientific and Industrial ResearchLahorePakistan
  3. 3.Department of Plant Pathology, Faculty of Agricultural Sciences and TechnologyBahauddin Zakariya UniversityMultanPakistan

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