Cellulase Production by Penicillium citrinum using Brewer’s Spent Grain and Pineapple Peels as Cheap, Alternate Substrates

Cellulase Production by Penicillium citrinum using Brewer’s Spent Grain and Pineapple Peels as Cheap, Alternate Substrates

Loading document ...
Loading page ...


Author(s): Olaoluwa Oyedeji, Opeyemi O. Ojekunle

Download Full PDF Read Complete Article

DOI: 10.18483/ijSci.1513 155 481 74-83 Volume 7 - Jan 2018


Cellulases are major group of enzymes with wide-ranging industrial and biotechnological applications. The high cost of cellulase production is a major factor limiting its industrial applications in cellulose bioconversions, hence the need to develop low-cost production systems for this enzyme. Cellulose-rich plant biomass which may be agricultural or industrial in origin exists abundantly as organic wastes which are detrimental to the environment. This study evaluated cellulase production by Penicillium citrinum isolated from deteriorating orange fruits, using brewer’s spent grain and pineapple peels as cheap, alternate substrates. Cellulase titres 3.82 ± 0.136 U/mL and 1.405 ± 0.151 U/mL were produced by the fungus, using pineapple peels and brewer’s spent grain as substrates, respectively, under submerged fermentation. Maximum cellulase production by P. citrinum occurred with the use of pineapple peels as substrate, after 72 h fermentation period, with the use of pineapple peels at a concentration of 1.5%w/v and peptone as the best nitrogen source. The optima pH and temperature for the production of cellulase by the fungus was found to be 6.0 and 50 oC, respectively. Findings from this study indicated the potential use of pineapple peels as cheaper, alternative substrate for the production of cellulase thus mitigating its hazardous effect on the environment as pollutant. P. citrinum was able to grow and produce good levels of cellulase using solely pineapple peels as low-cost substrate, at high temperature of 50 oC, making this strain and this low-cost agro-industrial residue worthy of further investigation and potentially feasible for a wide range of biotechnological applications.


Cellulase, Cellulose, Penicillium citrinum, Agro-Industrial Wastes, Brewer’s Spent Grain, Pineapple Peels


  1. Abo-State, M.A.M., Hammad, A.I., Swelim, M. and Gannam, R.B (2010). Enhanced Production of Cellulases by Aspergillus spp isolated from Agricultural Wastes by Solid State Fermentation. American Eurasian Journal of Agriculture and Environmental Sciences, 8(4):402-410.
  2. Abou-Taleb, K.A.A., Mashhoor, W.A., Nasr, S.A., Sharaf, M.S. and Abdel-Azeem, H.H.M (2009). Nutritional and environmental factors affecting cellulose production by two strains of cellulolytic bacilli. Australian Journal of Basic and Applied Sciences, 3:2429-2436.
  3. Acharya, S. and Chaudhury, A (2012). Bioprospecting Thermophiles for Cellulase Production: A Review. Brazilian Journal of Microbiology, 2012:844-856.
  4. Adsul, M.G., Ghule, J.E., Singh, R., Shaikh, H., Bastawdea, K.B., Gokhale, D.V. and Varma, A.J (2004). Polysaccharides from bagasse: applications in cellulase and xylanase production. Carbohydrate Polymer, 57:67-72.
  5. http://dx.doi.org/10.1016/j.carbpol.2004.04.001
  6. Akinyele, B.J., Fabunmi, A.O. and Olaniyi OO (2013). Effect of variation in growth parameters on cellulases activity of Trichoderma viride NSPR006 cultured on different wood-dusts. Malaysian Journal of Microbiology, 9(3):193-200.
  7. Ali, S., Sayed, A., Sarker, R.T. and Alam, R (1991). Factors affecting cellulase production by Aspergillus terreus. World Journal of Microbiology and Biotechnology, 7:62-66.
  8. Asgher, M., Ahmad, Z. and Iqbal, H.M.N (2013). Alkali and enzymatic delignification of sugarcane bagasse to expose cellulose polymers for saccharification and bioethanol production. Industrial Crops and Products, 44:488-495.
  9. Azzaz, H.H., Murad, H.A., Kholif, A.M., Hanfy, M.A. and Abdel Gawad, M.H (2012). Optimization of Culture Conditions Affecting Fungal Cellulase Production. Research Journal of Microbiology, 7(9):23-31.
  10. Bakare, M.K., Adewale, I.O., Ajayi, A.O., Okoh, A.I. and Shonukan, O.O (2005). Regulatory mutations affecting the synthesis of cellulase in Pseudomonas fluorescens. African Journal of Biotechnology, 4(8):838-843.
  11. Balaraju, K., Park, K., Jahagirdar, S. and Kaviyarasan, V (2010). Production of cellulase and laccase enzymes by Oudemansiella radicata using agro wastes under solid state and submerged conditions. Research in Biotechnology, 1:21-28.
  12. Bhat, M.K (2000). Cellulase and related enzymes in biotechnology. Biotechnology Advances, 18:355-383.
  13. Blanch, H.W., Simmons, B.A. and Klein-Marcuschamer, D (2011). Biomass deconstruction to sugars. Biotechnology Journal, 6:1086-1102.
  14. Bon, E.P.S. and Ferrara, M.A (2007). Bioethanol production via enzymatic hydrolysis of cellulosic biomass on "The role of agricultural biotechnologies for production of bioenergy in developing countries", FAO seminar, Rome.2007.
  15. Brijwani, K., Oberoi, H.S. and Valdani, P.V (2010). Production of a cellulolytic enzyme system in mixed culture solid state fermentation of soybean hulls supplemented with wheat bran. Process Biochemistry, 45: 20-128.
  16. Chandra, M.S., Viswanath, B. and Reddy, B.R (2007). Cellulolytic enzymes on lignocellulosic substrates in solid state fermentation by Aspergillus niger. Indian Journal of Microbiology, 17:323-328.
  17. Chen, M., Xia, I. and Xue, P (2007). Enzymatic hydrolysis of corncob and ethanol production from cellulosic hydrolysate. International Journal of Biodeterioration and Biodegradation, 59:85-89.
  18. Cosgrove, D.J (2005). Growth of the plant cell wall. Natural Review of Molecular and Celularl Biology, 6:850-861.
  19. Devanathan, G., Shanmugan, A., Balasubramanian, T. and Manivannan, S (2007). Cellulase production by Aspergillus niger isolated from coastal mangrove debris. Trends in Applied Sciences Research, 2:23-27.
  20. Eveleigh, D.E., Bok, J.D., El-Dorry, H.S., El-Gogary, S., Eliston, K., Goyal, A., Waldron, C., Wright, R. and Wu, Y.-M (2005). Cellulase lessons revealed through the microbe’s perspective. Applied Biochemistry and Biotechnology, 51-52(0):169-177.
  21. Fang, X., Yano, S., Inoue, H. and Sawayama, S (2009). Strain improvement of Acremonium cellulolyticum for cellulase production by mutation. Journal of Bioscience and Bioengineering, 107:256-261.
  22. Gautam, S.P., Budela, P.S., Pandey, A.K., Jamaluddin, A.M.K. and Sarsaiya, S (2010). Optimization of the medium for the production of cellulase by Trichoderma viride using submerged fermentation. Journal of Environmental Sciences, 4(1):656-665.
  23. Ghose, T.K (1987). Measurement of cellulase activities. Pure and Applied Chemistry 59:257-268.
  24. Gupta, R., Gigras, P., Mohapatra, H., Goswami, V.K. and Chauhan, B (2008). Microbial α-amylases. A Biotechnological Perspective. Process Biochemistry, 38(11):1599-1616.
  25. Hafiz, M.N.I., Muhammad, A., Ishtiaq, A. and Shahbaz, H (2010). Media optimization for hyperproduction of carboxymethyl cellulase using proximally analysed agro-industrial residues with Trichoderma harzianum under SSF. International Journal of Agriculture Science and Veterinary Medicine, 4(2):47-55.
  26. Han, S.O., Yukawa, H., Inui, M. and Doi, R.H (2003). Regulation of expression of cellulosomal cellulase and hemicellulase genes in Clostridium cellulovorans. Journal of Bacteriology, 185:6067-6075.
  27. Hikaru, N., Katsunori, O., Ryota K., Kubota, T., Onodera, T., Ochiai, N., Omata, N., Ogasawara, W., Okada, H. and Morikawa, Y (2008). Characterisation of the catalytic domain of Trichoderma reesei endo glucanase I, II and III expressed in Escerichia coli. Applied Microbiology and Biotechnology, 81:681-689.
  28. Howell, J.A. and Mangat, M (1978). Enzymatic deactivation during cellulose hydrolysis. Biotechnology and Bioengineering, 20:847-863.
  29. Iqbal, H.M.N., Kyazze, G. and Keshavarz, T (2013). Advances in valorization of lignocellulosic materials by biotechnology: An overview. BioResource, 8:3157-3176.
  30. Irshad, M., Anwar, Z., But, H.I., Afroz, A., Ikram, N. and 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. BioResource, 8:145-157.
  31. Kathiresan, K. and Manivannan, S (2006). Cellulase production by Penicillium fellutanum isolated from coastal mangrove rhizosphere soil. Research Journal of Microbiology, 1(5):438-442.
  32. Kim, S. and Dale, B.E (2004). Global potential bioethanol production from wasted crops and crop residues. Biomass and Bioenergy, 26(4):361-375.
  33. Klein-Marcuschamer, D., Oleskowicz-Popiel, P., Simmons, B.A. and Blanch, H.W (2011). The challenge of enzyme costs in the production of lignocellulosic biofuels. Biotechnology and Bioengineering, 109(4):1083-1087.
  34. https//doi:10.1002/bit.24370.
  35. Krishna, C. (1999). Production of bacterial cellulases by solid state bioprocessing of banana wastes. Bioresource Technology, 69(3):231-239.
  36. Lee, R.L., Willem, H.V.Z., John, E.M. and Mark, L (2005). Consolidated bioprocessing of cellulosic biomass: an update. Current Opinion in Biotechnology, 16:577-583.
  37. Liu, J. and Yang, J (2007). Cellulase production by Trichoderma koningi AS3.4262 in solid state fermentation using lignocellulosic waste from vinegar industry. Food Technology and Biotechnology, 45:420-425.
  38. Lynd, L.R., Weimer, P.J. and Van Zyl, W.H (2002). Pretorius IS. Microbial Cellulase Utilisation: Fundamentals and Biotechnology. Microbiology and Molecular Biology Reviews, 66:506-577.
  39. Mabrouk, E.M. and Ahwany, M.D (2008). Production of mannanase by Bacillus amyloliquifaciens 10A1 cultured on potato peels. African Journal of Biotechnology, 7(8):1128.
  40. Mandels, M. and Weber, J (1969). Exoglucanase activity by microorganisms. Advances in Chemistry, 95:391-414.
  41. Milala, M.A., Shugaba, A., Gidado, A., Ene, A.C. and Wafar, J.A (2005). Studies on the use of agricultural wastes for cellulose enzyme production by Aspergillus niger. Research Journal of Agricultural and Biological Sciences, 1(4):325-328.
  42. Miller, G.L (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31:426-428.
  43. Mrudula, S. and Murugammal, R (2011). Production of cellulose by Aspergillus niger under submerged and solid state fermentation using coir waste as a substrate. Brazilian Journal of Microbiology, 42:1119-1127.
  44. Mussato, S.I., Fernandes, M., Mancilha, I.M. and Roberto, I.C (2008). Effects of medium supplementation and pH control on lactic acid production from brewer’s spent grain. Biochemical Engineering Journal, 40:437-444.
  45. Narasimha, G., Sridevi, A., Buddola, V., Subhosh, C.M. and Rajasekhar, R.B (2006). Nutrient effects on production of cellulolytic enzymes by Aspergillus niger. African Journal of Biotechnology, 5:472-476.
  46. Nascimento, R.P., Junior, N.A., Pereira, Jr. N., Bon, E.P.S. and Coelho, R.R.R (2009). Brewer’s spent grain and corn steep liquor as substrates for cellulolytic enzymes production by Streptomyces malaysiensis. Letters in Applied Microbiology, 48:529-535.
  47. Nipa, M.N., Sultana, S. and Hakim, M.A (2006). Induction of cellulose biosynthesis by cellobiose octaacetate in Aspergillus humicola. Microbiology Journal, 23:174-176.
  48. Odeniyi, O.A., Onilude, A.A. and Ayodele, M.A (2009). Production characteristic and properties of cellulase/polygalacturonase by a Bacillus coagulans strain from a fermenting palm-fruit industrial residue. African Journal of Microbiology Research, 3(8):407-417.
  49. Ogel, Z.B., Yarangumeli, K., Du, H. and Ifrij, J (2001). Submerged cultivation of Scytalidium thermophilum on complex lignocellulose biomass. Enzyme and Microbial Technology, 28:689-695.
  50. Omojasola, P.F., Jilani, O.P. and Ibiyemi, S.A (2008). Cellulase Production by Some Fungi Cultured on Pineapple Waste. Nature and Science, 6(2):64-79.
  51. Paengkoum, S., Wanapal, M. and Paengkoum, P (2013). Effects of pineapple peel and rice straw ratios as basal roughage in dairy cow. World Academy of Science, Engineering and Technology, 7(1):7-8.
  52. Pham, T.H., Quyen, D.T. and Nghiem, N.M (2010). Optimization of endoglucanase production by Aspergillus niger VTCC-F021. Australian Journal of Basic and Applied Sciences, 4(9):4151-4157.
  53. Rani, D.S. and Nand, K (2004). Ensilage of pineapple processing water for methane generation. Waste Management, 24:523-528.
  54. Ravindran, C., Naveenan, T. and Varatharajan, G (2010). Optimization of alkaline cellulase production from marine derived fungi, Chaetomium sp., using agricultural and industrial wastes as substrates. Botanica Marina, 53(3):275-282.
  55. Reddy, G.P.K., Narasimha, G., Kumar, K.D., Ramanjaneyulu, G., Ramya, A., Shanti Kumari, B.S. and Reddy, B.R (2015). Cellulase production by Aspergillus niger on different lignocellulosic cubstrates. International Journal of Current Microbiology and Applied Science, 4(4):835-845.
  56. Riswan, S.B.A., Muthuvelayudham, R. and Viruthagiri, T. 92012). Statistical optimization of nutrients for production cellulase & hemicellulase from rice straw. Asian Journal of Biochemistry and Pharmaceutical Research, 2:154-174.
  57. Rubin, E.M (2008). Genomics of cellulosic biofuels. Nature, 454(14):841-845.
  58. Sarao, L.K., Arora, M. and Sehgal, V.K (2010). Use of Scopulariopsis acremonium for the production of cellulose and xylanase through submerged fermentation. African Journal of Biotechnology, 4(14):1506-1510.
  59. Shoichi, T., Xoighi, K. and Hiroshi, S (1985). Cellulase production by Penicillium purpurogenum. Journal of Fermentation Technology, 62:127-127.
  60. Singh, A (1999). Engineering enzyme properties. Indian Journal of Microbiology, 39(2):65-77.
  61. Sonjoy, S.B., Bex, K. and Honston, H (1995). Cellulase activity of Trichoderma reesei (RUT-C30) on municipal solid waste. Applied Biochemistry and Biotechnology, 51-52(1):145-153.
  62. Stojceska, V. and Ainsworth, P (2008). The effect of different enzymes on the quality of high-fibre enriched brewer’s spent grain breads. Food Chemistry, 110:865-872.
  63. Xavier, S. and Lonsane, B.K (1994). Factors influencing fungal degradation of total soluble carbohydrates in sugarcane-pressmud under solid state fermentation. Process Biochemistry, 16:435-440.
  64. Xiros, C., Topakas, E., Katapodis, P. and Christakapoulos, P (2008). Evaluation of Fusarium oxysporium an enzyme factory for the hydrolysis of brewer’s spent grain with improved biodegradability for ethanol production. Industrial Crops Production, 8:213-224.
  65. Youssef, G.A. and Berekaa, M.M (2009). Improved production of endoglucanase enzyme by Aspergillus terreus; Application of Plackett-Burman design for optimization of process parameters. Biotechnology, 8(2):212-219.

Cite this Article:

International Journal of Sciences is Open Access Journal.
This article is licensed under a Creative Commons Attribution 4.0 International (CC BY 4.0) License.
Author(s) retain the copyrights of this article, though, publication rights are with Alkhaer Publications.

Search Articles

Issue June 2024

Volume 13, June 2024

Table of Contents

World-wide Delivery is FREE

Share this Issue with Friends:

Submit your Paper