Studies of the Nutritional, Environmental Effects and Repressive Nature of Simple Sugars on the Production of endo-β-mannanase by Aspergillus flavus PT7 on Solid State Fermentation

Main Article Content

Ukponobong E. Anita
Nsikak U. Stephen
Abiodun A. Onilude
Inimfon A. Ibanga


Aims: The importance of nutritional and environmental factors in the production of microbial enzymes cannot be overemphasized. Hence, endo-β-mannanase production was systematically studied in a step-wise approach of building up on the experimentally observed conditions favouring the production of this enzyme in Aspergillus flavus PT7.

Place and Duration of Study: Department of Microbiology, University of Ibadan, Nigeria, between January 2018 and December 2018.

Methodology: Thirty-eight (38) fungal isolates obtained were screened for mannolytic ability using standard method. The highest producer of endo-β-mannasase was subjected to various production conditions by adjusting the nutritional and environmental factors in view of optimizing the production of this enzyme in the isolate Aspergillus flavus PT7.

Results: Copra meal was the highest inducer of mannanase production in the isolate at enzyme activity of 85.86±3.93 U/gds. Production increased to 94.54±0.42 when all forms of extraneous nitrogen sources were excluded from the production medium. pH 5.0, temperature 30°C, moisture content at 100% v/w and inoculum size of 8.0% v/w led to the increase in production by 44% (enzyme activity of 153.24±5.69 U/gds) in 5 days of incubation. Allowing the production set up additional two (2) days led to production increase with a recorded enzyme activity of 170.34±4.35 U/gds. Production of endo-β-mannanase in A. flavus PT7 was observed to be inductive as the presence of simple sugars like glucose, galactose, arabinose and xylose led to extended lag period in the production of the enzymes by the isolate.

Conclusion: Production of endo-β-mannanase by Aspergillus flavus PT7 was successfully optimized in a step-wise and systematic experimental study of the nutritional and environmental growth conditions of the isolate.

Aspergillus flavus PT7, optimization, fermentation, agro waste

Article Details

How to Cite
Anita, U. E., Stephen, N. U., Onilude, A. A., & Ibanga, I. A. (2019). Studies of the Nutritional, Environmental Effects and Repressive Nature of Simple Sugars on the Production of endo-β-mannanase by Aspergillus flavus PT7 on Solid State Fermentation. Journal of Advances in Biology & Biotechnology, 21(4), 1-12.
Original Research Article


Girio FM, Fonseca C, Carvalheiro F, Durate LC, Marques S, Bogel-Lukasik R. Hemicelluloses for fuel ethanol: A review. Bioresources Technology. 2010;13:4775-4800.

Reid JSG, Edwards ME, Dickson CA, Scott C, Gidley MJ. Tobacco transgenic lines that express fenugreek galactomannan galactosyltransferase constitutively have structurally altered galactomannans in their seed endosperm cell walls. Plant Physiology. 2003;131:1487–1495.

Petkowicz CLO, Reicher F, Chanzy H, Taravel FR, Vuoug R. Linear mannan in the endosperm of Schizolobium amazonicum. Carbohydrate Polymer. 2001;44:107-112.

Dhawan S, Kaur J. Microbial mannanases: An overview of production and applica-tions. Critical Reviews in Biotechnology. 2007;27:197–216.

Talbot G, Sygusch J. Purification and characterization of thermostable ß-mannanase and α-galactosidase from Bacillus stearothermophilus. Applied Environmental Microbiology. 1990;56: 3505-3510.

Takahashi R, Kusakabe L, Maekawa A, Suzuki T, Murakami K. Some properties of extracellular mannanase. Japanese Journal of Tropical Agriculture. 1983;27: 140.

Zakaria MM, Yamamoto S, Yagi T. Purification and characterization of an endo-1, 4-β-mannanase from Bacillus subtilis KU-1. FEMS Microbiol Lett. 1998; 158:25-31.

Sachslehner A, Haltrich D. Purification and some properties of a thermostable acidic endo-1,4-ß-mannanase from Sclerotium (Athelia) rolfsii. Universität fϋr Bodenkultur (BOKU), Austria. 1999;177:47-55.

Kurakake M, Komaki T. Production of β-mannanase and β-mannosidase from Aspergillus awamori K4 and their properties. Current Microbiology. 2001;42: 377–380.

Ferreira HM, Filho EXF. Purification and characterization of a mannanase from Trichoderma harzianum strain T4. Carbohydr Polym. 2004;57:23-29.

Ademark P, Larsson M, Tjerneld F, Stålbrand H. Multiple galactosidases from Aspergillus niger: Purification, characteri-zation and substratem specificities. Enzyme and Microbial Technology. 2001; 29:441-448.

Zhang J, He, ZM, Hu K. Purification and characterization of β-mannanase from Bacillus licheniformis for industrial use. Biotechnol Lett. 2000;22:1375-1378.

Haung XP, Monk C. Purification and characterization of a cellulase from a newly isolated thermophilic aerobic bacterium Caldibacillus cellulovorans gen. nov. sp. World Journal of Microbiology and Biotechnology. 2004;20:85-92.

Santiago SDN, Gonzalez CR, Almendarez BG, Fernandez FJ, Jurado AT, Ochoa SH. Physiology, morphological, and mannanase production studies on Aspergillus niger UAM-GS mutants. Electronic Journal of Biotechnology. 2007; 9:1.

Fawole MO, Oso BA. Laboratory manual of microbiology. Spectrum Books Ltd. Ibadan, Nigeria. 2004;1-48.

White TJ, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylo-genetics. In: PCR protocols: A guide to methods and applications. Edited by Inns MA, Gelfrand DH, Sninsky JJ, Withe TJ. Academic Press, New York. 1990;315- 322.

Sae-Lee N. The production of fungal mannanase, cellulose and xylanase using palm kernel meal as a substrate. Walailak Journal of Science and Technology. 2007;4:67-82.

Rashid SA, Ibrahim D, Omar IC. Mannanase production by Aspergillus niger USM F4 via solid substrate fermentation in a shallow tray using palm kernel cake as a substrate. Malaysian Journal of Microbiology. 2012;8(4):273-279.

Mabrouk MEM, El Ahway AMD. Production of ß-mannanase by Bacillus amylolequifaciens 10A1 cultured on potato peels. African Journal of Biotechnology. 2008;7(8):1123-1128.

Chantorn ST, Buengsrisawat K, Pokeseam A, Sombat T, Dangpram P, Jantawon K, Nitisinprasert S. Optimization of extracellular mannanase production from Penicillium oxalicum KUB-SN2-1 and application for hydrolysis property. Songklanakarin Journal of Science and Technology. 2013;35(1):17- 22.

Kote NV, Patil AG, Mulimani VH. Optimization of the production of thermostable endo-beta-1,4 mannanases from a newly isolated Aspergillus niger gr and Aspergillus flavus gr. Applied Biochemistry and Biotechnology. 2009; 152(2):213-223.

Pandey A. Solid-state fermentation. Biochemical Engineering Journal. 2003;13: 81–84.

Ramachandran S, Patel AK, Nampoothiri KM, Francis F, Nagy V, Szackacs G, Pandey A. Coconut oil cake-a potential material for production of a-amylase. Bioresource Technology. 2004;93(2):169-174.

Ibrahim D, Puspitaloka H, Rahim RA, Hong LS. Characterization of solid state fermentation culture conditions for growth and mananase production by Aspergillus niger USM F4 on rice husk in tray system. British Biotechnology Journal. 2012;2(3): 133-145.

Kumar RS, Shankar T, Anandapandian KTK. Characterization of alcohol resistant yeast Saccharomyces cerevisiae isolated from Toddy. International Research Journal of Microbiology. 2011;2(10):399-405.

De Ioannes P, Peirano A, Steiner J, Eyzaguirre J. An a-L-arabinofuranosidase from Penicillium purpurogenum: production, purification and properties. J Biotechnol. 2000;76:253–258.

Koseki T, Miwa Y, Mese Y, Miyanaga A, Fushinobu S, Wakgi T, Shoun H, Matsuzawa H, Hashizume K. Mutational analysis of N-glycosylation recognition sites on the biochemical properties of Aspergillus kawachii á-L-arabinofuran-osidase 54. Biochimica et Biophysica Acta (BBA). 1760;9:1458–1464.

Fritz M, Ravanal MC, Braet C, Eyzaguirre J. A family 51 á-L-arabinofuranosidase from Penicillium purpurogenum: purifica-tion, propertiesand amino acid sequence. Mycologia Research. 2008;112(8):933- 942.

Manpreet S, Sarwat S, Sachi D, Pankaj S, Banerjee UC. Influence of process parameters on the production of metabolites in solid-state fermentation. Malaysian Journal of Microbiology. 2005; 1(2):1-9

Wanderley KJ, Torres FAG, Moraes LMP, Ulhoa CJ. Biochemical characterization of α-amylase from the yeast Cryptococcus flavus. FEMS Microbiology Letters. 2004; 231(2):165–169

Jahangeer S, Khan N, Jahangeer S, Sohail M, Shahzad S, Ahmad A, Ahmed Khan S. Screening and characterisation of fungal cellulases isolated from the native environmental source. Pakistan Journal of Botany. 2005;37(3):739-748.

Singh C, Ahuja N, Batish M, Capalash N, Sharma P. Biobleaching of wheat straw-rich-soda-pulp with alkalophilic laccase from γ-Proteobacterium JB: Optimization of process parameters using response surface methodology. Bioresource Techno-logy. 2003;99:7472-7479.

Sachslehner A, Haltrich D, Nidetzky B, Kulbe KD. Production of hemicellulose and cellulose-degrading enzymes by various strains of Sclerotium rolfsii. Applied Biochemistry and Biotechnology. 1997;63-65:189-201.