Biosynthesis of Ascorbic Acid by Aspergillus Flavus and Aspergillus Tamarii Immobilized in Afzelia Africana Matrix


  • Temitope Temitayo Banjo Federal University of Agriculture, Abeokuta
  • Sarafadeen Olateju Kareem Federal University of Agriculture, Abeokuta
  • Temitope Olukayode Popoola Federal University of Agriculture, Abeokuta
  • Oluseyi Akinloye Federal University of Agriculture, Abeokuta



Ascorbic acid, Immobilization, Afzelia Africana, Matrix, Aspergillus flavus


A novel matrix for the immobilization of ascorbic acid produced by Aspergillus flavus and Aspergillus tamarii was reported. Spores of A. flavus and A. tamarii were immobilized on Afzelia africana matrix cross-linked with glutaraldehyde (2.5%) and the effects of Afzelia africana gel concentration (9–13%), spore load (100–500 mg/100 ml), bead size (2–7 mm) and bead number (2–10) on ascorbic acid yield were determined. The immobilized fungi were cultured in a liquid fermentation medium containing BSG (0.6% w/v) for ascorbic acid production for 144 h. The ascorbic acid produced was quantified titrimetrically. The statistical analysis of the effects of gel concentration, spore load and bead size on ascorbic acid production showed no significant difference at p>0.05. However, there was significant difference in the effect of bead number on ascorbic acid production at p<0.05. Ascorbic acid yield of 8.5 g/L and 7.5 g/L was produced by Aspergillus tamarii and Aspergillus flavus respectively using 9 beads at 96 h of fermentation. The immobilized Aspergillus tamarii and Aspergillus flavus retained activities of 72% and 70% respectively after five repeated cycle and also exhibited increased activities over the free cells. This study shows the potential of Afzelia africana as a novel matrix for enhanced ascorbic acid production.

Author Biographies

Temitope Temitayo Banjo, Federal University of Agriculture, Abeokuta

Institute for Human Resources and Development. Lecturer

Sarafadeen Olateju Kareem, Federal University of Agriculture, Abeokuta

Microbiology and Associate Professor

Temitope Olukayode Popoola, Federal University of Agriculture, Abeokuta

Microbiology and Professor

Oluseyi Akinloye, Federal University of Agriculture, Abeokuta

Biochemistry and Professor


Abdel-Naby, M. A., Sherif, A. A., El-Tanash, A. B. and Mankarios, A. T. 1999. Immobilization of Aspergillus oryzae tannase and properties of the immobilized enzyme. Journal of Applied Microbiology. 87(1): 108–114.

Adebayo, S.F. and Ojo, O.C. 2013. Nutrient composition and functional properties of Afzelia africana Seed. Journal of Environmental Science. Toxicology and Food Technology. 6(5): 01–03.

Ahmad, R. and Sardar, M. 2015. Enzyme immobilization: An overview on nanoparticles as immobilization matrix. Biochemistry and Analytical Biochemistry. 4(2): 178.

Ajibola, V.O., Babatunde, O.A. and Suleiman, S. 2009. The effect of storage method on the Vitamin C content in some tropical fruit juices. Trends in Applied Sciences Research. 4(2): 79–84.

Alsarra, I. A., Neau, S. H. and Howard, M. A. 2004. Effects of preparative parameters on the properties of chitosan hydrogel beads containing Candida rugosa lipase. Biomaterials. 25(13): 2645–2655.

Association of Vitamin Chemists. 1996. Methods of vitamin assay. New York: Interscience. 306–312.

Bayraktar, E. and Mehmetoglu, Ü. 2000. Production of citric acid using immobilized conidia of Aspergillus niger. Applied Biochemistry and Biotechnology. 87(2): 117–125.

Beshay, U. 2003. Production of alkaline protease by Teredinobacter turnirae cells immobilized in Ca-alginate beads. African Journal of Biotechnology. 2(3): 60–65

Betigeri, S.S. and Neau, S.H. 2002. Immobilization of lipase using hydrophilic polymers in the form of hydrogel beads. Biomaterials. 23(17): 3627–3636.

Brett, N.D. Benefits of Vitamin C. Retrieved March 3, 2017.

Ejikeme P.M, Obasi, L.N. and Egbuonu, A.C.C. 2010. Physico-chemical and toxicological studies on Afzelia africana seed and oil. African Journal of Biotechnology. 9(13): 1959–1963.

Elnashar, M.M. Low-cost foods and drugs using Immobilized enzymes on biopolymers. Retrieved September 3, 2017. From

Hancock, R. D., Galpin, J.R. and Viola, R. 2000. Biosynthesis of L-ascorbic acid (vitamin C) by Saccharomyces cerevisiae. FEMS Microbiology Letters. 186: 245–250.

Higdon, J. Vitamin C. Retrived September 16, 2017.

Huh, W.K., Lee, B.H., Kim, S.T., Kim, Y.R., Rhie, G.E., Baek, Y.W., Hwang, C.S., Lee, J.S. and Kang, S.O. 1998. D-Erythroascorbic acid is an important antioxidant molecule in Saccharomyces cerevisiae. Molecular Microbiology. 30(4): 895–903.

Ivanova, V., Petrova, P. and Hristov, J. 2011. Application in the ethanol fermentation of immobilized yeast cells in matrix of alginate/magnetic nanoparticles, on chitosan-magnetite micro particles and Cellulose-coated Magnetic nanoparticles. International Review of Chemical Engineering. 3: 289–299.

Jaiswal, N. and Prakash, O. 2011. Immobilization of soybean ?-amylase on gelatin and its application as a detergent additive. Asian Journal of Biochemistry. 6(4): 337–346.

Jegannathan, K. R., Chan, E. S. and Ravindra, P. 2009. Physical and stability characteristics of Burkholderia cepacia lipase encapsulated in ?-carrageenan. Journal of Molecular Catalysis B: Enzymatic. 58(4): 78–83.

Kareem, S.O., Oladipupo, I.O., Omemu, A.M. and Babajide, J.M. 2012. Production of Citric acid by Aspergillus niger immobilized in Detarium microcarpum matrix. Malaysian Journal of Microbiology. 9(2): 161–165.

Kareem, S.O., Adio, O.Q. and Osho, M.B. 2014. Immobilization of Aspergillus niger F7–02 lipase in polysaccharide hydrogel beads of Irvingia gabonensis matrix. Enzyme Research. 2014: 1–7.

Lee, D.H., Park, C.H., Yeo, J.M. and Kim, S.W. 2006. Lipase immobilization on silica gel using a cross-linking method. Journal of Industrial and Chemical Engineering.12(5): 777–782.

Li, Y. and Schellhorn, H.E. 2007. New developments and novel therapeutic perspectives for Vitamin C. Journal of Nutrition. 137: 2171–2184.

Lupulescu, A., 1993. The role of vitamins A, B Carotene, E and C in Cancer Cell Biology. International Journal for Vitamin and Nutrition Research. 63: 3–14.

Park, J.K. and Chang, H.N. 2000. Citric and gluconic acid production from fig by Aspergillus niger using solid state fermentation. Journal of Microbiology and Biotechnology. 25: 298–304.

Sawada, S. I. and Akiyoshi, K. 2010. Nano-encapsulation of lipase by self-assembled nanogels: induction of high enzyme activity and thermal stabilization. Macromolecular Bioscience.10(4): 353–358.

Shivani, P., Khushbu, P., Faldu, N., Thakkar, V. and Shubramanian, R.B. 2011. Extraction and analysis of Jatropha curcas L. seed oil. African Journal Biotechnology. 10(79): 8210–18213.

Talekar, S. and Chavare, S. 2012. Optimization of immobilization of α-amylase in alginate gel and its comparative biochemical studies with free α-amylase. Recent Research in Science and Technology. 4: 1–5.




How to Cite

Banjo, Temitope Temitayo, Sarafadeen Olateju Kareem, Temitope Olukayode Popoola, and Oluseyi Akinloye. 2017. “Biosynthesis of Ascorbic Acid by Aspergillus Flavus and Aspergillus Tamarii Immobilized in Afzelia Africana Matrix”. Food and Applied Bioscience Journal 6 (1):39-52.



Food Processing and Engineering