Bisher, A., Alsaman, M., Abdelaa, A., Mahrous, H. (2021). Optimization of thermostable inulinase production from Aspergillus niger NRRL 3122, purification, and characterization. Research Journal of Applied Biotechnology, 7(1), 15-29. doi: 10.21608/rjab.2021.303287
Ahmed Bisher; Mahmoud Alsaman; Asmaa Abdelaa; Hoda Mahrous. "Optimization of thermostable inulinase production from Aspergillus niger NRRL 3122, purification, and characterization". Research Journal of Applied Biotechnology, 7, 1, 2021, 15-29. doi: 10.21608/rjab.2021.303287
Bisher, A., Alsaman, M., Abdelaa, A., Mahrous, H. (2021). 'Optimization of thermostable inulinase production from Aspergillus niger NRRL 3122, purification, and characterization', Research Journal of Applied Biotechnology, 7(1), pp. 15-29. doi: 10.21608/rjab.2021.303287
Bisher, A., Alsaman, M., Abdelaa, A., Mahrous, H. Optimization of thermostable inulinase production from Aspergillus niger NRRL 3122, purification, and characterization. Research Journal of Applied Biotechnology, 2021; 7(1): 15-29. doi: 10.21608/rjab.2021.303287
Optimization of thermostable inulinase production from Aspergillus niger NRRL 3122, purification, and characterization
Department of Industrial Biotechnology, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Sadat City, Egypt
Abstract
Inulinase is a versatile glycoside hydrolase enzyme that targets the -2, 1 linkage of fructopolymers. The objective of the study was to produce inulinase using a low-cost carbon source from Aspergillus niger NRRL 3122, which was successful in producing a high titer of inulinase on agave. The Plackett-Burman design and Box-Behnken design were used to optimize the production of inulinase, which resulted in a high inulinase titer of 2170.22 U/ml, which is 2.83 times higher than the screening. The optimal levels of agave, NaNO3, and KCl were found to be 10 g/L, 4 g/L, and 0.3 g/L, respectively. The molecular weight of the enzyme was around 50 KDa. The enzyme showed maximum performance at 50°C and pH 6.0, and temperature stability up to 70°C, while pH stability was observed between 4-6. The pure inulinase could only hydrolyze inulin and sucrose, and not cellobiose and soluble starches. The Km and Vmax values for inulin were found to be 0.76 mg/mL and 100,000 U/mg, respectively.