Enzymatic hydrolysis of naringin by the action of naringinase is one of the standard practices adopted in the citrus fruit juice industry for debittering. In the present study, a submerged fermentation condition was optimized for producing naringinase from Aspergillus niger van Tieghem MTCC 2425. As per Placket–Burman design, pH (3–5), incubation temperature (26–30 °C), and inducer concentration (12–18 g·L−1) were the most important factors influencing the naringinase production. Naringin from citrus waste was used as an inducer. A rotatable central composite design was employed on these three variables and the numerical optimization predicted that fermentation at 29.8 °C, pH 4.7, and inducer concentration of 14.9 g L−1 would yield a maximum naringinase activity of 545.2 IU g−1. During partial purification, ion exchange chromatography led to a 9.92-fold increase in enzyme activity resulting a specific activity of 5460 IU g−1 with an activity recovery of 17%. As reflected by SDS–PAGE profile, the partially purified naringinase showed the molecular weight bands of 10–20, 65, and 80 kDa, respectively. The purified form of enzyme showed optimum stability at pH 5 and 50 °C. The naringinase activity was completely retained up to 150 days when stored at 4 °C.
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The authors express their sincere gratitude to the Department of Biotechnology, Government of India for financial support during the research.
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Borkar, V., Chakraborty, S. & Gokhale, J.S. Fermentative Production of Naringinase from Aspergillus niger van Tieghem MTCC 2425 Using Citrus Wastes: Process Optimization, Partial Purification, and Characterization. Appl Biochem Biotechnol 193, 1321–1337 (2021). https://doi.org/10.1007/s12010-020-03385-9
- Debittering enzyme
- Response surface methodology
- Numerical optimization
- Thermal stability
- Fungal enzyme