Document Type : Original Article
Authors
1
Chemistry of Natural and Microbial Products Department, Pharmaceutical and Drug Industries Research Institute, National Research Center (NRC), Dokki, Cairo, Egypt.
2
Agricultural Microbiology Department, National Research Center, Cairo, Egypt.
3
Chemistry of Natural and Microbial Products Department, Pharmaceutical and Drug Industries Research Institute, National Research Center (NRC), Dokki, Cairo, Egypt
4
Center of Excellence, Encapsulation and Nano biotechnology Group, Chemistry of Natural and Microbial Products Department, Pharmaceutical and Drug Industries Research Institute,, National Research Center, El Behouth Street, Cairo 12622, Egypt.
Abstract
Glucose tolerant β-glucosidases have attracted considerable attention due to their important roles in various biotechnological processes. Penicillium sp. NRC24, a new fungal isolate was used to produce soy glycosides-hydrolyzing β-glucosidase. The ability of β-glucosidase to convert soy glucosides into aglycone forms, as well as its activity on p-NPG, was investigated. Penicillium sp. NRC24 β-glucosidase was partially purified with 8.04-fold and final recovery of 45.7%. This enzyme exhibited a glucose-tolerant activity of 60.4% while the partially purified enzyme showed 85.6% activity at 0.5 M of glucose. Moreover, the produced β-glucosidase could tolerate ethanol up to 60%. β-glucosidase retained all its initial activity at 37 °C in presence of 15-60 % ethanol at pH 5.0. The immobilization and characterization of the produced β-glucosidase are described. The efficacy of β-glucosidase covalently immobilized on alginate/polysorbate complex was 70.1%. The impact of pH, and thermal stability, on the free and immobilized β-glucosidase activities and hydrolysis of soybean extract were evaluated. The optimum pH remained at 5.0 after immobilization, while the optimum temperature leveled up to 50 °C in comparison to the free enzyme, which remained at 37 °C. This is the first report to our knowledge on the use of Penicillium sp. NRC24 for the production, characterization, and immobilization of β-glucosidase, which can hydrolyze genistein and daidzein into their aglycone forms.
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