Purification and characterization of α-galactosidases from Penicillium griseoroseum for efficient soymilk hydrolysis.

Autor: Falkoski DL; Department of Biochemistry and Molecular Biology, Federal University of Viçosa, Viçosa, MG, 36570-000, Brazil; Department of Natural Science, Federal University of São João del-Rei, São João del-Rei, MG, 36301-160, Brazil., de Rezende ST; Department of Biochemistry and Molecular Biology, Federal University of Viçosa, Viçosa, MG, 36570-000, Brazil., Guimarães VM; Department of Biochemistry and Molecular Biology, Federal University of Viçosa, Viçosa, MG, 36570-000, Brazil., de Queiroz MV; Department of Biochemistry and Molecular Biology, Federal University of Viçosa, Viçosa, MG, 36570-000, Brazil., Almeida MN; Department of Natural Science, Federal University of São João del-Rei, São João del-Rei, MG, 36301-160, Brazil. Electronic address: malmeida@ufsj.edu.br.
Jazyk: angličtina
Zdroj: Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2024 Dec 10; Vol. 737, pp. 150905. Date of Electronic Publication: 2024 Oct 24.
DOI: 10.1016/j.bbrc.2024.150905
Abstrakt: Soybean utilization is limited by the presence of raffinose oligosaccharides (RFO), which are not digested by humans and cause gastrointestinal discomfort. This study explores the potential of α-galactosidases from Penicillium griseoroseum for RFO hydrolysis in soymilk. Two distinct α-galactosidase enzymes, designated α-Gal1 and α-Gal2, were purified using a combination of ion-exchange chromatography and native polyacrylamide gel electrophoresis. Both enzymes exhibited characteristics of multimeric proteins and displayed similar biochemical properties. Optimal activity was observed at a pH range of 4.5-5.0 and a temperature range of 40-45 °C. Notably, α-Gal1 demonstrated high thermostability with a half-life of 16 h at 40 °C. The α-galactosidases displayed different substrate affinitiesfor the substrates ρ-NP-αGal, o-NP-αGal, rD-raffinose, d-stachyose, and mD-melibiose. The Michaelis-Menten constant (Km) values for α-Gal1 were 1.06, 1.31, 28.74, 19.88, and 4.77 mmol/L, respectively, while those for α-Gal2 were 0.8, 1.26, 30.46, 21.74 and 5.01 mmol/L, respectively. Both α-Gal1 and α-Gal2 were strongly inhibited by metal ions (Ag⁺, Cu 2 ⁺, Fe 2 ⁺, and Hg 2 ⁺) and moderately inhibited by d-melibiose. Importantly, both enzymes efficiently hydrolyzed RFOs, achieving complete d-stachyose elimination from soymilk after a 6-h incubation. These findings propose the promising application of these α-galactosidases in industrial soymilk production, potentially enhancing its nutritional value and alleviating gastrointestinal issues in consumers.
Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Almeida, M. N. reports equipment, drugs, or supplies was provided by Federal University of São João del-Rei. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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