Purification and Characterization of Class III Lipase from a White-Rot Fungus Pleurotus ostreatus.

Seiji Nakagame, Hu Minagawa, Nagi Motegi

Applied biochemistry and biotechnology 2023 Feb

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Pleurotus ostreatus is an edible white-rot fungus with lignocellulosic biomass degrading enzymes that have been studied extensively. However, until now, lipolytic enzymes from P. ostreatus, which degrade extractives in lignocellulosic biomass, have not been purified and characterized. In this study, P. ostreatus was inoculated into the rapeseed oil containing culture to induce lipase. The lipase in the culture broth was successfully purified to homogeneity by chromatographic methods. The molecular weight of the purified lipase was 27 kDa, and its optimal pH and temperature were 5.0 and 30 °C, respectively. The purified lipase showed high activity with the substrates 4-methylumbelliferyl (4-MU) decanoate (C10:0) and 4-MU oleate (C18:1), and no activity with 4-MU acetate (C2:0) and 4-MU butyrate (C4:0). The amino acid sequences and substrate specificities of the purified lipase suggested that it belonged to class III. Kinetic parameters measurements (Km and Vmax) showed that 4-MU palmitate had a high affinity for the purified lipase, and it was the substrate most efficiently hydrolyzed by the purified lipase. © 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Seiji Nakagame, Hu Minagawa, Nagi Motegi. Purification and Characterization of Class III Lipase from a White-Rot Fungus Pleurotus ostreatus. Applied biochemistry and biotechnology. 2023 Feb;195(2):1085-1095