Differential scanning calorimetry (DSC) was used as an efficient and rapid tool in studying the conformational transitions between the folded and unfolded structures of cellulolytic enzymes. The thermal properties of two crude hydrolytic enzyme cocktails containing extracellular cellulases from Trichoderma longibrachiatum DIBAF-10 were analyzed and compared with three commercial cellulase preparations. Differences in the thermal behavior of fungal cellulases in the liquid phase, freeze-dried state, liquid formulations in sodium citrate buffer (pH 4.8), and contact with cellulose, carboxymethyl cellulose, and cellobiose were evaluated. DSC profiles of cellulases from the DIBAF-10 strain provided important thermodynamic information about the thermal stability of the included proteins. Crude enzyme cocktails underwent a reproducible and irreversible exothermic aggregation phenomenon at 52.45 +/- 0.90 degrees C like commercial beta-glucosidase. Freeze-dried and resuspended in a sodium citrate buffer, cellulases from T. longibrachiatum showed an endothermic peak dependent on buffer and enzyme concentration. In the enzyme-substrates systems, a shift of the same peak was recorded for all substrates tested. The thermal analysis of freeze-dried cellulase samples in the range of 20-150 degrees C gave information on the denaturation process. In conclusion, we demonstrated that DSC is a cost-effective tool for obtaining "conformational fingerprinting " of crude fungal cellulase preparations.[Graphics]

Differential scanning calorimetry (DSC) as a tool for studying thermal properties of a crude cellulase cocktail / Di Matteo, Paola; Luziatelli, Francesca; Bortolami, Martina; Mele, MARIA LUISA; Ruzzi, Maurizio; Russo, Paola. - In: CHEMICAL PAPERS. - ISSN 1336-9075. - (2023). [10.1007/s11696-022-02658-3]

Differential scanning calorimetry (DSC) as a tool for studying thermal properties of a crude cellulase cocktail

Paola Di Matteo
Primo
;
Martina Bortolami;Maria Luisa Mele;Paola Russo
Ultimo
2023

Abstract

Differential scanning calorimetry (DSC) was used as an efficient and rapid tool in studying the conformational transitions between the folded and unfolded structures of cellulolytic enzymes. The thermal properties of two crude hydrolytic enzyme cocktails containing extracellular cellulases from Trichoderma longibrachiatum DIBAF-10 were analyzed and compared with three commercial cellulase preparations. Differences in the thermal behavior of fungal cellulases in the liquid phase, freeze-dried state, liquid formulations in sodium citrate buffer (pH 4.8), and contact with cellulose, carboxymethyl cellulose, and cellobiose were evaluated. DSC profiles of cellulases from the DIBAF-10 strain provided important thermodynamic information about the thermal stability of the included proteins. Crude enzyme cocktails underwent a reproducible and irreversible exothermic aggregation phenomenon at 52.45 +/- 0.90 degrees C like commercial beta-glucosidase. Freeze-dried and resuspended in a sodium citrate buffer, cellulases from T. longibrachiatum showed an endothermic peak dependent on buffer and enzyme concentration. In the enzyme-substrates systems, a shift of the same peak was recorded for all substrates tested. The thermal analysis of freeze-dried cellulase samples in the range of 20-150 degrees C gave information on the denaturation process. In conclusion, we demonstrated that DSC is a cost-effective tool for obtaining "conformational fingerprinting " of crude fungal cellulase preparations.[Graphics]
2023
differential scanning calorimetry; cellulase activity; cellulase stability; trichoderma longibrachiatum; thermal stability
01 Pubblicazione su rivista::01a Articolo in rivista
Differential scanning calorimetry (DSC) as a tool for studying thermal properties of a crude cellulase cocktail / Di Matteo, Paola; Luziatelli, Francesca; Bortolami, Martina; Mele, MARIA LUISA; Ruzzi, Maurizio; Russo, Paola. - In: CHEMICAL PAPERS. - ISSN 1336-9075. - (2023). [10.1007/s11696-022-02658-3]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1670805
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