Effect of Organic Solvents on the Activity, Stability and Secondary Structure of asclepain cI, Using FTIR and Molecular Dynamics Simulations.

Autor: Origone AL; Laboratory of Bromatology, Faculty of Chemistry, Biochemistry and Pharmacy (FQByF), National University of San Luis, Chacabuco N° 917, San Luis, Argentina.; Institute of Applied Physics (INFAP) - Technological Scientific Center of San Luis - National Council of Scientific and Technique Research (CONICET), Ejército de los Andes N° 950, Block II, 2nd Floor, 5700, San Luis, Argentina., Hissi EGV; Physical-Chemistry Area, FQByF, National University of San Luis, Ejército de los Andes 950, 5700, San Luis, Argentina. egvega@gmail.com.; Multidisciplinary Institute of Biological Research (IMIBIO) - CONICET, Ejército de los Andes N° 950, 5700, San Luis, Argentina. egvega@gmail.com., Liggieri CS; Plant Protein Research Center (CIProVe), National University of La Plata, Calle 47 y 115, La Plata, Argentina., Camí GE; Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario, Suipacha N° 531, Rosario, Santa Fe, Argentina.; Faculty of Engineering and Chemistry, Av. Pellegrini N° 3314, Rosario, Santa Fe, Argentina., Illanes A; School of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso, Avenida Brasil N° 2085, Valparaiso, Chile., Barberis SE; Laboratory of Bromatology, Faculty of Chemistry, Biochemistry and Pharmacy (FQByF), National University of San Luis, Chacabuco N° 917, San Luis, Argentina. soniaebarberis@gmail.com.; Institute of Applied Physics (INFAP) - Technological Scientific Center of San Luis - National Council of Scientific and Technique Research (CONICET), Ejército de los Andes N° 950, Block II, 2nd Floor, 5700, San Luis, Argentina. soniaebarberis@gmail.com.
Jazyk: angličtina
Zdroj: The protein journal [Protein J] 2024 Jun; Vol. 43 (3), pp. 487-502. Date of Electronic Publication: 2024 Mar 07.
DOI: 10.1007/s10930-024-10182-4
Abstrakt: The present study aims at understanding the effect of organic solvents on the specific proteolytic activity and operational stability of asclepain cI in aqueous-organic media, using correlations between geometrical and structural parameters of asclepain cI. These correlations were determined by molecular dynamics (MD) simulations and the secondary structure of the enzyme validated by Fourier-transform Infrared (FTIR) spectroscopy. Asclepain cI exhibited significantly higher catalytic potential in 29 of the 42 aqueous-organic media tested, composed by 0.1 mM TRIS hydrochloride buffer pH 8 (TCB) and an organic solvent, than in buffer alone. Asclepain cI in water-organic miscible systems showed high FTIR spectral similarity with that obtained in TCB, while in immiscible systems the enzyme acquired different secondary structures than in buffer. Among the conditions studied, asclepain cI showed the highest catalytic potential in 50% v/v ethyl acetate in TCB. According to MD simulations, that medium elicited solvation and flexibility changes around the active center of asclepain cI and conducted to a new secondary structure with the active center preserved. These results provide valuable insights into the elucidation of the molecular mechanism of asclepain cI tolerance to organic solvents and pave the way for its future application for the synthesis of peptides in aqueous-organic media.
(© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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