Papain-Catalyzed Hydrolysis of N_α-Benzoyl–arginine–p-nitroanilide in an Aqueous–Organic Medium

ACS Omega, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 23, 2025

Enzyme-based biosensors have emerged as an effective alternative, providing simplicity, high sensitivity, and the capability to detect multiple residues. However, despite their widespread use, limited studies examined how organic solvents inhibit these sensors. This study investigates enzymatic reactions structure of selected model enzyme, papain, a protease derived from Carica papaya, in presence various solvents. Enzyme activity was monitored through hydrolysis Nα-benzoyl-arginine-p-nitroanilide (BAPNA), with resulting yellow product, p-nitroaniline, measured at wavelength 430 nm. The experiments incorporated 10% (v/v) concentration dimethyl sulfoxide (DMSO) ensure solubility BAPNA. Results showed that methanol ethanol increased K m value while causing little change V max, which negatively impacted enzyme's catalytic efficiency. In contrast, acetonitrile (ACN) behaved reversible mixed-competitive inhibitor exhibiting lower millimolar IC50 values. Furthermore, emission maximum shift wavelengths increasing concentrations ACN suggested tryptophan residues within enzyme were slightly more buried. Molecular dynamics simulations BAPNA-papain complex cosolvent environments containing water, DMSO, indicated could act alongside BAPNA solvent polarity influence binding papain. These findings provide valuable insights for application biosensor technologies.

Language: Английский

Selective Acetylation of Unprotected Thioglycosides and Fully Unprotected Monosaccharides with Candida antarctica Lipase-B

ACS Omega, Journal Year: 2025, Volume and Issue: unknown

Published: May 7, 2025

Language: Английский