RT info:eu-repo/semantics/article
T1 The role of polymeric chains as a protective environment for improving the stability and efficiency of fluorogenic peptide substrates
A1 Arnáiz Alonso, Ana
A1 Guembe García, Marta
A1 Delgado-Pinar, Estefanía
A1 Valente, Artur J. M.
A1 Ibeas Cortes, Saturnino
A1 García Pérez, José Miguel
A1 Vallejos Calzada, Saúl
K1 Actuators
K1 Sensors and biosensors
K1 Química orgánica
K1 Chemistry, Organic
AB We have faced the preparation of fully water-soluble fluorescent peptide substrate with long-term environmental stability (in solution more than 35 weeks) and, accordingly, with stable results in the use of this probe in determining the activity of enzymes. We have achieved this goal by preparing a co-polymer of the commercial N-vinyl-2-pyrrolidone (99.5% mol) and a fluorescent substrate for trypsin activity determination having a vinylic group (0.5%). The activity of trypsin has been measured in water solutions of this polymer over time, contrasted against the activity of both the commercial substrate Z-L-Arg-7-amido-4-methylcoumarin hydrochloride and its monomeric derivative, prepared ad-hoc. Initially, the activity of the sensory polymer was 74.53 ± 1.72 nmol/min/mg of enzyme, while that of the commercial substrate was 20.44 ± 0.65 nmol/min/mg of enzyme, the former maintained stable along weeks and the latter with a deep decay to zero in three weeks. The ‘protection’ effect exerted by the polymer chain has been studied by solvation studies by UV–Vis spectroscopy, steady-state & time resolved fluorescence, thermogravimetry and isothermal titration calorimetry.
PB Springer Nature
YR 2022
FD 2022-05
LK http://hdl.handle.net/10259/6685
UL http://hdl.handle.net/10259/6685
LA eng
NO "La Caixa" Foundation, under agreement LCF/PR/PR18/51130007. We also gratefully acknowledge the financial support provided by FEDER (Fondo Europeo de Desarrollo Regional) through Programa Operacional Factores de Competitividade (COMPETE), and the Spanish State Research Agency (PID2020-113264RB-I00/AEI/10.13039/501100011033). Centro de Química de Coimbra acknowledges Fundação para a Ciência e Tecnologia for financial support (Projects UIDB/00313/2020, UIDP/00313/2020). E.D.-P. thanks the “Concurso de Estímulo ao Emprego Científico” for the junior contract CEECIND/04136/2018.
DS Repositorio Institucional de la Universidad de Burgos
RD 24-nov-2024