2024-03-29T14:16:02Zhttps://riubu.ubu.es/oai/requestoai:riubu.ubu.es:10259/66852022-06-24T11:00:39Zcom_10259_4313com_10259_5086com_10259_2604col_10259_4314
The role of polymeric chains as a protective environment for improving the stability and efficiency of fluorogenic peptide substrates
Arnáiz Alonso, Ana
Guembe García, Marta
Delgado-Pinar, Estefanía
Valente, Artur J. M.
Ibeas Cortes, Saturnino
García Pérez, José Miguel
Vallejos Calzada, Saúl
Actuators
Sensors and biosensors
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.
2022-05-30T12:37:10Z
2022-05-30T12:37:10Z
2022-05
info:eu-repo/semantics/article
http://hdl.handle.net/10259/6685
10.1038/s41598-022-12848-4
2045-2322
eng
Scientific Reports. 2022, V. 12, 8818
https://doi.org/10.1038/s41598-022-12848-4
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-113264RB-I00/ES/DISEÑO, SINTESIS Y CARACTERIZACION DE POLIMEROS INTELIGENTES. APLICACION DE SENSORES QUIMICOS POLIMERICOS EN CONTROL Y SEGURIDAD ALIMENTARIA Y EN EL AMBITO SANITARIO
info:eu-repo/grantAgreement/FCT//UIDB%2F00313%2F2020
info:eu-repo/grantAgreement/FCT//UIDP%2F00313%2F2020
http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
Atribución 4.0 Internacional
Springer Nature