A TECHNIQUE TO IMMOBILIZE TRYPSIN ON STAINLESS STEEL IN ORDER TO PREVENT BIOFILMS FORMATION
LUCICA BARBES 1*, MICHELE SALMAIN 2, GABRIELA STANCIU 1 (1)OVIDIUS UNIVERSITY; FACULTY OF PHYSICS, CHEMISTRY, ELECTRONICS
AND PETROLEUM TECHNOLOGY ; CHEMISTRY DEPARTMENT; 124, MAMAIA BLVD, 900527, CONSTANTA, ROMANIA
(2)SUPERIOR NATIONAL SCHOOL OF CHEMISTRY, CHEMISTRY AND BIOCHEMISTRY OF MOLECULAR COMPL
The molecular mechanisms governing the formation of biofilms by the colonization of solid surfaces with living organisms such as bacteria, algae and eukaryotic cells, are poorly known, in spite of their importance in the medical devices, the food-processing industry and the marine equipments. The aim of this work was to explore different pathways to attach in a covalent manner commercial trypsin to stainless steel coupons to create a thin layer of active enzyme. Different reagents such as functionalized trialkoxysilanes compounds [γ-(aminopropyl)trietoxysilane] has been use to create a reactive molecular layer to attach proteins. The surfaces were characterized after each treatment by Fourier transform infrared reflection-absorption spectroscopy (FT-IRRAS technique) to determine the molecular composition of the films. Finally, the efficiency of the different ways immobilization technique have been assessed by enzymatic assays with selected substrate (N-benzoyl-L-arginine-ethyl-ester â BAEE), then on microbial cultures generating biofilms. The immobilized trypsin exhibited much higher relative activity than lysozyme in the similar conditions.
