Enzyme nanoparticles as bioactive compounds for biotechnological applications

GABRIELA GARRIDO PEROSSI; GUSTAVO H.C.VARCA; MARIANO GRASSELLI; MONICA BEATRIZ MATHOR; PATRICIA SANTOS LOPES; ADEMAR B.LUGÃO

Campos do Jordao

Encuentro; XII Encuentro de la Sociedad Brasilera de Pesquisas em Materiais; 2013

Sociedad Brasilera de Pesquisas em Materiais

Nanotechnology has allowed the development of nanostructured materials with distinct properties. Regarding enzymes, such systems seek the development of particles with preserved conformation and bioactivity [1]. In this work we report the development of bioactive enzyme nanoparticles, by the use of radiation induced protein crosslinking [2], for biotechnological applications. Papain, a cysteine protease, widely applied in textile, cosmetic and biomedical industries was used as a model enzyme. Papain was diluted in phosphate buffer in presence of ethanol and submitted to ɣ-irradiation (10 kGy) on a gammacell. The structural rearrangement led to the formation of particles ranging from 5-11nm as determined by Dynamic Light Scattering. Biological activity was determined using Nα-Benzoyl-DLarginine p-nitroanilide hydrochloride as substrate at 40oC (pH 7.0) and conformational changes were monitored by fluorescence and UV 4th derivate spectra. The absence of new peaks in the spectra indicated preserved conformation. Bioactivity measurements over time confirmed that the particles were biologically active after the process and less likely to undergo degradation processes. Applications of the developed nanoparticles are directed towards their use as stable papain particles for biotechnological purposes. Further experiments shall be carried out to evaluate the cytotoxic and allergenic potential of the system.