Banca de DEFESA: ISABELE ALBUQUERQUE ALCOFORADO FERREIRA

Uma banca de DEFESA de DOUTORADO foi cadastrada pelo programa.
STUDENT : ISABELE ALBUQUERQUE ALCOFORADO FERREIRA
DATE: 30/08/2022
TIME: 10:00
LOCAL: Remoto (Online)
TITLE:

COLLAGENASE PRODUCTION BY Rhizopus microsporus UCP 1296 AND ITS APPLICATION IN THE OBTAINING OF COLLAGEN PEPTIDES WITH BIOLOGICAL ACTIVITY

KEY WORDS:

Rhizopus microsporus, Collagenolytic Enzyme, Collagen, ATPS.

PAGES: 97
BIG AREA: Ciências Agrárias
AREA: Medicina Veterinária
SUMMARY:

The search for new enzymes is a constant challenge, mainly due to the need to develop more sustainable and financially viable production conditions. With the advancement of biotechnology, research aimed at the discovery of microbial enzymes has been developed mainly due to the favorable conditions of production in relation to other organisms. Enzymes that have high specificity and that can be used in small amounts are extremely interesting from a biotechnological point of view. Filamentous fungi have stood out in terms of the production of enzymes of industrial interest, especially collagenases, which are specific enzymes capable of degrading the triple helix of native or denatured collagen. In this context, a strain of the filamentous fungus Rhizopus microsporus (UCP1296) isolated from the soil of the Caatinga, an exclusively Brazilian biome, was selected for collagenase production. In this work, we used the submerged fermentation system (FS) to obtain the crude extract and aqueous two-phase system (ATPS) as a purification strategy. We used the gelatine culture medium as a source of carbon and nitrogen. The production of collagenolytic enzyme reached a peak after 120 hours of fermentation with 550 U/mL of collagenolytic enzyme, biomass of 0.42 g/L and specific collagenolytic activity of 808.23 U/mg. A factorial design was applied and as a result we obtained a 32% increase in enzyme production, equivalent to 727.50 U/mL of collagenolytic activity. Purification by ATPS was efficient for collagenase produced by R. microsporus UCP 1296. The highest values of yield and partition coefficient were obtained in the factorial design with PEG 8000 g/mol at 12.5% (m /m) of concentration, pH = 8 and phosphate concentration at 10.0% (m/m). The parameters, optimum pH and temperature, as well as the influence of inhibitors were determined for the characterization of the purified enzyme. In this context, the optimum pH was 8.0 and the optimum temperature was 37 °C. Regarding inhibitors, the enzyme showed partial inhibition towards ethylenediaminetetraacetic acid (EDTA) and iodoacetic acid (IAA), thus, it may have portions of metallo and cysteine proteases. The results suggest that the enzyme produced presents itself as a promising biotechnological product with applicability in several areas.

BANKING MEMBERS:
Presidente - 384972 - ANA LUCIA FIGUEIREDO PORTO
Interna - 1508562 - KEILA APARECIDA MOREIRA
Externa à Instituição - JUANIZE MATIAS DA SILVA BATISTA - UFRPE
Externo à Instituição - ROMERO MARCOS PEDROSA BRANDÃO COSTA
Externo à Instituição - THIAGO PAJEÚ NASCIMENTO - UFPI