Produção de Glicose Oxidase (E.C. 1.1.3.4) por fungos isolados da Floresta Amazônica

Abstract

Glucose oxidase (Gox) has several industrial applications. It is believed that there are diversified fungal species that have the production capacity of this enzyme and most are unexplored. The objective of this work was to investigate a production of Glucose Oxidase (EC 1.1.3.4) by fungi isolated from soil samples from the Amazon Forest. Soil fungi were isolated from the Adolpho Ducke Forest Reserve, located in the city of Manaus-Amazonas. To select good GOx producers, such as submitted in submerged bioprocess. The isolates highlighted in the production of the enzyme were submitted to kinetic tests where the enzyme production, biomass and substrate consumption by isolates were evaluated. Univariate and multivariate assays performed to optimize the production process. An enzymatic production for the semi-continuous process. The crude enzyme was purified by precipitation processes, dialysis ion exchange chromatography. The potential of this enzyme in the elaboration of a Glucose quantification kit through calibration curves, the regression coefficient, was investigated as biochemical characteristics and GOx (optimum temperature, optimum pH, stability and ionic effect in the enzymatic activity. However, only seven were identified as potential producers of this enzyme, being five of the genus Aspergillus and two of the genus Penicillium, with values ranging from 14.93 U/mL – 35.54 U/mL. Kinetics showed that the Aspergillus LMM01 isolate was the best producer of the enzyme, and this was used isolated in the subsequent experiments. Experiments with variants demonstrated with the greatest effect on the production of GOx were PH and agitation. A result for GOx (7.74 U/mL) was obtained, for 7 days, use the semi-continuous. After purification process, an activity and the enzyme improved 938%, a temperature of 40 ° C and pH 6.0 were the optimum for a catalytic action of the enzyme and stability presented as its main industrial needs. Fe+++, Pb++, Ag+, Hg++ and Cu++ ions showed significant inhibition of GOx activity. In the present study, a Pearson correlation of the developed Kit and the commercial Kit was strong positive, with value of r = 0.9978. The results are compatible with a literature where the genera Aspergillus and Penicillium stand out as the main industrial producers of GOx.