Skopuleriopsis koningii enzyme prospection using proteome: Recombinant production of enzimes with biotecnological potential and evaluation of Antifungal

Abstract

The biotechnological industry has growing around the world, including Brazil. However, research and development national funding is under the satisfactory level. The production of enzymes is an important biotechnological branch given that represents high economic gains and is in expansion. Microorganisms are the most applied in enzyme production, given its low cust of manipulation and high yield of the process. Peptidases are the most produced enzymes and show a large range of applications. Considering the Skopulariopsis koningii enzyme prospection, we propose a proteomic study based on differential expression inducing processes. The organism will be induced in different medium composition. The induction will be verified with 1D and 2D electrophoresis gel. The selected proteins will be sequenced in mass spectrometer to identification. We have two aims: collagenolitic peptidases prospection and prospection of proteins related to antifungal response. We also intend to recombinantly produce these enzymes in the yeast Pichia pastoris, under methanol induction. After the purification, enzymatic characterizations will be done, as substrate specificity, kinetic parameters, temperature and pH influence and other relevant tests. The enzymes might show activity close to the endogens. We expect find collagenases with potential to application in pharmaceutical industry. Moreover, the description of metabolic pathway response to antifungal resistance can be enlightening and open a way to depth studies.