Bioprospecção de enzimas microbianas a partir de bactérias isoladas do alimento larval de abelhas sem ferrão

Abstract

The search for more sustainable alternatives for industrial processes has become the focus of much research that aims to replace the use of chemical products, which pose risks to the environment and require large amounts of natural resources, with biodegradable inputs extracted from nature. From this perspective, microbial enzymes become an interesting option as they present greater efficiency and stability at extreme pHs and high temperatures, when used in industrial processes. The objective of the present work was to identify a possible biotechnological potential for the production of microbial amylase and cellulase with possible industrial applications, by evaluating 20 bacteria isolated from the larval food of stingless bees native to Brazil, belonging to the Collection of Microorganisms Isolated from Stingless Bees ( CoMISBee) from the Federal University of Uberlândia. First, enzyme-producing bacteria were screened using solid medium enriched with 1% soluble starch or carboxymethyl cellulose (CMC), and bacteria with the best enzymatic indexes (IE) were inoculated and incubated in liquid medium also enriched with the aforementioned substrates. to give rise to the crude enzyme extract. Subsequently, the crude enzyme extract was quantitatively evaluated for its ability to hydrolyze soluble starch and CMC using the DNS test (3,5-dinitrosalicylic acid). Ta19 and Ta35 bacteria showed higher IE values, with Ta19 bacteria producing enzymes capable of hydrolyzing soluble starch and Ta35 bacteria producing enzymes that degraded both soluble starch and CMC. The methodology used as the basis for this study was adapted to use a smaller amount of reagents and other laboratory supplies by replacing eppendorf with PCR microplates and the water bath with a thermocycler. The incubation time and temperature parameters for the enzymatic test were chosen according to the literature. It is concluded that further studies are needed to clarify the real potential of microbial enzymes when evaluating their activity using different variations in temperature, pH and incubation time.