BIOSYNTHESIS OF INDUSTRIAL ENZYMES BY FREE AND IMMOBILIZED Alicyclobacillus spp IN DIFFERENT MATRICES AND THE USE OF ULTRAFILTRATION IN THE ENZYMES CONCENTRATION

Abstract

The biosynthesis of amylase and collagenase, produced by A. acidocaldarius and A. sendaiensis respectively, were studied, and different matrices evaluated for the microorganisms immobilization and enzymes production optimization, such as loofa sponge, alginate-sponge and alginate, followed by concentration through ultrafiltration. Using a wheat bran substrate, the amylase enzyme displayed enzymatic activity of 0.45 U mL-1 and optimum temperature and pH conditions of 75 °C and pH 3.0, respectively. Thermal stability was in the range of 55 to 60 °C. The apparent Km and Vmax were 3.2 mg mL-1 and 0.5 U mL-1, respectively. The production of collagenase by A. sendaiensis was carried out with potato dextrose broth substrate and the activity obtained was 7.2 U mL-1. For amylase, the best results were obtained from immobilization in loofa sponge and the use of ultrafiltration (0.67 U mL-1) and for the collagenase extract, from the free biomass and ultrafiltration (13.6 U mL-1). The use of an ultrafiltration system enabled an average increase of 54% in the activity of both enzymes. Therefore, Alicyclobacillus are capable of producing enzymes of industrial interest, with the possibility of economically viable application of the substrate, and the use of immobilization and ultrafiltration produced positive results.

Keywords

• Alicyclobacillus
• amylase
• cell immobilization
• collagenase
• ultrafiltration