Isolation and characterization of lactic acid bacteria proteases from Bekasam for use as a beef tenderizer

Abstract

Background and Objective: Proteases are important enzymes and have high economic value due to their wide applications in the food industry as a meat tenderizer. Protease use in the food industry necessitates an understanding of the capabilities and influencing factors of these enzymes to accelerate enzymatic reactions. This study aimed to isolate and characterize the proteases of lactic acid bacteria (LAB) from Bekasam. Methodology: The samples were obtained from the third, fifth, seventh, ninth and eleventh day of fermentation to isolate the proteolytic LAB. Characterization of proteases includes the incubation time, casein substrate concentration, optimum temperature and pH, metal ion contents and stability. The LAB with the highest protease activity is identified molecularly and isolated through 16S rDNA sequencing and phylogenetic analysis based on the Neighbor Joining method. Results: The results showed that the best isolate was BAF-715 because it had the highest protease activity (18.84 U mL–1) at 40 h of incubation. The optimum activity of this protease on a casein substrate at 2.5% occurred at an incubation temperature of 40EC at pH 7 and in the presence of Mg2+ and Mn2+ (5 mM) as activators. Based on molecular DNA identification, the BAF-715 isolate is determined to be Pediococcus pentosaceus. Conclusion: A protease produced by Pediococcus pentosaceus showed the highest proteolytic activity, making it the best protease for application as a beef tenderizer.