Production of Partially Purified Collagenase from Bacillus licheniformis and It’s Use to Tenderize Aged Buffalo Meat

Abstract

This study aimed to isolate and identify the Bacillus licheniformis bacteria that produces the enzyme collagenase. Due to the selective nature of this enzyme in breaking down and degrading collagen, it was used to tenderize aged buffalo meat. The bacterial isolates were screened using a selective medium and then grown on collagen-agar medium to identify enzyme-producing isolates. Thirteen enzyme-producing isolates were obtained based on the formation of a transparent corona around the colony. Biochemical assessments of the bacterial isolates were carried out using the ABIS online program. The isolates BL1–BL9, BL12 and BL13 were found to be related to B. licheniformis with a percentage match of 98%, while BL10 and BL11 were related to Bacillus pumilus and Bacillus subtilis with a percentage match of 95% and 95.3%, respectively. The enzyme activity of the isolates varied, with the highest activity reaching 200.71 units.ml-1 for isolate BL12. The enzyme of this isolate was chosen to complete the other tests. The optimal time required to obtain the highest enzymatic activity was found to be 20 h, which amounted to 210.25 units.ml-1, with a specific activity of 135.64 units.ml-1. The enzyme was partially purified using ammonium sulphate at a saturation rate of 20–70% as it showed an enzymatic activity and specificity of 355.81 units.ml-1 and a specific activity of 423.58 units.ml-1. Partially purified collagenase was used to tenderize aged buffalo meat. The results showed that treating aged buffalo meat with the enzyme solution for different immersion durations improved the pH, water-holding capacity (WHC) and cooking yield, with an increase in the concentration of hydroxyproline and a significant percentage of soluble collagen; the treated samples showed the best properties with a 60-min immersion duration.

Keywords

• Bacillus
• Biochemical test
• Buffalo meat
• Collagenase
• Tenderization