Purification and Characterization of Cellulase from Lactobacillus for Enhanced Essential Oil Extraction

The objective of the study is to isolate and identify lactic acid bacteria Lactobacillus from various local sources and to purify, screen, identify, and assess their efficiency in producing the Cellulase enzyme. The most effective isolates for producing the Cellulase enzyme were identified as fish isolate 3. Purification was accomplished through sequential ammonium sulfate precipitation (40-80%) and dialysis, achieving a specific activity of 59.51 U.mg-1. This was followed by gel filtration chromatography using a Superdex 200 column on an ÄKTA Pure 25 apparatus, resulting in a purification fold of 20.39 and a yield of 11.49%. The molecular weight of Cellulase was determined to be 36 kDa using sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. The optimum pH for both the purified and partially purified enzyme activity and stability was found to be 5. The optimum temperature for enzyme activity was 55°C for both the completely purified and partially purified enzymes. The temperature range for enzyme stability was between 25-45°C for the partially purified enzyme and between 25-55°C for the completely purified enzyme. Furthermore, the partially purified enzyme was utilized to enhance the efficiency of extracting essential oils from Star Anise (Illicium verum).