Efficacy Of Protease Research Articles

Evaluation of enzymatic degradation of ochratoxin A by protease and lipase produced by lactobacillus isolated from dairy products

Contamination with ochratoxin A (OTA) is widespread in many foods and it is necessary to look for modern and specialized methods to remove OTA. The current study aimed to produce protease and lipase from Levilactobacillus brevis and Lactobacillus plantarum, respectively, which were isolated from Egyptian soft cheese, and evaluate them on enzymatic degradation of OTA in phosphate buffered saline (PBS) after treatment with doses ranging from 50 to 200U/ml for 6, 2 and 18h at 37 °C. The results showed that 60–80 % saturation ammonium sulfate gave the highest protease activity (1295.95 U/ml) and specific activity (3570.11), while lipase recorded high-value activity (15608.2 U/ml) and Specific activity (40021.1) at 40-60% ammonium sulfate. Partially purified using Sephadex G -100 showed only one peak for protease and lipase at fraction 55 and 32, respectively. The results indicated that using protease and lipase at 200 U/ml gives the highest degradation for OTA after incubation for 18h were 82.6 and 77.3%, respectively. Enzymatic degradation of OTA indicated that the efficacy of protease and lipase for degradation of OTA increases with the duration of incubation and the dose of the enzyme.

Screening and optimization of candidate alkaline protease for dehairing potential from marine Bacillus paramycoides M2

Alkaline proteases are active from neutral to alkaline pH range and have extensive applications in detergent and leather industries. In the present research, bacteria isolated from marine water samples were screened for proteolytic activity. Among the isolates, M2 showed maximum proteolysis with a clear zone when cultured on skim milk agar plates at 37°C for 24 h. Molecular identification using 16S rRNA sequencing and phylogenetic analysis revealed that M2 has sequence identity (99.93%) to Bacillus paramycoides. SEM analysis was carried for determining the morphology of M2 and also for enzyme treated skin. FAME analysis using GCMS was performed for the determination of fatty acids in the strain. The selected isolate was inoculated into protease production medium under submerged fermentation conditions at 37ºC for 48 h with a constant agitation of 120 rpm. Protease activity was determined under varying conditions of pH, incubation temperature, carbon and nitrogen sources, metal ions and NaCl (1- 5%) using casein as substrate. The isolate M2 utilized molasses and peptone as carbon and nitrogen sources for better alkaline protease production at 40°C and pH 10 under optimal conditions. The dehairing experiments with M2 alkaline protease revealed dehairing efficacy of protease over chemical treatment. Hence, extracellular alkaline protease from M2 isolate could find potential application in leather processing industries and can be exploited commercially.

Evaluation of Ethylenediaminetetraacetic Acid, Tetrasodium Salt Dihydrate (EDTA)-Tween 20 Treatment versus Protease Digestion of Formalin-fixed Tissue Sections for Detection of Bovine Respiratory Syncytial Virus Antigen in Infected Ovine Lung

The efficacy of protease and ethylenediaminetetraacetic acid, tetrasodium salt dihydrate (EDTA)-Tween 20 in unmasking bovine respiratory syncytial virus (BRSV) antigens in formalin-fixed lung tissue was compared using avidin-biotin immunoperoxidase procedure. Tissues were taken from experimentally infected lambs. BRSV antigen stained in both techniques. Treatment with EDTA-Tween 20 resulted in more intense staining of BRSV infected cells, more uniform cytoplasmic staining, less non-specific background, and superior cellular detail in comparison to protease digestion.