Antibiotic resistance as become one of the largest worldwide anxieties. īacterial diseases become a serious problem in public health. Bacillus species were specific producers of extracellular protease. Among the different sources of proteases, bacterial protease was the most significant compared with animal, fungi, and plant protease. Proteases are produced in animals, plants, bacteria, archaea, and viruses. These enzymes are of wide distribution, and they perform significant biological processes. Proteolysis considers one of the most important biological reactions. The potential Bacillus proteases can promote new industry as antimicrobial agents. All pathogenic bacteria were totally inhibited with 10 μl of crude enzyme. It was found high effect on inhibiting the growth of pathogenic bacteria using 5 μl of crude enzyme with specific enzyme activity 73, 76, and 92 (units/ml)/(mg protein/ml) for B. However, the maximum bacterial growth and enzyme activity for B. subtilis were reached to the maximum at 5% NaCl. In addition, it was found that the enzyme activity and bacterial growth of B. licheniformis gave the highest growth and activity at neutral pH. cereus strains were increased at alkaline medium. ![]() The enzyme activity and bacterial growth of B. cereus and its activity gradually increased up to 45☌. licheniformis up to 37☌ then decreased at 45☌. It was found a considerable increase in the level of both of protease activity (units/ml) and bacterial growth (colony-forming units/ml) of the cultures that were directed by the B. subtilis showed resistance to Aztreonam only. licheniformis recorded high sensitivity (71%) against most antibiotics, in addition, B. The isolates were identified as Bacillus subtilis, Bacillus licheniformis, and Bacillus cereus. The sensitivity of the tested strains Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae against the extracellular crude protease enzyme produced from the three isolated bacteria and different antibiotics was done. The genetic identification was done using 16S rRNA gene sequencing. API biochemical kit was used to characterize the three isolates using some selective media. Skim milk nutrient agar medium was used for protease screening for several colonies by streaking method at 37☌. Three samples were collected from extreme environment, Jazan, KSA. The bottom test tube shows a gelatin medium after chilling that has a semi-solid state indicating that gelatin hydrolysis did not occur (gelatin hydrolysis negative).Biochemical and molecular characterization of proteolytic bacterial strains isolated from Jazan region, KSA with the application as an antibacterial agent. The top test tube shows a gelatin medium after chilling that has a more liquid form indicating that gelatin hydrolysis occurred (gelatin hydrolysis positive). Those species that do not hydrolyze gelatin will not have an effect on the gelatin and it will maintain its semi-solid state after chilling.įigure 1: Results from two gelatin hydrolysis tests. After incubation of the bacteria in the medium that hydrolyze gelatin, the medium will no longer form a semi-solid state, even after chilling. When bacteria that have this enzyme are inoculated into a nutrient medium containing gelatin, they will produce a gelatinase enzyme and break down the gelatin. To determine if a bacterial species hydrolyzes gelatin (and therefore has the a gelatinase gene and produces a gelatinase enzyme), the gelatin hydrolysis test is used. As such, the gelatin hydrolysis test is used to differentiate between bacteria that do and do not hydrolyze gelatin, and therefore is useful for characterizing and identifying bacterial species. Only those species of bacteria that do have a gelatinase gene are capable of producing a gelatinase and are capable of breaking down gelatin and using it as a nutrient source. ![]() Not all species of bacteria have a gene for a gelatinase enzyme. Therefore, it must first be broken down ( gelatin hydrolysis) by exoenzyme proteins called gelatinases. Gelatin provides a rich source of amino acids and peptides for bacteria, but its structure is too large to be transported inside the cell directly. When chilled on ice, gelatin forms cross-links to itself to create a semi-solid state (gelatin makes Jello! have its unique form and texture). Gelatin is a protein derived from collagen, a connective tissue of animals.
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