Microbial cells produce a variety of enzymes. These enzymes are the biological catalysts for the biochemical reactions, leading to microbial growth and respiration, as well as to produce fermentation products. Enzymes are used for a variety of purposes. They are employed in three major fields.
Amylase is an enzyme, which is produced by the microorganisms, which has many applications in today's market. Amylase is required in digestion of carbohydrates into smaller units and eventually converting them into even smaller units such as glucose. It is also involved in inflammatory reaction, such as those caused by the release of Histamine and similar substances. A number of digestive enzymes including amylase are required to produce Fructose in large quantities. Many detergents also contain enzymes such as Amylase to remove stains.
[...] Our aim was to collect the sample rich in Bacillus sp., for this soil sample was collected from the basement of our lab. Sample was collected by using sterile gloves, scalpel and placed in sterile container. SERIAL DILUTION AGAR-PLATING METHOD After collecting the samples serial dilution was carried out to isolate bacteria from soil. For this 10ml sterile distilled water was taken in a test tube. To this 1gm of finely pulverized, air dried soil was added. The tube was vigorously vortexed for 3 minutes to obtain uniform suspension of microorganisms. [...]
[...] The slides were rinsed with distilled water and decolouriser was slowly applied drop by drop from one end of the smear for 1 min. The slides were washed with distilled water and counter stained with saffranin for 30 seconds. The slides were rinsed in distilled water, air-dried and observed under oil immersion (100x) objective of the microscope. Pure culture: After identification, Bacillus subtilis was sub cultured on starch agar medium to get a pure culture. Approximately 15ml of molten Starch Agar media was poured into sterilized Petri plates and allowed to solidify. [...]
[...] The cellophane tapes were removed from the slides; white petroleum jelly is carefully removed with the help of blotting paper. The bulldog clips were removed µl of enzyme sample was mixed in 10 µl of tracking dye in an eppendofs tube µl of buffer was added to the above mixture. This mixture was incubated for 10 minutes at 600C in a water bath. After incubation the mixture was loaded into a well. The glass slide along with the sample in the gel was tightly clamped to the electrophoresis setup. [...]
[...] Inoculate the tube containing MRBP broth with E.coli Incubate the tubes for 35'C for 48 hrs Add 5-10 drops of Methyl Red indicator to each tube Observe the change in color of Methyl Red for Methyl Red test The Methyl Red indicator in the pH range of 4 will remain red, which is an indication of positive test while turning of Methyl Red to yellow is a negative test. RESULT: E.coli tubes showed a positive Methyl Red test. VOGES-PROSKAVER TEST AIM: To indicate the organism produces acetyl methyl carbinol. PROCEDURE: 1. Inoculate the test tubes containing MRVP Broth with E.coli 2. Incubate the tube at 35'C for 48 hrs Remove 2ml of the culture from the above tubes and place it in a test tube Add 0.6 ml of alpha napthol solution and 0.2 ml of [...]
[...] The organisms used for the commercial production of Amylase are bacteria such as Bacillus subtilis, some fungal species such as Aspergillus sp., Bacillus subtilis can be cultured by submerged state fermentation method. In case of submerged state fermentation liquid broth is used to provide nutrition to the growing bacteria. Amylase activity in a given sample can be confirmed by using DNS method in which DNS reacts with the reducing sugars to give red color. Amylase activity can be measured by measuring the O.D of the test solution. [...]
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