After completion of the reaction, the mixture was poured into crushed ice. The aqueous mixture was extracted (3 × 10 mL) EtOAc The combined organic extracts were dried (MgSO4) and the solvent was evaporated to give the desired product. (5-nitroiminotetrazole-1-yl)-acetic acid: yield: 0.28 g (57%); white crystals; m.p 160-163°C; IR (KBr): 3544, 3467, 3016, 2968, 1736, 1634, 1582, 1486,1338, 1308, 1224 cm-1; 13C NMR (75 MHz (DMSO-d6)): 167, 151,
Gelatine hydrolysis test The tubes containing sterilized gelatine liquefaction medium was stab inoculated with fresh culture of bacterial isolates and incubated at 37±1o C for 24 h. After incubation the tubes were placed in refrigerator at 40 c for 30 min. Then these tubes were kept in ice cubes containing beaker and observed for Gelatine liquefaction by the bacterial isolates. The gelatine liquefied tubes were considered as positive reaction (Aneja, 2001). Composition of Gelatine liquefaction medium Peptone 5g Beef extract 3g Sodium chloride 5g Gelatine 120g Agar 20g Distilled water 1000ml PH 7 IV. Casein hydrolysis test The sterilized plates containing skim milk agar medium were point inoculated with fresh bacterial cultures and incubated
We started of by adding a small piece of liver to an empty test tube and placing just enough water to cover it. After this we placed it in a hot bath for five minutes and after letting it cool we tested to see its reaction rate with hydrogen peroxide, it received a one because of the very little reaction occurring. Then we got four more empty test tubes and filled two of them with hydrogen peroxide and two of them with equal amounts of liver. Then we placed a test tube with hydrogen and another test tube with liver into an ice bath and placed the remaining into a warm bath for three minutes. After this we tested the reaction rates by pouring the respective hydrogen peroxide and live (warm with warm and cold with cold).
Then, 3.7 µl of 10 mM Fluorescein Maleimide was added in equal quantities in wild type, mutant and purified ERAB protein, incubated at RT for 10 minutes. Similarly, 4.1 µl of 100 mM DDT was added in the three proteins respectively, incubated for 20 minutes again. And 3 µl of unmodified wt protein was transferred into a 1.5 ml tube as an additional sample. Then, 10.3 µl of 5 x SDS sample buffer was added to each sample and boiled for 5 minutes at 95 oC. Samples were cooled on ice for 1 min, spin down and now they were ready to load on the
actococcus lactis ssp. lactis C2 and bacteriophages c2, ml3, sk1 were obtained from T. R. Klaenhammer (North Carolina State University, Raleigh, NC). Both bacterial Lactococcus lactis ssp. lactis C2 stains and bacteriophages c2, ml3, sk1 were stored in microcentrifuge tubes at -80℃ refrigerator to keep frozen until use. 3.3 Preparation of M17 medium, skim milk, Bottom agar, Top agar and CaCl2 solution M17 medium, skim milk, Bottom agar, Top agar and CaCl2 solution were all prepared in lab 201 in Garrugus building (University of Kentucky).
The P2 tube was then placed in the refrigerated centrifuge at a speed of 15,000g for 30-60 minutes at 4°C. 1ml of the homogenisation buffer was added to the P1 pellet and was vortexed to resuspend it. The supernatant was then removed from the P2 tube and placed into a micro tube labelled ‘S’. 1 ml of the homogenisation buffer was added to the P2 pellet and placed on ice. The pellet was then resuspended again by adding small quantity of glass beads and it was vortexed vigorously until the pellet has disappeared from the bottom of the
Clear serum samples were separated by centrifugation at 3000 r.p.m. for 20 min. and then transferred into Eppendorf tubes and stored in a deep freeze (-20oC) for biochemical analysis. However determination of glucose was carried out on fresh serum samples. Determination of Biochemical parameter Serum glucose, total protein, albumin, globulin, urea, creatinine and uric acid were analyzed by Kone lab 60 Auto analyzer in Al Shifa Hospital Clinical Chemistry Laboratory.
ISOLATION, IDENTIFICATION OF STREPTOKINASE PRODUCING STREPTOCOCCUS SPECIES AND PRODUCTION OF STREPTOKINASE ABSTRACT: The objective of the study is to identify a potent streptococcal species producing streptokinase enzyme from different samples. Various food samples and soil samples were collected and analyzed for the potent streptococcal strain. They were confirmed for streptococcus species by biochemical characterization. Further, they were screened for the streptokinase activity. Among them curd sample and bore soil sample showed good activity and they were taken for further analysis and production process.
In manufacturing industries, lubricants are being used extensively as it plays important roles in the machining process. The function of lubricants in machining processes acts as a coolant where it lubricates the tool-workpiece interface to reduce friction at low cutting speed or high cutting speed. Furthermore, lubricant consequently helps to prolong the machine tool life. Thus, to ascertain the suitability of vegetable oils as lubricants, the properties of vegetable oils will be analysed. 2.1 EXCELLENT LUBRICITY Vegetable oils are divided into two broad chemical categories which are triesters and monoesters (Biresaw et al., 2003).
This medium was centrifuged in a cooling centrifuge 5,000 rpm at 4°C to remove bacterial cell and residual waste. The protein content and activity of the enzyme solution were measured before salting-out with ammonium sulphate. The whole enzyme solution was then kept for about 20 minutes in an ice bath. Fractional precipitation began after adding solid ammonium sulphate slowly with stirring the ice-cold enzyme solution until the desired saturation of ammonium sulphate was reached. The solution was left over night at refrigerator and rotated for 15 minutes at 5,000rpm in a refrigerated centrifuge.