The purpose of continuous enzyme assay to study B-galactosidase is to determine the rate of enzyme action (B-galactosidase) at different substrate concentrations of o-Nitrophenylgalactoside (ONPG). The experiment is conducted by adding of the enzyme into the substrate preparation quickly and measure the change in absorbance of the reactants as a function of time. By plotting the
Agitation rpm level tannase production Agitation rate was at a range of 50 to 250 rpm level was chosen to determine the optimal rate. It was found that an agitation rate of 100 rpm at pH 6.0 and 37°C maximum yield of tannase production of 3.12 U/ml (Fig.4) it was also noted that an increase in agitation speed above100 rpm resulted in a drastic fall in tannase enzyme production. The agitation speed below 100 rpm level resulted in an inadequate mixing of the broth towards of the broth. Towards the later stages of growth. Effect of carbon and nitrogen sources In the present study, addition of sugars as a potential carbon source did not have any positive effect on extracellular tannase enzyme production.
The active fractions were pooled and extensively dialyzed against distilled water. • The active elutes with highest peaks were tested for antimicrobial activity and stored at -20°C until required for further use and considered to be purified protein. (Experiment was out sourced from Biotech Company and help of Dr Rakesh is duly acknowledged.) 3.9.4 Estimation of protein content The protein concentrations of crude, ammonium sulfate precipitate and purified extract were determined by the Folin-Lowry assay (Lowry et al., 1951) using Bovine serum albumin (2 mg/ml)) as a standard. The absorbance at 540 nm was used to estimate protein content in aqueous, saturated fractions and purified extract.
Catalase reacts with hydrogen peroxide, binding onto it and breaking it down into the less toxic water and oxygen. The equation for this reaction is the following: 2 H2O2 = 2 H20 + 2 O2 This experiment will use 1% catalase solution and 3% hydrogen peroxide solution, both diluted into water so the reaction slows down. Temperature will be controlled in this experiment to change the reaction speed of the enzyme and the substrate, this is what the experiment is looking at. The effect of the temperature will be determined by how much gas is released in two minutes, which will change the pressure inside the test tube and will be measured by a gas
To represent the blood of a person with ketoacidosis, we used a liquid that had a high pH level. Using the Lugol’s Iodine test, we investigated the effect of an environment with a high pH on the function of enzymes. In our experiment we added 15 drops of starch (amylose), 15 drops of an enzyme that breaks up starch (amylase), and 15 drops of the high pH liquid into a test tube. Next, we added 15 drops of the Lugol’s Iodine solution into the same test tube. We measured how long it took for the high pH test tube to change color versus how long our control took.
To calculate RMSd we used PTRAJ from the AMBER package and then we compared the conformation of the enzyme after the simulations at the 0.1 ps interval.  2.8.2. The Hydrogen Bond Analysis Hydrogen bonds formed between residues of the protease and between residues and water molecules, were analyzed using PTRAJ. Only bonds with a distance less than 3.5 Å with the angle of interaction greater than 140 ° were considered. The output file gave us the results which hydrogen bonds were formed, with which occupancy, distance and angle.
3.Mechanism of threonine protease Threonine proteases use the secondary alcohol of their N-terminal threonine as a nucleophile to perform catalysis. ( Brannigan,etal1995) The threonine must be N-terminal since the terminal amine of the same residue acts as a general base by polarising an ordered water which deprotonates the alcohol to increase its reactivity as a nucleophile. ( Ekici, OD 2008) Catalysis takes place in two
The components required for prokaryotic elongation process includes ,the initiation complex described above, aminoacyl-tRNAs, a set of three soluble cytosolic elongation factors (EF-Tu, EF-Ts, and EF-G in bacteria), and GTP. During the first step of elongation cycle the next aminoacyl-tRNA binds to the ribosomal A site. The appropriate aminoacyl-tRNA associates with a complex of GTP-bound EF-Tu resulting in formation of aminoacyltRNA–EF-Tu–GTP complex. It binds to the ribosomal A site with simultaneous hydrolyzed of GTP and an EF-Tu–GDP complex is released from the 70S ribosome. The EF-Tu–GTP complex is then regenerated in a process requiring EF-Ts and GTP.
Low temperatures, can cause enzymes to slow down and decrease their rate of interaction with substrates. The structure of an enzyme are chains of amino acids, and have a specific shape that allow chemicals to react with the enzyme. Enzymes are natural atoms that altogether speed up the rate of essentially all of the chemical reactions that take place inside cells. A lab was conducted to test the effects of different diets on enzyme
The reactor was pressurized with nitrogen up to 5 barg to allow the reaction taking place at increased temperature (95oC) while maintaining methanol in liquid phase. Fig. 6 shows biodiesel yields at various catalyst loadings for the transeterification of palm oil with methanol at