The nucleophilic attack pushes the carbonyl electrons onto the carbonyl oxygen, which forms a short-lived intermediate. The third step is where the oxyanion electrons reform the bond with the aromatic amino acid. Then the bond between the carboxyl-terminus of the amino acid and the n-terminus of the residue is cleaved and its electrons are used to take out the hydrogen of the nitrogen on the Histidine 57. The c-terminal side of the polypeptide is free to dissociate form the active site. Step four is basically just where water can now enter and bind to the active site through hydrogen bonding, which is between the hydrogen atoms of water and the Histidine-57 nitrogen.
The Binding interaction of complex pyridoxine 5 phosphate oxidase – Nitroimidazooxaine conformations including hydrogen bond and the bond length are analyse by chimera software. 3. Result and Disscussion Docking AnalysisResult : Pyridoxine 5 Phosphate oxidase ,, the macromolecule and the ligand molecule were subject to docking analysis by using Autodock 4.2 .Molecular docking simulations were conducted with this software 10 docking runs were performed. Gird parameter were set as mentioned earlier and the grid spacing point was 0.375 angstoms . After the simulations were complete ,the docked structures were analyzed and the interactions were seen.
ABSTRACT NRC-04, a novel antimicrobial peptide derived from skin mucous secretions of flat fish winter flounder, shows a broad spectrum of antimicrobial activity. In order to understand the conformational change of NRC-04 in different types of membrane, our team did experiments on NRC-04 with negatively charged bacterial surface membrane mimetic micelles sodium dodecyl sulphate(SDS), zwitterionic eukaryotic middle membrane mimetic micelles dodecylphosphocholine(DPC), gram-negative bacteria outer membrane mimetic micelles Lipopolysaccharide(LPS) and bacterial inner membrane mimetic micelles 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol(POPG). Fluorescence test shows that the C-terminus tryptophan residue of NRC-04 interacts with the hydrophobic
The alkaline phosphatase has a lower enzymatic activity when pH 7 and 7.5 Tris-HCl were used; whereas the enzyme has a higher enzymatic activity when pH 8, 8.5 and 9 Tris-HCl were used, with the enzymatic activity at pH 9 being the highest. This shows that the alkaline phosphatase works optimally at alkaline condition such as pH 9 (Kaslow, n.d). The results of the graph show that alkaline phosphatase work less optimally at lower pH, this may due to at lower pH, the positively-charged amino side chain of the peptide sequence of alkaline phosphatase will release the hydrogen ions, therefore causing the tertiary structure of the enzyme altered, which will causes conformation change in alkaline phosphatase’s active site. Thus, the substrate p-nitrophenyl phosphate will experience some difficulties to bind to the binding site on active site of alkaline phosphatase, therefore decreasing the enzyme activity when pH becomes
Once iodine was dropped onto the circle labeled “ Saliva”, it transformed into a white/ yellow Colour, due to it granting the starch to break down properly, it transformed white as a result of there being no starch, hence it executed its main action. Once iodine was dropped onto the circle, which was labeled “HCL+ saliva”, was divided into two colours, white and navy blue, this arose because the starch did not get broken down. The enzyme got denatured with the extension of hydrochloric acid of hydrochloric acid. The acid found in hydrochloric acid obtains a low level oh pH (2), causing it not to be broken down, rather to be denatured, confirms amylase can’t continue it’s activity without a particular high amount of acid, (that is found in stomach acid). My hypothesis is incorrect due to the fact that my prediction is
A: Isolate HHHHHHH Method: Ni-Affinity Chromatography Reason: Six histadine amino acids at the end of the protein can bind to nickel very tightly. Nickel can bind to agarose bead very tightly. Use this strong affinity column we can isolate our protein, which has seven histadine amino acids. Buffer condition: His-binding buffer: • 50 mM Tris-HCl (pH8.0) • 5 mM Imidazole • 100 mM NaCl • 0.1 mM EDTA • 1 mM PMSF made fresh His-wash buffer: • 50 mM Tris-HCl (pH8.0) • 300 mM NaCl • 15 mM Imidazole • 0.1 mM EDTA • 1 mM PMSF made fresh is-elution buffer: • 50 mM Tris-Cl (pH8.0) • 50 mM NaCl • 300 mM Imidazole • 0.1 mM EDTA • 1 mM PMSF made fresh Procedure: 1. Ni-Agarose Beads Preparation: 1L mixture proteins will need 25 ml of beads.Transfer beads into a column.
Two small additional peaks at δ = 0.8 and δ = 1.6 were found may be due to impurities present. 1H-NMR spectrum of PHA isolated from glucose or molasses media indicated characteristic signals of PHB, namely a doublet at 1.26 ppm, which is attributed to the methyl (CH3) group coupled to one proton while a doublet of quadruplet at 2.51 ppm due to the methylene (CH2) group adjacent to an asymmetric carbon atom bearing a single proton. The third signal at 5.25 ppm, which was attributed to the methine (CH) group. 1H-NMR is a very sensitive method for determining the domain size and miscibility, which is difficult to identify by conventional microscopic or thermal analysis (Kichise et al., 2002).The values of the chemical shifts as well as the assignments of the 1H-NMR signals, which appeared in the spectra are in agreement with results obtained by Kichise et al. (2002) and typically identical to peaks of the authentic PHB sample produced from Aldrich Company, which clearly shown that the extracted biopolymer from the B.thuringienesis in this study was poly-3-hydroxybutyric acid.
For ascorbate peroxidase assay extraction 241 buffer was supplemented with 1.0 mM ascorbic acid. The homogenate was centrifuged at 242 15,000×g for 15 min at 40C, and the supernatant was used as a crude enzyme extract. 243 Spectrophotometric determinations were performed using UV visible spectrophotometer 244 (UV-1700, Shimadju, Japan). 245 2.11.2. Estimation of superoxide dismutase (SOD) activity 246 SOD activity was estimated by its ability to catalyse NBT to formazan at 560nm 247 according to the method of Beyer and Fridovich (40).
Alkaline phosphatases (ALP), members of the phosphomonoesterase family, hydrolyze the oxygen-phosphorus bond of organophosphates using metal ions to release an inorganic phosphate under alkaline conditions.1,2 These enzymes are dimeric metalloenzymes containing two Zn2+, one Mg2+, and a serine residue in the active site of each monomeric subunit, in both prokaryotes and higher eukaryotes.2,3 Studies have shown that the three divalent cations are essential for enzymatic activity to catalyze the formation of an alcohol and an inorganic phosphate (Figure 1). In E. coli, the zinc ions are positioned to activate the serine and water for nucleophilic attack and ultimate cleavage of the bond, in addition to holding the phosphate moiety of the substrate. The magnesium ion has been suggested to stabilize the transferred phosphoryl group by a water molecule, using a separate mechanism by which the zinc ions function.3 Although ALPs are found widely in nature and function similarly on the biochemical level, there are locational