Protein Synthesis Lab

The more acidic a substance is the less oxygen it will produce when going through a chemical reaction. During the Lab “How Do Changes in pH Levels Affect Enzymes Activity”, the researcher conducted an experiment to test the effects that an acidic, neutral, and a base substance will have when combine it with hydrogen peroxide. The data table shows that HCL (acidic substance) barley produced any oxygen at all when it was combining with Hydrogen Peroxide. The pH level for HCL was 2.5; this level indicates that the substance was very acidic. When the H2O and NaOH were tested they produced more bubbles than HCL. NaoH produced a little more bubbles than HCL. The pH that NaoH produced was a 9, which is a base. H2O produced more bubbles than both substances;

The use of multiple test tubes and Parafilm was used for each experiment. Catechol, potato juice, pH 7 phosphate buffer, and stock potato extract 1:1 will be used to conduct the following experiments: temperature effect on enzyme activity, the effect of pH on enzyme action, the effect of enzyme concentration, and the effect of substrate concentration on enzyme activity. For the temperature effect on enzyme activity, three test tube were filled with three ml of pH 7 phosphate buffer and each test tube was labels 1.5 degrees Celsius, 20 °C, and 60 °C. The first test tube was placed in an ice-water bath, the second test tube was left at room temperature, and the third test tube was placed in approximately 60°C of warm water. After filling the test tubes with three ml of the

The effect that increasing Na+Cl- concentration had on osmotic pressure was that the pressure also

These results shown from this experiment led us to conclude that enzymes work best at certain pH rates. For this particular enzyme, pH 7 worked best. When compared to high levels of pH, the lower levels worked better. The wrong level of pH can denature enzymes; therefore finding the right level is essential. The independent variable was the amount of pH, and the dependent being the rate of oxygen. The results are reliable as they are reinforced by the fact that enzymes typically work best at neutral pH

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Hold the IKI spray bottle 25 - 30 cm away from the paper towel, and mist with the IKI solution.

In this lab, the organism that we have been working with is the bacterium, Serratia marcescens. S. marcescens is a member of the Enterobacteriaceae family, and tends to grow in damp environments. S. marcescens is an ideal bacterium to work with in the lab because it reproduces quicker than other bacterium. This bacterium produces a special pigment called prodigiosin, which is red in color. The prodigiosin pigment is intensified when S. marcescens is grown at higher densities. During our experiment, temperature, pH, salinity concentration and oxygen requirements were tested on S. marcescens to measure their optimal growth and prodigiosin production.

The results from the experiment determining the effects of pH on enzyme activity show that as the independent variable, pH, increases the dependent variable, percentage transmission, decreases. This is shown in the results as at the lowest pH, pH 4.0 the average percentage transmission is at its highest at 97%. At the highest pH, pH 8.0 the average transmission is 78.5%. This is also supported from the graph as it produces a negative gradient showing that as the percentage transmission will decrease with an increasing pH. This happens because the enzyme trypsin acts on the gelatine. Therefore as the pH increases towards the optimum pH more jelly will be broken down by the enzyme,

Therefore this experiment was to determine that lobsters in various salinities will osmoconform to their environment. In order to test that lobster's osmoconform, we had to extract approximately 1.0 ml hemolymph from their hemocyannin on the ventral first section of the pre-branchial region. The hemolymph was spun for three minutes in a microcentrifuge and the serum was then tested on an osmometer, which determined the osmolarity of the hemolymph. The results substantiated the hypothesis, in that, lobsters internal osmoles fluctuate with the salinity of the external environment. The two lobsters in the low salinity tank had the lowest osmolarity 0.746 osmoles; the two lobsters in the normal salinity had 0.873 osmoles. The last tank with the highest salinity had the lobsters with the highest osmolarity at 1.445 osmoles. Therefore our data suggests that lobster's osmoconform, with respect to the salinity of their environment by readjusting their intracellular solute concentration to prevent swelling or dehydration because the osmolarity of their hemolymph dictates that of the environment.

Hope Spivey Gate City High School Block 4 Introduction Enzymes cause havoc and speed up reactions, but why exactly does this occur? Enzymes are biological molecules, proteins, that act as catalysts and help complex reactions occur everywhere in life.

Enzymes are proteins that catalyze (speed up) biological reactions in an organism by lowering the activation energy of a reaction. They do this by either straining the bonds in a molecule so that is easier to break up or by placing separate molecules/elements close to each other so that bonds are formed. Enzyme activity is influenced by an array of different factors such as enzyme concentration, substrate concentration, temperature, pH and inhibitor concentration. All of these affect the rate of reactions of enzymes and some such as temperature, inhibitors and pH can under circumstances cause enzymes to become permanently affected. Catalase is an enzyme found in almost all organisms on earth exposed to Oxygen.

Background Research Enzymes are tertiary and quaternary structures of proteins and biological catalysts that accelerate the rates of chemical reactions. Enzymes are significant in that they are not permanently changed in the process and may be reused. Enzymes speed up chemical reactions by lowering the activation energy needed to start a chemical reaction. The process of speeding up a chemical reaction via the use of enzymes begins when a substrate, the substance (reactant) that an enzyme acts upon, binds to the active site of an enzyme. The active site is the region on an enzyme where substrate binds to and undergoes an accelerated chemical reaction; it is where catalysis occurs.

Enzyme synthesis is regulated and controlled via regulatory proteins that are repressed or induced, repression is associated with anabolism while catabolism is seen with inducers. Another component is that of promoter and effector. The effector molecule is one that works with the repressor, altering the affinity for the operator in the controlling region of the gene (McClean 2017). The promoter is described as the binding site for the RNA polymerase (KhanAcademy). This regulation occurs at the transcriptional level (UW-Milwaukee, 2017).

Proteins belongs to diverse class of macromolecules made of hundreds or thousands of smaller units called amino acids that provide structure and support to cell (1). These proteins are encoded by specific genes to carry out the cell cycle progression (3). CDK2 gene encodes a protein which belongs to Ser/Thr protein kinase family (3). This complex is necessary for G2/M and G1/S phase conversions of cell cycle in eukaryotes (4). Cyclin dependent 2 kinase, a protein encoding gene is associated with several diseases such as breast cancer and star syndrome (1). Cdk2 was better understood in S. cerevisiaeas model organism with 75 targets of cdk2 controlling transcriptional regulation, replication, morphogenesis and segregation (1). CDK2 is well known to bring some dynamic effects in the 3D representation of proteins (2).

Protein purification is a process that can be employed to separate a single protein from a larger starting material which may be anything from an organ to a cell. Isolating a purified protein from a larger fraction enables further analysis such as determination of amino acid sequence, potential biological function, and even evolutionary relationship. (Cuatrecasas 1970) In this experiment, the enzyme lactate dehydrogenase will be purified, this enzyme is found extensively in human cells and catalyzes the conversion of lactate to pyruvate, an essential part in energy production. LDH is a key part of anaerobic energy production especially within glycolysis in which LDH catalyzes the conversion of the reverse reaction, pyruvate to lactate, generating NAD+ from NADH, reproducing the oxidized form of the coenzyme which can be used for oxidative respiration. (Markert 1963) Due to the fact that number of purification steps correlates with the purity of the protein multiple purification techniques will be used to isolate a pure form of LDH. LDH will be isolated from a larger “cytosol” fraction collected from a homogenized rat liver in a previous fractionation exercise. Of the procedures that will be used to isolate and purify proteins from a larger fractionate are a set of techniques collectively known as chromatography. These techniques all have the same premise, in that they consist of a stationary phase, also known as the