Chainobead Lab

Chainobead lab
Part 1AAnalysis questions: 1. How many "chainobeads" was your enzyme able to make per minute in the 0 –
15 second interval? Our enzyme was able to make 6 chainobeads in the 0–15 interval. 2. How many
"chainobeads" was your enzyme able to make per minute in the 60 – 120 second interval? Our enzyme was able
to make 49 chainobeads in the 60–120 intervals. 3. Did your enzyme's rate change over time? How does this
compare to a real enzyme? The enzyme's rate did change over time. This compares to a real enzyme because an
enzyme's job is to speed up the reactions and as time allotted. That did happen since the enzyme in our lab was
able to make more chainobeads as time progressed. 4. Graph 5. Table Chainobead Construction Time Part A 15
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9. Is Chainobead polymerase a catabolic enzyme or an anabolic enzyme? Are the enzymes that digest your food
catabolic enzymes or anabolic enzymes? Chainobead polymerase is an anabolic enzyme because it's
constructing complex molecules from simpler ones. The enzymes that digest my food are catabolic because
they need to break down the food into smaller molecules so that I may retain my energy. Part 2 1. An enzyme is
a macromolecule that acts as a catalyst and also speeds up reactions. 2. A catalyst is a chemical agent that
speeds up a reaction without being consumed by the reaction. 3. A substrate attaches to an active site on an
enzyme. 4. 5. Surcase is an enzyme that catalyzes the hydrolysis of the disaccharide sucrose into its two
monosaccharides: glucose and fructose. 6. Sucrose alters yeast by yeast using sugar as fuel for its fermentation
process since sucrose is a sugar. 7. The common suffix found at the end of most biological enzymes is as. 8. If
an enzyme is present, it also lowers the activation energy needed to get the reaction started. 9. Denaturation of
proteins is the process in which a protein unravels and loses its native shape, becoming biologically inactive.
10. Three things that can denature an enzyme are pH, temperature, environmental conditions, and salt
concentrations. 11. The purpose of placing ht test tubes in a hot water bath is to stimulate the temperature of the
human
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Ultrasound Accelerated Syntheis of Novel Benzimidazole...
Objective: Present research work is carried out for rapid development of some biological active new
heterocyclic moieties. Method: A novel series of 3–substituted–1–[1–(toluene–4–sulfonyl)–1H–benzoimidazol–
2–yl]–propen–1–one (3a–3j) were synthesized by tosylation of benzimidazole chalcones (2a–2j) using
ultrasound in lesser time with higher yields. All the synthesized compounds were characterized by Elemental
analysis, IR spectra, 1H, 13C NMR and Mass spectroscopy. The newly synthesized compounds were screened
for their α–glucosidases and antibacterial activity. Result: Biological evaluation of the compounds (3a–3j)
revealed that most of them are good α–glucosidases inhibitors and possess significant antimicrobial activity
compared to their respective standards. Discussion: Amongst (3a–3j), compound 3h showed moderate inhibition
of glucoamylase and compound 3j with compelling inhibition of α–amylase. Compound 3g and 3e displayed
excellent antibacterial activity against all tested strains and admirable antifungal activity against Candida
albicans respectively. INTRODUCTION Glycosidases play an essential role in numerous biological recognition
processes and their function or dysfunction has been implicated in a number of different disease states, leading
to an interest in the screening of different glycosidases inhibitors (Naoki et al., 2000; Terry et al., 2003). α–
Glucosidases (EC 3.2.1.20) constitute a group of exo–acting glycoside hydrolases of diverse specificity that

Catalase on Hydrogen Peroxide Essay
Abstract
The aim of this study was to test the rate of reactivity of the enzyme catalase on hydrogen peroxide while
subject to different concentrations of an inhibitor. The hypothesis was that hydrogen peroxide will be broken
down by catalase into hydrogen and oxygen, where a higher concentration of inhibitor will yield less oxygen,
resultant of a lower rate of reaction. Crushed potato samples of equal weight were placed in hydrogen peroxide
solutions of various temperatures. The results showed that less gas was produced as the concentration of the
inhibitor rose. This Is because more enzymes were inhibited, and so less active sites were available for reaction.
Reasearch and rationale
Hypothesis:
Catalase will break down hydrogen ... Show more content on Helpwriting.net ...
Planning I will be studying the rate of catalase activity on hydrogen peroxide while varying the amount of
inhibition, which should influence the rate of the reaction, and thus the amount of oxygen, observed in a given
time. The concentration of the inhibitor will therefore be the independent variable, while the amount of oxygen
will be the dependent variable. There should a smaller volume of oxygen observed as the concentration of the
enzyme increases. As I am comparing two variables to each other, it would be wise to calculate the correlation
of the two variables. To calculate the correlation, I should use Spearman's rank correlation coefficient. To find a
suitable correlation, I should use at least 8 samples. After plotting a scattergraph, I will proceed to find out the
correlation if the correlation looks reasonable enough. Variables Temperature: Temperature is an extremely
important variable to consider, as it can affect the rate of reaction in a number of ways. If the temperature of a
system is increased, more molecules will reach the activation energy and the rate of reaction will increase. The
number of collisions will also increase. As enzymes must collide with substrates, an increase in temperature and
thus kinetic energy will result in more collisions occurring in a given time. Increasing the temperature will also
increase the heat of the molecules.

MIF and Autoinmmune Liver Disease
MIF and Autoimmune Liver Disease
MIF is constitutively expressed in low amounts by hepatocytes and has been shown to be weakly expressed
within sinusoidal lining cells as well as biliary epithelial cells of the liver [1, 2]. Previous studies have described
an increase in MIF levels being associated with inflammatory hepatic disorders such as cirrhosis and
hepatocellular carcinoma [3], however, to date little is known about MIF and its implications in the
pathogenesis of autoimmune liver disease. A recent study by Assis and colleagues evaluated changes in the
levels of both circulating MIF and its soluble receptor (CD74) in sera to determine if clinical manifestations
correlated with the inflammatory profile observed in autoimmune hepatitis (AIH) and primary biliary cirrhosis
(PBC) [4]. Immuno–histological analysis levels of MIF and CD74 protein expression, although varied between
mild and severe cases, were more abundantly expressed in liver sections in both AIH and PBC patients
compared to control specimens. Serological data collected from patients in this study also confirmed that both
AIH and PBC cohorts had significantly higher levels of circulating MIF compared to healthy controls. Further
analysis revealed however that PBC patients had higher concentrations of the MIF soluble neutralising receptor
CD74 in contrast to both AIH and control cohorts. This finding in respect to disease progression between PCB
and AIH cohorts is suggested to correlate with the

Succinate Dehydonate Lab Report
Hypothesis: We hypothesize that reagent Malonate will inhibit Succinate Dehydrogenase (SDH) activity.
Background: Malonate, also known as propanedioate, is a classic example of a competitive inhibitor. Malonate
has a molecular structure that is similar to succinate, so it has the ability to bind to the active site of succinate
dehydrogenase and inhibit enzyme activity. However, malonate is sufficiently different from succinate that it
cannot be dehydrogenated. [1] Malonate working as an inhibitor to succinate oxidation is a popular occurrence.
Succinate is oxidized to fumarate during the Kreb cycle and the reagent's (malonate) inhibitory effect slows or
halts that important step from taking place.[2] A competitive inhibitor had a similar molecular ... Show more
content on Helpwriting.net ...
"MALONATE INHIBITION OF OXIDATIONS IN THE KREBS TRICARBOXYLIC ACID CYCLE."
Jurimetrics Journal 10.2 (1969): 73–77.The Journal Of Biological Chemistry. AFFINITY SITES, 1949. Web. 8
Oct. 2014. <http://www.jbc.org/content/178/1/241.full.pdf>. [3] "Biomolecules:Enzymes." Competitive
Inhibition. Chempages Netorials, n.d. Web. 06 Oct. 2014.
<http://www.chem.wisc.edu/deptfiles/genchem/netorial/modules/biomolecules/modules/enzymes/enzyme5.htm>.
[4]"Inhibitors." Chemistry Explained. Foundations and Applications, n.d.Web. 10 Oct. 2014.
<http://www.chemistryexplained.com/Hy–Kr/Inhibitors.html>. [5]Fernandez–Gomez, Francisco J., Maria F.
Galindo, Maria Gómez–Lázaro, Victor J. Yuste, Joan X. Comella, Norberto Aguirre, and Joaquín Jordán.
"Abstract." National Center for Biotechnology Information. U.S. National Library of Medicine, 17 Jan. 2005.
Web. 08 Oct. 2014. <http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1576031/#bib3>. [6]Fernandez–Gomez,
Francisco J., Maria F. Galindo, Maria Gómez–Lázaro, Victor J. Yuste, Joan X. Comella, Norberto Aguirre, and
Joaquín Jordán. "Abstract." National Center for Biotechnology Information. U.S. National Library of Medicine,
17 Jan. 2005. Web. 08 Oct. 2014.

Essay On Substrate Specificity Of Enzymes
Substrate Specificity of Enzymes
A substrate is a substance that the enzyme acts on. When an enzyme binds to its substrate at the active site of
the enzyme, an enzyme–substrate complex is created. While in the complex, chemical reactions takes place and
products are made.
Enzymes have specific shapes that determine their functions.
The shapes accommodate very specific molecules.
Enzymes change shape after chemical reactions. There different energy changes with the reactions due to the
state of the enzyme.
The active site also changes shape when the substrate binds to it, thus allows for the site to have an induced fit.
Catalysis in the Enzyme's Active Site
In the active site of the enzyme, the substrate bonds with weak bonds such as ... Show more content on
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The normal temperature for enzymes is between 35 and 40 degrees celsius.
Most enzymes function at around a 6–7 pH levels.
Some enzymes such as pepsin, which is an acid in the stomach, is activated at a pH of 2.
Cofactors
Cofactors such as ions and metals are nonprotein helpers to enzymes.
A cofactor that is an enzyme is known as a coenzyme.
Enzyme Inhibitors
Competitive inhibitors are inhibitors that block substrates from binding to the active sites of enzymes.
They make the enzyme less effective.
Noncompetitive inhibitors are inhibitors that bind to the outside of an enzyme far away from the active, to
change its shape and reduce its effectiveness.
If inhibitors bond covalently to the enzyme, then the inhibitors are permanent.
For example, in poisons such as DDT, antibiotics also use such inhibitors to kill bacteria.
Although it is used to regulate cellular enzymatic activity, inhibition is harmful.
The Evolution of Enzymes
The function and diversity of enzymes is due to gene mutations. If a gene of an enzyme changes, the protein
will have one or more different amino acids which can impact an enzyme's activity or allow it to bind to
different substrates.
Enzyme – A protein
Catalyst – Speeds up chemical reactions.
Activation Energy – Energy required to produce a chemical reaction.

Substrate – The reactant.
Enzyme/Substrate Complex – A substrate's interaction with an enzyme.
Active Site – The site of an

Enzymes Lab Report
Introduction Enzymes are an important aspect of many physiological processes because they act as catalysts to
speed up reaction rates. Some metabolic reactions have competitive and non–competitive inhibitors that can
hinder reaction rates. Competitive inhibitors target the active site of an enzyme, resulting in an equal Vmax, but
different Km. Noncompetitive inhibitors, on the other hand, bind to an allosteric site and change the shape of
the active site, resulting in the same Km and a lower Vmax (Griffin, 2017, Lab 3). Vmax is defined as the
maximum velocity when an enzyme is used in a reaction. This occurs when all substrates are bound to enzyme
(Vmax, 2008). Km is the substrate value at half of Vmax (Worthington Biochemical Corporation, ... Show more
These volume amounts remained the same in runs one through three. Once the cuvetes entered the
spectrophotometer and the reading began, 10ul of β–lactamase was added. While examining results from run
one, the next run was prepared. Both run two and three were generally set up equal to run one, the only
difference being the dilution of β–lactamase each time. In an eppendorf vial, 150ul of PBS was mixed with
150ul of the enzyme were mixed together. This diluted β–lactamase to half of its original concentration. The
second dilution was prepared using 150ul of the first diluted enzyme and another 150ul of PBS. The new
enzyme used for run three as then ¼ of the original concentration. For run four, the volume of PBS in each
cuvete was reduced by 10 ul to allow room in the original total volume for the addition of 10ul of the inhibitor,
clavulanate. The cuvetes were prepared otherwise as normal. The same enzyme from run three was used in run
four, but it was not added until after the cuvetes were already in the spectrophotometer. After all four runs were
complete, the station was cleaned and the data analysis began. Results These data were first entered into excel
for analysis. For the first run, the Vmax was 0.242 with a Km value of 37.745 (Excel cells J11, J13 on Rate
Calculations sheet). The highest average rate for run one was 0.1207 (Excel cell F6), this did not reach or
exceed the Vmax value

Explain The Significance Of Reaction Coupling In Living...
7. How do enzymes catalyze chemical reactions (don't just say "they lower the activation energy", give me
specific mechanisms). Enzymes catalyze chemical reactions by lowering the activation energy. To be more
specific, the enzymes have a pocket called an active site where substrates, which are the reactants of the
reaction, can bind to. Once they are bound, the complex is called an enzyme–substrate complex. The catalytic
action of the enzyme converts the substrate to products by stretching, stressing, and bending bonds to break
them and make it easier to get to the transition state where the molecules can form new bonds. The amount of
free energy needed to go forth with the reaction decreases, as a result.
8. Explain the significance of reaction coupling in living systems. How is it used, and what does it allow living
systems to do that they would not be able to do otherwise? Reaction coupling in living systems is the act of
exergonic reactions and endergonic reactions working together. Exergonic reactions release free energy that will
fuel endergonic reactions. Living systems will be able to be self sufficient and not rely on other substances or
processes besides it's only as well as reuse and recycle energy throughout the body to be more efficient.
9. Why does the activation energy of many reactions in living systems need to be reduced for living systems to
function? The activation energy of many reactions in a living system need to be reduced for living systems

Rate Of Enzymatic Activity Lab Report
Abstract
The purpose of this experiment was to observe and determine the rate of enzymatic activity in the presence of a
substrate. Starch was used as a substrate and was placed in cuvettes in increasing increments of concentration.
Potassium iodine (KI) was used as an indicator of starch concentration. A separate series of cuvettes was
observed with the same concentrations but with an addition of amylase. A spectrophotometer measured the
absorbance of each concentration and the enzymatic activity was determined. It was hypothesized that the rate
of enzymatic activity would increase as the substrate concentration increased.
Introduction
Enzymes are biological catalysts that speed up the rate of a chemical reaction, without being absorbed.
Enzymes temporarily bind to substrates in order to lower the activation energy. Activation energy is threshold
that must me crossed in order for a reaction to occur before successful collisions give rise to products (Hardin,
2012). Several factors influence the rate at which enzymatic activities occur, such as the concentration of
substrate molecules, temperature, inhibitors, and pH. Enzyme kinetics is the study of the rate at which enzymes
work, which is a function of the ... Show more content on Helpwriting.net ...
The same quantities that were measured in the A series were added to the corresponding B cuvettes. In addition,
100 μL of amylase was added to each B cuvette. KI was then added to each cuvette in order to see the
concentration of starch based on the pigmentation intensity. A spectrometer was used to measure the absorbance
of starch in each sample. The instrument was zeroed out using the one of the cuvettes with 0% starch
concentration. Starting with 2A, each A cuvette was placed individually into the spectrometer in increasing
order with the absorbance rate recorded after each placement. This was then repeated with the B series.

Chemotherapy Research Paper
Introduction
Enzyme inhibition is a process that can be used in the treatment of many diseases. These are disease that rely on
enzymes for structural binding. If a drug can prevent this binding from occurring, essentially we can prevent a
disease from progressing. Cancer is a disease in which unhealthy cells replicate as such a quick rate that they
overwhelm the body's normal healthy cells and eventually results in the death of the patient (Chemoth.com,
2017).
Chemotherapy and Topoisomerase Inhibition
Chemotherapy is used in the treatment of cancer. Various types of chemotherapy exist, but here we will discuss
the specific type of chemotherapy that works as Topoisomerase
Inhibitors. These can inhibit Topoisomerase I, II or both. These two enzymes ... Show more content on
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This is because the inhibition of topoisomerase in these cells also kills them. Patients with lowered while blood
cell counts are more prone to infections and lowered red blood cell counts makes patients anemic. Diarrhea,
nausea and other gastrointestinal issues are also side effects of camptothecin as it also affects DNA binding in
cells of the gastrointestinal system. Camptothecin is a competitive inhibitor since it binds with the enzyme
molecule in order to prevent it from binding the DNA back together, essentially, it is in competition with the
enzyme molecule(OpenStax College, 2013).
Conclusion
Overall, enzyme inhibitors can be very helpful in dealing with diseases but because they target specific enzymes
indiscriminately, they can also be harmful as in the case of
Camptothecin. Whether these are used or not depends on the gains that can be received versus the loss. In the
case of cancer treatment, it is a risk that should be taken as the end result of untreated cancer is almost certainly
death.
Topoisomerase Inhibition
References
Chemoth.com, (2017), Types of Chemotherapy Agents and Regimens, Retrieved from
http://chemoth.com/types/topoisomerase Willey C. D., Bonner J. A., (2012), Interaction of Chemotherapy and

Advantages And Disadvantages Of Arbutin
Arbutin Arbutin, from the dried leaves of bearberry, blueberry, cranberry, and pear trees, is a naturally β–D–
glucopyranoside and derivative of hydroquinone. (20, 21, 24) The mechanism of suppressing melanin
production is to competitively inhibit tyrosinase enzyme at binding site and DHICA (5,6–dihydroxyindole–2–
caboxylic acid) and also inhibit melanosome maturation. (21, 24, 25) Arbutin can cause paradoxical
hyperpigmentation if high concentration is used.(26, 27) Deoxyarbutin has shown a promising skin lightener.
(25) Alpha–arbutin are widely use, 20–fold more potent and stronger inhibitory activity than arbutin.(21, 25)
Aloesin Aloesin is a derivative of aloe vera plant.(21) Competitive inhibitor at binding site of tyrosinase and
DOPA (3,4–dihydroxyphenylalanine) oxidase is its major mechanism in dose–dependent manner.(21, 24)
Ellagic acid Ellagic acid is . It is tyrosinase copper chelation.(22) The study by A. Dahl et al.(28) was compared
0.5% ellagic acid and 0.1% salicylic acid with 4% hydroquinone in hyperpigmenatation and dark spots. They
have shown that no adverse ... Show more content on Helpwriting.net ...
synthesized N–nicotinoyl dopamine, a newly niacinamide derivative, which significantly reduce skin
pigmentation.(34) Licorice extracts (Available concentration of glabridin: 0.5%) Licorice extracts are derived
from Glycyrrhiza Glabra Linnera and Glycyrrhiza uralensis.(24, 26) Licorice extracts properties are like two–
face of coin, there are many active compounds that may inhibit or stimulate melanogenesis. Glabridin, a
polyphenolic isoflavonoid, is the main ingredient in licorice extract which is tyrosinase inhibitor and anti–
inflammatory compound.(21) In addition, it has a role in dispersing the melanin by liquiritin and decrease free
radical formation.(20, 24, 26) Mulberry Mulberroside F, as the active compound, is derived from dried
mulberry root bark (Morus alba L) which can inhibit tyrosinase activity and scavenge superoxide in
melanogenesis process.(21,

Taking a Loot at the Urease Enzyme
Introduction to Urease Enzyme: Urease is an enzyme which catalyzes the hydrolysis of Urea into ammonia and
carbamate; on later stage carbamate spontaneously yield carbonic acid and one more molecule of ammonia.
Urease enzymes are included in super family of amidohydrolases and phosphotriestreases. It required binding
of two nickel ions for its proper activity therefore known as Nickel dependent Metalloenzyme. (NH2)2CO
+H2O → NH3 + NH2COOH NH2COOH +H2O → H2CO3 + NH3 Figure 1: Hydrolysis of Urea into Carbonic
acid and ammonia by Urease In 1926 James Summer proposed that Urease is a globulin type of protein and it
was first time that an enzyme was crystallized from Canavalia ensiformis (jack bean). Urease occurs in different
organisms including plants, some bacteria, invertebrates and fungi. Bacterial source of urease enzyme:
Lactobacillus ruminis, Lactobacillus fermentum, Lactobacillus reuri and Klebsiella aerogenes. Fingal source of
urease is Rhizopus oryzae. There is 40% urease extracted from filamentous fungus. It can cause urinary stones,
gastric ulceration, pyelonephritis and other diseases. It is found in solution part of cytoplasm called cytosol. It
helps different organism for nitrogenous source production. In organisms, urea can't be used as nitrogen source
but Urease hydrolyzed urea into ammonia and carbonic acid which can be used as nitrogen source. Structure of
Urease: Function of Urease X–ray structure from Protein Data Bank:

Design Experiment
Design Experiment: Enzyme Inhibitors.
Research question: What is the effect of adding lead nitrate solution on the activity of amylase enzyme?
Aim: To test the effect of adding nitrate solution on the activity of amylase.
Background Information:
Inhibitors are molecules which repress or prevent another molecule from engaging in a reaction. They are
substances that attach themselves onto an enzyme and reduce or prevent the enzyme's ability to catalyse
reactions. Competitive Inhibitors are inhibitors that occupy the active site of an enzyme or the binding Site of a
receptor and prevent the normal substrate or ligand from binding. An active site is a region on the surface of an
enzyme to which substrates bind and which catalyzes a ... Show more content on Helpwriting.net ...
The exact volumes were measured out with the help of a syringe.
Materials:
1. Syringe
2. 6 test tubes
3. Stop watch
4. Water bath
5. White spotted tile
6. 5% amylase solution
7. Iodine solution
8. Lead nitrate crystals.
9. 6 boiling tubes.
10. A pipette.
Procedure:
1. The water bath was fixed at 40˚C.
2. The six boiling tubes were labelled A–F and then each was filled 10cm3 of starch solution with the help of a
syringe.
3. Different quantities of lead nitrate were then added to the 6 test tubes labelled 1–6. The quantities were:
0.00gm; 0.10gm; 0.20gm; 0.30gm; 0.40gm and 0.50gm.
4. In the test tube 10gms of 5% amylase was then added.
5. The contents of each test tube were then transferred to the boiling tubes i.e. the contents of test tube 1 were
transferred to boiling tube A and so on.
6. The boiling tubes were then placed in a water bath with a fixed temperature at 40˚C.
7. The reaction was allowed for 20 minutes.
8. A drop of iodine was added on to the spotting tile.
9. After 20 minutes the boiling tubes were removed from the water bath and using a pipette a drop was
withdrawn from each test tube and placed on the white tile containing the iodine solution.
10. The effect of the drop of liquid on the iodine was then noted.
Change of color of the solution on the iodine solution.
Mass of lead nitrate (±0.01 grams) Color of the solution
0.00 Light Green
0.10 Light Green

0.20 Light Green
0.30 Dark

Pharmacology Of Pain Essay
Pathophysiology and pharmacology of pain remain complex despite of the recent advancements in pain
molecular and neurobiology (1, 2). Current findings suggest several new drug targets such as transient receptor
potential (TRP) channels, voltage–gated potassium, sodium channels, imidazoline receptors, cholecystokinin
(CCK) receptors and novel strategies like designer receptor G protein coupled receptors (DREADDs) and
optogenetics for the pain management (3–5). Enzymes of the endocannabinoid cascade such as N–
Acylphosphatidylethanolamine–selective phospholipase D (NAPE–PLD), Fatty acid amide hydrolase (FAAH),
Diacylglycerol lipase (DAGL) α, α, β–Hydrolase domain containing 4, (ABHD–4), ABHD–6, ABHD–12 and
Monoacylglycerol lipase (MAGL) and the ... Show more content on Helpwriting.net ...
CDOCKER is a grid–based molecular docking method which employs CHARMM force fields. The grid
extension was set to 10 Å. Addition of hydrogen atoms to the structure and all ionisable residues were set at
their default protonation state at a neutral pH. For each chalcone, ten ligand binding poses were ranked
according to their CDOCKER energies. Input site sphere dimensions were set to –11.2454, 19.8898, –8.42488
and 11.7564 respectively and binding interactions were

Absorbance Experiment With Turnip Peroxidase
Introduction
Enzyme can be defined as a protein molecule that is a biological catalyst. (Ophardt, 2003) Catalyst increases the
speed of a reaction but does not have to use anything to help increase the speed. An enzyme can be determined
by their properties. Enzyme are a substrate specific, substrate connects to an enzyme at the location of an active
site. Enzyme is not used in a reaction and enzyme function in a good condition at the optimum temperatures and
pH. (Ahmez2005) Peroxidase is a type of enzyme which is used in the experiment.
The type of peroxidase is used is called turnip peroxidase. Turnip peroxidase is made up of Guaiacol and
hydrogen peroxide. The reactants to the product are turnip peroxidase or called tertraguaiacol and water. The
color of the react is brown. In the experiment was conducted there were baseline experiment, temperature, pH,
10X substrate, Inhibitor, and half the amount of enzyme.
The baseline experiment is the control group. If the time increases over a period a time then turnip hydrogen
peroxidase will increase absorbance.
The temperature can have a major impact on an enzyme. According to Campbell Biology author Reece etc.
2011 "The enzyme reaction will increase as the temperature increase with the increasing
temperature....substrates collide with active sites more frequently when the molecules move rapidly."(Reece etc
2011) Every enzyme hits its optimal temperature the reaction will be at its highest point.(Reece etc. 2011) When
the

What Is Angiotensin-Converting Enzyme Inhibitors?
Angiotensin–Converting Enzyme Inhibitors (ACE Inhibitors) are a group of drugs that belong to the
antihypertensive drug class. There are currently 10 ACE inhibitors that are available. ACE inhibitors can be
taken either by themselves or can be combined with a calcium channel blocker or a thiazide diuretic to enhance
the treatment This group of drugs is often used as a first line of treatment to treat both heart failure and
hypertension. Some of these drugs include; benazepril (Lotensin), captopril (Capoten), enalapril (Vasotec), and
fosinopril (Monopril) (Lilley, p. 348). The medical diagnosis to take ACE inhibitors includes hypertension and
can be used as an adjunct for heart failure. These drugs are considered to be the drug of choice ... Show more
All ACE inhibitors cross the placenta (Vallerand, p 164), which can cause injury or death to a fetus if taken
while pregnant. Also if a patient has a baseline potassium level of 5 mEq/L, they may be recommended not to
take ACE inhibitors due to the risk of causing hyperkalemia(Lilley, p. 349). ACE inhibitors should be used
cautiously in those with any renal or hepatic impairment, hypovolemia, or hyponatremia (Vallerand, p. 165).
ACE inhibitors can cause excessive hypotension when used with diuretics or other antihypertensive drugs. Also,
they should be used cautiously when taking potassium supplements. NSAID's can potentially block the
antihypertensive effects of the ACE inhibitor and should be avoided. Potential food interactions can occur due
to food dramatically decreasing the absorption of certain ACE inhibitors including captopril and meoxipril.
These drugs should be taken at least one hour before meals to ensure proper absorption (Vallerand, p. 165).
Some common side effects that are associated with taking ACE inhibitors include; nonproductive cough,
hypotension, taste disturbances, diarrhea, nausea, and vomiting (Vallerand, p. 165)). Significant effects they can
have on the central nervous system include, headaches, mood and behavior changes, fatigue and dizziness.
More dangerous reactions include angioedema, hyperkalemia, and renal

Peroxidase Enzyme Lab Report
Introduction Cells are the building blocks of life. Life itself would not be possible without cells and the actions
they carry out. Hundreds of biological and chemical reactions take place in the cell every second. Most of the
reactions in a cell use enzymes to speed up the reaction. An enzyme is a protein catalyst used by living
organisms to increase the rate of biological reactions (Freeman et. al. 2016, p90). A catalyst brings substrates
together in a precise orientation that makes reactions more likely. Enzymes have an "active site," which is
where the reactants bind to the enzyme. The active site is where catalysis occurs. The reactants of the enzyme
are called the substrates. Enzymes are extremely effective at catalyzing reactions because ... Show more content
on Helpwriting.net ...
The first part of the experiment measured the effects temperature has on the enzyme activity. Our hypothesis for
part one is that peroxidase activity is affected by temperature. One can predict that if the temperature of the
environment around the enzyme increases, then the enzyme activity will increase. With that being said, there is
an optimal temperature for peroxidase, so at some point the peroxidase will decrease in activity once it exceeds
it's optimal temperature. The second part of the experiment measured the effects of inhibition and how it
influenced enzyme activity. A hypothesis for part two would be that peroxidase activity is affected by an
addition of an inhibitor. If hydroxylamine is added to the reaction mixture, then the breakdown of hydrogen
peroxide will decrease. Since hydroxylamine is an inhibitor, one can predict that the rate of peroxidase activity
will decrease and the hydrogen peroxide concentration in the mixture will be much

Factors Affect The Chemical Rate Of A Chemical Reaction
Introduction:
Enzymes are a protein that replicates a catalyst which helps chemical reactions begin to move without the
enzymes being changed in the process. Enzymes work to complete only one task. But there are a variety of
different enzymes in the body that each one has their specific task to complete. Something that enzymes do is
they are used in making foods, used for digestion. Many of the chemical reactions in the body have a bunch of
energy in them when they happen, which means it can occur in a faster amount of time. Enzymes speed up a
reaction by lowering the activation energy. An inhibitor makes the reaction slow down. In this experiment we
are testing how three factors affect a chemical reaction. We will be representing different objects or materials as
the enzyme. We will use three objects or body part: a taped hand, a hand with a tennis ball in it, and a rendered
hand. The importance of this experiment is to see how three different elements affect the chemical rate of a
chemical reaction.
Materials and Method:
In this lab we needed supplies to help us conduct this experiment to make it successful. The materials we used
were 100 pennies, which represent the substrates. We also needed a hand to help us represent the active site.
The person in this experiment was the enzyme. The tennis ball in their hand is represented by second enzyme
we were testing. We used tape to help us with the third enzyme trial. We used a stopwatch to help us record how
long we had to

The Benefits Of Irreversible Enzyme Inhibitors
Enzyme inhibitors are molecules that bind to enzymes to decrease their activity. Many drugs are enzyme
inhibitors because by blocking the enzymes activity it can kill pathogens (shpanskaya and Melechko, 2008)
Enzyme inhibitors assist in altering the catalytic action of an enzyme in order to slow down or stop catalysis
(Worthington biochemical corporation, 2015). Inhabitation can either be reversible or irreversible. During
irreversible inhabitation the inhibitor detaches slowly from its targeted enzyme as it is tightly bound to the
enzyme by covalent or noncovalent bonds. Important drugs such as penicillin are irreversible inhibitors. The
penicillin covalently modifies the enzyme transpeptidase which prevents synthesis of bacterial cell walls
consequently killing off the bacteria (Biochemistry, 2002)
As opposed to irreversible inhibition, reversible inhibition is a rapid detachment of the enzyme inhibitor
because the inhibitors bind to the enzymes with non–covalent bonds. Unlike irreversible inhibitors, reversible
inhibitors do not have chemical reaction and can be broken down easily by methods such as dilution
(shpanskaya ... Show more content on Helpwriting.net ...
This is a hormone that whose main function is to constrict blood vessels. This constriction increases the amount
of energy needed to pump blood around the body and can cause high blood pressure. This can be dangerous for
the heart if it had been weakened by heart attack. By blocking the production of Angiotensin II with ACE
inhibitors this, in turn prevents constriction and lowers blood pressure (American Heart Association, 2015).
ACE inhibitors also help to stop angiotensin I from converting into angiotensin II. By doing this there is a
reduction in dilation of the blood vessels. There is also a reduction of the amount of water returned to the blood
by the kidneys which ultimately leads to decreased blood pressure (Kenny,

Competitive Inhibition Lab Report
Competitive Inhibition is usually temporary, and the Inhibitor eventually leaves the enzyme. This means that the
level of inhibition depends on the relative concentrations of substrate and Inhibitor, since they are competing for
places in enzyme active sites. The inhibitor (I) binds the free enzyme reverse to form enzyme inhibitor complex
(EI) that is catalytically inactive and cannot bind the substrate. The competitive inhibitor reduces the
availability of free enzyme for the substrate binding. The maximal rate of the reaction is not changed, but higher
substrate concentrations are required to reach a given maximum rate, increasing the apparent Km. Thus, the Km
of the normal reaction is increased to a new Km (aKm) as a function of the inhibitor concentration, ... Show
more content on Helpwriting.net ...
Malonate prevents succinate from reacting with the enzyme, an effect due to the similarity between the
structures of the two compounds. This blockage causes accumulation of succinate.
(ii) Cyanide is an extremely effective, reversible inhibitor of cytochrome oxidase. Cyanide binds to the
cytochrome c, inhibiting the passage of electrons to O₂. Rate of oxygen uptake decline, and if add ascorbate
electrons the rate of oxygen uptake increases. Cyanide is a respiratory inhibitor; it blocks cytochrome oxidase
(complex 4) and prevents both coupled and uncoupled respiration with all substrates, including NADH,
succinate and ascorbate + TMPD. When we add the cyanide substrate to isolated mitochondria we see a change
in the rate of oxygen consumption followed by a steady state.
Since cyanide inhibit the transfer of electrons from cytochrome a3 to the final electron acceptor, oxygen, thus
electrons cannot be passed along by the preceding cytochromes and coenzyme Q. Instead, the electrons from
SDH–FADH2 can be picked up by an artificial electron acceptor, ascorbate which donates electrons specifically
to cytochrome

Lab Report On The Effect Of Substrate Concentration On...
Introduction
The purpose of this lab report is to investigate the effect of substrate concentration on enzyme activity as tested
with the enzyme catalase and the substrate hydrogen peroxide at several concentrations to produce oxygen. It
was assumed that an increase in hydrogen peroxide concentration would decrease the amount of time the paper
circle with the enzyme catalase present on it, sowing an increase in enzyme activity. Therefore it can be
hypothesised that there would be an effect on catalase activity from the increase in hydrogen peroxide
concentration measured in time for the paper circle to ride to the top of the solution.
Enzymes are natural catalysts that work from the ability to increase the rate of reaction by decreasing the
activation energy of a reaction. (Blanco, Blanco 2017) An enzyme can do this 10^8– to 10^10 fold, sometimes
even 10^15 fold. (Malacinsk, Freifelder 1998) The substrate will momentarily bind with the enzyme making the
enzyme–substrate complex, of which the shape of the substrate is complimentary to the shape of the active site
on the enzyme it is binding with. There are two main theories as to how an enzymes and substrates interact, the
lock–and–key model and induced fit theory. The lock–and–key model suggests that the enzyme has a specific
shape that fits the substrate and only that substrate. The induced fit theory says the active site and substrate are
able to change shape or distort for the reaction to take place with (Cooper,

A Study On The System
Caspase–1, found in Homo sapiens (Human) has been extensively studied. The PBD code for this enzyme is
3E4C. Using this PBD code, and the Uniprot Knowledge Base, the entry of the studied enzyme was found:
P29466. From this accession code, a homologous protein with 48.6% similarities was found. This homologous
protein was called Caspase–4 isoform alpha (Uniprot accession number: H9Z2M5) found in Macaca mulatta
(Rhesus macaque). Using the BLAST results, an alignment (Figure 1) between chosen sequences were
analyzed.
The active site function was the same for both. Based on Fig. 1, the active site for Caspase–1 is His237 and
Cys285 and for Caspase–4 isoform alpha the active site is His209 and Cys257. For Caspase–1, the active site
residues His209 is important because as a base it demonstrates nucleophilic behavior when attracting a proton
from Cys285. Thus, His209 acts as a proton shuttle. For the active site residue, Cys285, it acts as a nucleophile;
this cysteine is important for cleaving the peptide bond in proteins that contain aspartic acid. To further
characterize the enzyme Caspase–1, the properties of the protein were compared to the homologous protein
Caspase–4 isoform alpha (Table 1).
For caspases, their usual role as an enzyme is to cleave peptides in their mechanism and trigger apoptosis.
However, the specific enzyme chosen, caspase–1 is important to inflammatory response. The general reaction is
pro–interleukin–1β + H2O mature pro–interleukin–1β. It is a small

Ellman's Spectrophotometric Analysis Of Rivastigmine :...
All the chemicals used in the synthesis were of analytical grade purity and were purchased from Sigma–Aldrich
(India). Rivastigmine was obtained as a gift sample from Sun Pharmaceutical Industries Ltd (Silvassa, India).
Melting point of the synthesized analogues was determined by using Stuart melting point apparatus and were
uncorrected. Equimolar (0.01mol) quantity of NaCNO in 25ml of warm water was added with continuous
stirring, the reaction mixture was allowed to stand for 4 h and the product was obtained by filtration, washed
with water, dried in an oven below melting point and recrystallized from ethanol to afford key intermediate–1
.The precipitate was obtained by filtration, washed with water, dried in an oven below melting point and
recrystallized from ethanol to afford key intermediate–2 .Equal moles of intermediate–2 (0.456g, 0.003mol) in
5ml of ethanol mixed with equal moles of the different aldehyde or ketone was refluxed for 2hrs and glacial
acetic acid was added to adjust the pH of the reaction between 5–6. The solid obtained after cooling was filtred,
dried and crystallized from 95% ethanol to afford compounds .Ellman's spectrophotometric analysis [16] was
used to determine IC50 values. This method is based on the reaction between synthetic substrate
acetylthiocholine iodide (ATChI) and 5,5–dithio–bis–(2–nitrobenzoicacid) (DTNB) to produce a yellow colour
(5–mercapto–2–nitrobenzoicacid) which was detected by Colorimeter. Determination of IC50 values was
performed by recording the rate of increase in the absorbance at 412 nm for 5 min.
Stock solution of AChE was obtained by dissolving AChE in 0.1 M phosphate buffer (pH 8). The final solution
for assay consisted of 0.1 M phosphate buffer (pH 8.0) with the addition of 340 mM 5, 5–dithio–bis (2–
nitrobenzoic acid) (DTNB), 0.02 unit/mL of AChE and 550 mM of substrate (acetylthiocholine iodide, ATChI).
Different concentrations of test compounds (inhibitors) between 20% and 80%) were selected in order to obtain
inhibition of the enzymatic activity. From the inhibitors (synthesized analogues) solution (50 µL), increasing
concentrations of the inhibitors were added to the assay solution and were pre–incubated for 20 min at 37 oC
with the enzyme followed by

Succinate Dehydrogenase Lab Report
Introduction Results
Succinate dehydrogenase is an enzyme found in the mitochondrial inner membrane. The enzyme catalyzes the
reaction of oxidizing its substrate, succinate, into fumarate via the removal of hydrogen ions from succinate.
This oxidation is vital in the Krebs cycle.
The krebs cycle is the second step in aerobic respiration of cells, which takes place in the matrix of the
mitochondria of eukaryotic cells. This process is to oxidize pyruvate. figure 1: krebs cycle Succinate fumarate
Figure 2: oxidation of succinate
Method
One gram of liver was sliced into pieces, then 10ml of homogenization buffer was added to it. The mixture of
the liver and homogenization buffer was then placed into a homogenizer to make the liquid slurry. Once the
slurry was made it was placed into a 50ml falcon tube to then be placed in the centrifuge. The slurry was
centrifuged for 2 minutes at 2000rpm. Once the spin in the centrifuge was complete, the slurry had separated,
the most dense particles to the bottom of the tube forming sediment and the lighter (liver succinate
dehydrogenase enzyme) also known as the supernatant. The supernatant was extracted from the falcon tube and
placed into a test tube. The tube was then kept at a low temperature, (in an ice bath) until it was required for
use.
8 test tubes were then labelled A–H, 1ml of DCPIP was added to each of the test tubes. 2ml of NaHCO3 was
added tubes A through G.

Acetylcholinesterase Enzyme Inhibitors
Acetylcholinesterase enzyme Inhibitors, preventing acetylcholine degredation through inhibition of
Acetylcholinesterase enzyme. That would cause the accumulation of acetylcholine in synaptic cleft therefore
prolong the neurotransmitter action. Acetylcholinesterase enzyme inhibitors action can be reversible and
irreversible. The reversibility or irreversibility of the inhibitory action depends on the functional group available
in the agent.
Carbamates are derived from carbamic acid, the structure activity relation of carbamates are:
X: can be Sulphur (Thiocarbamate) or oxygen
R1 and R2 : Substitute can by alkyl or hydrogen
R3 : mostly should be organic substitution and sometimes metal
Carbamtes has been used as theraputic agents for ... Show more content on Helpwriting.net ...
Mechanism of action of organophosphorus compounds used in pesticides and nerve agents is the same however,
the compounds in pesticides are developed to be more specific on insects than warm–blooded organisms, such
compound are thio forms of OP , while nerve agents contain oxo form of Op. In addition OP, acetylcholine
esterase inhibitors used in insecticides degrades and hydrolysis rapidly once comes in contact to sunlight, air
and soil. However, in case of prolong and or repeated exposure to the Ops insecticides can be very toxic and
may cause the same result as acute exposure.
After exposing to organophosphorus compounds, prevention of serine aging and release of phosphate group is a
vital step for survival. Hydroxylamine (NH2OH) is a potent nucleophilic compound which is able to cleave
phosphate esters, therefore it helps hydrolyzing phosphorylated Ach Esterase enzyme. That leads to develop
hydroxylamine derivatives such as; oxime and followed by pralidoxim as a antidote for phosphorylated
acetylchilone esterase enzyme. Pralidoxim chloride is the only known antidote.
The treatment can be in form of parasymatholytics administration; such as

The Role Of Bioenergetics On Disease And Use Of Small...
Role of Bioenergetics in Disease and use of Small Molecule Therapeutics
Name
Institutional Affiliations
Role of Bioenergetics in Disease and use of Small Molecule Therapeutics
Introduction
The study of bioenergetics includes and not limited to study of biological membranes incurred in energy
conversion and transfer. In particular, the study concentrates on structures acquired using X–ray
craystallography, molecular mechanisms of the photosynthesis processes, bacteria respiration, mitochondrial,
transport, motility and oxidative phosphorylation. Furthermore, areas of structural biology, spectroscopy,
molecular modelling and biophysics of the system applications are not left out while studying the specific
chemical process of a disease. Bioenergetics further spans in the biology of mitochondrial that embodies
biomedicine, features of mitochondrial disorders and energy metabolism (Zheng et–al, 2010, p.519).
Alzheimer's disease, Parkinson's disease, aging, cancer and diabetes are among the well–known
neurodegenerative illnesses studied under bioenergetics and use of small molecule therapeutics.
Small molecule therapeutics is one of the scientific techniques designed to help visualize the magnanimity of
genomics data which is prodigious in the process of making drugs. When this technique is used, genomics data
can yield random number of proteins produced in a disease tissue. By understanding the role played by
bioenergetics in a particular

Peroxidase Enzyme Lab
The Change in Enzyme Activity Affected by Inhibitors
Gilliam, M., Morton, D., Osornio Padron, J., Simmons, A.,
Bahle, T. Lab Section Wednesday: 08/31/16
Introduction: Enzymes are important proteins that control certain most reactions in our cells (Vodopich, 2008).
Not all enzymes are alike, as they all have a different structure and functions. An example of an enzyme is
Peroxidase. Enzymes react with subtracts when it connects in the appropriate active site. Temperature, pH
levels, and inhibitors affect the activity of enzymes. There are two different kinds of inhibitors that affect the
activity of enzymes. These two inhibitors are competitive inhibitors and non–competitive inhibitors.
Competitive inhibitors act like substrates but instead of causing a reaction it blocks the active site. In order to
measure absorption of light, the dye Guaiacol is used. Using the dye is a usefully method for evaluating the
activity of enzymes (Doerhe, 1997). Guaiacol changes color to from clear to a brown color because it is
oxidized by oxygen (Vodopich, 2008). The hydroxylamine is the competitive inhibition, which competes with
the substrate hydrogen peroxide. The enzyme used in this experiment is turnip extract which is the peroxidase.
The hypothesis is that the competitive ... Show more content on Helpwriting.net ...
From the table provided in the lab manual, nine solutions are prepared in a separate test tube. Each solution
varied from a combination of distilled water, guaiacol, hydrogen peroxide (3%), turnip extract, and
hydroxylamine. When preparing the solution, it is crucial that each tube is prepared and test one at a time before
preparing the next test tube. Once a test tube is prepared, it is immediately placed in the spectrophotometer to
measure light absorption. After the lid is closed, the light absorbed is recorded every thirty seconds for a total of
five

Aconitase Research Paper
Aconitase is one of the enzymes vital for the human. This enzyme is classified under the group of lyases. To be
more specific, it functions as the hydro–lyases, breaking down the bond linking the carbon and oxygen through
the dehydration. (NCBI, 1963) It also can perform the hydration under certain circumstances. Thus, aconitase is
also known as citrate hydro–lyase and aconitate hydratase. (Kremer, 2013) The main metabolic pathway
aconitase participates will be the aerobic respiration, especially the citric acid cycle of it. The cycle is also
known as the Krebs cycle. Aconitase can catalyze the citrate to isocitrate in stage two of the cycle. (Voet, Voet,
& Pratt, 2008) The mechanism involved in this process is actually a dehydration–rehydration ... Show more
Both types of the aconitase can catalyze the interconversion of citrate and isocitrate. However, for the cytosolic
aconitase, it can accelerate the reaction in the cytoplasm as an alternative, unlike the mitochondrial aconitase
which proceeds the reaction inside of the mitochondrion. In addition to that, the cytosolic aconitase has the
specific feature of balancing the number of acetyl–CoA and NADPH synthesized during the citric acid cycle.
As the result of this, the other metabolisms can use these two compounds to generate other essential
compounds, such as amino acids and fatty acids. Even though, the cytosolic aconitase can only be active in
catalysis when the level of the iron atoms in the cytosol is high under normal circumstances. Otherwise, the
cluster with four iron atoms and four sulfur atoms will be detached from the cytosolic aconitase, losing the role
of being an enzyme. (McDowall,

Optimal Dh And Temperature Conditions Lab Report
Optimal pH and Temperature Conditions Comparative studies show that the optimum pH of LDH from rabbit
muscle ranges from pH 6.2 to 7.8 [6]. A pH–rate profile graph in a study by Fritz [6] reflects the results
obtained in Figure 1, where the optimum pH was found to be 7.5 for LDH extracted from rabbit muscle. This is
further supported by the optimal pH of rabbit muscle LDH obtained by another study [7], in which the pH is
determined to be 7.5. Thus the results obtained in Figure 1 fall consistent with literature values. In a study by
Buhl et al., the optimal temperature of LDH from skeletal muscle of aa human was 37oC[4], suggesting that
denaturation characteristics of the enzyme would be observed beyond this temperature condition [1]. ... Show
The low Vmax could be due to an incomplete reaction. However, despite the large difference in Vmax between
experimental and literature values, Figure 3 reflects similar data obtained in similar studies [9]. Michaelis–
Menten graphs depicted by Figure 4 and Figure 5 show that all concentrations of oxalic acid (0M, 0.01M and
0.03M) and oxamic acid (0M, 0.0200M and 0.0600M) ) seemingly saturate at the highest lactate concentration
which was 95mM. In both figures, LDH saturates the fastest in the absence of inhibitors, followed by 0.01M
oxalic acid and 0.0200M oxamic acid, and 0.03M oxalic acid and 0.0600M oxamic acid having the slowest
reaction rate respectively shown in Figure 4 and Figure 5. These results reflect the Michaelis–Menten graphs
reproduced by experiments under similar conditions as explained by Powers et al [9]. As predicted, Km and
Vmax values obtained from the Michaelis Menten graphs in Figure 4 and Figure 5 do not correspond with Km
and Vmax values obtained in the Lineweaver–Burk plot of LDH in Figure 3. The Vmax obtained in Figure 4
and Figure 5 were both 10.6µM/min in the absence of inhibitor whereas the Vmax in the absence of inhibitor
was 0.34 µM/min in Figure 3. Lineweaver–Burk plots allow for an accurate derivation of an enzyme's Km and
Vmax through an extrapolation of reciprocal values as opposed to the Michaelis–Menten graph which indicates
a direct correlation of an enzyme's substrate

Lab Report On Peroxidase
Introduction Enzymes are macromolecules that act as a catalyst, and it's a chemical agent that accelerates the
reaction without being consumed by the feedback or the results (Campbell and Reece, 2005). After the
adjustment by the enzymes, the chemical movement through the pathways of metabolism will become awfully
crowded because many chemical reactions are taking a long time (Campbell and Reece, 2005). There are two
kinds of reactions in nature. The first one is Catabolic reaction and the second one is Anabolic reaction.
Catabolic reactions are large molecules that are broken up into smaller molecules (Ahmed, 2013). Anabolic
reactions are small molecules that join to make larger molecules, like polymerization (Ahmed, 2013). If you ...
Show more content on Helpwriting.net ...
W. John Albery (1976) stated that the "improvement in the catalytic efficiency of enzymes, compared with
simple organic molecules, is separated into three broad types of alteration to the Gibbs free–energy profile"
(Albery, 1976). The experiment that the class worked on was about peroxidase. Peroxidase is part of the
enzyme group that presents most living organisms (Ahmed, 2013). Peroxidase interferes with the removal of
hydrogen peroxide (Ahmed, 2013). Hydrogen peroxide is a toxic product that have normal metabolism before it
causes any cell damages (Ahmed, 2013). Peroxidase has two substrate and both of them must present a reaction
(Ahmed, 2013). One of the two substrate is H2O2 and other one just depends on the organism or the cell type
(Ahmed, 2013). The substrate that the class uses is turnip extract. In the class there were five experiments to do
but the class were assigned into groups and each group were going to do two experiment. The names of the
experiments are: Baseline, Temperature, and pH. The first experiment was Baseline and for that experiment we
needed to get three tubes but one of the tubes were already done so the only thing was left is to do test tube two
and three and put it together than put it in the spectrophotometer 20. The hypothesis for this experiment

Selective Inos Inhibitor
Specific Aims
We propose to modify the recently discovered natural product terpenoids to develop a selective iNOS inhibitor
with improved in vivo properties.
Specific Aim #1: To confirm and enhance bonding of terpenoids to iNOS. A SILAC experiment will allow the
action of the small molecule on iNOS, eNOS, and nNOS to be confirmed. SAR studies will then be performed
to enhance selectivity towards iNOS and improve pharmacological properties.
Specific Aim #2: To study the effects of iNOS inhibition on angiogenesis. The development of vessels from
human umbilical stem cells in the presence of either one of our terpenoids, a traditional iNOS inhibitor, a
genetic knockdown, or a control will be studied to determine the effect of iNOS inhibition ... Show more
Provided the natural product does not simply quench nitric oxide, further testing will be performed.
Alternatively, if the natural product does show activity towards nitric oxide, it will be modified to a moiety less
sensitive to the radical before further testing is performed. To test for the selectivity of these molecules SILAC
testing will be performed. Using the computer dock data from the terpenoid paper, a bead will be attached to the
small molecule near the isopropyl group, which does not seem to be involved in any major interactions in the
active site. This bead placement will need to be confirmed experimentally by testing the nitric oxide production
of cells in the presence of the compound and the compound with the bead are similar. After the location of bead
placement is determined, one set of cells will be grown with the small molecule in heavy medium and another
control set of cells in regular medium with no compound. After incubation, the proteins that have attached to the
small molecule can be determined through mass spectroscopy will be

Investigating The Reaction Rate And Specific Activity Of...
Polyphenol Oxidase Analysis
By Natasha Patel
Miracosta College
1 Barnard Dr, Oceanside, CA 92056
Abstract
Our goal in this experiment is to determine the reaction rate and specific activity of the protein extract. Also, the
purpose of this lab is to characterize Vmax and Km for catechol and L–Dopa. Along with, determining whether
the mode of inhibition by PTU is competitive or non–competitive? We performed a spectrometry analysis in
order to determine the reaction rate of PPO, specific activity, and substrate– enzyme relationships among L–
Dopa, catechol, and PTU. The reaction rate of polyphenol oxidase determined to be 0.04932957 μmin/min. By
incoporating Beer's Law, the specific activity of our protein extract was 0.2517127 μm/mgmin.In order to
determine Vmax and Km for catechol and L–dopa we had to take averages from 3 dfferent groups. The Vmax
for L–dopa determined to be 14.27, the ½ Vmax was 7.134, and the Km value was 0.485. The Vmax for
catechol determined to be 10.00, ½ Vmax was 5.33, and the Km value was 0.38. The mode of inhibition by
PTU determined to be non– competitive. The graph shows a rough hyperbolic trend with the slop increasing
and then slowly making a curve towards the end for the catechol substrate concentration. However, the reaction
with catechol and PTU portrayes a flat line. This is due to the fact that the increasing amount of substrate
concentration had no effect on the reaction at all. Therefore, PTU was inhibiting the

Evaluation Of Glucosylceramide Synthase ( Gcs )
Abstract
Glucosylceramide synthase (GCS) is an enzyme that catalyzes the first step in the biosynthesis of
glucosylceramide (GlcCer)–related glycosphingolipids (GSLs). In the present report we have used atom–based
3D–QSAR method to analyze the structural aspect on a series of iminosugar derivatives as potent inhibitors of
glycosylceramide synthase. In this approach the experimental dataset was divided into training and test sets and
the model was chosen based on randomized trial distribution which has high correlation factors. A ligand–based
pharmacophore and atom–based 3D–QSAR studies were carried out on a set of 65 inhibitors of GCS. After the
QSAR studies, a five–point pharmacophore with two hydrogen bond acceptors (A), two hydrophobic group (H)
and one ring aromaticity (R) was obtained. The pharmacophore hypothesis yielded a 3D–QSAR model with
good partial least square statistics results. The training set correlation has these partial least square factors
(R2=0.92, SD = 0.178, F = 843.9, P = 1.12e–15). The test set correlation has these partial least square factors
(Q2 = 0.25, RMSE = 0.665, Pearson–R =0.54).
After the QSAR studies, docking study is being carried out. The docking study will reveal the binding
orientation of the active ligands on the protein which will be completed in BTP–phase II.
The result for QSAR and docking studies will provide structural characteristics of the active ligands and also
give detailed information about binding features which can useful

How Enzymes Affect Our Body
Enzymes are biologically active proteins that serve as catalysts; they have the ability to speed up chemical
reactions. Without a catalyst many biochemical reactions will be carried out too slowly for the body to benefit.
All sorts of enzymes occur naturally in our bodies, some help digest food. An enzyme such as amylase is
present in saliva which breaks down starch into smaller maltose molecules. Without an enzyme holding the
large molecule in position, this reaction will be extremely slow. Every reaction requires certain amount of
energy to form. Enzymes lower the activation energy (amount energy required to be put into the reaction to
begin). It does this by applying pressure to bonds that require s the molecule to break, or bring the molecules
closer to bond. The molecules or chemicals the enzymes work on are called substrates, substrates then bind to
the enzyme active site, once they bond to the active site, the enzyme will change shape to fit the substrate, and
this is known as the induced fit model. All enzymes are specific to particular molecules or chemicals and are
usually named after their substrate 'ase'. For example lactase is an enzyme that breaks down lactose.
Allosteric enzymes differ from non–allosteric enzymes. Allosteric enzymes at least have more than one binding
site, where as regular enzymes have one or more site that they can bind to. Unlike allosteric enzymes, if a non–
allosteric enzyme binds to an enzymes active site this does not increase or

Cu466 Analysis
A new compound CU466 is a potential potent inhibitor of Dihydrofolate Reductase to treat methotrexate–
resistant breast cancer
Hayfa Akoubi *
*From the department of Biology, Concordia University, Montreal, Quebec H4B 1R6, Canada
Background: cancer has developed mechanisms to survive and resist drugs including methotrexate, a DHFR
inhibitor
Results: CU466 is a stronger inhibitor of DHFR because its positive charge interacts with leucine 22 on DHFR.
Methotrexate resistance in cancer is mediated by DHFR gene amplification
Conclusion: CU466 is a good alternative to treat methotrexate resistant breast cancer
Significance: understanding the mechanism of drug resistance in cancer leads to developing more potent drugs.
Cancer is among those ... Show more content on Helpwriting.net ...
However, the majority of known anti–cancer drugs target normal cells as well. Methotrexate, formerly known
as amethopterin, is an antimetabolite used in the treatment of some forms of cancer (Shacter and Law, 1956). It
inhibits the function of dihydrofolate reductase (DHFR) by tightly binding to the enzyme. Following this slow
interaction between methotrexate and DHFR, an enzyme–NADPH–inhibitor complex is formed (Stone et al.,
1984). Therein, DHFR is rendered unable to catalyze the NADPH–dependent reduction of dihydrofolate to
tetrahydrofolate (Anderson, 2017). Tetrahydrofolate is an important methyl group shuttle in the de novo
synthesis of purine and pyrimidine nucleotides and some amino acids (Barbara and Hiroshi, 2002). Therefore,
eliminating it from cells is a good therapeutic strategy against cancer cells, as it would limit the de novo
synthesis of purines and pyrimidines (Lane and Fan, 2015). Inhibiting the function of DHFR by its competitive
inhibitor methotrexate is one such good therapeutic strategy, as it would disable DHFR from converting
dihydrofolate into tetrahydrofolate. However, some types of cancer cells had acquired some forms of resistance
against methotrexate (Hans et al., 2011). Equally important, methotrexate should be administered in well
established low doses, because it affects

Classic Galactosemia Type 1
Classic Galactosemia, Type 1, is a complex disorder and the exact pathophysiology has is controversial.
However, it is most commonly accepted that the main factor is the accumulation of galactose–1–phosphate,
gal–1P, which is due to the impairment of galactose–1–phosphate uridylytransferase, GALT. This reaction uses
the GALT enzyme as a part of the Leloir pathway which enables the body to process galactose. The GLAT
enzyme itself belongs to the histidine triad super family and is a member of branch III. This enzyme shows
specific nucleoside monophosphate activity and is a homodimer with each monomer containing a single domain
comprised of 6 α– helices and a β–sheet which is formed by 13 antiparallel and 1 parallel strand. The
mechanism of this enzyme is described as having ping–pong kinetics with the following two steps. In the first
step the active site histidine attacks R–phosphorus of UDP–glucose which displaces glucose–1–phosphate and
forms a covalent intermediate. The second step involves the previously formed intermediate reacting with
galactose–1–phosphate to displace the histidine and produce UDP–galactose. (Facchiano, 103–104). A
deficiency in the GALT enzyme leads to Galactosemia which can present with the following acute symptoms:
"jaundice, cataracts, vomiting, diarrhea, hepatomegaly, sepsis, and neonatal death" (McCorvie, 1) with the
possibility of long term problems without treatment. Currently, restriction of galactose is able to help reduce the
acute

Prediction By Komputer Assisted Technology : Lab Analysis
In the materials and methods section; ligand preparation, target protein identification and preparation, molecular
descriptors calculation, ADME (Absorption, Distribution, Metabolism and Excretion) and TOPKAT (Toxicity
Prediction by Komputer Assisted Technology) analysis were carried out according to the previously reported
method as briefly stated below.
Ligand preparation
Chemical structures of the ligands i) asperyellone [CID101600052]; ii) asperenone [CID5368642]; iii)
hydroasperyellone [CID561143]; iv) CHEMBL1715716 [CID49859207] and v) CHEMBL2152350
[CID71458428] were downloaded PubMed (www. pubmed.com) database. The ligands were drawn in
ChemBioDraw Ultra 12.0 (www.cambridgesoft.com) and subsequently molecular mechanics (MM2) ... Show
ADME and TOPKAT analysis
Both ADME and TOPKAT analysis were performed using Discovery Studio® 3.1 (Accelrys, San Diego, USA).
ADME analysis was performed using six descriptors such as human intestinal absorption (HIA), aqueous
solubility (AS), blood brain barrier (BBB), cytochrome P450 2D6 (CYP2D6), plasma protein binding (PPB)
and hepatotoxicity (HT). As for the TOPKAT analysis, five descriptors were used which includes aerobic
biodegradability (AB), Ames mutagenicity (AM), ocular irritancy (OI), skin irritancy (SI), skin sensitization
(SS) and oral toxicity (OT) in rat (LD50 in g/Kg of body weight).
Docking studies Docking studies were performed on the protein crystal structures of HMGR, HNE, SQS,
tyrosinase and XO obtained from Protein Data Bank (PDB) using the CDOCKER protocol under the protein–
ligand interaction section in Discovery Studio® 3.1 (Accelrys, San Diego, USA). In general, CDOCKER is a
grid–based molecular docking method that employs CHARMM force fields. A protein was firstly held rigid
while the ligands were allowed to flex during the refinement.

The Enzyme Peroxidase : Lab Report
The Enzyme Peroxidase Case Study Lab Report
Autumn Vick
I. ABSTRACT:
The purpose of this experiment is to learn the effects of a certain enzyme (Peroxidase) concentration, to figure
out the temperature and pH effects on Peroxidase activity and the effect of an inhibitor. The procedure includes
using pH5, H202, Enzyme Extract, and Guaiacol and calibrating a spectrophotometer to determine the effect of
enzyme concentration. As the experiment continues, the same reagents are used with the spectrophotometer to
determine the temperature and pH effects on Peroxidase activity. Lastly, to determine the effect of an inhibitor
on Peroxidase, an inhibitor is added to the extract. It was found that an increase in enzyme concentration also
caused an increase in the reaction rate. The reaction rate of peroxidase increases at 40oC. Peroxidase performed
the best under pH5 and declined as it became more basic. The inhibitor (Hydroxy–lamine) caused a decline in
the reaction rate. The significance of this experiment is to find the optimal living conditions for Peroxidase.
This enzyme is vital because it gets rid of hydrogen peroxide, which is toxic to living environments.
II. INTRODUCTION: There are thousands of chemical reactions that occur in a cell at every moment. These
chemical reactions do not occur randomly, they are highly under the control of biological catalysts called
enzymes. Most of these enzymes are proteins. These proteins have certain primary structures directed by

How The Enzymes Affect A Substance?
Enzymes are complex proteins that function as biological catalysts, which act by increasing the rate of
biochemical reactions to alter a substance (known as a substrate) in metabolic processes, such as respiration and
digestion. In order for enzymes to function at their best, they need their environment to be at certain conditions
for the chemical reactions to take place. The environment where the enzymes will function at their best is called
'optimum conditions' and will vary from enzyme to enzyme, depending on the chemical reaction.
Denaturation is where the proteins of an enzyme unfold and the structure of an enzyme irreversibly changes so
that it cannot perform its intended function. An example of non–specific enzyme inhibitors would be
temperature and pH. If the conditions are too acidic or too alkaline, or the temperature is too high, the structure
of the enzyme will denature. If the active site of an enzyme is altered, then the substrate cannot fit in to the
enzyme and the chemical reaction will not take place.
The conditions of the environment the enzyme is in can have a huge effect on the enzymes performance and the
rate of reaction. Some of these conditions include temperature, pH, substrate concentration and product
concentration.
Temperature – Reducing temperature can slow down the rate of reaction as there is a lower amount of kinetic
energy, leading the enzymes and substrate to collide less often, meaning that the reactions will not take place..
Conversely,

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