EnzymeLAB

Instructions (because of difficulties we are experiencing with the simulation,  expect this guide to be modified) 

EnzymeLAB (available at https://learntech.imsu.ox.ac.uk/enzlab/

  • Simulate the behaviour of an enzyme over a range of substrate concentrations, at various pH values, and in the presence/absence of an inhibitor. 
  • Simulated enzymes obey Michaelis-Menten kinetics, as described above. 
  • Faster than a real time enzyme assay so it allows you to spend time designing your experiment and try our lots of different setting. 
  • Note: You must include the enzyme reference number in your report.
  • It mimics real life! You need to eat between enzyme assay, and have tea (The simulation is from University of Oxford!)

First you need to choose a sensible enzyme volume. Did you get the specific activity (mmolmin-1mg-1) and the enzyme concentration (mgmL-1). You know the maximum rate measurable is 10 µmolmin-1. This should enable you to calculate the enzyme volume needed.Next! Substrate concentration – Hint! start with high substrate concentration at pH 7, so the enzyme will be near saturated and this gives an estimate of VmaxFinding a rough value for the optimum pH –plot activity versus pH.Finding Km and Vmax – take values of [S] varying by at least 10 fold, until rate drops below 0.5×Vmax. You will need about half the rates below 0.5×Vmax and half above.

  1. Find optimal enzyme amount (once set this will not need to be changed)
  2. Figure out a sensible substrate range (how are you planning this range?)
  3. Now you are ready to determine Km and Vmax
    1. The accuracy will depend on the spread of data points and number of data points. 
    1. to get reasonable values for x (mean) and  (standard deviation), need to repeat measurements at least 5 times
    1. Generally it is better to take values at a large number of [S] values rather than repeats at a smaller number of [S] values
  4. And you can check how pH affects Km and Vmax
  5. Determine the optimal pH
  6. Determine the determine Km and Vmax again at the optimal pH
  1. Using this link do the experiment and complete the following: you will have to use he unsecure web page to complete and use this software (advanced settings then proceed to link). https://learntech.imsu.ox.ac.uk/enzlab/
  2. Answer questions as you are directed, keep original formatting. The formatting to use is directly below in the table with grey, blue and red typing.
  3. Refer to the “BCEM2201 figure making guide” when constructing your figures, and figure caption! (below instructions on page 4)
  4. There are video links below to the class lectures that have some explanation also to help with the software experiment.
1. What is the optimal pH of the enzyme? (10 points)
Insert a Figure (including figure caption) here showing V0vs pH.
 In a short paragraph, explain the process of acquiring the data set (amount of substrate used, number of trials), and its significance.
2. Calculate the values of V0 at or close to Vmax and at or close to Vmax10 at the optimal pH, quoting your answers as mean +/- standard error. Comment on your answer. (10 points) Note after the lab on Sept 30, I got totally confused with this question when I tried to clarify it for you, I am sorry.    The formula of course to calculate is   V0 = Vmax or V0 = Vmax10 of course you do not need to use the MM equation… and the rest is to report individual values from the individual trials and the average, as seen below.
Insert a table here (including figure caption) showing the raw V0values from the 5 trials as well  as mean of  V0values  +/- standard deviation. 
In a short paragraph, explain the process of acquiring the data set (amount of substrate used, number of trials), and its significance.
3. Calculate the values of Km and Vmax you obtained for this enzyme at its optimal pH or pH7, and indicate how you derived them. (20 points)
Insert a table here (including figure caption) showing Km and Vmax as determined by the simulation’s iterative fitting method. 
Insert the Double reciprocal plot (Lineweaver-Burk plot, (include figure caption)) you generated from the raw dataset.  
4. Calculate the specific activity of this enzyme at its optimal pH or pH 7 in units of mmol product min-1 (mg of enzyme)-1, showing your working clearly. (10 points)
Type your answer here

https://lh3.googleusercontent.com/z0mIrG_uCk-iRx1Gf3ASdaAVDGdsQcepKvzyGTHmURQZhVhxSeDX1ar_7H4ERGWcZ_9T0FDBlrFPHqYAVbPE5HtGx_PXFNlJh7fAntDSSFiOqtnR6u1_7Aa3nDXVkT0Yu9SrL6rE
Title: Should capture the essence of the figure (experiment), Do not simply restate the axis labels with a “versus” written in between. For lab reports the figure title is part of the caption.

Caption: Should be concise but comprehensive. It should describe the data shown, draw attention to important features contained within the figure, and may sometimes also include interpretations of the data.

Labels: use to label different plots on the same graph. Only use if it is needed to aid the interpretation of the figure.

Axis titles: describe the dependent and independent variable of the experiment.  Do not forget ti have the units included.

In a very plain language, a good figure Caption answers the following questions with one short sentence each.

What did you measure?

           How did you measure it, what was the methodology?

           What did you get, summarise the main results of the figure?

           What does it mean, explain results if necessary?

Keep in mind every figure should “stand on its own” itmust be sufficiently clear, well-labelled, and described by its caption to be understood by your intended audience without reading the lab write up. 

Questions frequently arise about how much methodology and what results  to include in the caption. Figures are part of lab reports ( posters, manuscripts). Detailed methodology will be described in the Methods section. Specific results will be reported in the results text with a reference to the applicable Table or Figure. For BCEM 2201 worksheets imagine that your figures belong in a full lab report and report only the main result of the figure in the caption. 

How to fit the linear part of the calibration curve on excel

When you make a calibration curve, sometimes towards the higher concentrations the curve will plateau. that part should be omitted when you find the line of best fit, but still, it should be presented on the figure. 

Here is how you can manage to fit only the datapoint on the linear region with the line of best fit: 

The set highlighted represents the orange points on the figure, and it was generated by “doubling up the A measurement of the linear points. Now the orange dot can be fitted with a linear threadline. 

Make sure when you add the threadline you set intercept as 0 (this step of course assumes when you measured the A of your standard you used the blank appropriately and zeroed the spec.) If your line is not trending toward zero, it is an indication not blinking properly, or of a wrongly prepared blank. 

https://lh4.googleusercontent.com/XryFg1CdpNy7lmDhgLhSmBR26bMu21jQuA8Ezl7DHywu21E65vIiiv-jcZ3raKAu_dgXnx9QXC4ZlgHJd0WDHCjdEcHY7OcjVMcMlw_9JoHNXtk3bpbeXkjGwGNr_2GmakGw_T01
https://drive.google.com/file/d/1ofI-i9mhsuQUE_MQyBCSiBjNcXfvvWm7/view?ts=5f7956c4
Research Paper 101
Calculate your paper price
Pages (550 words)
Approximate price: -

Reasons to trust Research Paper 101

On Time Delivery

We pride ourselves in meeting the deadlines of our customers. We take your order, assign a writer but allow some more time for ourselves to edit the paper before delivering to you. You are guaranteed a flawless paper on a timely manner...

24x7 Customer Live Support

Our team at Research Paper 101 is committed to handling your paper according to the specfications and are available 24*7 for communication. Whenever you need a quick help, you can talk to our writers via the system messaging or contact support via live chat and we will deliver your message instantly.

Experienced Subject Experts

Online Experts from Research Paper 101 are qualified both academically and in their experiences. Many are Masters and Phd holders and therefore, are qualified to handle complex assignments that require critical thinking and analyses...

Customer Satisfaction

We offer dissertation papers as per students’ wishes. We also help craft out the best topics and design concept papers. By ordering with us, you are guaranteed of defending and making through those hard faced professors in the defense panel!

100% Plagiarism Free

We at Research Paper 101 take plagiarism as a serious offence. From the start, we train our writers to write all their papers from scratch. We also check if the papers have been cited appropriately. Our website also has a tool designed to check for plagiarism that has been made erroniusly. In essense, the paper you get will be 100% legit...

Affordable Prices

We understand that being a student is very challenging, some students balance between work and studies in order to survive. We therefore offer pocket friendly rates that are very competitive in the market.

Try it now!

Calculate the price of your order

Total price:
$0.00

How it works?

Follow these simple steps to get your paper done

Place your order

Fill in the order form and provide all details of your assignment.

Proceed with the payment

Choose the payment system that suits you most.

Receive the final file

Once your paper is ready, we will email it to you.

Our Services

No need to work on your paper at night. Sleep tight, we will cover your back. We offer all kinds of writing services.

Essays

Essay Writing Service

No matter what kind of academic paper you need and how urgent you need it, you are welcome to choose your academic level and the type of your paper at an affordable price. We take care of all your paper needs and give a 24/7 customer care support system.

Admissions

Admission Essays & Business Writing Help

An admission essay is an essay or other written statement by a candidate, often a potential student enrolling in a college, university, or graduate school. You can be rest assurred that through our service we will write the best admission essay for you.

Reviews

Editing Support

Our academic writers and editors make the necessary changes to your paper so that it is polished. We also format your document by correctly quoting the sources and creating reference lists in the formats APA, Harvard, MLA, Chicago / Turabian.

Reviews

Revision Support

If you think your paper could be improved, you can request a review. In this case, your paper will be checked by the writer or assigned to an editor. You can use this option as many times as you see fit. This is free because we want you to be completely satisfied with the service offered.

error: