Saturday, November 16, 2013

Cellular Respiration Lab



Purpose: 
In this lab, we wanted to test the whether germinated seeds and non-germinated seeds would respire, and if so, the rate at which they do so.  Also, we wanted to see how temperature impacted rate of respiration as well.  

Introduction: 
Cellular respiration is the process in which organic molecules are broken down in order to create energy for an organism.  Cell respiration occurs aerobically, which mean that oxygen is necessary for the process to occur.  Cell respiration also occurs in the mitochondria.  Before respiration begins  glucose is broken down into two molecules of pyruvate in a process called glycolosis.  If oxygen is present, then respiration will occur next.  But if oxygen isn't present, fermentation will occur.  When oxygen is present, the two pyruvate molecules are converted to AcetylCoA, which is a coenzyme.  One molecule of CO2 is released in this stage.  Next, the citric acid cycle (or Kreb's cycle) occurs.  In this stage of respiration,  2 molecules of CO2 are released as a byproduct of the breakdown of pyruvate. Two pyruvates are present, so each molecule creates one ATP and 2 molecules of CO2.  Next, the pyruvates undergo oxidative phosphorylation.  In this stage, the electrons carried by NADH (NADH collects extra electrons from molecules throughout respiration) we released onto the electron transport chain, and are subsequently carried down to increasingly more electronegative molecules.  The spillover of extra electrons in this stage fuels the transport of H+ ions across the membrane of the mitochondria, creating a large difference in energy.  When the H+ ions travel across the membrane to once again, Chemiosmosis happens.  The movement of H+ ions across the membrane to a lower concentration create energy for the synthesis of ATP.  32 to 34 ATP are created in this stage.  Overall, 2 ATP are created during glycolosis, 2 ATP are created during the citric acid cycle, and 32 to 34 ATP are created during oxidative phosphorylation.  A way to see if respiration occurs is to monitor the output of CO2 in organisms, because CO2 is emitted in both the conversion of pyruvate to AcetylCoA and the citric acid cycle.
Germination of seeds is the process in which seeds become active and able to grow.  This happens because seeds that are dehydrated intake water, and therefore, enzymes that need water to function are able to do so once again.  A germinated seed can have shoots coming off of it, and a non- germinated seed looks shriveled and water-less.
                                                 
                                                        Non-germinated seeds
                                               
                                                            Germinated seeds

Methods:
First, we collected germinated and non-germinated pea seeds.  In order to germinate the peas, we soaked them in water overnight.  Then weproceeded to test the level of CO2 emitted from several different seed types.  Here's how we did it:
Next, we separated 25 germinated, 25 non-germinated, and 25 glass beads (this was used for a control) into separate containers.

Second, we tested each of those materials for the presence of CO2 over a ten minute time span.


Third, we put the 25 germinated peas into ice-cold water for approximately ten minutes, in order of I test the affect of tempurature of germinated peas on levels of CO2 output.  We then tested the cold peas for presence of CO2 as well.



Lastly, we created graphs showing the CO2 output over ten minutes.



Data:
       




Graphs and Charts:

                                                          CO2 Release vs. Time Graph
    



Discussion: 
In this lab, we tested cellular respiration in different types of seeds. We did this by calculating the amount of CO2 emitted by the seeds, since CO2 is a product of cellular respiration. Our group tested peas that were germinated at room temperature, peas that were germinated in ice water, and dormant seeds. We also used glass balls as a control group. Our results showed that at 0.80 ppm/s, peas germinated at room temperature emitted the most carbon dioxide, showing that they have the highest rate of cellular respiration. The germinated peas in ice water had a lower rate, at 0.71 ppm/s, showing that cellular respirations is still occurring in lower temperatures, just at a lower rate. The dormant peas were drastically lower, with a rate of 0.15 ppm/s. Our control group of glass beads had a rate of 0.10 ppm/s, showing that a small amount of carbon dioxide could have been coming from other places, such as the water trapped in the bottle, it could've been leaking into the bottle, or it could have been trapped in the bottle from the previous test. Overall, our results showed that a germinated cell has a higher rate of cellular respiration than a non-germinated seed, as well as there is a direct relationship between temperature and respiration: as temperate goes up, so does respiration; as temperature goes down, so does respiration. 

Conclusion: From this experiment we can conclude that the non-germinated seeds do not respire, as shown by the very little amount of CO2 that was being given off by the non-germinated seeds. As for the germinated seeds we found that when the seeds were at room temperature that they respired at the highest rate.  The cold seeds had a lower amount of CO2 then the room temperature seeds. In summary when the temperature is higher the respiration of CO2 is higher than those of seeds in colder environments and that non-germinated seeds do not respire.

References: 
http://plantsinmotion.bio.indiana.edu/plantmotion/earlygrowth/germination/germ.html
Campbell Biology, 9th edition


Monday, November 4, 2013

Enzyme Catalysis Lab


Purpose:
The purpose of this experiment is to understand the functions of enzymes and how pH changes the reaction rate. This can be done by observing the enzyme catalase in the break down of hydrogen peroxide (H2O2) into water (H2O) and oxygen gas (O2), and measuring the amount of oxygen generated and the rate of the enzyme-catalyzed reaction.

Introduction:
Enzymes are proteins that act as catalysts in biochemical reactions; catalysts affect the rate of a chemical reaction. In enzyme-catalyzed reactions, the substrate binds to the active site of the enzyme, and the activation energy of the reaction is lowered so that the produced can be formed more easily. Ways in which enzyme action can be affected include salt concentration, temperature, pH, and activations/inhibitors. 

Methods:
The main point in this lab was to understand enzyme activity. We did this by collecting baseline trials of the rate of reaction of the uncatalyzed reactions. Next we would use the same process to collect data of the catalyzed reactions, after we had added the enzyme Catalase. Then we compared the trials of the two types of reactions to one another and observed what happened to the rate of reaction.

Data:

                     
 

Graphs:


                              



Discussion:
 In this lab, we tested the affect of denaturing enzymes and how that relates to the rate of reaction between two substances.  The substances we tested were catalase,which is an enzyme, and hydrogen peroxide, which is a substrate.  In order to denature the enzyme, we added an acid, H2SO4.  The results we got indicated a direct curve, with a few exceptions.  When aci was added after 90 seconds, there was a slight decrease in rate of decomposition, which did not fillow the trend of increasing rate of reaction.  Also, at 180 seconds, there was a slight decrease as well which did not follow the trend.  This could be caused by a few things.  One, we may have put the acid in slightly too early, which would have caused this decrease.  Two, we may have contaminated the beaker with leftover acid from other experiments and the denaturing could have occurred too early, causing less decomposition to occur.  But, on average, we observed that as an enzyme is left with a substrate for a longer time, more decomposition occurs.  Therefore, length of time the substrate is exposed to the enzyme and amount of decomposition are directly related.  

Conclusion:
We can conclude that concentration of enzymes directly related to rate of decomposition of H2O2.
References:

Campell Biology, 2009