Enzyme project data and results- Duaa, Audrey, Elena

Elena M, Audrey M, and Duaa K

Question

How do abiotic or biotic factors influence the rates of enzymatic reactions (chemical reactions that are assisted by enzymes)?

Background

Enzymes speed up chemical reactions by lowering activation energy (that is, the energy 

needed for a reaction to begin). In every chemical reaction, the starting materials (the 

substrate(s) in the case of enzymes) can take many different paths to forming products. 

For each path, there is an intermediate or transitional product between reactants and 

final products. The energy needed to start a reaction is the energy required to form that 

transitional product. Enzymes make it easier for substrates to reach that transitional 

state. The easier it is to reach that state, the less energy the reaction needs. 

Enzymes are biological catalysts. They are large protein molecules, folded so that they 

have very specifically shaped substrate binding sites. These binding sites make substrates 

go into the transition state. To catalyze the reaction, several regions of the binding site 

must be precisely positioned around the substrate molecules. Any change in the shape of 

the overall folded enzyme molecule can change the shape of the binding site. 

The optimum reaction conditions are different for each enzyme. The correct 

environmental conditions, proper substrates, and, often, particular cofactors associated 

with an enzyme are needed. In some instances, the optimum conditions can be deduced 

fairly accurately based on the following:

• The organism from which the enzyme is derived
• The part of the organism in which the enzyme functions
• The environmental conditions in which that organism lives

Take the example of lactase, an enzyme that catabolizes (breaks down) the disaccharide sugar lactose into two monosaccharides, glucose and galactose. In humans, lactase is found mostly in the small intestine, where the pH is around 7. It would be reasonable to hypothesize that human lactase is optimally active at pH 7 and at 37°C (normal human core body temperature in degrees celsius). Free-living decomposer fungi in soil also produce lactase. However, soil pH usually is between 5 and 6.5. As could be predicted, the purified enzyme from a common soil fungus has a pH optimum of 5.5. The main enzyme for this lab, peroxidase, is found in many different forms, with optimum pHs ranging from 4 to 11 depending on the source and optimum temperatures varying from 10 to 70°C.

Purpose

In this experiment you will investigate the effect of environmental factors on the enzyme hydrogen peroxidase. This enzyme is found in all aerobic (using oxygen) cells and functions to decompose hydrogen peroxide into O2(g) and H2O. The specific environmental factors you will test (as a class) are temperature, pH, substrate concentration, and enzyme concentration. Your team will select one of these factors (variables) to test and report on.

Materials

• 5 to 10 grams of freshly picked ripgut bromegrass (Bromus diandrus) blades (about 1 handful)
• Digital balance (scale)
• Mortar and pestle
• Distilled water
• 3 100-liter glass or plastic beakers
• 1 mL or 5 mL syringe
• Hydrogen peroxide
• 1 Paper towel square (for filtration)
• Glass test tubes
• Test tube rack or holder
• Small plastic ruler
• Safety glasses

Depending on which environmental factor you choose to investigate, some of the following items will be needed for your experiment:

• Acid solutions with pH values between 2 and 6
• Alkaline solutions with a pH between 8 and 12
• pH test strips
• Ice
• Large plastic beaker (for ice bath)
• Hot water
• Large plastic beaker (for hot water bath)
• Thermometers

Procedure

This part will be determined by the students conducting the experiment. For an overview of the general scientific experimentation and research process, see the flow diagram below. Use the worksheet that follows to write out in detail the hypothesis you are seeking to test, the materials you will use, and the steps you will follow to conduct your experiment.

Enzyme Lab Worksheet

Hypothesis: If changing the temperature level affects how the enzymes function, then increasing the temperature of the ripgut brome grass juice and peroxide will speed up the reaction, while decreasing the temperature will slow the reaction compared to the room temperature control.

Independent Variable: The temperature

Dependent Variable: The height of the foam in cm. 

Controlled Variables: Grass juice and water.

Justification of hypothesis: We think this because when the enzymes come into contact will different temperatures then the enzymes will either speed up or slow down depending on the water temperature.

Why did you choose this as your hypothesis?

We wanted to observe the overall reaction of the temperature differences that would either speed up or slow down the process of the catalyst. 

Materials (Your Team’s Experiment):

• 8.62 ml grams of grass
• 30  milliliters of water
• Test tubes
• Test rack 
• Paper towels 
• Guacamole thing (mortar and pestle)
• Syringe 
• Thermometer 
• Ruler

Procedure: Our group will be mixing a certain amount of distilled water and grass to make grass juice. We will then add different temperature mixtures to see whether the reaction goes faster or slower, and what the reaction is. 

Summary: 

     For our experiment we put the grass juice in the water and overall concluded that the temperature does affect the way the enzymes work. We know this because during our experiment we tested three different temperatures of water. This data showed that the room temperature water went the slowest in terms of how fast the bubbles appeared. While the cold slowed down our reaction time of the bubbles compared to the heated peroxide. Lastly, the fastest was the heated water which dramatically speed up the reaction time of the bubbles. 

Detailed Steps:

     We started our experiment by grinding up 8.62 grams of grass and in total added 30 milliliters of water to the grass. By mashing the two together in a mortar and pestle it formed a juice substance as (we referred to it as grass juice). Next we lined up 9 test tubes on a rack, having the room temperature, cold, and heated water each grouped in one section by temperature. After we put approximately 2 milliliters of grass juice  and 1 milliliter of hydrogen peroxide into each test tube. As we put the hydrogen peroxide in the cold temperature test tubes we watched the reaction. At 0 degrees celsius, 30 seconds in the foam/bubbles in the test tube had grown 0.2 centimeters. After 60 seconds at 0 degrees celsius the bubbles/foam moved up 0.8 centimeters, and after 90 seconds 1 centimeter. Lastly after 120 seconds (2minutes) the foam/bubbles came up to 1.5 centimeters. Next we tried the room temperature water (24 degrees Celsius). As we out the hydrogen peroxide into the test tube we began to watch the bubbles/foam. We observed the reaction from the room temperature test tubes and it showed that after 30 seconds the foam/bubbles was at 0.3 centimeters. Next after 60 seconds it was grown 0.5 centimeters, and after 90 seconds it had moved 1 centimeter. Finally after 120 seconds (2 minutes) it had moved to 1.5 centimeters. The last test we did was with the heated water; the temperature from that was 40 degrees celsius. For our first test after dumping the peroxide in we turned on the watch and watched it move up 0.5 centimeters. Soon 60 seconds passed and we watch is go up 1 centimeter, and after 90 seconds it had gone up 2 centimeters. In closing at 120 seconds (2 minutes) it had gone up to 2.3 centimeters. After we had gotten all of our data from the experiment we washed out all of the materials we had used for the lab.

Data and Results:

(This section should include at least one graph!)

	24 Degrees Celsius	0 Degrees Celsius	40 Degrees Celsius
30 seconds	0.3 cm	0.2 cm	0.5 cm
60 seconds	0.5 cm	0.8 cm	1 cm
90 seconds	1 cm	1 cm	2 cm
120 seconds	1.5 cm	1.5 cm	2.3 cm
max	4.5cm	4cm	7cm

https://drive.google.com/drive/folders/1V4TXAwrsEv1NV1UG2E-fR3BU6lnqodcD

Conclusions:

Overall, the heated peroxide mixed with the grass juice created foam faster rather than the room temperature peroxide and the cold peroxide. The cold peroxide mixed with the grass juice had a relatively same reaction rate as the room temperature, but the room temperature succeeded the colder peroxide after 2 minutes, so we can conclude that the cooler the temperature the slower the reaction will be.

 To start, the heated peroxide was to reach 7 cm, which was the tallest amount of foam. The heated hydrogen peroxide mixed with the grass juice had increased an average of 0.575 cm per 30 seconds.  The heated hydrogen peroxide sped up the reaction, we can tell this from the fact that more foam was able to form faster over the given amount of time. 

The room temperature experiment ( our control ) reached a maximum foam height of 5 cm and had an average reaction rate of 0.56 cm per 30 seconds.  

The cold hydrogen peroxide reached up to 4 cm. The average rate per 30 seconds for the cooled peroxide mixed with the grass juice was 0.4 cm per 30 seconds. The cooled hydrogen peroxide slowed down the reaction, we can tell this because the average rate of reaction is slower than the room temperature and the greatest height of the reaction is lower. 

  In closure, our experiment data has shown that when water is heated, cold, or just at room temperature, the temperature has an affect on how the enzymes are performing their job and how fast and how much of a reaction will occur.  When increasing the temperature the enzyme, reaction rate increases. When decreasing the temperature the enzyme, reaction rate will mildly decreases. We know that if we were to increase the temperature too much the enzyme will be denatured, but in this experiment we know that the enzyme was not denatured at 40 degrees celsius, but would most likely become denatured if we were to increase the temperature any more. 

Post Assignment #4: Audrey Malone

     Some abiotic factors that our broccoli plants are by our rocks, dirt, and the fence that lines the garden bed. The broccoli plants depend on dirt due to the fact that dirt has nutrients that the plants depend on for survival. Another abiotic factor that is important is water. Plants rely on water for survival because water is crucial for photosyntheis that creates energy that the plants rely on to survive . Lastly the fence is used to keep other things out of the garden, so the broccoli plants don't get stepped on by humans. Some biotic factors that our broccoli plants rely on are worms. Worms are important because they decompose dead plants and animals giving nutrients back into the soil. After the plants absorb the nutrients in the soil helping plants grow. Another biotic factor are pollinators, pollinators are helpful because when the broccoli begins to flower then, the pollen from other plants will spread on the broccoli plant so it will blossom.
     I know that our broccoli plants are engaged in competition with weeds. We know this because they are competing for water and nutrients in the soil. The winner is decided through these by competitions by which plant lives and which plant dies. The determination is complicated because sometimes a plant can continue to grow even if it's losing nutrients or the plant could continue to grow, but end up being very small and malnourished. Another type of relationship that occurs with the broccoli is parasitism. This occurs if a slug is eating the leaves of the broccoli plant. Therefore the broccoli plant is getting nothing out of the relationship making it parasitism.
     Lastly our broccoli plant is involved in secondary succession. It seems like a primary producer(autotroph) because the broccoli plant can produce it's own food by using photosynthesis. Photosynthesis is based on water and  carbon dioxide . It is also secondary succession because the dirt and other plants were already growing in that ecosystem making it secondary succession. All in all I hope our broccoli plants continue to grow.

Post Assignment#3: Audrey Malone

     Our small broccoli plants have changed throughout time in many different ways. For example they have grown in the sense that they have gained some bigger leaves and grown a little taller. Although they haven't changed too much in their size their leaves have gained some darker shades of green on their leaves. Also physically they have thicker stems and will continue to grow. We now have a total of two healthy broccoli plants living in the garden.
     To begin our plants take part in the water cycle because as we water our plants they absorb the water from in the roots and preform transpiration. Also they can get water from other places for example they can get water from rain. Although there has not been much although that is a possibility. Also they could get water from other people who have watered the broccoli. Overall water is important to the plants because they use the water to do photosynthesis. Although the broccoli can also collect water from the leaves of the plants which drip down into the soil and the plants are able to absorb the water taken in by the roots, and broccoli can collect water from being watered by humans as well.
     Next the plants contribute to the carbon cycle by practicing photosynthesis. Where they take in the carbon dioxide and use the carbon for glycerol which later becomes ATP and a by product of this process is oxygen which humans use to breathe. Lastly our plants are helping the nitrogen cycle because they take in nitrogen through the soil and use it for nutrients. Without plants in the ecosystem the carbon dioxide in the air would most likely pile up and become fatal to humans. An example is fossil fuels contributing carbon dioxide to the air, and because of that the air is polluted with the carbon dioxide becoming dangerous.
     Next, our broccoli plants take part in the nitrogen cycle everyday. The nitrogen cycle is important because it is part of amino acids. Plants play an important role by collecting nitrogen from the soil. Also when the broccoli plants die they will release nitrogen back into the soil when they decompose. Therefore the nitrogen will be recycled, so the nitrogen cycle can start again.
     All in all our broccoli plants contribute to not only just for food later on, but also now through the many cycles they participate in, and what they contribute to the environment. As you can see not only our broccoli plants, but all other plants are important to the ecosystem we have created in our garden.

Post assignment #3 : Elena

Over the course of the week our two little seedlings are growing up into healthy broccolis. So far one of them has grown pretty tall and has two stems that are attached. Both seedlings have grown a little thicker and a little taller. They have grown many leaves that are heart shaped, making it easy to tell them apart from weeds. The seedlings are a light green color. We hope that our little brocs will continue their growth to become healthy big broccolis.

Out plants participate in the movement of water in the biosphere by absorbing water through their roots. This process is called root uptake. Our seedling’s leaves also catch rain and the water droplets are evaporated via: transportation. The water that our plant absorbs helps it with photosynthesis, growing and developing. The water gives the plant nutrients to grow. This has allowed our plant to grow taller and bigger leaves.

Our plants participate in the movement of carbon in the biosphere by absorbing it through its leaves and using the carbon and water for photosynthesis. Our plant uses the carbon for photosynthesis to get the nutrients, which has allowed it to grow new leaves and get bigger. Usually plants will take in carbon dioxide but when plants die they loose all the carbon that they had stored and is released into the atmosphere.

Our plants participate in the Nitrogen cycle.  Bacteria in the soil converts the nitrogen to ammonia, the ammonia is then changed into nitrates which can then be taken up by the plants. The plants have gotten to grow bigger stems and bigger leaves which are taking to a heart shape. After the plant has absorbed the ammonium and nitrates using it for nutrients a process called ammonification happens. Bacteria in the soil will convert the nitrogen nutrients back into ammonia, from there bacteria will also convert the ammonia to nitrogen gas, this process is called denitrification. 

Post assignment #4 : Elena

Some abiotic factors that my plant depends on is water, sunlight, carbon dioxide, dirt, and nitrogen. A plant needs soil to grow in as well as get nutrients from it. Our plant needs water, sunlight, and carbon dioxide for photosynthesis, and a plant needs nitrogen for nutrients.  My plant also depends on some biotic factors.  Some of those biotic factors being other plants that compete with my plant for space, and water, bacteria which turns nitrogen into ammonia for our plant to absorb as nutrients, and bugs that might land on or even eat our plant.  

I know that my plants are engaged in competition with other plants for space in the soil, nutrients, sunlight, and water because other plants may be growing faster than mine. Or plants will start growing up for more sunlight rather than out. 

With the struggle of competition winners and losers are determined by what plants survive and grow nice and tall and healthy. And by what plants die or are stunted. Not always is it clear to see what plants “win” or “ lose”. Sometimes all the plants are genuinely similar.  Different plants grow to different standards.  It is hard to say who won or lost if you are comparing multiple species.  But in competition you can have one plant that grew taller and another plant that grew wider and spread out more.  There are different characteristics of a plant that shows that both won but in different categories. The taller one got more sunlight, and the more spread out one managed to get more water and nutrients from the soil. 

Not only do plants interact with each other, bacteria also has a relationship with plants.  In the soil the bacteria turns the nitrogen into ammonia and then into nitrates which the plant then uses for nutrients. 

In the garden there seems to be a secondary succession. There was a disturbance in the ecosystem, taking the weeds out of the area and shifting the soil. Then new life started growing from the soil.

Post Assignment #4 - Alia Latimer

There are several abiotic factors that contribute to our plants survival, and generally help it thrive. Many of these factors are aspects that make up the climate, such as the perception the plant receives as part of the water cycle. This can be received through natural means, such as rain or a river, but in our case it mostly comes through us watering our plants. Another would be the temperature, our plants being affected by how hot or cold the area is. 

One of the easiest ways I know that our plants are involved in competition is the fact that we have to weed the area fairly frequently. Before we started using the area there were lots of grass, and other volunteer plants, and even after we weeded the area, we still have to weed the area every couple days to keep other plants from sprouting in our little plot. So while our little plants may not have much competition because of our teams intervention, they still must compete to get the most water, space and sunlight.

In this sort of environment the winners and looses are determined by who gets to survive. Our plants are currently the winners, and the weeds that we have torn up are the looses. Of course it can be hard to know for sure who exactly wins, because often both organisms are injured or effected in some way. There also could be some unintended consequences, like the soil being diseased and that eventually killing all the plants that grow there, and spreading it to the others in that species.

While we can't see too much interaction, we have found evidence in the form of a worm. The worm would be These worms convert the organic material into the soil that the plant uses to grow up big and strong. There is also evidence of a predator prey relationship in the fact that we found a bite mark in one of the leaves. Obviously our little plants have been preyed on by some sort of insect, and there for has a relationship to the insect.

Since the soil still remains, there is definitely no primary secession, however there still is secondary secession at play. While there may have been no natural disaster, we did uproot the majority of the plant life that was living that area. We then brought in a new species that has been populating the area, along with some of the previous species that were inhabiting the area.

Post Assignment #3 - Alia Latimer

Over all our little seedlings haven't changed much. While it is true that their leaves have gotten longer, their stalks have gotten bigger, and just their overall size has increased there hasn't been much other change. It is important to note that the leaves have started to change in shape, and there are more leaves, that is where the changes stop. I do think that there will be more changes t occur, but currently they remain a pale green color.

In terms of the water cycle our little plants interact with it in the same way i assume most plants do. Obviously they are taking in the water that we give them and using it to grow and thrive, which then goes back into the environment through transpiration. There is also the less direct interaction of when the morning dew collects on the leaves on the plants, evaporation off later in the day, or being used by an insect or animal as a source of water. The plants also receive water on occasion from rain, which despite being scarce would have still been an interaction with the water cycle. These interactions have helped our broccoli to thrive and grow. As we saw in a previous attempt the water is essential to the growth of the plant throughout it's life, and especially in the earliest stages. Water is essential for all life and these little plants are not exception, which is why we have to make sure that they are constantly engaged in the water cycle through receiving water.

Plants as a variety have one of the most essential parts in the carbon cycle, which is recycling the carbon into oxygen. Our broccoli are responsible for taking in the carbon put out into the atmosphere by fires, human machinery, and animal respiration, and turn it back into breathable oxygen. They store up this carbon and use it for growing, only returning the carbon back to the earth through being burned, being eaten, or dying. Obviously our plants have been using the carbon to grow, since they have been getting larger slowly.

Plants again have a similar niche in the nitrogen cycle that they do in the carbon cycle. After bacteria  breaks down the nitrogen from the air into the soil, the plants use it to grow through assimilation. The nitrogen is then passed from the plants into the natural flow of energy in the food chain. So obviously as the plants live in the soil they will be taking in nitrogen, and when insects or humans eat them they will be passing it on.