David Vejar, Vincent Baker, Michael Phan, Richard Rutledge Ms.

Chulapatrcheevin Biology HL 2 2 October 2013 The Effects of Salinity/Salt Concentrations on Enzymatic Activity Research Question: How is the rate of enzymatic activity affected by various external factors? Introduction: Enzymes are responsible for expediting chemical reactions, acting as catalysts. As catalysts, the enzymes lower the activation energy required for the reaction to occur. The surface area of reactions in also increased within the Enzyme-Substrate Complex. However, enzymes can only function in optimal conditions, meaning that factors such as concentration, pH, temperature, and salinity. These factors affect the operational faculties of the enzyme and can be optimized to either increase or decrease the speed of the reaction. In the context of our lab, we will be working with salt concentration and at the specific levels where the enzyme will denature or catalyze the reaction. However, the type of enzyme/substrate can play a role in determining the rate at which the complex denatures, especially when considering high vs. lower amounts of salt. For catalase and salinity, higher concentrations of salt will not enable the reaction to occur at the proper rate since the R groups of the proteins attract to each other and denature the protein, while too little salt will denature the protein since the Sodium and Chloride ions will interfere with the bonds of the tertiary proteins, which therefore does not allow the reaction to occur. However, in between too high of a molality and too low of a molality, the enzyme-substrate complex will be able to produce products from reactant (in this case, catalase will produce the maximum hydrogen peroxide possible). Hypothesis:___________________________________________________________________ ______________________________________________________________________________ ______________________________________________________________________________ ______________________________________________________________________________ IV: Salt Concentration (in %) DV: Enzymatic activity Operational Definition: Time (in seconds) Materials: Tweezers Pipette Filter Paper Disks Hydrogen Peroxide (H2O2) Salt Solution

Catalase Solution Large, Clear Plastic Cups Wells (if available)

Procedure: Day 1: 1. Obtain 10mL of catalase using a pipette and place into the well. 2. Obtain x% salt solution from glass beakers located in the station and mix with the catalase in the well. a. Note: If a well is not available, it is ok to mix within one of the plastic cups as well. 3. Obtain 50mL of hydrogen peroxide and pour into one of the plastic cups. 4. Dip one of the filter paper discs within the mixed solution. 5. Place the soaked disc inside the cup filled with hydrogen peroxide. 6. Observe the disc until it rises to the surface of the hydrogen peroxide and record qualitative data (such as bubble formation, rate of appearance, etc.). 7. Record the amount of time it takes for the disc to rise. 8. Repeat steps 2-6 two more times for the same concentration. 9. Repeat steps 2-7 for each concentration of catalase + salt solution. Day 2: 1) 2) 3) 4) 5) 6) Obtain 10mL of the Catalase Solution and place into well/cup. Obtain 10mL of unknown A and mix with the catalase. Obtain 50mL of hydrogen peroxide and pour into the plastic cup Dip one of the filter paper discs within the unknown concentration of salt + catalase. Place the soaked disc inside the cup filled with hydrogen peroxide. Observe the disc until it rises to the surface of the hydrogen peroxide and record qualitative data (such as bubble formation, rate of appearance, etc.). 7) Record the amount of time it takes for the disc to rise. 8) Repeat steps 2-7 two more times for the chosen unknown concentration. 9) Repeat steps 2-8 for unknown B 10) Based on the results from day one, predict the percent concentration of salt within both unknowns.

Data Table:
Time it takes for a paper disc dipped in a catalase + salt solution to float to the surface of an H2O2 solution. % Salt 0.02 0.06 0.1 0.14 0.16 Unknown A Unknown B Trial 1 Trial 2 Trial 3 Average Qualitative Data

Based on the information obtained, unknown solution A has ______% concentration of salt and unknown B has ______% of salt.