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Seung Soo (Jason) Lee 002213-065

Internal Assessment – Investigating the Relationship between Concentration of Sodium Chloride and the Rate of Reaction of Enzyme Amylase

Research Question: How will changing the percentage of sodium chloride concentration affect the rate of reaction of enzyme amylase, measured using the absorbance of starch and iodine with a spectrophotometer.

Introduction: Amylase is an enzyme that is involved in the human digestive process. Found in both the human pancreas and the human saliva, amylase breaks down starch into sugar so that large molecules can be easily digested1. Like all enzymes, amylase must be kept in a certain condition in order to function properly. In this experiment, the effect of sodium chloride concentration on the rate of reaction of amylase will be investigated with the use of starch and iodine. When starch is mixed with iodine, the coils of beta amylose molecules found in starch trap iodine, causing the mixture to turn into a shade of blue-black. 2 When starch is broken down into glucose, however, the monosaccharide does not react with iodine. Therefore, glucose does not change color even when it’s mixed with iodine. Correspondingly, when drops of amylase are inputted into a blueblack mixture of starch and iodine, the starch molecules will be broken down into glucose molecules, causing the mixture to turn colorless. Thus, the rate of reaction of amylase correlates to the absolute value of the rate of change in absorbance of the solution. A rapid decrease in the absorbance of the blue-black color equates to a high rate of reaction of amylase, whereas a slow decrease in absorbance signifies a low rate of reaction. In this experiment, an external variable of sodium chloride will be manipulated into the amylase enzyme to determine the effect the concentration of sodium chloride on the rate of reaction of amylase. Rate of Reaction = │

1

"Amylase." Wikipedia. N.p., n.d. Web. 12 Jan 2011. <http://en.wikipedia.org/wiki/Amylase>. 2

Senese, Fred. "How Does Starch Indicate Iodine?." N.p., 15 Feb 2010. Web. 6 Jan 2011. <http://antoine.frostburg.edu/chem/senese/101/redox/faq/starch -as-redox-indicator.shtml>.


Seung Soo (Jason) Lee 002213-065

Hypothesis: As aforementioned, amylase, like all enzymes, must be kept under a certain set of conditions in order to function properly. Factors such as pH level, temperature, and salt concentration could all denature the enzyme and decrease its activity. . When a substrate can no longer bind to the active site of an enzyme due to its conformational change, the enzyme activity and the rate of reaction of the enzyme drops significantly. For instance, a high concentration of sodium chloride would alter the electrostatic interactions between charged amino acids, causing conformational change in the enzyme and destroying its active site.3 Furthermore, the presence of sodium chloride will only have little impact on the enzyme structure unless the sodium chloride concentration is very high, when it could completely denature the enzyme. Therefore, an enzyme should experience an exponential decrease in its rate of reaction as the concentration of sodium chloride is increased Rate of Reaction of Amylase, Abss-1 Concentration of Sodium Chloride, %

Figure 1: Prediction of the Effect of Sodium Chloride Concentration on Rate of Reaction of Amylase Enzyme

Thus, the hypothesis for this experiment is that if the sodium chloride concentration is increased, then the rate of reaction of amylase will decrease. A high concentration of sodium chloride will denature the enzyme amylase and, as a result, it will no longer be able to break down starch into glucose. The figure above demonstrates that the average rate of change in absorbance will undergo an exponential decrease as the concentration of sodium chloride is increased.

3

"Rule of Protein Structure." N.p., n.d. Web. 6 Jan 2011. <http://users.rcn.com/jkimball.ma.ultranet/BiologyP ages/D/DenaturingProtein.html>.


Seung Soo (Jason) Lee 002213-065

Variables:

Variable

Description

Units / range

Method of Measuring / Manipulating

Independent

Concentration of sodium chloride

%

The independent variable will be manipulated by a process of serial dilution, from 20% concentration of sodium chloride to 10%, 10% to 5%, 5% to 1%, and 1% to 0.1%.

Dependent

Rate of reaction of amylase

│ (ΔAbss-1)

Controlled

This will be measured with a spectrophotometer and Logger Pro. Because amylase breaks down starch into glucose, and glucose does not react with iodine, the enzyme activity of amylase will lower the blue-black absorbance of starch+iodine. Therefore, the rate of decrease in absorbance over time correlates to the absolute value of the rate of reaction of amylase. The change in absorbance will be measured from 0-20 seconds, and the rate of reaction can be calculated by finding the slope of the absorbance vs. time graph. The uncertainty can be considered negligible.

Concentration of starch & iodine

%

This will be kept constant by using the same mixture created through steps 1-3 of procedures for every trial. (0.05% starch + 300μl of iodine)

Amount of solutions inside the cuvette

μl

For every trial, 2.5ml of starch & iodine solution and 500μl of sodium chloride & amylase solution is put inside the cuvette.

Temperature

°C

Temperature is kept constant by conducting the experiment at room temperature (about 25 °C) for every trial.

Table 1: List of Variables


Seung Soo (Jason) Lee 002213-065

Apparatus and Materials:        

      

Electronic balance (±0.001g) 100 cm3 & 10 cm3 volumetric flasks 10 cm3 pipette (±0.02 cm3) 1000 μl & 50 μl micropipettes 3 cm3 cuvettes 2 cm3 micro tubes Microcentrifuge Five small beakers for serial dilution

1 medium sized beaker Sodium Chloride Iodine Starch Hot plate Vernier Spectrophotometer Logger Pro

Procedures: Preparation of 0.05% starch mixed with iodine 1. 0.05g of starch and 100cm3 of distilled water are poured into a medium sized beaker. 2. The beaker is placed on a hot plate, and then stirred several times using a plastic stirrer until a homogenous solution is made. 3. 300μl of iodine is put into the starch solution. It is stirred several times using a plastic stirrer until a blue-black solution is made.

Preparation of sodium chloride of various concentrations (serial dilution) 8 cm3 distilled water

3

5 cm distilled water

5 cm3

20%

5 cm3

10%

2 cm3

5%

9 cm3 distilled water

1 cm3

1%

Figure 2: Serial Dilution of Sodium Chloride Solution

0.1%


Seung Soo (Jason) Lee 002213-065 4. 2g of sodium chloride and 10cm3 of distilled water is poured into a small beaker. 5. The beaker is stirred several times using a plastic stirrer until a homogenous solution is made, creating a 20% sodium chloride solution. 6. 5 cm3 of the obtained solution is transferred into another small beaker using a 10 cm3 pipette, and another 5 cm3 of distilled water is added into the beaker. The beaker is then stirred using a stirrer until a homogenous solution is made, creating a 10% sodium chloride solution. 7. The serial dilution of sodium chloride is continued, according to the layout in figure 2, to obtain 5%, 1%, and 0.1% concentrations of sodium chloride solutions.

Conducting the experiment 8. The Vernier spectrophotometer is calibrated using distilled water. Then, the wavelength at which to measure the absorbance is determined using the maximum wavelength of the blueblack mixture of starch and iodine. 9. 450μl of 20% sodium chloride solution is put inside a micro tube using a 1000μl micropipette. 50μl of amylase solution is added into the micro tube using a 50μl micropipette. The micro tube is then placed inside a microcentrifuge so that the solution will mix together. 10. Step 9 is repeated three times for all concentrations of sodium chloride, creating three mixtures of sodium chloride and amylase for each of the five variables. 11. 2.5cm3 of the starch & iodine mixture is put into a 3cm3 cuvette using a micropipette. 500μl of the sodium chloride & amylase mixture is added into the cuvette using a micropipette. 12. The solution is squeezed in and out three times using the micropipette to ensure that amylase spreads throughout the starch solution. After mixing three times, the “start” button on Logger Pro is clicked. Steps 11 and 12 are performed with the cuvette placed inside the spectrophotometer to minimize error. 13. The rate of change in absorbance of the mixture is measured using Logger Pro for 20 seconds. 14. Steps 11-13 are repeated for triplicate trials for all five concentrations of sodium chloride.


Seung Soo (Jason) Lee 002213-065

Data Collection:

Qualitative Data: ď Ź

Even with the naked eye, one could observe the disappearance of color inside the cuvette, from a dark, blue-black coloration to a clear, colorless state.

Quantitative Data:

*** Refer to the Appendix for a complete table of raw data from Logger Pro.


Seung Soo (Jason) Lee 002213-065

Data Processing: Rate of Decrease in Absorbance / ΔAbss-1

Sodium Chloride Concentration /% 20.0

Trial 1

Trial 2

Trial 3

-0.001240

-0.001280

-0.001223

10.0

-0.001517

-0.001299

-0.001402

5.0

-0.001812

-0.001523

-0.0012304

1.0

-0.001836

-0.001703

-0.001714

0.1

-0.001845

-0.001985

-0.001931

Control (no sodium chloride): -0.002830 Table 2: Rate of Decrease in Absorbance for All Trials5 Sodium Chloride Concentration / %

Calculation

Average Rate of Reaction (±Standard Deviation)6 / ΔAbss-1

20.0

0.001248 ± 0.000029

10.0

0.001406 ± 0.000109

5.0

0.001668 ± 0.000204

1.0

0.001751 ± 0.000074

0.1

0.001920 ± 0.000071

Control (no sodium chloride): 0.002830 ΔAbss-1 Table 3: Calculation of Average Rates of Reaction

**Because the rate of reaction must be a positive value, the average rate of reaction was taken as an absolute value. 4

This value was neglected in data processing because it was considered as an outlier.

5

The rate of decrease in absorbance was determined by finding the slope of absorbance vs. time graph

using linear regression on Logger Pro software. 6

The processing of standard deviation is shown in table 4


Seung Soo (Jason) Lee 002213-065

Data Presentation:

LEGEND Run 4: Control (no sodium chloride) Run 5: 20% sodium chloride Run 10: 10% sodium chloride Run 11: 5% sodium chloride Run 14: 1% sodium chloride Run 19: 0.1% sodium chloride

Figure 4: Graph of Raw Data from Logger Pro7

7

Slopes of lines that have values closest to the average slope value for each concentration of sodium chloride is shown in boxes.


Seung Soo (Jason) Lee 002213-065

Average Rate of Reaction / Î&#x201D;Abss-1

Effect of Sodium Chloride Concentration on the Rate of Reaction of Amylase 0.00185

0.0016 y = -3E-05x + 0.0018 R² = 0.914 0.00135

0.0011 0

5

10

15

Sodium Chloride Concentration / % Figure 5: Graph of Average Rates of Reaction against Concentration of Sodium Chloride8 9 8 9

Vertical error bars represent standard deviation for triplicate trials. Though they are difficult to discern, horizontal error bars represent the absolute uncertainty of sodium chloride concentration.

20


Seung Soo (Jason) Lee 002213-065

Uncertainties: Standard Deviation: Rate of Reaction of Amylase / ΔAbss-1

Average / ΔAbss-1 (±Standard Deviation)

Sodium Chloride Concentration /%

Trial 1

Trial 2

Trial 3

20.0

-0.001240

-0.001280

-0.001223

0.001248 ± 0.000029

10.0

-0.001517

-0.001299

-0.001402

0.001406 ± 0.000109

5.0

-0.001812

-0.001523

-0.00123010

0.001668 ± 0.000204

1.0

-0.001836

-0.001703

-0.001714

0.001751 ± 0.000074

0.1

-0.001845

-0.001985

-0.001931

0.001920 ± 0.000071

Table 4: Standard Deviation at Different Concentrations of Sodium Chloride

Example of Standard Deviation Calculation: [Sodium Chloride Concentration] = 20%

≒ 0.000029 Same calculations were done for 10%, 5%, 1%, and 0.1% sodium chloride concentrations.

10

This value was neglected in data processing because it was considered as an outlier.


Seung Soo (Jason) Lee 002213-065

Uncertainty due to dilution of glucose solution: *Uncertainty due to 10cm3 pipette = ±0.02 cm3 Concentration of Glucose / %

20.000

Volume of sodium chloride solution added / cm3 – 3

Uncertainties Volume of distilled Total percentage 3 water added / cm error for concentration of glucose / % – – 3

Absolute uncertainty for concentration of glucose / % –

10.000

5.00 ± 0.02cm = 5.00 ± 0.4%

5.00 ± 0.02cm = 5.00 ± 0.4%

±0.80

0.008

5.000

5.00 ± 0.02cm3 = 5.00 ± 0.4%

5.00 ± 0.02cm3 = 5.00 ± 0.4%

±0.80

0.004

1.000

2.00 ± 0.02cm3 = 2.00 ± 1%

8.00 ± 0.02cm3 = 8.00 ± 0.25%

±1.25

0.013

0.100

1.00 ± 0.02cm3 = 1.00 ± 2%

9.00 ± 0.02cm3 = 9.00 ± 0.22%

±2.22

0.011

Table 5: Uncertainty for Concentration of Glucose Solution

Sodium Chloride Concentration (±Uncertainty) / %

Average Rate of Reaction (±Standard Deviation) / ΔAbss-1

20.000

0.001248 ± 0.000029

10.000 ± 0.008

0.001406 ± 0.000109

5.000 ± 0.004

0.001668 ± 0.000204

1.000 ± 0.013

0.001751 ± 0.000074

0.100 ± 0.011

0.001920 ± 0.000071

Table 6: Combined Uncertainties for Independent & Dependent Variables


Seung Soo (Jason) Lee 002213-065

Conclusions: The hypothesis was supported by the results to the extent that an increase in sodium chloride concentration decreased the rate of reaction of enzyme amylase. However, the decrease in the rate of reaction was not exponential; rather, the relationship between NaCl concentration and average rate of reaction was pretty linear. As sodium chloride concentration increased, the average rate of reaction decreased at a fairly constant rate. Furthermore, once extrapolated, the graph in figure 5 demonstrates that the rate of reaction will be 0 when the sodium chloride concentration is at 60%. From this data, one could conclude that the enzyme amylase will completely cease to catalyze reactions at NaCl concentration of 60%.

Average Rate of Reaction / Î&#x201D;Abss-1

In the hypothesis, it was stated that a slight presence of sodium chloride will not affect the rate of reaction of amylase significantly, but as the concentration of sodium chloride increases, the enzyme will undergo a rapid decrease in its rate of reaction. This is due to the fact that, as more sodium chloride ions are present in amylase, the ions associate with oppositely charged groups in the enzyme protein, increasing protein hydration and denaturing the enzyme.11 Contrary to the hypothesis, where the rate of reaction was predicted to undergo a slight decrease up until a certain concentration of sodium chloride, then a rapid decrease as the concentration is at a level high enough to denature the enzyme, the graph below displays the fact that even 0.1% of sodium chloride was enough to largely decrease the rate of reaction of amylase. Although the 0.1% sodium chloride did not completely denature amylase, it was still enough to cause the greatest decrease in the rate of reaction of amylase.

Effect of Sodium Chloride Concentration on the Rate of Reaction of Amylase 0.003 0.0025 0.002 0.0015 0.001 0

5

10

15

20

Sodium Chloride Concentration / % Figure 6: Graph Demonstrating the Relationship between Sodium Chloride Concentration and Rate of Reaction, Including the Control 11

"Protein Denaturation." N.p., n.d. Web. 7 Jan 2011. <http://class.fst.ohio state.edu/FST822/lectures/Denat.htm>.


Seung Soo (Jason) Lee 002213-065

Evaluation: Overall, the results of this experiment seem fairly accurate and reliable. There are no striking outliers – except for the one value shown in table 2 – and although the standard deviations are bit sizeable for some values, they are not critical enough to negate the conclusions drawn. As the model in figure 5 represents, the relationship between sodium chloride concentration and the rate of reaction of amylase is clearly a negative correlation. On a separate note, while it is true that the best-fit line in figure 5 is a linear one, the best-fit line for the graph in figure 6 would more likely be an exponential one. Going back to one of the conclusions drawn, the relationship shown in figure 6 represents an exponential decrease because of the fact that the control is also included in the graph. The jump from 0% sodium chloride to 0.1% sodium chloride is largely significant – more significant than any of the other increases in sodium chloride concentration. Thus, such results encourage the next experiment to, perhaps, incorporate an even smaller concentration of sodium chloride. The results of this experiment support the idea that a miniscule NaCl concentration such as 0.1% was still significant enough to disrupt the electrostatic bonds within the enzyme. In order to observe the effect of NaCl concentration on the activity of enzyme more efficiently, it would be apt to utilize even more miniscule concentrations of sodium chloride. The sizeable nature of the standard deviation could be caused by the discrepancy created by human error. Although a standard was set at the beginning of the experiment, to mix the amylase and starch in the cuvette – in & out using the micropipette three times – then pressing “start” on Logger Pro, this process posed the biggest error throughout the experiment. The time taken between the moment when enzyme amylase was put into the cuvette – thus starting to interact and break down starch – and the moment when the “start” button was clicked varied, though only by little, for every trial. Furthermore, mixing the solutions in the cuvette three times – and taking up time in the process – may have been a bad idea, for that time could have been sufficient for the enzyme to do all of its work. Moreover, another problem during the procedures could have occurred with the mixing of amylase with sodium chloride. Because 15 separate micro tubes had to be filled one by one, and then mixed through the microcentrifuge one by one, some of the amylase solutions in the micro tubes had longer time to interact with sodium chloride. This could have meant longer time for the sodium chloride to denature the enzyme, thus lowering its rate of reaction. Though it’s not certain, this could have been another source of error in the experiment. Overall, however, this investigation was successful in terms of the accuracy of its results. The increased presence of sodium chloride did lower the enzyme activity of amylase, as predicted in the hypothesis, and as accepted as a scientific fact. Although improving on minor errors could strengthen the investigation, the experiment successfully produced consistent and reliable data, leading up to a solid conclusion.


Seung Soo (Jason) Lee 002213-065

Improving the Investigation: Error

Impact

Improvement

Time discrepancy between the moment amylase is inserted into the cuvette and Logger Pro reading is started

It could have allowed more time for amylase to break down starch in some trials than in others, causing differences in rate of reaction from trial to trail and increasing standard deviation

There are a few ways to improve this error. One way would be to get a help of another person, allowing him to press the “start” button on Logger Pro as soon as the amylase is mixed three times. Another method would be to simply take out the mixing process, and start the Logger Pro reading as soon as amylase is inserted into the cuvette.

Time discrepancy in the amount of time sodium chloride was allowed to interact with amylase between each trial

It allowed more time for the sodium chloride in some micro tubes to denature amylase than in other micro tubes, allowing the possibility for further decrease in the enzyme activity for amylase used in some trials compared to other trials.

All 15 micro tubes could be incubated for an allotted amount of time – around 30-40 minutes – to equalize the amount of time that sodium chloride is allowed to interact with amylase.

450μl of sodium chloride solution was inputted into the micro tube, while only 50μl of amylase solution was inputted, causing imbalance

As mentioned in the conclusion, the results demonstrate a huge decrease from no sodium chloride to 0.1% sodium chloride. This suggests that too much sodium chloride was incorporated throughout the experiment, compared to the amount of amylase. The abundance of sodium chloride could have disrupted the enzyme activity of amylase too much.

The amount of sodium chloride solution and the amount of amylase solution could be balanced, to about 250μl each used for every trial. This change could perhaps produce results that are closer to those that were hypothesized.

10 cm3 pipette used during serial dilution of sodium chloride

Decreased precision & increased range of uncertainty

Since only about 1.5ml of each sodium chloride concentration was necessary for the experiment, a micropipette could have been used to perform the serial dilution, which would have lowered the range of uncertainty.

Table 7: Ways to Improve the Investigation


Seung Soo (Jason) Lee 002213-065

Appendix: 20% Trial 1 Time (s)

20% Trial 2 Time (s)

0

Abs 614.6nm 0.325344

1

20% Trial 3 Time (s)

0

Abs 614.6nm 0.332057

0.323938

1

2

0.322158

3

10% Trial 1 Time (s)

0

Abs 614.6nm 0.313663

0.329859

1

2

0.328055

0.320197

3

4

0.319295

5

10% Trial 2 Time (s)

0

Abs 614.6nm 0.324849

0

Abs 614.6nm 0.328899

0.312331

1

0.324127

1

0.322839

2

0.310414

2

0.327022

2

0.323331

0.326182

3

0.308982

3

0.32371

3

0.323634

4

0.325192

4

0.306824

4

0.317347

4

0.318245

0.317086

5

0.321327

5

0.30595

5

0.316302

5

0.317459

6

0.316004

6

0.322082

6

0.303372

6

0.311371

6

0.315669

7

0.31459

7

0.32031

7

0.301999

7

0.312996

7

0.315408

8

0.31396

8

0.319333

8

0.301567

8

0.310745

8

0.314665

9

0.312072

9

0.317908

9

0.299804

9

0.309092

9

0.313663

10

0.310708

10

0.315929

10

0.298299

10

0.309422

10

0.311851

11

0.309642

11

0.314813

11

0.297477

11

0.309973

11

0.310488

12

0.308286

12

0.313737

12

0.296372

12

0.305913

12

0.309019

13

0.306788

13

0.312848

13

0.294631

13

0.303481

13

0.307299

14

0.305986

14

0.311556

14

0.29378

14

0.302649

14

0.305913

15

0.304786

15

0.31034

15

0.29332

15

0.301747

15

0.305404

16

0.303916

16

0.309459

16

0.292472

16

0.300774

16

0.304278

17

0.302866

17

0.308469

17

0.291767

17

0.298908

17

0.303553

18

0.302396

18

0.307774

18

0.290852

18

0.298586

18

0.302758

19

0.301495

19

0.306788

19

0.29029

19

0.297549

19

0.301639

20

0.301098

20

0.305658

20

0.289237

20

0.296479

20

0.300631

10% Trial 3 Time (s)

5% Trial 1 Time (s)

0

Abs 614.6nm 0.32863

1

5% Trial 2 Time (s)

0

Abs 614.6nm 0.339073

0.327137

1

2

0.324811

3

5% Trial 3 Time (s)

0

Abs 614.6nm 0.322687

0.333645

1

2

0.329398

0.322271

3

4

0.323028

5

1% Trial 1 Time (s)

0

Abs 614.6nm 0.346723

0

Abs 614.6nm 0.320573

0.320762

1

0.342346

1

0.320573

2

0.316787

2

0.34045

2

0.31675

0.328362

3

0.314999

3

0.341792

3

0.312109

4

0.328055

4

0.314108

4

0.339938

4

0.310892

0.320875

5

0.322536

5

0.31115

5

0.333917

5

0.30781

6

0.32178

6

0.320988

6

0.31012

6

0.332986

6

0.30635

7

0.31847

7

0.31832

7

0.31034

7

0.331864

7

0.302613

8

0.315743

8

0.316675

8

0.306642

8

0.331285

8

0.301711

9

0.314293

9

0.316302

9

0.305113

9

0.330668

9

0.299194

10

0.311814

10

0.313515

10

0.303843

10

0.329744

10

0.299266


Seung Soo (Jason) Lee 002213-065 11

0.311556

11

0.312183

11

0.3033

11

0.328515

11

0.296978

12

0.31012

12

0.310561

12

0.301567

12

0.327787

12

0.294701

13

0.308506

13

0.308359

13

0.299445

13

0.326144

13

0.292014

14

0.306605

14

0.307189

14

0.298514

14

0.325039

14

0.290817

15

0.306532

15

0.306059

15

0.299338

15

0.3239

15

0.289623

16

0.304641

16

0.304097

16

0.298192

16

0.323255

16

0.288467

17

0.303952

17

0.303807

17

0.296408

17

0.322498

17

0.287106

18

0.303264

18

0.302469

18

0.295625

18

0.321667

18

0.286131

19

0.302721

19

0.30135

19

0.294276

19

0.321365

19

0.285609

20

0.301387

20

0.300092

20

0.294063

20

0.320235

20

0.286235

1% Trial 2 Time (s)

1% Trial 3 Time (s)

0

Abs 614.6nm 0.337622

1

0.1% Trial 1 Time (s)

0

Abs 614.6nm 0.337035

0.334461

1

2

0.332212

3

0.1% Trial 2 Time (s)

0

Abs 614.6nm 0.342465

0.33485

1

2

0.33256

0.33009

3

4

0.327749

5

0.1% Trial 3 Time (s)

0

Abs 614.6nm 0.341121

0

Abs 614.6nm 0.326678

0.340529

1

0.340253

1

0.323066

2

0.337152

2

0.335317

2

0.321252

0.329898

3

0.333762

3

0.331246

3

0.318695

4

0.327864

4

0.331169

4

0.328208

4

0.316712

0.325192

5

0.325915

5

0.329475

5

0.326831

5

0.31333

6

0.323824

6

0.324241

6

0.326946

6

0.324925

6

0.311224

7

0.321365

7

0.322271

7

0.325687

7

0.322536

7

0.308872

8

0.319859

8

0.320423

8

0.323407

8

0.320047

8

0.307189

9

0.317721

9

0.317908

9

0.321554

9

0.317983

9

0.305258

10

0.315743

10

0.316302

10

0.319145

10

0.316339

10

0.302938

11

0.314331

11

0.314776

11

0.317684

11

0.314145

11

0.301423

12

0.31333

12

0.313293

12

0.316228

12

0.312737

12

0.300056

13

0.311888

13

0.311851

13

0.314702

13

0.310819

13

0.29812

14

0.310782

14

0.310488

14

0.313218

14

0.309679

14

0.296194

15

0.309422

15

0.308579

15

0.311298

15

0.308469

15

0.294595

16

0.308213

16

0.307372

16

0.31012

16

0.306642

16

0.29279

17

0.30635

17

0.306532

17

0.308909

17

0.304822

17

0.291696

18

0.304931

18

0.304822

18

0.30708

18

0.303409

18

0.290044

19

0.303011

19

0.303988

19

0.306205

19

0.302252

19

0.288992

20

0.302541

20

0.302685

20

0.30504

20

0.300056

20

0.287315


Seung Soo (Jason) Lee 002213-065 Maximum Absorbance Wavelength (nm) 400

Abs 0.140983

Wavelength (nm) 523.68

403.5

0.143298

407

Control (No NaCl) Abs

Time (s)

0.269512

Wavelength (nm) 629.44

0.506694

0

Abs 614.4nm 0.356633

526.6

0.276394

632.4

0.505749

1

0.351468

0.144298

529.52

0.284868

635.36

0.502435

2

0.352439

410.5

0.143858

532.44

0.293012

638.32

0.499854

3

0.345206

414

0.144867

535.36

0.303899

641.28

0.497412

4

0.341121

417.5

0.145449

538.28

0.313062

644.24

0.493168

5

0.34116

421

0.144269

541.2

0.322568

647.2

0.492372

6

0.337113

424.5

0.15575

544.12

0.332483

650.16

0.488485

7

0.337543

428

0.16144

547.04

0.34552

653.12

0.482216

8

0.329129

431.5

0.168923

549.96

0.356925

656.08

0.480191

9

0.327175

435

0.171384

552.88

0.367604

659.04

0.474983

10

0.323634

438.5

0.177023

555.8

0.378477

662

0.47441

11

0.321214

442

0.18106

558.72

0.389967

664.96

0.469595

12

0.318058

445.5

0.185164

561.64

0.402134

668

0.464303

13

0.314479

449

0.184681

564.56

0.413991

671

0.461757

14

0.312368

452.5

0.164762

567.48

0.424809

674

0.455878

15

0.311999

456

0.184657

570.4

0.435809

677

0.451873

16

0.309202

459.5

0.184207

573.32

0.446248

680

0.448952

17

0.306168

463

0.183719

576.24

0.456774

683

0.445418

18

0.304133

466.5

0.19358

579.16

0.46569

686

0.438446

19

0.303011

470

0.198735

582.08

0.47381

689

0.43383

20

0.301639

473.5

0.205575

585

0.48009

692

0.428318

477

0.209701

588

0.487579

695

0.425969

480.5 484

0.213411 0.216026

590.96

0.490925

698

0.42161

593.92

0.492981

701

0.416259

487.5

0.218792

596.88

0.501117

704

0.411412

491

0.221762

599.84

0.504044

494.5

0.224311

602.8

0.507599

498

0.227361

605.76

0.511082

501.5

0.232694

608.72

0.510439

505

0.235745

611.68

0.51301

508.5

0.240517

614.64

0.514248

512

0.245664

617.6

0.512766

514.92

0.253401

620.56

0.510628

517.84

0.258245

623.52

0.509945

520.76

0.264801

626.48

0.508682

Abs


Amylase enzyme reaction