Transmittance between an amylase and starch solution

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Table 1: Color and Percent Transmittance between an amylase and starch solution with a pH of 6.0 tested in 3 minute intervals

Time

Color

Percent (%)Transmittance

Average Rate per minute

3

Dark Navy

0

6

Dark Navy

0

0

9

Dark Navy

0

0

12

Brown

0.2

0.0667

15

Dark Orange

8.2

2.6667

18

Orange

28.3

6.7

21

Orange

62.3

11.3333

24

Orange

67.4

1.7

27

Bright Orange

79.1

3.9

30

Bright Orange

85.3

2.0667

33

Bright Orange

87.7

0.8333

36

Bright Orange

90.2

-0.6333

39

Bright Orange

88.3

-0.6333

42

Bright Orange

83

-1.7667

45

Orange

74.4

-2.8667

48

Bright Orange

88.4

4.6667

51

Bright Orange

100

3.8667

Average rate=percent transmittance at t 2 - percent transmittance at t1 / t 2 - t 1

Figure 1. Amount of light transmitted through a pH of 6.0 over time in 3 minute intervals

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Table 2: Color and Percent Transmittance between an amylase and starch solution with a pH of 7.0 tested in 3 minute intervals

Table 2

Time

Color

Percent (%) Transmittance

Average Rate per minute

3

Dark Navy

0

6

Dark Navy

0

0

9

Blue/Brown

0.1

0.0333

12

Dark Orange

3.5

1.1333

15

Orange

21.3

5.9333

18

Light Orange

67.1

15.2667

21

Light Orange

58.8

-2.7667

24

Light Orange

72.3

4.5

27

Bright Orange

76.3

1.3333

30

Bright Orange

81.1

1.6

33

Bright Orange

77.6

-1.1667

36

Bright Orange

85.5

2.6333

39

Bright Orange

73.4

-4.0333

42

Bright Orange

91.2

5.9333

45

Bright Orange

73.8

-5.8

48

Bright Orange

86.8

4.3333

51

Bright Orange

95.4

2.8667

54

Bright Orange

97.6

0.7333

57

Bright Orange

98.9

0.4333

60

Bright Orange

98.8

-0.0333

63

Bright Orange

91.8

-2.333

66

Bright Orange

100

2.7333

Average rate=percent transmittance at t 2 - percent transmittance at t1 / t 2 - t 1

Figure 2. Amount of light transmitted through a pH of 7.0 over time in 3 minute intervals

Table 3: Color and Percent Transmittance between an amylase and starch solution with a pH of 7.0 tested in 3 minute intervals

Table 3

Time

Color

Percent (%) Transmittance

Average Rate per minute

3

Dark Navy

0

6

Dark Navy

0

0

9

Blue/Brown

0.2

0.667

12

Dark Orange

10.1

3.3

15

Orange

33.8

7.9

18

Light Orange

48.6

4.9333

21

Light Orange

56.4

2.6

24

Light Orange

77.6

7.0667

27

Bright Orange

78.2

0.2

30

Bright Orange

83.7

1.8333

33

Bright Orange

80.1

-1.2

36

Bright Orange

88.5

2.8

39

Bright Orange

67.3

-7.0667

42

Bright Orange

82.2

4.9667

45

Bright Orange

78.2

-1.3333

48

Bright Orange

85.5

2.4333

51

Bright Orange

94.8

3.1

54

Bright Orange

82.4

-4.1333

57

Bright Orange

100

5.8667

Average rate=percent transmittance at t 2 - percent transmittance at t1 / t 2 - t 1

Figure 3. Amount of light transmitted through a pH of 7.0 over time in 3 minute intervals

Table 4: Color and Percent Transmittance between an amylase and starch solution with a pH of 8.0 tested in 3 minute intervals

Table 4

Time

Color

Percent (%) Transmittance

Average Rate per minute

3

Dark Navy

0

6

Dark Navy

0

0

9

Dark Brown

0.1

0.0333

12

Dark Brown

0.2

0.0333

15

Dark Orange

4.9

1.5667

18

Orange

19.4

4.8333

21

Orange

34.4

5

24

Orange

27.4

-2.3333

27

Orange

48.9

7.1667

30

Light Orange

58.1

3.0667

33

Light Orange

62.3

1.4

36

Light Orange

72.9

3.5333

39

Orange

64.6

-2.7667

42

Orange

78.7

4.7

45

Orange

73.5

-1.7333

48

Orange

74.1

0.2

51

Light Orange

80.8

2.2333

54

Bright Orange

87.3

2.1667

57

Bright Orange

88.5

0.4

60

Bright Orange

87.1

-0.4667

63

Bright Orange

87.8

0.2333

66

Bright Orange

90

0.7333

69

Bright Orange

90.5

0.1667

72

Bright Orange

82.4

-2.7

75

Bright Orange

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78.9

-1.1667

78

Bright Orange

76.4

-0.8333

81

Bright Orange

93.1

5.5667

84

Bright Orange

94

0.3

87

Bright Orange

94.1

0.0333

90

Bright Orange

100

1.9667

Average rate=percent transmittance at t 2 - percent transmittance at t1 / t 2 - t 1

Figure 4. Amount of light transmitted through a pH of 8.0 over time in 3 minute intervals

Table 5: Color and Percent Transmittance between an amylase and starch solution with a pH of 8.0 tested in 3 minute intervals

Table 5

Time

Color

Percent (%) Transmittance

Average rate per minute

3

Dark Navy

0

6

Dark Navy

0

0

9

Dark Navy

0

0

12

Dark Navy

0

0

15

Dark Navy

0

0

18

Dark Navy

0

0

21

Dark Navy

0

0

24

Dark Navy

0.1

0.0333

27

Dark Brown

0.1

0

30

Dark Orange/Brown

0.1

0

33

Orange/Brown

0.4

0.1

36

Dark Orange

1.4

0.3333

39

Dark Orange

3.5

0.7

42

Dark Orange

11.1

2.5333

45

Dark Orange

10.5

-0.2

48

Dark Orange

16.2

1.9

51

Dark Orange

21.3

1.7

54

Dark Orange

26.4

1.7

57

Dark Orange

32.7

2.1

60

Dark Orange

33.5

0.2667

63

Orange

45.7

4.0667

66

Orange

46.3

0.2

69

Orange

42.1

-1.4

72

Orange

64.8

7.5667

75

Orange

52.5

-4.1

78

Orange

54.3

0.6

81

Orange

54.6

0.1

84

Orange

53.8

-0.2667

87

Orange

50

-1.2667

90

Orange

55.5

1.8333

93

Light Orange

65.7

3.4

96

Light Orange

62

-1.2333

Average rate=percent transmittance at t 2 - percent transmittance at t1 / t 2 - t 1

Figure 5. Amount of light transmitted through a pH of 9.0 over time in 3 minute intervals

Table 6. Total Rate transmittance per minute of different pH groups.

Varying pH

Rate transmittance per minute

pH 6.0

1.961 (% transmittance/min)

pH 7.0

1.515 (% transmittance/min)

pH 7.0 (control)

1.754 (% transmittance/min)

pH 8

1.111 (% transmittance/min)

pH 9

0.646(% transmittance/min)

Total Rate = total transmittance / ending time

Figure 6. Percent Transmittance of an amylase starch solution in 3 minute intervals with varying pH

Starch Hydrolysis Discussion

This experiment was conducted to show if varying the pH in an environment will change the rate of the reaction between starch and amylase. The pH of the starch amylase solution was the independent variable, and the amount of light being transmitted through the solution after adding iodine was the dependent variable. The tables and figures determine the reaction rate by plotting the percentage of light transmitted versus time measured to get an equation. The null hypothesis of varying the pH of the enzyme solution will not affect the reaction between starch and amylase is in incorrect according to tables 1 -5. The varying pH groups in a solution had different rates according to table 6. The results showed that the first pH to reach 100% was the pH of 6.0. According to table 1 it took 51 minutes for the pH 6.0 solution to reach 100% transmittance. The solution of pH 7.0 took 57 minutes to reach 100% transmittance according to table 3. Only 6 minutes from the pH of 6.0. According to table 2 the second pH of 7.0 (control) showed 100% transmittance at 66 minutes. The solution of pH 8.0 took 90 minutes to reach 100% transmittance according to table 4, most likely because it is more basic. The pH of 9.0 was either a dark navy or dark orange for most of the experiment. The solution of pH of 9.0 did not reach 100% transmittance. According to table 5 the pH of 9.0 solution took a total of 96 minutes to reach a transmittance of only 62 %.The pH of 9.0 solution ran out before it reached 100%. Pertaining to the data in figure 5 it took almost 40 minutes before much light was transmitted for pH of 9.0. The solution of pH 9.0 had the slowest rate of the 5 varying pH solutions; the rate was 0.646 according to table 6. The pH with the fastest rate was pH 6.0 with a rate of 1.961.

Citing the data from figure 6 the pH 0f 9.0 had the lowest percent transmittance. Not too far behind was the pH of 8.0. The solutions with a pH of 6.0, 7.0 and the control group were close in percent transmittance.

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Conclusion

In conclusion the hypothesis for the starch, amylase experiment was correct. Yes varying the pH of the enzyme solution will affect the rate of the reaction between starch and amylase. The 5 different ph groups, pH 6.0,7.0, 7.0 (control) 8.0 and 9.0 used in this experiment all had different reaction rates and different times when the percent transmittance reached 100%.It was discovered that the data dropped a few times. This could have been because the calibration was not completed enough. This also could have been because cuvettes may have had finger marks and not properly wiped off. Another question to be proposed would be, does varying the pH in different solutions change the reaction rate

between starch and amylase?