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**Volume of inoculum in bioreactor: 100mL
**

Volume of Terrific Broth: 900mL

Bottle

Time(hr)

pH

T (OC)

pO2

OD

M1

M2

CDW,

X

(g/ML

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Time (hr)

0

1

2

0

1

2

3

4

6

8

10

12

14

16

18

20

22

24

7.03

7.05

6.96

7.20

7.51

8.05

8.25

8.38

8.44

8.44

8.43

8.42

8.39

8.36

8.33

36.70

37.06

37.04

37.00

37.04

37.00

37.01

37.00

37.00

37.00

37.00

37.00

37.00

37.00

37.00

39.9

51.6

46.0

74.8

63.4

61.0

44.5

55.5

66.0

70.4

72.2

73.2

73.5

70.9

67.2

0.327

1.160

2.280

2.555

2.733

2.814

2.806

2.743

2.711

2.705

2.715

2.701

2.697

2.703

2.708

Log OD

-0.4855

0.0645

0.3579

0.977

1.018

1.020

1.002

1.024

0.981

1.055

0.965

0.986

0.977

0.996

0.965

1.028

0.995

1.021

1.009

1.081

1.079

1.075

1.113

1.077

1.132

1.037

1.054

1.072

1.073

1.024

1.098

1.085

1.097

)

0.032

0.063

0.056

0.073

0.089

0.096

0.077

0.072

0.068

0.095

0.077

0.059

0.070

0.090

0.076

8781 0.6118 .3 4 6 8 10 12 14 16 18 20 22 24 0.4326 Time (hr) 0 1 2 3 4 6 8 10 12 14 16 18 Ln (X/Xo) 0 0.0881 0.6774 0.8247 1.4493 0.8781 0.4321 0.4366 0.4382 0.7538 1.4074 0.4315 0.4331 0.4481 0.4318 0.0229 1.4309 0.4338 0.5596 0.8109 0.0986 0.

8 f(x) = 0.13 R² = 0.5 f(x) = 0.6 ln (X/Xo) Ln(X/Xo) 0.8650 The graph of OD vs Time 3 2.05x + 1.5 0 0 5 10 15 20 25 30 Time (hr) Figure 1: The graph of optical density against time The graph of ln(X/Xo) vs Time 0.5 Optical Density (OD) Linear (OD) 1 0.20 22 24 0.5 2 2.9 R² = 0.4 Linear (Ln(X/Xo)) 0.3 2 OD 1.0341 0.2 0 0 0.6 0.5 1 1.28x + 0.5 Time (hr) Figure 2: The graph of ln (X/Xo) against time .7828 1.

12 0.04 CDW Linear (CDW) 0.06 f(x) = 0x + 0.The graph of ln X vs Time 0.12 0.07 R² = 0.06 Ln X Linear (Ln X) 0.02 0 0 5 10 15 20 25 30 Time (hr) Figure 4: The graph of cell dry weight against time .12 0.04 0.1 0.07 R² = 0.02 0 0 5 10 15 20 25 30 Time (hr) Figure 3: The graph of ln X against time The graph of CDW vs Time 0.08 ln X f(x) = 0x + 0.08 Cell Dry Weight (g) 0.1 0.12 0.

96x + 51.The graph of pO2 vs time 80 f(x) = 0.44 60 pO2 pO2 40 Linear (pO2) 20 0 0 5 10 15 20 25 30 Time (hr) Figure 5: The graph of pO2 against time .73 R² = 0.

The slope represent μ max of the reaction. slope of the curve is 0. So the concentration of cell is just simply dividing the CDW with 1 mL of media.Calculation 1 To calculate the TB used in this experiment: Prepared 1000 mL of TB.0007. Thus net growth rate for the TB broth is 0. Thus maximum growth rate for the TB broth is 0. It is state on the bottle of TB. Example: 0. From the figure 3. The slope represent μ net of the reaction.2798. 100 mL for inoculation and 900 mL for bioreactor. 4 To calculate the value of μnet : The graph of ln X against time is plotted. 3 To calculate the μmax : The graph of ln X/Xo against time is plotted. slope of the curve is 0. 5 To calculate the mass doubling time.032 g/mL The other value is calculated by using same method of calculation. 47. 2 To calculate the cell concentrations. From the figure 2. X: 4mL of media is transferred into centrifuge tube. X = = 0.2798 hr .0007 hr-1.6 g of TB is needed with 4mL of glycerol as a carbon source.032 g 1mL Cell concentrations.2798 hr-1. td : The equation used is: t d= ¿ ln2 μmax ln2 −1 0.

17 7.967 1.71 35.995 1.836 2.045 Dried Centrifuge Tube + Sample m2 0.750 2.63 8. Other group result: No.005 0.75 38.022 0.2 13 20 37.087 1.01 8.11 35.986 1.7 7 8 37.01 7.017 0.005 0.8 15 24 37.3 14 22 37.01 7.785 2. X (g/L) 0.985 1.019 0.004 1.019 0.5 9 12 36.9 3 2 37.4 Table 1: Temperature.027 1.980 1.3 6 6 37.909 2.978 0.1 5 4 37.020 0.3 11 16 36.020 0.349 2.1 10 14 37.020 0.16 19. Time (h) Absorbanc e Real OD Empty Centrifuge Tube m1 1 2 3 4 5 6 7 8 9 10 11 12 0 1 2 3 4 6 8 10 12 14 16 18 0.005 0.969 0.022 0.030 0.00 7.57 28.345 m2 – m1 Cell Dry Weight. pH reading and PO2 reading of the bioreactor during 24 hours.997 0.019 0.803 2.98 8.02 7.966 1.3 8 10 37.384 1.506 2.030 .93 14.038 0.4773 hr We have been told to compare the results between our group and another group that perform the LAB 2.006 0.024 0.017 0.010 0.2 4 3 37.009 0.014 0.988 0.00 6.6 12 18 37.70 49.688 2.90 9.5 2 1 37.00 7.76 45.05 11.00 7.968 0.989 1.955 2. Time (h) Temperature (oC) pH Reading PO2 (%) 1 0 37.985 0.006 0.00 8.106 1.313 2.003 1. No .019 0.00 8.08 23.00 8.00 8.010 0.¿ 2.005 0.009 0.98 8.020 0.41 7.022 0.46 20.018 0.01 8.

4789 ln X -5.2983 -5.5 0 0 5 10 15 Time 20 25 30 .4393 0. Coli based on 24 hour time using TB broth.977 0.3642 0.043 0.012 0.1823 0.5878 0.4816 1.1466 ln (X/Xo) 0 0 0.035 0.3990 0.8167 -3.9120 -3.1300 0.4294 0.4637 0.987 1.13 14 15 20 22 24 2.2983 -5.89 R² = 0.964 3.3862 1.4719 0.3350 1.9633 -3.06x + 1.015 0.4157 0.052 1.4476 0.4527 0.035 0.9459 2.9120 -3.46 3 2.9633 -3.2238 1.3862 1.972 1.6931 1.6052 -4.043 Table 2: The optical density and cell dry weight of bacteria E.3350 1.5 f(x) = 0.7917 1.4739 0.4706 0.0986 1.967 2.1160 -4.7105 -4.0745 -3.017 0.3524 -3.1518 Real OD vs Time 3. Time (hr) 0 1 2 3 4 6 8 10 12 14 16 18 20 22 24 Log OD -0.5066 -4.4448 0.5 1 0.4723 0.020 0.5 2 Real OD 1.015 0.1997 -3.

07x .78 1.ln (X/Xo) vs Time 2.78 -5 -6 Time 20 25 30 .4.5 ln ( X/X0 ) 1 0.5 0 0 5 10 15 20 25 30 Time ln X vs Time 0 0 5 10 15 -1 -2 ln X -3 -4 f(x) = 0.31 R² = 0.98 R² = 0.5 2 f(x) = 0.07x + 0.

02 0.83 R² = 0.11x + 11.05 0.04 f(x) = 0x + 0.39 PO2 30 20 10 0 0 5 10 15 Time 20 .78 0.03 CDW 0.01 0 0 5 10 15 20 25 30 25 30 Time PO2 vs Time 60 50 40 f(x) = 1.01 R² = 0.CDW vs Time 0.

To calculate the mass doubling time. Thus maximum growth rate for the TB broth is 0. td t d= ¿ : ln2 μmax ln2 −1 0. The slope of ln X against time represent for the TB broth is 0. To calculate the μmax : The slope of ln X/Xo against time represent μ max of the reaction.0743 hr-1. μ 2.3290 hr net of the reaction. Thus net growth rate .0743 hr ¿ 9.CALCULATION 1. 3.0743 hr-1.

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