Internal assessment

Candidate name : Sofía Rodríguez Zambrano

Candidate number: 2026145

Total of pages :

Enzymatic activity of lactase of the production of glucose at

temperatures of 35, 36, 38, 39 & 40 ºC.
INTRODUCTION:

Enzymes also known as biological polymers are the most diverse produced by living organisms, whose main
function is to act as a catalyst (rate increase) and speed up a chemical reaction (Metabolism includes all the
chemical activities that keep organisms alive). Because of being proteins they own a very stable original
structure, however, there are some external factors that affect their optimal activity and configuration, these
are pH, by changing electrical charge of proteins; Temperature because ability of proteins at some level of
heath; and cofactors: required by almost a third of the known enzymes and thus called coenzymes. Lactose (a
disaccharide or chain of two saccharides or sugars that will go divided in simple Monosaccharides Glucose
and Galactose after Enzyme activity final outcome), is found in Milk and Dairy products, it cannot be
absorbed by our intestines and it can be converted into glucose and galactose via the enzyme lactase; The
material on which the enzyme will act is called the substrate. The enzyme attaches to the substrate molecule
at a specific location called the active site. When the enzyme has attached to the substrate, the molecule is
called the enzyme-substrate complex. For example, the sugar found in milk is called lactose. With the aid of
the enzyme, lactase, the substrate, lactose, is broken down into two products, glucose and galactose by
Catalysis which is the process of modifying a chemical reaction with the use of a catalyst. Lactase is obtained
from Kluyveromyces lactis, or a type of yeast that grows naturally in milk. In humans, the average internal
temperature is 37.0 °C, and although a fever (pyrexia) could be considered when the body temperature is
above the normal temperature such as 38, 39, and 40 C°. In different circumstances is considered to be
hypothermia when the body temperature is below the normal one, such as a 36 and 35 C°.

The goal of this investigation is to determine the effects on glucose production in the lactase activity on its
substrate: Lactose. In my personal opinion, a lot of people consumes delactosed milk and don’t even know
why it is called like that, I was one of those persons, and it’s important to people to understand whether what
is lactase and why is it so important to take care of our health and not exceed or not reach the temperature of
37 ° C in the human body.
RESEARCH QUESTION:

To what extent the temperatures of 35,36,38,39 and 40°C effects the enzymatic activity of lactase on the

production of glucose that is derived from the interaction of lactase and lactose with a PH of 7?

HYPOTHESIS:

If the temperature of 37°C gets increased or decreased, then the production of glucose derived from the

interaction between lactase and lactose will decrease.

VARIABLES

Table 1. Variables

Type of variable CONTROLLED INDEPENDENT DEPENDENT

Variable PH AMOUNT OF TEMPERATURE PRODUCTION

LACTOSE OF GLUCOSE

Unit It does not have mg/dL °C mg/dL

MATERIALS AND PROCEDURE:

I. Materials:

● Lactase enzyme activity with data analysis simulator

https://exchange.iseesystems.com/public/jondarkow/mg/dL/index.html#page4

II. Procedure:

A. First I get into the simulator

B. Second, I put the following values:

1. 500 mg/dL of Lactose

2. PH of 7
 3. Temperature of 35 °C

C. Then, I ran the simulator 10 times.

D. Finally, I register the data on a table.

E. Repeat the steps B, C & D replacing the temperature with 36,38,39 & 40 °C

RAW DATA:

Lactose 500 mg, PH 7, Temperature 35 º C (with an uncertainty of ±1)

Glucose Production
Try
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE
2 106.00 181.00 168.00 199.00 80.00 166.00 162.00 216.00 87.40 127.00 149.24
4 258.00 291.00 294.00 336.00 190.00 334.00 246.00 375.00 199.00 308.00 283.10
6 326.00 412.00 400.00 399.00 314.00 393.00 359.00 426.00 275.00 396.00 370.00
8 406.00 459.00 451.00 441.00 395.00 418.00 423.00 457.00 346.00 447.00 424.30
10 426.00 475.00 474.00 475.00 450.00 460.00 458.00 482.00 412.00 469.00 458.10
12 447.00 481.00 481.00 485.00 474.00 482.00 471.00 489.00 456.00 483.00 474.90
14 468.00 488.00 488.00 490.00 480.00 492.00 483.00 494.00 468.00 491.00 484.20
16 482.00 492.00 494.00 493.00 486.00 495.00 490.00 496.00 484.00 495.00 490.70
18 489.00 495.00 497.00 496.00 492.00 496.00 494.00 497.00 492.00 497.00 494.50
20 493.00 496.00 498.00 497.00 496.00 498.00 497.00 498.00 496.00 498.00 496.70

Lactose 500 mg, PH 7, Temperature 36 º C (with an uncertainty of ±1)

Glucose Production
Try
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE
2 129.00 191.00 103.00 95.70 160.00 159.00 164.00 193.00 192.00 200.00 158.67
4 272.00 325.00 242.00 251.00 300.00 330.00 325.00 317.00 304.00 285.00 295.10
6 366.00 390.00 362.00 375.00 365.00 385.00 380.00 370.00 390.00 378.00 376.10
8 432.00 428.00 421.00 435.00 424.00 419.00 432.00 441.00 451.00 438.00 432.10
10 466.00 463.00 465.00 466.00 467.00 450.00 468.00 468.00 474.00 464.00 465.10
12 476.00 480.00 477.00 475.00 480.00 471.00 487.00 485.00 485.00 478.00 479.40
14 483.00 489.00 488.00 485.00 487.00 483.00 491.00 490.00 492.00 488.00 487.60
16 492.00 494.00 495.00 494.00 492.00 490.00 495.00 495.00 496.00 492.00 493.50
18 495.00 497.00 497.00 496.00 497.00 495.00 498.00 498.00 498.00 495.00 496.60
20 497.00 498.00 498.00 497.00 498.00 498.00 499.00 499.00 499.00 497.00 498.00

Lactose 500 mg, PH 7, Temperature 38 º C (with an uncertainty of ±1)

 Glucose Production
Try
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE
2 203.00 206.00 95.60 174.00 202.00 209.00 139.00 155.00 156.00 128.00 166.76
4 292.00 330.00 249.00 279.00 364.00 241.00 342.00 317.00 307.00 394.00 311.50
6 342.00 394.00 350.00 389.00 412.00 420.00 337.00 397.00 394.00 384.00 381.90
8 409.00 440.00 415.00 447.00 439.00 457.00 393.00 444.00 424.00 434.00 430.20
10 451.00 464.00 437.00 473.00 468.00 476.00 437.00 471.00 457.00 458.00 459.20
12 472.00 479.00 466.00 483.00 481.00 486.00 461.00 485.00 469.00 470.00 475.20
14 483.00 484.00 482.00 489.00 491.00 492.00 480.00 492.00 482.00 486.00 486.10
16 491.00 491.00 492.00 494.00 493.00 495.00 486.00 496.00 491.00 492.00 492.10
18 495.00 494.00 496.00 495.00 496.00 496.00 493.00 97.00 495.00 495.00 455.20
20 497.00 497.00 497.00 498.00 497.00 498.00 496.00 498.00 497.00 497.00 497.20

Lactose 500 mg, PH 7, Temperature 39 ºC (with an uncertainty of ±1)

Glucose Production
Try
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE
2 153.00 211.00 90.70 125.00 113.00 133.00 110.00 154.00 120.00 184.00 139.37
4 320.00 339.00 253.00 302.00 273.00 291.00 345.00 292.00 305.00 276.00 299.60
6 391.00 403.00 339.00 382.00 361.00 365.00 381.00 385.00 404.00 392.00 380.30
8 440.00 446.00 411.00 413.00 423.00 425.00 436.00 430.00 446.00 434.00 430.40
10 469.00 468.00 443.00 451.00 448.00 463.00 468.00 459.00 468.00 456.00 459.30
12 486.00 481.00 466.00 472.00 464.00 480.00 483.00 476.00 481.00 475.00 476.40
14 491.00 487.00 485.00 482.00 478.00 484.00 490.00 484.00 488.00 484.00 485.30
16 494.00 493.00 491.00 487.00 485.00 491.00 494.00 491.00 492.00 492.00 491.00
18 497.00 497.00 494.00 493.00 493.00 496.00 496.00 494.00 496.00 496.00 495.20
20 499.00 498.00 496.00 496.00 496.00 498.00 498.00 496.00 498.00 498.00 497.30

Lactose 500 mg, PH 7, Temperature 40 ºC (with an uncertainty of ±1)

Glucose Production
Try
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE
2 179.00 138.00 177.00 156.00 118.00 135.00 83.40 129.00 115.00 174.00 140.44
4 314.00 278.00 271.00 285.00 199.00 243.00 250.00 262.00 219.00 274.00 259.50
6 395.00 351.00 381.00 374.00 255.00 328.00 312.00 348.00 338.00 327.00 340.90
8 442.00 408.00 427.00 399.00 333.00 375.00 363.00 409.00 393.00 388.00 393.70
10 461.00 439.00 456.00 449.00 405.00 414.00 417.00 447.00 424.00 434.00 434.60
12 469.00 462.00 472.00 460.00 453.00 446.00 453.00 460.00 460.00 451.00 458.60
14 483.00 478.00 481.00 478.00 466.00 463.00 473.00 471.00 480.00 471.00 474.40
 16 488.00 485.00 489.00 487.00 478.00 471.00 482.00 480.00 487.00 481.00 482.80
18 494.00 491.00 492.00 493.00 487.00 479.00 487.00 489.00 492.00 488.00 489.20
20 496.00 495.00 495.00 496.00 497.00 489.00 492.00 494.00 496.00 493.00 494.30
The preceding tables shown, represent the production of glucose of the lactase enzyme activity with the

simulator, done with temperatures of 35,36,38,39 & 40 °C. The biggest value of these table was 498 mg/Ld

nevertheless the difference with the other tables was not prominent.

DATA PROCESSING:

For knowing the uncertainties for this type of values (average of glucose production) , it is necessary to use

the formula of the standard deviation:

σ =√ ❑
The following tables are the values of the raw data with standard deviation:
Lactose 500 mg, PH 7, Temperature 35 º C (with an uncertainty of ±1)
Glucose Production
Try STANDART
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE DEVIATION
2 106.00 181.00 168.00 199.00 80.00 166.00 162.00 216.00 87.40 127.00 149.24 46.80928683
4 258.00 291.00 294.00 336.00 190.00 334.00 246.00 375.00 199.00 308.00 283.10 59.98416458
6 326.00 412.00 400.00 399.00 314.00 393.00 359.00 426.00 275.00 396.00 370.00 49.51318567
8 406.00 459.00 451.00 441.00 395.00 418.00 423.00 457.00 346.00 447.00 424.30 35.19169725
10 426.00 475.00 474.00 475.00 450.00 460.00 458.00 482.00 412.00 469.00 458.10 22.94655627
12 447.00 481.00 481.00 485.00 474.00 482.00 471.00 489.00 456.00 483.00 474.90 13.51090095
14 468.00 488.00 488.00 490.00 480.00 492.00 483.00 494.00 468.00 491.00 484.20 9.484490263
16 482.00 492.00 494.00 493.00 486.00 495.00 490.00 496.00 484.00 495.00 490.70 5.01220732
18 489.00 495.00 497.00 496.00 492.00 496.00 494.00 497.00 492.00 497.00 494.50 2.718251072
20 493.00 496.00 498.00 497.00 496.00 498.00 497.00 498.00 496.00 498.00 496.70 1.567021236

Lactose 500 mg, PH 7, Temperature 36 º C (with an uncertainty of ±1)

Glucose Production
Try STANDART
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE DEVIATION
2 129.00 191.00 103.00 95.70 160.00 159.00 164.00 193.00 192.00 200.00 158.67 38.03185945
4 272.00 325.00 242.00 251.00 300.00 330.00 325.00 317.00 304.00 285.00 295.10 31.60326987
6 366.00 390.00 362.00 375.00 365.00 385.00 380.00 370.00 390.00 378.00 376.10 10.25724676
8 432.00 428.00 421.00 435.00 424.00 419.00 432.00 441.00 451.00 438.00 432.10 9.757618106
 10 466.00 463.00 465.00 466.00 467.00 450.00 468.00 468.00 474.00 464.00 465.10 6.100091074
12 476.00 480.00 477.00 475.00 480.00 471.00 487.00 485.00 485.00 478.00 479.40 5.059644256
14 483.00 489.00 488.00 485.00 487.00 483.00 491.00 490.00 492.00 488.00 487.60 3.134042473
16 492.00 494.00 495.00 494.00 492.00 490.00 495.00 495.00 496.00 492.00 493.50 1.900292375
18 495.00 497.00 497.00 496.00 497.00 495.00 498.00 498.00 498.00 495.00 496.60 1.264911064
0.816496580
20 497.00 498.00 498.00 497.00 498.00 498.00 499.00 499.00 499.00 497.00 498.00 9

Lactose 500 mg, PH 7, Temperature 38 º C (with an uncertainty of ±1)

Glucose Production
Try STANDART
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE DEVIATION
2 203.00 206.00 95.60 174.00 202.00 209.00 139.00 155.00 156.00 128.00 166.76 38.76192863
4 292.00 330.00 249.00 279.00 364.00 241.00 342.00 317.00 307.00 394.00 311.50 48.53234889
6 342.00 394.00 350.00 389.00 412.00 420.00 337.00 397.00 394.00 384.00 381.90 28.98064488
8 409.00 440.00 415.00 447.00 439.00 457.00 393.00 444.00 424.00 434.00 430.20 19.66836151
10 451.00 464.00 437.00 473.00 468.00 476.00 437.00 471.00 457.00 458.00 459.20 14.03012632
12 472.00 479.00 466.00 483.00 481.00 486.00 461.00 485.00 469.00 470.00 475.20 8.715248451
14 483.00 484.00 482.00 489.00 491.00 492.00 480.00 492.00 482.00 486.00 486.10 4.557045827
16 491.00 491.00 492.00 494.00 493.00 495.00 486.00 496.00 491.00 492.00 492.10 2.766867463
18 495.00 494.00 496.00 495.00 496.00 496.00 493.00 97.00 495.00 495.00 455.20 125.8621821
20 497.00 497.00 497.00 498.00 497.00 498.00 496.00 498.00 497.00 497.00 497.20 0.632455532

Lactose 500 mg, PH 7, Temperature 39 ºC (with an uncertainty of ±1)

Glucose Production
Try STANDART
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE DEVIATION
2 153.00 211.00 90.70 125.00 113.00 133.00 110.00 154.00 120.00 184.00 139.37 36.62349125
4 320.00 339.00 253.00 302.00 273.00 291.00 345.00 292.00 305.00 276.00 299.60 29.12120266
6 391.00 403.00 339.00 382.00 361.00 365.00 381.00 385.00 404.00 392.00 380.30 20.18277594
8 440.00 446.00 411.00 413.00 423.00 425.00 436.00 430.00 446.00 434.00 430.40 12.41146423
10 469.00 468.00 443.00 451.00 448.00 463.00 468.00 459.00 468.00 456.00 459.30 9.475699916
12 486.00 481.00 466.00 472.00 464.00 480.00 483.00 476.00 481.00 475.00 476.40 7.260241808
14 491.00 487.00 485.00 482.00 478.00 484.00 490.00 484.00 488.00 484.00 485.30 3.860051813
16 494.00 493.00 491.00 487.00 485.00 491.00 494.00 491.00 492.00 492.00 491.00 2.905932629
18 497.00 497.00 494.00 493.00 493.00 496.00 496.00 494.00 496.00 496.00 495.20 1.549193338
20 499.00 498.00 496.00 496.00 496.00 498.00 498.00 496.00 498.00 498.00 497.30 1.159501809

Lactose 500 mg, PH 7, Temperature 40 ºC (with an uncertainty of ±1)

 Glucose Production
Try STANDART
Minute 1 2 3 4 5 6 7 8 9 10 AVERAGE DEVIATION
2 179.00 138.00 177.00 156.00 118.00 135.00 83.40 129.00 115.00 174.00 140.44 31.20296282
4 314.00 278.00 271.00 285.00 199.00 243.00 250.00 262.00 219.00 274.00 259.50 33.2908062
6 395.00 351.00 381.00 374.00 255.00 328.00 312.00 348.00 338.00 327.00 340.90 39.95122026
8 442.00 408.00 427.00 399.00 333.00 375.00 363.00 409.00 393.00 388.00 393.70 31.47856413
10 461.00 439.00 456.00 449.00 405.00 414.00 417.00 447.00 424.00 434.00 434.60 19.02746553
12 469.00 462.00 472.00 460.00 453.00 446.00 453.00 460.00 460.00 451.00 458.60 8.058122196
14 483.00 478.00 481.00 478.00 466.00 463.00 473.00 471.00 480.00 471.00 474.40 6.669999167
16 488.00 485.00 489.00 487.00 478.00 471.00 482.00 480.00 487.00 481.00 482.80 5.573747991
18 494.00 491.00 492.00 493.00 487.00 479.00 487.00 489.00 492.00 488.00 489.20 4.366539438
20 496.00 495.00 495.00 496.00 497.00 489.00 492.00 494.00 496.00 493.00 494.30 2.406010991
It is necessary to mention that the values of the deviation decrease , in all of the tables because any

enzyme can be used once. According to this information, a small percentage of the substrate is not used.

PROCESS INFORMATION :
CONCLUSION:

Coming back to the research question To what extent the temperatures of 35,36,38,39 and 40°C effects the

enzymatic activity of lactase on the production of glucose that is derived from the interaction of lactase

and lactose with a PH of 7?. It is noticeable that the hypothesis previously established : If the temperature

of 37°C gets increased or decreased, then the production of glucose derived from the interaction between

lactase and lactose will decrease. cannot be supported. Because the average of all the trials is similar. In an

experiment in a Chilean nutrition journal article, the effect of temperature on protein denaturation in whole

and skim milk with lactose was evaluated.It can be seen that the denaturation of proteins in whole milk,

showed a significant increase between temperatures. Treatments at 100 ° C and 130 ° C .Between

temperatures at 110 ° C , 120 ° C and 130 ° C the denaturation was gradual, presenting a significant

difference when compared to the control group at 25 ° C and heat treatment at 100 ° C .This could be the
reason why the difference of the tables was not perceptible, with the temperature of 35,36,38,39 & 40 °C

and obviously with this temperatures the enzyme could not denature.

EVALUATION:

The probably problem that could happen is that the collection of is not an exact because the simulator does

not include data that was verified, it is possible that if this experiment had been carried out in a laboratory,

there would be changes or more variables.Also the collect of data could have been change because because

they were not all done on the same day.

IMPROVEMENTS:

This investigation can be improve by also comparing with a regular temperature of 37°C , and using the PH

of different parts of the human body, not only the small intestine.As well in my point of view if the

independent variable was PH I think that this investigation could be more extensive.Finally it would have

been better, if the experiment with which it was compared in the conclusion had been similar in terms of

temperature, however it was the only one from a reliable source on lactase with changes in temperature.

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http://www.ehu.eus/biomoleculas/index.htm

Gómez, E. (2019, agosto 8). Consumo de leche y otros productos lácteos: gustos y preferencias. Recuperado

2 de junio de 2020, de https://www.wearetesters.com/wat-open-data/consumo-de-leche-y-otros-productos-

lacteos-gustos-y-preferencias

Khan Organization . (s. f.). Enzymes review (article) | Enzymes. Recuperado 2 de junio de 2020, de

https://www.khanacademy.org/science/high-school-biology/hs-energy-and-transport/hs-enzymes/a/hs-

enzymes-review
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Recuperado 4 de junio de 2020, de https://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-

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