Effect Of Whey Protein Powder On Total Protein Biology Essay

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Whey proteinhas been shown to increases oil quality and water capacity. (Sharrattet aI., 1959) This experimentwas designed to determinewhetherpea plant samples would exhibitsimilar effects when treatedwith whey. The test

reagent was diluted into two different treatments (lx and O.lx), and a control group was treatedwithdistilled water. The plants were wateredfive days a week for the first week with 15 mL of distilled water. The next two weeks, the two treatments

were nourishedwith 15 mL of the whey dilutions and the control with 15 mL of distilled water. At the end of the grow period, a Bradford Assay was performedto determinethe overall proteinconcentrationof the pea plants as it pertainedto the fresh weight of the plant. The pea plant leaf samples(control, lx, and O.lx) had average fresh weights of 8.00 mg, 21.33 mg, and 92.67 mg, respectively.Their average absorbancevalues in the spectrometer were as follows: 0.159, 0.111, and

0.157, respectively.A one-way analysis of variance(ANOVA)was performedto determinethedifference of the numbersin the threedata sets. A p-value of around

0.009 was achieved, leading to the acceptanceof the null hypothesis.This means

that whey proteinhad a no effect on plant height growth, fresh weight, or protein concentration.

Introduction

Whey proteinis used commerciallyas a supplementfor athletesand active human beingswho want to build protein.It is widely used by weight lifters and workoutfiends in shakes andother types of drinks designedto increase

performancein sportsand workouts.It is a source of branchedamino acids that fuel muscles andstimulatethe synthesisof proteins(Krissansen,2007). In past experiments,it has been shown thatwhey proteinis particularlysuccessful in increasingoverall proteincontent in plants. Sharratt,Peterson,and Calbert showed in their 1959 experimentthat whey increasedthe quality of soil aggregationin the treatments.Good soil aggregationallows for a greateramountof waterto enter at

the surface of the soil as well as a bigger capacity for waterin general, and also providesbetteraeration.(Sharrattet at,1959) Their study showed that whey proteinwas particularlysuccessfulin treatmentof bluegrass. This study reasoned that the increasein bluegrassyields and height weredue in part to the fact that whey increasedthe level of nitratesin the soil, which are essentialto plantgrowth. Nitrogen is an importantcomponentin plant growthas plants use it to make proteins.Plants get about 90% of theirnitrogenin the nitrateform. (Crawford,

1995)

The questionfor this experimentwas whetheror not whey protein(as it appearsin supplementalform for humans)would affect pea plant growth with respectto plant height, fresh weight, and proteinconcentration.In this study, there were two treatments of plant growth, and one control. Each treatmentand the control consistedof four differentpea plants, grown separatelyfrom one another. The treatmentswere as follows: 1X and O.lX. The 1X treatmentcontainedthe recommendeddosage of whey proteinpowderfor human consumptionmixed with

distilled water. The O.lX treatmentwas a dilution of the first treatment by a factor of ten. It was predictedthat the first treatment (1X) would have too much protein

powder and would inhibit plant growth, fresh weight, and proteinconcentration (Sharrattet al., 1959). It was also predictedthat the second treatmentwould show increasedgrowthas comparedto the control and the first treatment. The null hypothesiswas that whey proteinwould have no effect on plant height growth, fresh weight, or proteinconcentration.

Methods

Stock solutionsof the whey proteintest reagentwere first made in lx and O.lx concentrates.The lx concentratewas the normal amountof powder recommendedfor humanconsumptionand the O.lx concentratewas a dilution of

the original by a factor often.The pH of the lx solution was found to be 5.97, and for the O.lx the pH was 5.99. Each treatmentand the control had a total of 4 pea plants, grown independently of one another.For the first week, all the subjects were

watereddaily with 15 mL of distilled water.For the second week, the lx and O.lx treatmentswerenourisheddaily with 15mL of their specific reagent. At the end of the grow period, the plant leaveswere weighed for their fresh weight values. A Bradford Assay for total protein was performednext.A standardcurve (Figure 1) was producedusing known concentrations of bovine serum albumin(BSA)and Bradford reagent. The absorbancesof these known concentrations were used to make thestandardcurve. When the absorbanceof an unknownsample is calculated using a spectrometer,thatvalue is plugged into the equationof the standardcurve to give the proteinconcentration.A one-wayanalysis of variance(AN OVA) was performedwith the proteinconcentrations of the treatments andthe control.

Control

O.lX

lX

Sample

A

B

C

A

B

C

A

B

C

Absorbance

(A=594)

0.253

0.074

0.15

0.218

0.167

0.085

0.019

0.073

0.019

FreshWeight

(mg)

8

4

12

85

101

92

17

31

16

Protein

Concentration

[mg/gFW)

61.0125

35.685

24.115

4.95

3.13

1.7824

2.15647

4.54258

2.29125

Average Protein Concentration

40.2708333

3.28746667

2.99676667

Stand.Dev.

18.8713745

1.58966011

1.34040873

)

Table1: Absorbance, freshweight,andprotein concentration of peaplantsamples.

Averageprotein concentration andstandard deviation arealso shown.

'1

1.8

1.6

1.4

Bradford Assay Standard Curve

forBSA

- --- ---

-~--------- Y- = 0.031x

R2 = 0.668

•aJ 1.2 •

..~ 1

•~ 0.8 o0.6

(/)

~ 0.4

0.2

o

• -----

- -- ---r---

o 20 40 60

Protein Concentration (JA.g/mL)

-----

Figure 1: Standardcurve of known proteinconcentrations versusabsorbanceat 594 nm. The responseof an unknownproteinsample to CBBGdye was comparedto the responseof known proteinconcentrations to determinethe proteincontentof the unknownsample. When the absorbancevalue is plugged into the equationof the trend line, a concentrationof proteinis given in ug/ml,

Average ProteinConcentration

60 ----

eo50<, - --- -

eo

:::1..

40C

0

'ra

I-.

4-'

C

uCJ.) 30

c

u0

.ca:;

4-'

100

20 --- -

1-0

0..

0

control O.IX IX

Solution Concentration

- ---

Figure 2: Average proteinconcentrationof pea plant samples. Tick marks indicate standarddeviationof the data set. When treatedwith test reagentin separate dilutions, pea plants showed relatively low proteinlevels as comparedto the control group.

Results/Conclusion

It was found throughexperimentation that the both the Ix and O.lx solutions wereineffectivein aiding pea plant growth, fresh weight, and proteinconcentration. The cause of this is due to the fact that whey proteindrink mix has many globular proteinsthat inhibit the pea plantsability to get the adequatenitratesit needs from the soil in order to successfully grow. The size of thewhey mix proved to be too

large for the pea plant seeds or roots to absorbit. The one-way ANOVAperformed

produceda p-value of 0.0089; because this value is less than 0.05, the null

6

hypothesiscan be accepted. Whey proteinpowderhas no effect on plant growth with respectto height, fresh weight, and total protein.

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