Data sheet

Module 2: BROMELAIN EXTRACTION AND CHARACTERIZATION

Group/ Subgroup:
Name of student 1: ________________________ Student ID: ___________
Name of student 2: ________________________ Student ID: ___________

Signature of lab instructor:

o Calculate the concentration of protein (µg/ml) in each of the five samples that make up
the standard curve and draw a calibration curve of Bradford assay.

Table 1. Bradford protein assay standard curve

Vol of BSA Vol of Protein OD595nm Corrected
Test tubes 0.1 mg/ml deionized concentration OD595nm
(mL) water (mL) (µg/ml) (∆OD595nm)

Blank 0 10

1 1 9

2 2 8

3 3 7

4 4 6

5 5 5

o Calculate the protein concentration in samples (I-II) using the standard curve.

Table 2. Determination of protein concentration in unknown samples

Protein
OD595nm OD595nm OD595nm OD595nm concentration Dilution Actual protein
calculated factor
Samples (1) (2) (3) (Average) ∆OD595nm concentration
based on the
(µg/ml)
standard curve
(µg/ml)

Solution
(I)

Solution
(II)
 Data sheet
Module 2: BROMELAIN EXTRACTION AND CHARACTERIZATION

o Caculate the enzyme activity in samples (I-II)

Table 3. Determination of bromelain activity in unknown samples

Solution (I) Solution (II)
Samples
Blank Tyrosine Sample Blank Tyrosine Sample

OD620nm

OD620nm (Average)

∆OD620nm

Dilution factor

Amount of tyrosine (µg) in

0.5 mL solution used in
“tyrosine” batch
Amount of tyrosine (µg)
produced in “sample” batch

Bromelain activity (UI/ml)

o Show the results of protein and enzyme activity assays from the protein purification in
Table 4.

Table 4. Bromelain purification table

Bromelain
Total Total Specific Protein
Volume Purification activity
Step protein activity activity recovery
(ml) fold recovery
(mg) (UI) (UI/mg) (%)
(%)
Homogenate /
(crude extract) (I)
Solvent
precipitation (II)