Total volume = 132.5 mL
Isolation of Lysozyme A
Isolation of Lysozyme B
Isolation of Lysozyme C
Isolation of Lysozyme D
State of Tyrosine in Ovalbumin
*1mL of Ovalbumin sample from week 1 was added with 9mL of water
Table 1:
Spectrum Diagram Obtained
Figure 1
Figure 2:
Figure 3:
Week 3
Concentration of standard protein containing Bovine Serum Albumin (BSA) = 10mg/mL
Table 2: Different amount of protein contains in different sample
Table 3: Respective data of dilution, absorbance, suitability of spectroscopy analysis, mass of protein and protein concentration with different types of protein used.
Table 4: Records of results including volume, total mass and percentage obtained for each protein
*Conalbumin and Ovalbumin protein were obtained from the dialysis reading in week 1’s result table
* Volume Lysozyme A, B, C, D were recorded in week’s result table
Calculations:
From the standard curve, the equation Absorbance = 0.0402(mass of BSA);
Mass of Conalbumin (diluted 1/10) = 0.147/0.0402
= 3.657 mg
Mass of Ovalbumin (diluted 1/10) = 0.177/0.0402
= 4.403 mg
Mass of Lysozyme A (diluted 1/10) = 0.085/0.0402
= 2.114 mg
Mass of Lysozyme A (undiluted) = 0.064/0.0402
= 1.592 mg
Mass of Lysozyme C (diluted 1/10) = 0.009/0.0402
= 0.224 mg
Mass of Lysozyme D (diluted 1/10) = 0.008/0.0402
= 0.199 mg
Mass of Lysozyme D (undiluted) = 0.048/0.0402
= 1.194 mg
Mass of Lysozyme B (diluted 1/10) = 0.150/0.0402
= 3.731 mg
Mass of Lysozyme B (diluted 1/100) = 0.122/0.0402
= 3.035 mg
Mass of Egg White (diluted 1/30) = 0.438/0.0402
= 10.896 mg
From Table 4, 12mL of conalbumin was isolated,
Thus the total protein mass in 12mL conalbumin = 46.0 X 12
= 552.0 mg proteins.
Percentage of conalbumin isolated from egg white 1 = (552/18408.6) X 100%
= 2.999 % isolated
In 1mL of 1/10 diluted conalbumnin, concentration of protein is 4.60mg/mL
In 1mL of undiluted conalbumin, concentration of protein = 4.60 X 10
= 46.0mg/mL
Week 4
Table 5: Preparations on different lysozyme samples respectively for determination of lysozyme activity
Table 6: Absorbance reading of lysozyme A, B, C, and 1/10 D
Figure 1: Graph of Absorbance Reading vs. Time for Lysozyme A, B, C and D
Table 7:
Table 8:
DISCUSSION
CONCLUSION
REFERENCES
Chawla, H. S., 2002, ‘Chapter 14: Basic Techniques’, Introduction to plant biotechnology (2nd ed.), Science Publishers, USA, pp. 173-180.
Bertino, A. J. & Bertino, P. N., 2008, ‘Chapter 7’, Forensic Science: Fundamentals and Investigations (1st ed.), Nelson Education, Canada, pp. 179.
Carter, M. & Shieh, J. C., 2010, Guide to research techniques in neuroscience, Elsevier Inc, London.
PRE-LABORATORY QUESTIONS
-
The movement of molecules in paper electrophoresis is based on the charge of the molecule. A positively charged molecule moves towards the cathode and the negatively charged molecules moves towards the anode (Chawla, 2002).
- This technique is only useful for the separation of small charged molecules such as amino acids and small peptides. Proteins are very large in size and do not separate effectively in paper electrophoresis.
-
5 moles = 5 x 10-6 mol
10 mM = 10 x 10-3 M
Volume of sample containing 5 moles of aspartate = 5 x 10-4 L
Biochemistry (BTH2741) Experiment 5