Objective: The aim of this study was to evaluate the protein digestibility and analyze the residual enzymatic activity of lysozyme.
Methods: Protein digestibility was evaluated hydrolyzing the protein with pepsin at pH 1.2, 2.0, and 3.2 during 60, 90, and 120 minutes of incubation.
These hydrolysates were analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and reversed-phase high-performance liquid
chromatography. Residual enzymatic activity was evaluated with the spectrophotometric method at 450 nm.
Results: Lysozyme was totally hydrolyzed with pepsin at pH 1.2. At pH 2.0, lysozyme was partially hydrolyzed and at pH 3.2 hydrolysis was absent at
all times of the assay.
Conclusions: Lysozyme was hydrolyzed with pepsin at low pH. Residual enzymatic method can be used to determine the grade of hydrolysis in
Keywords: Lysozyme, Enzymatic hydrolysis, Muramidase activity, Antimicrobial activity.

Author Biographies

Wilman Carrillo, Universidad Técnica de Ambato
Facultad de Ciencia e Ingeería en Aliementos
Jessica Tubón, Universidad Técnica de Ambato
Facultad de Ciencia e Ingeería en Alimentos


1. Jollès P, Jollès J. What’s new in lysozyme research? Always a model
system, today as yesterday. Mol Cell Biochem 1984;63(2):165-89.
2. You SJ, Udenigwe CC, Aluko RE, Wu J. Multifunctional peptides from
egg white lysozyme. Food Res Int 2010;43:848-55.
3. Memarpoor-Yazdi M, Asoodeh A, Chamani T. A novel antioxidant and
antimicrobial peptide from hen egg white lysozyme hydrolysates. J
Funct Foods 2012;4:278-86.
4. Mine Y, Ma F, Lauriau S. Antimicrobial peptides released by
enzymatic hydrolysis of hen egg white lysozyme. J Agric Food Chem
5. Mine Y, Yang M. Recent advances in the understanding of egg
allergens: Basic, industrial, and clinical perspectives. J Agric Food
Chem 2008;56(13):4874-900.
6. Callewaert L, Michellies C. Lysozymes in the animal kingdom. J Biosci
7. Carrillo W, García-Ruiz A, Recio I, Moreno-Arribas MV. Antibacterial
activity of hen egg white lysozyme modified by heat and enzymatic
treatments against oenological lactic acid bacteria and acetic acid
bacteria. J Food Prot 2014;77:1732-9.
8. Cegielska-Radziejewska R, Lesnierowski G, Kijowski J. Antibacterial
activity of hen egg white lysozyme modified by thermochemical
technique. Eur Food Res Technol 2009;228:841-5.
9. Achouri A, Boye JI. Thermal processing, salt and high pressure
treatment effects on molecular structure and antigenicity of sesame
protein isolate. Food Res Int 2013;53:240-51.
10. Osborne NJ, Koplin JJ, Martin PE, Gurrin LC, Lowe AJ, Matheson MC,
et al. Prevalence of challenge-proven IgE-mediated food allergy using
population-based sampling and predetermined challenge criteria in
infants. J Allergy Clin Immunol 2011;127(3):668-76.e1-2.
11. Sicherer SH. Epidemiology of food allergy. J Allergy Clin Immunol
12. Orruño E, Morgan MR. Resistance of purified seed storage proteins
from sesame (Sesamum indicum L.) to proteolytic digestive enzymes.
Food Chem 2011;128:923-9.
13. Laemmli UK. Cleavage of structural proteins during the assembly of
the head of bacteriophage T4. Nature 1970;227(5259):680-5.
14. Martos G, Contreras P, Molina E, López-Fandiño R. Egg white
ovalbumin digestion mimicking physiological conditions. J Agric Food
Chem 2010;58(9):5640-8.
15. Shugar D. The measurement of lysozyme activity and the ultra-violet
inactivation of lysozyme. Biochim Biophys Acta 1952;8(3):302-9.
16. Ibrahim HR, Matsuzaki T, Aoki T. Genetic evidence that antibacterial
activity of lysozyme is independent of its catalytic function. FEBS Lett
17. Thomas K, Aalbers M, Bannon GA, Bartels M, Dearman RJ,
Esdaile DJ, et al. A multi-laboratory evaluation of a common in vitro
pepsin digestion assay protocol used in assessing the safety of novel
proteins. Regul Toxicol Pharmacol 2004;39(2):87-98.
18. Polverino de Laureto P, Frare E, Gottardo R, Van Dael H, Fontana A.
Partly folded states of members of the lysozyme/lactalbumin
superfamily: A comparative study by circular dichroism spectroscopy
and limited proteolysis. Protein Sci 2002;11(12):2932-46.
19. Fu TJ, Abbott UR, Hatzos C. Digestibility of food allergens and nonallergenic

comparative study.
J Agric
Food Chem 2002;50(24):7154-60.
20. Ibrahim HR, Inazaki D, Abdou A, Aoki T, Kim M. Processing of
lysozyme at distinct loops by pepsin: A novel action for generating
multiple antimicrobial peptide motifs in the newborn stomach. Biochim
Biophys Acta 2005;1726(1):102-14.
169 Views | 275 Downloads
How to Cite
Carrillo, W., and TubónJ. “DIGESTIBILITY AND ENZYMATIC ACTIVITY IN VITRO OF HEN EGG WHITE LYSOZYME”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 9, no. 3, May 2016, pp. 376-8, https://innovareacademics.in/journals/index.php/ajpcr/article/view/11626.
Original Article(s)

Most read articles by the same author(s)