Increased reactive carboxyl and free alfa-amino groups from fish type I collagen peptides by Alcalase® hydrolysis exhibit higher antibacterial and antioxidant activities

This study aimed to generate low molecular weight peptides (LMWP) from fish collagen type I hydrolysed by increasing activity of Alcalase (R) from 0.0 to 12.0 (AU-A per 100 g) at 55 degrees C and 7.4 of pH for 3 h 40 min. The results showed that all enzyme activity levels caused 34-55 % reductions in protein recovery, 1.0-3.0 folds' increase in free alpha-amino groups and 1.7-3.2 folds' increase in carboxyl groups. Degree of hydrolysis ranged from 20 to 30 % with increasing enzyme activity. The number average molecular weight significantly reduced from 3200 g/mol in 0.0 AU-A per 100 g enzyme activity to 1151, 1398, 1175, 1040 and 1246 g/mol in 2.4, 4.8, 7.2, 9.6 and 12.0 AU-A per 100 g enzyme activities, respectively. Depending upon enzyme activity level, the produced LMWP with reactive carboxyl and amino end-groups exhibited 2.5- to 4.0-fold increases in antioxidant capacity and 1.0-3.5 log cfu/ml inhibition of four pathogen bacteria. Highest inhibition of 2.5 log cfu/ml in Escherichia coli was obtained from 2.4 AU-A per 100 g enzyme activity and 3.5 log cfu/ml in Listeria monocytogenes from 9.6 and 12.0 AU-A per 100 g enzyme activity levels. Infrared spectroscopy clearly identified reactive end-groups and showed remarkably differences in molar absorptivity of various molecular regions between non-enzyme and enzyme treated collagen type I molecule. A 9.6 and 12.0 AU-A per 100 g enzyme activity levels were found optimally effective to generate LMWP. In conclusion, LMWP exhibited high antioxidant and antibacterial activity due to increased functional reactive end-groups, and these bio-active peptides may have greater potentialities in various food and pharmaceutical applications.