In vitro trypsin digestion and identification of possible cross-linking sites induced by transglutaminase (TGase) of silver carp (Hypophthalmichthys molitrix) surimi gels with different degrees of cross-linking.

Surimi gels with different cross-linking degrees (18.52%, 34.67%, 62.87% and 79.11%) were prepared to identify the numbers and locations of lysine residues involved in TGase-induced cross-linking, and to reveal the quantity and location relationships among cross-linking degrees, cross-linking sites and digestion sites by using trypsin digestion, SDS-PAGE and LC-MS/MS methods. The results showed that with the increase in cross-linking degree from 18.52% to 79.11%, 1) the quantity of cross-linking sites gradually increased from 25 sites to 47 sites, 2) the main possible cross-linking domain moved from myosin head to rod, 3) the numbers of digestion sites first decreased from 1262 sites to 1194 sites, and then increased to 1302 sites, 4) the changes in the values of digestion sites were mainly concentrated in myosin rod and it was also the main region of digestion. This study can help exploring the relationship between enzymatic cross-linking and nutritional properties of food.