The combined enzyme-alkali treatment (E-AO) is currently employed in the processing of salted bovine omasal laminae due to its advantages of low alkalinity and mild treatment. This study was aimed at investigating the mechanism by which the E-AO treatment improves the swelling ratio and texture of salted omasal laminae. Urea and sodium dodecyl sulfate (SDS) were used to monitor the contributions of hydrogen bonds and hydrophobic interactions in the processing of omasal laminae. The results indicated that E-AO treatment improved water migration and moisture content, resulting in an increased swelling ratio. Meanwhile, the decreased total collagen content and increased soluble collagen improved the texture of omasal laminae. The expansion and destruction of protein structure were observed through microstructure analysis. This study elucidates the mechanisms underlying quality changes in omasal laminae, providing a theoretical foundation for production of bovine omasal laminae and other by-products containing smooth muscle and connective tissue.