Background: Textural deterioration is a serious problem in chilled fish flesh. Cysteine proteinases are proposed to participate in disintegration of collagen fibers during this process, while its mechanism remains elusive. In the present study, a cysteine proteinase was purified from grass carp muscle and identified by mass spectrometry, and its effect on structural changes of collagen fibers was investigated.
Results: During storage at 4 °C, cysteine proteinase activity in fillets increased to 1.53-fold at day 5 and maintained a high level later, and this variable was related to a decline in shear force and an increase in drip loss. A 29 kDa cysteine proteinase was purified through ammonium sulfate precipitation and column chromatography, and identified as cathepsin L. Cathepsin L caused collagen fibers to partly disintegrate into fibril bundles and individual fibrils at 48 h, while the triple helical structure of collagen molecules remained stable. Release of soluble proteins and glycosaminoglycans from cathepsin L-treated collagen fibers was time dependent, coinciding with a release of 4.12 ± 0.13% and 8.57 ± 0.03% at 48 h respectively. However, 0.85 ± 0.02% of hydroxyproline was freed from cathepsin L-treated collagen fibers at 48 h. Furthermore, scanning electron microscopy revealed that the inhibitory effect of cathepsin L could retard the destruction of intramuscular connective tissues (IMCTs).
Conclusion: These results indicated that cathepsin L might be involved in collagen fiber breakdown by degrading collagen-associated proteoglycans during textural deterioration of grass carp. © 2022 Society of Chemical Industry.
Keywords: cathepsin L; collagen fibers; grass carp; hydroxyproline; proteoglycans.
© 2022 Society of Chemical Industry.