A quantitative comparable study on multi-hierarchy conformation of acid and pepsin-solubilized collagens from the skin of grass carp (Ctenopharyngodon idella).

This work aimed to improve yield of collagen from the grass carp skin by employing different strategies (acid-acid method, pepsin-pepsin method and acid-pepsin method, denoted as A-A, P-P, A-P, respectively). And further to conduct quantitative characterization on structural properties, self-assembly kinetics and gelation properties of these collagens. Herein, a two-step collagen extraction method (pepsin-pepsin) was established with the high yield. Meanwhile, structural measurements of high-yield collagen (pepsin-soluble collagen, PSC) and acid-soluble collagen (ASC) indicated that both collagens maintained the typical triple helical conformation of collagen type I. Moreover, the fibrillogenesis tests of PSC and ASC at the various temperatures confirmed that self-assembly were the entropy-driven process. The gelation time of both ASC and PSC was determined by the dynamic time sweep at the different frequencies combined with Winter's criterion. The self-assembly kinetics results showed that fibrillogenesis rate for ASC solution was faster, and more liable to gelation relative to PSC. Mechanical measurements suggested that ASC showed the more resistance ability to deformation than PSC due to more complicated architecture, confirmed by higher fractal dimension. However, the equivalent typical assemblies of PSC to ASC at the various stages can still be expected via controlling incubation time or temperature under the guidance of Arrhenius equation. This study would provide some strategies for achieving maximum utilization of waste biomass and significant insights into the mechanisms underlying the quantitative differences in multiple hierarchy conformation (molecule, fibrillogenesis and hydrogel) of ASC and PSC, which may benefit for subsequent design, development and optimization of collagen-based hydrogels in biomedical industries.