This research aimed to establish an effective storage method to maintain the quality of processed sea cucumbers. In this study, sea cucumbers were stored by various methods including the storage of live sea cucumbers (seawater treatment, oxygen treatment, and ascorbic acid treatment) and the storage of dead sea cucumbers (frozen treatment). The sea cucumber quality was monitored after storage and boiling. The weightlessness rate and WHC of the frozen group increased to 86.96% ± 0.83% and 93.29% ± 0.32%, respectively. Frozen sea cucumbers shrunk with the meat’s textural properties deteriorated. During the live sea cucumber storage, the tissue protein degraded from day 3 to day 7 which led to the promotion of TVB-N. Among these, the oxygen group showed the smallest TVB-N increase from day 0 (3.78 ± 0.60 mg 100 g⁻¹) to day 7 (10.40 ± 0.12 mg 100 g⁻¹). The oxygen group exhibited the most moderate change in weightlessness rate (4.24% ± 0.45%) and the most moderate texture parameters decline, such as the hardness of 32.52%, chewiness of 78.98 ± 5.10 N, and adhesion of 0.84 ± 0.00. The oxygen method showed the best condition of sea cucumber after 5 days of storage.

Autolysis in sea cucumber has long been a threat to raw material storage and product processing. The involvement of endogenous cysteine protease in sea cucumber autolysis has been proved extendedly. However, as an essential part of the mechanism of autolysis, the role of its endogenous inhibitor has seldom been reported. To investigate the role of cysteine protease inhibitors in the early stage of hypoxic exposure-induced autolysis, a novel cystatin gene (SjCyt) belonging to the subfamily of cystatin C was cloned from Apostichopus japonicus by homology cloning and rapid amplification of cDNA ends. The affinity of SjCyt to cysteine protease (cathepsin L and cathepsin B) was investigated by molecular dynamics simulations. Pertinent metrics, including the root mean square deviation, radius of gyration, Gibbs free energy, binding free energy, and bond-forming frequency, showed that the conformation of SjCyt–SjCL was more stable and confirmed a stronger interaction of SjCyt with cathepsin L than with cathepsin B. Thus, cathepsin L (SjCL) was selected to further study its co-expression with SjCyt over a period of 9 h at an early stage of hypoxic exposure. Quantitative RT-qPCR revealed a ubiquitous transcriptional profile of SjCyt and SjCL in all the tested tissues, with the highest abundance in the dorsal epidermis, tube feet, and coelomocytes. Temporal transcription of them showed an overall up-regulated co-expression in the dorsal epidermis and tube feet. However, up-regulated SjCyt and down-regulated SjCL were observed at the protein level. Further immunofluorescence double labeling also found increased staining of SjCyt and SjCyt–SjCL complexes and decreased SjCL. Additionally, recombinant SjCyt was prepared and demonstrated an evident autolysis-inhibiting effect. The results of this study indicated that the anti-autolytic regulation of SjCyt functions at the very early stage of hypoxic exposure, exerting effects at both the transcriptional and translational levels. The above finding offers new insights into the mechanisms of sea cucumber autolysis.

Few studies are concerned with the effect of the conjugat protein on the bioactivities of the abalone gonad polysaccharide (AGP). In this study, a series of treatments, including raw material (female and male) defatting, extraction temperature (25–121 °C), proteolysis, ultrafiltration, and ethanol precipitation, was conducted to investigate the role of the conjugate protein on AGP anticoagulant activity. All AGP extracts significantly prolonged activated partial thromboplastin time (APTT) and thrombin time (TT). The strongest was observed in the female AGPs prepared at 50 and 121 °C. The most active is located at 30–300 kDa by ultrafiltration. After being exposed to neutral protease, quick shortening of APTT and TT was found in all AGPs. Further ethanol precipitating of found the longest APTT in the sediment, which contains most polysaccharides and proteins. Defatting lowered the activity of female AGP but increased that of males. Proteolysis also significantly weakened the clotting factor inhibition effect of the 50 °C female AGP, but heating seemed not affect the effect. Five fractions were obtained after the 50 °C female AGP was subjected to ion exchange column. Fraction V, with the highest protein and medium polysaccharide content, showed the strongest anticoagulant effect and was also much higher than AGSP, which was obtained by multi-step proteolysis. The findings supported positive effect of the conjugate protein in AGP anticoagulant activity.

In our preliminary work, a clam sauce prepared by fermentation with Aspergillus oryzae 3.042 (AO) exhibited desirable flavor and quality; however, the process was prolonged (exceeding 30 d), and a high salt concentration (6–15%) was necessary to prevent spoilage. Consequently, shortening production cycle and reducing salt content without compromising product quality became a new objective. Enzymatic hydrolysis has long been recognized as an efficient approach in seasoning production, with enzyme efficacy being a key competitive factor. Accordingly, an AO-derived aminopeptidase–protease complex (AOAP) was optimized and prepared as a preparatory step. In this study, AOAP was applied to hydrolyze clam meat to evaluate its potential for producing a seasoning base. A two-step enzymatic hydrolysis process was employed. In the first step, the highest hydrolysis degree (29.1%) was achieved using alkaline protease (AP). The resulting hydrolysate was subsequently subjected to secondary hydrolysis with AOAP, achieving a degree of hydrolysis as high as 49.8%. Sensory evaluation revealed a significant reduction in bitterness and enhancement of umami in the final hydrolysate, a finding corroborated by electronic tongue analysis. Further characterization via LC-MS and amino acid (aa) analysis showed that a substantial number of bitter and umami peptides were released following AP treatment; however, the number of these peptides was markedly reduced after a subsequent AOAP hydrolysis, with concurrent substantial changes in the peptide profile. In the two-step hydrolysate, umami peptides mostly contain 3–10 aa, whereas bitter peptides typically contain only 3–5 aa. The content of free aa increased from 369.17 mg/100 g in the control to 3026.25 mg/100 g in the two-step hydrolysate, half of which were bitter, indicating the debittering efficiency of AOAP. Electronic nose analysis revealed similar flavor profile and characteristic presence of nitrogen oxides in all hydrolysates. GC-MS analysis further demonstrated that, after combined enzymatic hydrolysis, the short-chain aldehydes and ketones responsible for the fishy odor in the raw material almost completely disappeared, while long-chain aldehydes with pleasant aromas were generated. These findings suggest that the secondary hydrolysis step using AOAP can effectively improve the overall flavor profile of the clam hydrolysate, which may support its potential applicability in seasoning production, though further optimization and scale-up validation are needed.

Collagen-derived products are widely applied in functional foods; however, limited information is available regarding how different preparation methods, particularly thermal degradation and enzymatic hydrolysis, affect their anti-aging efficacy and biological functions. In this study, sturgeon skin was used as raw material to prepare gelatin–peptide complexes via thermal degradation (GPC-TD) and enzymatic hydrolysis (GPC-EH), and their comparative anti-aging and biological effects were evaluated in D-galactose-induced aging mice. Female ICR mice were divided into eight groups: a blank control group (normal saline), an aging model group (D-galactose, 500 mg/kg), three GPC-TD and three GPC-EH groups (D-galactose supplemented with 100, 200, 400 mg/kg GPC-TD or GPC-EH). After eight weeks of administration, various physiological parameters were evaluated. Throughout the experiment, no statistically significant difference in body weight (BW) was observed among the groups; however, the blank and model groups consistently maintained the highest BW. The medium- and high-dose GPC-TD groups showed relatively faster weight gain, whereas the 100 mg/kg GPC-TD group and all three GPC-EH groups exhibited the slowest BW gain. Notably, the gastric indices of these latter groups were significantly lower than those of other groups (p < 0.05), which might be a key factor affecting BW gain. Behavioral tests revealed that the model group exhibited significantly reduced swimming speed and weakened nesting ability (p < 0.05), both of which were alleviated to varying degrees by treatment with GPC-TD and GPC-EH. Furthermore, both complexes markedly decreased malondialdehyde content in liver tissue (p < 0.05). Compared with the model group, high-dose GPC-TD and GPC-EH effectively increased acetylcholine content and inhibited acetylcholinesterase activity (p < 0.05). Masson staining revealed abnormal collagen fibers accumulation in certain tissues of model mice, a condition that was clearly ameliorated by GPC-TD and, to a greater extent, by GPC-EH. In addition, medium and high doses of both complexes significantly protected against D-galactose-induced loss of Nissl bodies in brain neurons; in the high-dose GPC-EH group, the density and number of Nissl bodies approached those observed in the blank group. These findings suggest that both GPC-TD and GPC-EH possess potential anti-aging effects, with GPC-EH exhibiting superior efficacy. This study provides theoretical support for consumers, the catering industry, and manufacturers in selecting appropriate processing techniques for the preparation of sturgeon skin GPC.