Identification, expression, and characterization of a marine-derived chitinase Ce0303 from Chitiniphilus eburneus YS-30 with exo- and endo-hydrolytic properties.

N-acetyl-oligosaccharides exhibit antioxidant and antibacterial activities. However, the low catalytic efficiency of chitinase on crystalline chitin hinders the eco-friendly production of N-acetyl-oligosaccharides. A marine-derived chitinase-producing strain Chitiniphilus eburneus YS-30 was screened in this study. The genome of C. eburneus YS-30 spans 4,522,240 bp, with a G + C content of 63.96 % and 4244 coding genes. Among the chitinases secreted by C. eburneus YS-30, Ce0303 showed the highest content at 19.10 %, with a molecular weight of 73.5 kDa. Recombinant Ce0303 exhibited optimal activity at 50 °C and pH 5.0, maintaining stability across pH 4.0-10.0. Ce0303 demonstrated strict substrate specificity, with a specific activity toward colloidal chitin of 6.41 U mg-1, Km of 2.34 mg mL-1, and kcat of 3.27 s-1. The specific activity of Ce0303 toward α-chitin was 18.87 % of its activity on colloidal chitin. Ce0303 displayed both exo- and endo-hydrolytic properties, primarily producing (GlcNAc)1-3 from colloidal chitin. The structure of Ce0303 includes one catalytic domain and two chitin-binding domains. Docking results revealed that the GlcNAc at -1 subsite formed two hydrogen bonds with conserved Trp380. The hydrolytic properties of Ce0303 will provide technical support for the comprehensive utilization of crustacean raw materials.

A comprehensive study of the colloidal properties, biocompatibility, and synergistic antioxidant actions of Antarctic krill phospholipids.

Excipient selection is crucial to address the oxidation and solubility challenges of bioactive substances, impacting their safety and efficacy. AKPL, a novel ω-3 polyunsaturated fatty acids (PUFAs) esterified phospholipid derived from Antarctic krill, demonstrates unique antioxidant capabilities and synergistic effects. It exhibits pronounced surface activity and electronegativity at physiological pH, as evidenced by a critical micelle concentration (CMC) of 0.15 g/L and ζ-potential of -49.9 mV. In aqueous environments, AKPL self-assembles into liposomal structures, offering high biocompatibility and promoting cell proliferation. Its polyunsaturated bond-rich structure provides additional oxidation sites, imparting antioxidant properties superior to other phospholipids like DSPC and DOPC. Additionally, AKPL augments the efficacy of lipophilic antioxidants, such as alpha-tocopherol and curcumin, in aqueous media through both intermolecular and intramolecular interactions. In sum, AKPL emerges as an innovative unsaturated phospholipid, offering new strategies for encapsulating and delivering oxygen-sensitive agents.

Versatile Bilayer Hydrogel for Wound Dressing through PET-RAFT Polymerization.

Multifunctional hydrogel-based wound dressings have been explored for decades due to their huge potential in multifaceted medical intervention to wound healing. However, it is usually not easy to fabricate a single hydrogel with all of the desirable functions at one time. Herein, a bilayer model with an outer layer for hydrogel wound dressing was proposed. The inner layer (Hm-PNn) was a hybrid hydrogel prepared by N-isopropylacrylamide and chitosan-N-2-hydroxypropyl trimethylammonium chloride (HACC), and the outer layer (PVAo-PAmp) was prepared by polyvinyl alcohols and acrylamide. The two hydrogel layers of the bilayer model were covalently connected with excellent interfacial strength by photoinduced electron/energy transfer-reversible addition-fragmentation chain transfer (PET-RAFT) polymerization. The outer layer exposed to the ambient environment exhibited good stretchability and toughness, while the inner-layer hydrogel adhered to the skin exhibited excellent softness, antibacterial activity, thermoresponsivity, and biocompatibility. In particular, the inner layer of a hydrogel demonstrated excellent antibacterial capability toward both Staphylococcus aureus as Gram-positive bacteria and Escherichia coli as Gram-negative bacteria. Cell cytotoxicity showed that the cell viability of all Hm-PNn layer hydrogels exceeds 80%, confirming that the hydrogels bear excellent biocompatibility. In vivo experimental results indicated that the Hm-PNn/PVAo-PAmp bilayer hydrogel has a significant effect on the acceleration of wound healing, which was demonstrated in a full-thickness skin defect model showing improved collagen disposition and granulation tissue thickness. With these results, the established multifunctional bilayer hydrogel exhibits potential as an excellent wound dressing for wound healing applications, especially for open and infected traumas.

Preparation process optimization, structural characterization and in vitro digestion stability analysis of Antarctic krill (Euphausia superba) peptides-zinc chelate.

In the study, a novel kind of peptides-zinc (AKP-Zn) chelate was obtained using the Antarctic krill (Euphausia superba) peptides (AKP) as raw material, the reaction was carried out with the mass ratio of the AKP to ZnSO4·7H2O of 1:2 at pH 6.0 and 60 °C for 10 min. The structure and composition of the AKP, including particle size, Zeta potential, molecular weight distribution, amino acid composition, microstructure and surface elemental composition, changed significantly after chelating with zinc. The result of Fourier transform infrared spectroscopy indicated that zinc could be chelated by carboxyl oxygen and amino nitrogen atoms of the AKP. Furthermore, compared with zinc sulfate and zinc gluconate, the AKP-Zn chelate was more stable at various pH conditions and the simulated gastrointestinal digestion experiment. These findings would provide a scientific basis for developing new zinc supplements and the high-value utilization of Antarctic krill protein resource.

Chitin from Antarctic krill shell: Eco-preparation, detection, and characterization.

Antarctic krill is a nutrient-rich crustacean that is one of the main species in the Antarctic ecosystem. Antarctic krill shell (AKS) can be used as raw materials to prepare chitin. In this study, lactic acid and dispase were used to prepare Antarctic krill chitin (AKC-1). Amino-monosaccharide contents of chitin samples were detected by pre-column PMP-HPLC method. Analytical instruments were conducted to determine characteristics of chitin samples. Results showed that the amino-monosaccharide content of AKS was 4.62 g/100 g (measured in D-glucosamine). The yield of AKC-1 was 5.49 g/100 g, and the amino-monosaccharide content was 80.90 g/100 g. AKC-1 showed smooth flakes, a porous surface, and α-chitin structural characteristics. The maximum degradation temperature (DTGmax) was 318.3 °C. The yield of deacetylated chitin (AKC-2) was 4.74 g/100 g, with deacetylation degree of 80.8%, viscosity average molecular weight of approximately 145.7 kDa, and amino-monosaccharide content of 97.06 g/100 g. The surface morphology of AKC-2 was similar to that of AKC-1, and the DTGmax was 311.5 °C. A mild, eco-friendly chitin preparation method and an amino-monosaccharide content detection method of raw material before chitin preparation are described in this study, which can provide technical support for comprehensive utilization of Antarctic krill resources.

Effects of Drying Methods on the Content, Structural Isomers, and Composition of Astaxanthin in Antarctic Krill.

Antarctic krill (Euphausia superba) is one of the important bioresources in Antarctic waters, containing many bioactives (e.g., astaxanthin), which have a highly potential value for commercial exploitation. In this study, the effects of processing methods on the content, structural isomers, and composition of astaxanthins (free astaxanthin and astaxanthin esters) were studied. Three drying methods, comprising freeze-drying, microwave drying, and hot-air drying, were used. Free astaxanthin (Ast), astaxanthin monoesters (AM), and astaxanthin diesters (AD) in boiled krill (control) and dried krill were extracted and analyzed using high-resolution mass spectrometry with ultraviolet detection. After the three processes, total astaxanthin loss ranged from 8.6 to 64.9%, and the AM and AD contents ranged from 78.3 to 16.6 and 168.7 to 90.5 µg/g, respectively. Compared to other kinds of astaxanthin esters, astaxanthin esters, which linked to eicosapentaenoic acid and docosahexaenoic acid, as well as the Ast, were more easily degraded, and AM was more susceptible to degradation than AD. All-E-astaxanthin easily transformed to the 13Z-astaxanthin than to the 9Z-astaxanthin during the drying process, but the proportions of optical isomers changed due to drying by no more than 5%. The results suggested that freeze-drying, low-power microwave drying (≤1 kW), and low-temperature hot-air drying (≤60 °C) are optimal drying methods for ensuring the quality of krill products.