Systematical Investigation on Anti-Fatigue Function and Underlying Mechanism of High Fischer Ratio Oligopeptides from Antarctic Krill on Exercise-Induced Fatigue in Mice.

High Fischer ratio oligopeptides (HFOs) have a variety of biological activities, but their mechanisms of action for anti-fatigue are less systematically studied at present. This study aimed to systematically evaluate the anti-fatigue efficacy of HFOs from Antarctic krill (HFOs-AK) and explore its mechanism of action through establishing the fatigue model of endurance swimming in mice. Therefore, according to the comparison with the endurance swimming model group, HFOs-AK were able to dose-dependently prolong the endurance swimming time, reduce the levels of the metabolites (lactic acid, blood urea nitrogen, and blood ammonia), increase the content of blood glucose, muscle glycogen, and liver glycogen, reduce lactate dehydrogenase and creatine kinase extravasation, and protect muscle tissue from damage in the endurance swimming mice. HFOs-AK were shown to enhance Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities and increase ATP content in muscle tissue. Meanwhile, HFOs-AK also showed significantly antioxidant ability by increasing the activities of superoxide dismutase and glutathione peroxidase in the liver and decreasing the level of malondialdehyde. Further studies showed that HFOs-AK could regulate the body's energy metabolism and thus exert its anti-fatigue effects by activating the AMPK signaling pathway and up-regulating the expression of p-AMPK and PGC-α proteins. Therefore, HFOs-AK can be used as an auxiliary functional dietary molecules to exert its good anti-fatigue activity and be applied to anti-fatigue functional foods.

Impact of L-arginine on the quality of heat-treated Antarctic krill: Influence of pH and the guanidinium group.

Antarctic krill suffers from severe water loss after heating, and its quality deteriorates, so it is in urgent need of a green and healthy improver. In this paper, the effects of L-arginine (L-Arg) soaking on the modification of the quality of heat-treated Antarctic krill and the structure of myofibrillar proteins (MPs) in Antarctic krill were investigated. The results showed that L-Arg had an ameliorating effect on heat-treated krill in a concentration-dependent relationship. The water-holding capacity of L-Arg-soaked krill was 1.41 times higher than that of sodium tripolyphosphate (STPP) at an equivalent concentration (80 mM). At 120 mM L-Arg soaked, L* and hardness of krill decreased to 58.31 and 334.81 g, while resilience and moisture content increased to 0.47 and 85.29 % after heating, respectively. The scanning electron microscopy (SEM) results revealed that the tissue state of the pH-corrected groups was better than the control, but not as well as that of the pH-uncorrected groups. pH and the guanidinium group in L-Arg both played roles in promoting the transition of MPs from disordered to ordered secondary structures. This transition reduced the exposure of hydrophobic and sulfhydryl groups in MPs, inhibited the protein aggregation and increased the solubility of MPs to 71.61 %, which ultimately improved the quality of heat-treated krill. It is worth noting that the pH effect had a primary influence on the observed effects, while the guanidinium group made a secondary contribution. These results could broaden the potential application of L-Arg as an improver of the quality of heat-treated krill.

Novel Ca-Chelating Peptides from Protein Hydrolysate of Antarctic Krill (Euphausia superba): Preparation, Characterization, and Calcium Absorption Efficiency in Caco-2 Cell Monolayer Model.

Antarctic krill (Euphausia superba) is the world's largest resource of animal proteins and is thought to be a high-quality resource for future marine healthy foods and functional products. Therefore, Antarctic krill was degreased and separately hydrolyzed using flavourzyme, pepsin, papain, and alcalase. Protein hydrolysate (AKH) of Antarctic krill prepared by trypsin showed the highest Ca-chelating rate under the optimized chelating conditions: a pH of 8.0, reaction time of 50 min, temperature of 50 °C, and material/calcium ratio of 1:15. Subsequently, fourteen Ca-chelating peptides were isolated from APK by ultrafiltration and a series of chromatographic methods and identified as AK, EAR, AEA, VERG, VAS, GPK, SP, GPKG, APRGH, GVPG, LEPGP, LEKGA, FPPGR, and GEPG with molecular weights of 217.27, 374.40, 289.29, 459.50, 275.30, 300.36, 202.21, 357.41, 536.59, 328.37, 511.58, 516.60, 572.66, and 358.35 Da, respectively. Among fourteen Ca-chelating peptides, VERG presented the highest Ca-chelating ability. Ultraviolet spectrum (UV), Fourier Transform Infrared (FTIR), and scanning electron microscope (SEM) analysis indicated that the VERG-Ca chelate had a dense granular structure because the N-H, C=O and -COOH groups of VERG combined with Ca2+. Moreover, the VERG-Ca chelate is stable in gastrointestinal digestion and can significantly improve Ca transport in Caco-2 cell monolayer experiments, but phytate could significantly reduce the absorption of Ca derived from the VERG-Ca chelate. Therefore, Ca-chelating peptides from protein hydrolysate of Antarctic krill possess the potential to serve as a Ca supplement in developing healthy foods.

The enormous repetitive Antarctic krill genome reveals environmental adaptations and population insights.

Antarctic krill (Euphausia superba) is Earth's most abundant wild animal, and its enormous biomass is vital to the Southern Ocean ecosystem. Here, we report a 48.01-Gb chromosome-level Antarctic krill genome, whose large genome size appears to have resulted from inter-genic transposable element expansions. Our assembly reveals the molecular architecture of the Antarctic krill circadian clock and uncovers expanded gene families associated with molting and energy metabolism, providing insights into adaptations to the cold and highly seasonal Antarctic environment. Population-level genome re-sequencing from four geographical sites around the Antarctic continent reveals no clear population structure but highlights natural selection associated with environmental variables. An apparent drastic reduction in krill population size 10 mya and a subsequent rebound 100 thousand years ago coincides with climate change events. Our findings uncover the genomic basis of Antarctic krill adaptations to the Southern Ocean and provide valuable resources for future Antarctic research.

Metals and metalloids in Antarctic krill and water in deep Weddell Sea areas.

The study on the concentration of trace elements in Antarctic krill and in water in the deep areas of the Atlantic sector of the Antarctic was performed. Concentrations of 22 trace elements were studied to determine their spatial distribution in krill, and to assess the accumulation ability of the krill against 8 of them. The trace elements concentration in krill diminished in the following order: Fe > Cu > Zn > Bа > B > Se > As > Cr > Ni > Ag > Li > Mn > V > Mo > Cd > Co > Hg > Be. Concentrations of Pb, Ti, Tl, Sb were below their detection limits. Concentration factors of trace elements by krill varied from n × 102 to n × 104. The Cu and As concentrations in dry krill exceeded their MPC. Concentrations of all trace elements in wet mass of krill were not exceeded established regulative values.

Improving the texture properties and protein thermal stability of Antarctic krill (Euphausia superba) by L-lysine marination.

BACKGROUND: The quality deterioration of Antarctic krill (Euphausia superba) after thermal processing limits its industrial application. This study sought to improve the texture characteristics of Antarctic krill after heat treatment through pre-soaking using l-lysine (Lys) solution and sodium tripolyphosphate (STPP). Moreover, the effects of Lys on heat-treated Antarctic krill were explored. RESULTS: Lys significantly reduced the cooking loss and improved the texture characteristics of Antarctic krill during heat treatment. The low-field nuclear magnetic resonance results showed that Lys reduced the water loss of Antarctic krill during heat treatment. Additionally, the surface hydrophobicity, Fourier transform infrared spectroscopy, and endogenous fluorescence spectroscopy results showed that Lys could inhibit the structural damage of Antarctic krill protein under the thermal denaturation condition and enhance the thermal stability of the protein. The scanning electron microscopy results showed that Lys could protect the structural integrity of Antarctic krill muscle fibers during heat treatment. CONCLUSION: The cooking loss in the Lys added groups was better than the sodium tripolyphosphate added group, and 2.0% Lys solution could minimize the cooking loss of Antarctic krill. The secondary and tertiary structures of the Antarctic krill protein were actively protected by Lys during heat treatment. Overall, the study will provide insights into the application of Lys in the food industry as a natural additive and an alternative to phosphate. © 2021 Society of Chemical Industry.

Purification, Identification, Activity Evaluation, and Stability of Antioxidant Peptides from Alcalase Hydrolysate of Antarctic Krill (Euphausia superba) Proteins.

For utilizing the largest source of marine proteins, Antarctic krill (Euphausia superba) proteins were defatted and hydrolyzed separately using pepsin, alcalase, papain, trypsin, and netrase, and alcalase hydrolysate (EPAH) showed the highest DPPH radical (DPPH·) and hydroxyl radical (HO·) scavenging activity among five hydrolysates. Using ultrafiltration and chromatography methods, fifteen antioxidant peptides were purified from EPAH and identified as Asn-Gln-Met (NQM), Trp-Phe-Pro-Met (WFPM), Gln-Asn-Pro-Thr (QNPT), Tyr-Met-Asn-Phe (YMNF), Ser-Gly-Pro-Ala (SGPA), Ser-Leu-Pro-Tyr (SLPY), Gln-Tyr-Pro-Pro-Met-Gln-Tyr (QYPPMQY), Glu-Tyr-Glu-Ala (EYEA), Asn-Trp-Asp-Asp-Met-Arg-Ile-Val-Ala-Val (NWDDMRIVAV), Trp-Asp-Asp-Met-Glu-Arg-Leu-Val-Met-Ile (WDDMERLVMI), Asn-Trp-Asp-Asp-Met-Glu-Pro-Ser-Phe (NWD-DMEPSF), Asn-Gly-Pro-Asp-Pro-Arg-Pro-Ser-Gln-Gln (NGPDPRPSQQ), Ala-Phe-Leu-Trp-Asn (AFLWA), Asn-Val-Pro-Asp-Met (NVPDM), and Thr-Phe-Pro-Ile-Tyr-Asp-Tyr-Pro-Gln (TFPIYDPQ), respectively, using a protein sequencer and ESI/MS. Among fifteen antioxidant peptides, SLPY, QYPPMQY and EYEA showed the highest scavenging activities on DPPH· (EC50 values of 1.18 ± 0.036, 1.547 ± 0.150, and 1.372 ± 0.274 mg/mL, respectively), HO· (EC50 values of 0.826 ± 0.027, 1.022 ± 0.058, and 0.946 ± 0.011 mg/mL, respectively), and superoxide anion radical (EC50 values of 0.789 ± 0.079, 0.913 ± 0.007, and 0.793 ± 0.056 mg/mL, respectively). Moreover, SLPY, QYPPMQY and EYEA showed strong reducing power, protective capability against H2O2-damaged plasmid DNA, and lipid peroxidation inhibition ability. Furthermore, SLPY, QYPPMQY, and EYEA had high stability under temperatures lower than 80 °C, pH values ranged from 6-8, and simulated GI digestion for 180 min. The results showed that fifteen antioxidant peptides from alcalase hydrolysate of Antarctic krill proteins, especially SLPY, QYPPMQY and EYEA, might serve as effective antioxidant agents applied in food and health products.

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.

The First Genome Survey of the Antarctic Krill (Euphausia superba) Provides a Valuable Genetic Resource for Polar Biomedical Research.

The world-famous Antarctic krill (Euphausia superba) plays a fundamental role in the Antarctic food chain. It resides in cold environments with the most abundant biomass to support the Antarctic ecology and fisheries. Here, we performed the first genome survey of the Antarctic krill, with genomic evidence for its estimated genome size of 42.1 gigabases (Gb). Such a large genome, however, is beyond our present capability to obtain a good assembly, although our sequencing data are a valuable genetic resource for subsequent polar biomedical research. We extracted 13 typical protein-coding gene sequences of the mitochondrial genome and analyzed simple sequence repeats (SSRs), which are useful for species identification and origin determination. Meanwhile, we conducted a high-throughput comparative identification of putative antimicrobial peptides (AMPs) and antihypertensive peptides (AHTPs) from whole-body transcriptomes of the Antarctic krill and its well-known counterpart, the whiteleg shrimp (Penaeus vannamei; resident in warm waters). Related data revealed that AMPs/AMP precursors and AHTPs were generally conserved, with interesting variations between the two crustacean species. In summary, as the first report of estimated genome size of the Antarctic krill, our present genome survey data provide a foundation for further biological research into this polar species. Our preliminary investigations on bioactive peptides will bring a new perspective for the in-depth development of novel marine drugs.

High Fischer ratio oligopeptides determination from Antartic krill: Preparation, peptides profiles, and in vitro antioxidant activity.

In this work, alcalase and flavorzyme were chosen as the hydrolase for preparing high Fischer ratio oligopeptides from Antarctic krill (Euphausia superba) (HFP) using sequential enzyme hydrolysis process, and their hydrolysis conditions were optimized using single factor experiment. According to the Fischer ratio, granular activated carbon of XHJ-200 (200 mesh) showed the best way to remove aromatic amino acids and its optimal parameters were pH value of 6.0, adsorption time of 2.5 hr, temperature of 25°C, and solid-liquid ratio of 1:20. The Fischer ratio and average molecular weight of HFP were 21.12 (>20) and 779.9 Da, respectively. In addition, the peptide profile of HFP was established using RP-HPLC and 23 oligopeptides isolated from HFP including 6 dipeptides, 9 tripeptides, 3 tetrapeptides, and 5 pentapeptides were identified using protein amino acid sequence analyzer and mass spectrum. Furthermore, HFP exhibited high radical scavenging activity, reducing power, and lipid peroxidation inhibition capability. PRACTICAL APPLICATIONS: High Fischer ratio oligopeptides (HFO) is a kind of small peptide mixture with the mole ratio of branched-chain amino acids (BCAA) to aromatic amino acids (AAA) higher than 20, which has drawn a great attention due to its great potential in clinical nutrition approaches for the treatment of liver disease when amino acid composition is out of proportion characterized by low levels of BCAA and high levels of AAA in the systemic blood. Antarctic krill is regarded as the largest animal protein resource for various food and pharmaceutical products. However, there is no report on the preparation of HFO of Antarctic krill (HFP). Therefore, the aim of the present study was to investigate the preparation process, peptide profiles and in vitro antioxidant activity of HFP. This study will potentially enhance the value-added utilization of Antarctic krill by making it an important raw material in the health-promoting functional products.

Eight antihypertensive peptides from the protein hydrolysate of Antarctic krill (Euphausia superba): Isolation, identification, and activity evaluation on human umbilical vein endothelial cells (HUVECs).

In this report, eight antihypertensive peptides were isolated from protein hydrolysate of Antarctic krill (Euphausia superba) using ultrafiltration and chromatography consecutively, and their sequences were identified as Trp-Phe, Tyr-Arg-Lys-Glu-Arg, Tyr-Arg-Lys, Val-Asp, Tyr-Lys-Asp, Phe-Gln-Lys, Phe-Ala-Ser, and Phe-Arg-Lys-Glu. The IC50 values of Trp-Phe (0.32 ±â€¯0.05 mg/mL) and Phe-Ala-Ser (0.15 ±â€¯0.02 mg/mL) on ACE inhibitory activity were significantly (p ≤ .05) lower than those of the other six peptides. Furthermore, Trp-Phe, Tyr-Arg-Lys, Phe-Gln-Lys, and Phe-Ala-Ser did not only increase the nitric oxide (NO) concentration and decreased the content of endothelin-1 (ET-1) in the medium of human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner after 24 h, but also significantly reversed the decreased production of NO in the presence of 0.5 µM norepinephrine and the effect of NE on ET-1 production. These results indicate that the isolated antihypertensive peptides can correct the endothelial cell dysfunction induced by norepinephrine.

Characterization of protease and effects of temperature and salinity on the biochemical changes during fermentation of Antarctic krill.

BACKGROUND: Despite their abundance, Antarctic krill are underutilized because of numerous difficulties in their commercial processing. Ideally, fermentation technology can be applied to transform them into a popular condiment. In addition to the exploration of protease properties, the present study aimed to evaluate proteinase activity, pH, amino nitrogen, and histamine formation during fermentation at different temperatures and salt treatments. RESULTS: Even though the activity of Antarctic krill protease reached a maximum at 40 °C and pH 7, it was stable at 30 °C and pH 7-9. Among the metal ions tested, Ca2+ , Mg2+ and K+ increased protease activity, in contrast to Zn2+ and Cu2+ . Within each treatment, the highest protease activity and amino nitrogen content, as well as the lowest histamine level, were observed on day 12 of fermentation. Treatment at 35 °C with 180 g kg-1 salt led to the production of maximum amino nitrogen (0.0352 g kg-1 ) and low histamine (≤0.0497 g kg-1 ). CONCLUSION: Krill paste fermented for 12 days at 35 °C with 180 g kg-1 salt exhibited the optimal quality and properties, suggesting an efficient method for fermentation of Antarctic krill and other aquatic resources. © 2017 Society of Chemical Industry.