Identification, Screening and Antibacterial Mechanism Analysis of Novel Antimicrobial Peptides from Sturgeon (Acipenser ruthenus) Spermary.

Fish is an important source of antimicrobial peptides. This study aimed to identify and screen antibacterial peptides with excellent antibacterial activity derived from sturgeon spermary peptides (SSPs) and to analyze their antibacterial activity and mechanism. Liquid chromatography-mass spectrometry/mass spectrometry methods were used to analyze and identify peptide sequences, computational prediction tool and molecular docking methods were used for virtual screening of antimicrobial peptides, and finally, candidate peptides were synthesized by solid-phase synthesis method. The results demonstrate that SSPs have excellent inhibitory activity against Escherichia coli with an inhibitory rate of 76.46%. Most parts of the SSPs were derived from the sturgeon (Acipenser ruthenus) histones, and the coverage of histone H2B was the highest (45%). Two novel peptides (NDEELNKLM and RSSKRRQ) were obtained by in silico prediction tools and molecular docking, which may interact with the DNA gyrase and dihydrofolate reductase of E. coli by forming salt bridges and hydrogen bonds. Compared to the individual peptides, the antibacterial effect was significantly improved by mixing the two peptides in equal proportions. Two novel peptides change the permeability of the E. coli cell membranes and may exert antimicrobial activity by inhibiting the metabolic process of the nucleic acids.

Silver Carp (Hypophthalmichthys molitrix) Scale Collagen Peptides-1 (SCPs1) Inhibit Melanogenesis through Downregulation of the cAMP-CREB Signaling Pathway.

The mechanism of silver carp scale collagen peptides (SCPs1) on melanogenesis and its mechanism of action were examined in mouse melanoma cells (B16). The cell viability and effects of SCPs1 on intracellular tyrosinase (TYR) activity and melanin, reactive oxygen species (ROS), glutathione (GSH) and cyclic adenosine monophosphate (cAMP) content were examined. The regulatory mechanism of SCPs1 on the cAMP response element-binding protein (CREB) signaling pathway was analyzed. The cell viability of the SCPs1 group was >80% (0.01-1 mg/mL) and the inhibitory rate of SCPs1 on B16 cell melanin increased in a dose-dependent manner. The highest inhibitory rate of SCPs1 on melanin content reaching 80.24%. SCPs1 significantly increased the GSH content and decreased the tyrosinase activity, as well as the content of ROS and cAMP. Western blot analysis showed that SCPs1 significantly inhibited melanocortin-1 receptor (MC1R) expression and CREB phosphorylation in the cAMP-CREB signaling pathway, leading to downregulation of microphthalmia-associated transcription factor (MITF) and the expression of TYR, TYR-related protein-1 (TRP-1) and TRP-2. SCPs1 also inhibited the expression of MC1R, MITF, TYR, TRP-1 and TRP-2 at the transcriptional level. Taken together, SCPs1 inhibited melanin synthesis through the downregulation of the cAMP-CREB signaling pathway. Fish-derived collagen peptides could potentially be applied in skin whitening products.

Isolation, Identification, and Biological Activity Analysis of Swim Bladder Polypeptides from Acipenser schrencki.

Swim bladder polypeptides (SBPs) of Acipenser schrencki were analyzed for their antioxidant activity and physicochemical properties. The results showed the optimal enzymatic conditions were alkaline protease with a solid-to-liquid ratio of 1:20, an incubation time of 4 h, a temperature of 55 °C, and an enzyme dosage of 5000 U/g. Three different molecular weight fractions (F1, F2, and F3) were obtained via ultrafiltration. F3 (912.44-2135.82 Da) showed 77.90%, 72.15%, and 66.25% removal of O2•-, DPPH•, and •OH, respectively, at 10 mg/mL, which was significantly higher than the F1 and F2 fractions (p < 0.05). F3 contained proline (6.17%), hydroxyproline (5.28%), and hydrophobic amino acids (51.39%). The UV spectrum of F3 showed maximum absorption at 224 nm. Peptide sequence analysis showed that F3 contained antioxidant peptides (MFGF, GPPGPRGPPGL, and GPGPSGERGPPGPM) and exhibited inhibitory activities on angiotensin-converting enzyme and dipeptidyl peptidase III/IV (FRF, FPFL and LPGLF). F3 was considered a good raw material for obtaining bioactive peptides.

Silver Carp (Hypophthalmichthys molitrix) Scales Collagen Peptides (SCPs): Preparation, Whitening Activity Screening and Characterization.

This study involves the preparation of scale collagen peptides (SCPs) with whitening activity from silver carp (Hypophthalmichthys molitrix) and their characterization and peptide sequence identification. In this article, scanning electron microscopy (SEM) was used to observe structure changes of sliver carp scales; enzymatic hydrolysis was optimized through protease screening and response surface optimization. The ultrafiltration was used to separate SCPs and the whitening activity was comprehensively evaluated using radical scavenging rate and tyrosinase-inhibiting activity, among others. An optimal component was characterized and identified using various modern spectral analysis techniques. The results showed that the surface of silver carp scales after decalcification was smooth and clear. The pepsin had the highest peptide yield and tyrosinase-inhibiting activity (90.01% and 82.25%, respectively). The optimal enzymatic hydrolysis conditions were an enzyme dosage of 16.1%, a solid-liquid ratio of 1:15.6 and a time of 4.9 h. The proportions of hydrophobic and basic amino acids in the peptide composition were 32.15% and 13.12%, respectively. Compared with SCPs2, SCPs1 (6096.68-9513.70 Da) showed better ·OH scavenging ability, tyrosinase-inhibiting activity and moisture absorption. SCPs1 was a macromolecular fragment of type I collagen with a triple helix structure, containing three peptide sequences with the potential for tyrosinase activity inhibition (AGPPGADGQTGQRGE, SGPAGIAGPAGPRGPAGPNGPPGKD and KRGSTGEQGSTGPLGMRGPRGAA). These results show that SCPs1 is a collagen peptide product with whitening potential.

Analysis of Water Distribution and Muscle Quality of Silver Carp (Hypophthalmichthys molitrix) Chunks Based on Electron-Beam Irradiation.

Electron-beam irradiation (EBI) is an efficient, safe, and nonthermal sterilization technique that is extensively used in food preservation research. Here we report the effects of different EBI doses (0, 4, 8 kGy) and preservation temperatures (room temperature [RT], 4 °C) on the muscle water distribution and muscle quality indices of silver carp chunks (SCCs). The highest entrapped water content was found in the 4-kGy-irradiated/4-°C-stored samples. The expressible moisture content (EMC) of the SCCs increased with increasing irradiation dose and was significantly lower in the RT group than in the 4 °C group. The irradiation dose and preservation temperature had no significant effect on the moisture content, whiteness value and protein content of SCCs (p > 0.05). When the irradiation dose reached 8 kGy, AV value, POV value and TVB value were significantly increased (p < 0.05). The myofibrillar protein content and actomyosin content of the SCCs in the 4 °C group was higher than that of the specimens in the RT group by 0.29−0.98 mg/mL (p < 0.05) and 36.21−296.58 µg/mL (p < 0.05), respectively. Overall, EBI treatment (4 kGy) and low-temperature preservation (4 °C) helped retain the muscle water content of the SCCs and preserve their quality, thereby endorsing the EBI treatment of silver carp products.

Antimicrobial activities and mechanism of sturgeon spermary protein extracts against Escherichia coli.

This study aimed to evaluate the antimicrobial activities and mechanism of sturgeon spermary protein extracts (SSPE) against Escherichia coli. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. Cell structural change was analyzed using scanning electron microscopy-energy dispersive X-ray spectrometry and transmission electron microscope. Moreover, pH, zeta potential, membrane potential, intracellular ATP concentrations and the interaction of SSPE with genomic DNA were analyzed. Results showed that molecular weight of SSPE is 13.4 kDa, the content of basic amino acids is the highest, in which arginine accounts for 73.2%. The MIC and MBC of SSPE for E. coli were 0.05 and 5 mg/mL, respectively. After SSPE treatment, cell membrane permeability changes, zeta potential decrease and genomic DNA lysis occurred in E. coli, which indicated it exerted bacteriostatic effects either independently or simultaneously by destroying the cell membrane and genomic DNA. These findings indicated that SSPE has potential to be a natural antiseptic.

Physicochemical Properties and Biological Activities of Silver Carp Scale Peptide and Its Nanofiltration Fractions.

To explore the physicochemical properties and biological functions of silver carp scale peptide (SCSP), its molecular-weight fractions SCSP-I, II, and III obtained by nanofiltration were assessed for their solubility, emulsibility, free radical scavenging ability, effect on the proliferation of mouse B16 cells. The results showed that the solubility of each fraction of SCSP was higher than 90%, SCSP-II and III were higher than 95%. The antioxidant powers on ⦁OH, O 2 - ⦁ and Fe3+ were ranked as SCSP-III > SCSP-II > SCSP-I > SCSP. All fractions of SCSP had no toxic or side effects in mouse B16 melanoma cells experiments in vitro. At a concentration of 0.01 mg/mL, the tyrosinase activity of B16 cells in the SCSP-II fraction was significantly lower than that of the α-arbutin (P < 0.05), at 65.37%. The molecular weight distribution of SCSP was 399-1404 Dalton and 13 peptide sequences were detected. Among them, SCSP-II contained many hydrophobic amino acids, and SCSP-III stood out for combining arginine with hydrophobic amino acids. This may be the reason why the low molecular-weight SCSPs show the strong antioxidant activity and strong tyrosinase inhibition. The work provides a data base for the development of SCSP and increases the possibility of its application.

Cobalt-60 and electron beam irradiation-induced lipid oxidation in largemouth bass (Micropterus salmoides).

BACKGROUND: Irradiation can cause lipid oxidation of fish. This study aimed to examine the effect of radiation (method, dose and dose rate) on the acid value (AV), peroxide value (PV), thiobarbituric acid reactive substances (TBARS) content and fatty acid profile of fresh and freeze-dried largemouth bass flesh. RESULTS: AV, PV and TBARS presented a dose-dependent increase in fish meat for both cobalt-60 (60 Co) and electron beam (EB) irradiation. With a 6 kGy dose of radiation, all measured indices in the 60 Co group were significantly higher than those in the EB group (P < 0.05 or P < 0.01). With a 3 kGy dose of radiation, AV, PV and TBARS in the 200 Gy min-1 dose rate group were significantly lower than those in the 2 and 80 Gy min-1 groups (P < 0.05). After 60 Co irradiation, AV, PV and TBARS in most fresh samples were significantly higher than those in freeze-dried samples (P < 0.01). And 60 Co irradiation decreased the unsaturated fatty acid (UFA) content in fresh samples and increased the UFA content in freeze-dried samples. Our study indicated that 60 Co irradiation, particularly at a low dose rate, accelerated lipid oxidation in fish meat. A large amount of muscle moisture enhances the amount of UFA loss in fish meat during 60 Co irradiation. CONCLUSIONS: A low dose (3 kGy) of EB irradiation, a high dose rate (200 Gy min-1 ) of 60 Co irradiation or freeze-drying treatment can alleviate the lipid oxidation of largemouth bass meat. © 2020 Society of Chemical Industry.

Arachis hypogaea L. stem and leaf extract improves the sleep behavior of pentobarbital-treated rats.

This study was conducted to evaluate the sedative effects of Arachis hypogaea L. stem and leaf extract (AHSLE) and determine its effect pathways through γ-aminobutyric acid (GABA)-gated channels on male Sprague-Dawley rats treated with pentobarbital. AHSLE was obtained from 98°C water (3 h, extracted twice). AHSLE and flumazenil (a GABA type A receptor antagonist) were administered to the rats orally, whereas pentobarbital sodium and muscimol (a GABA type A receptor agonist) were administered intraperitoneally (i.p.). The results demonstrated that AHSLE decreased sleep latency and increased sleep time in pentobarbital-treated rats (50 mg/kg, i.p.). The coadministration of AHSLE and muscimol (0.05 mg/kg) significantly increased sleep time and reduced sleep latency in pentobarbital-treated rats and these actions were significantly antagonized by flumazenil at a dose of 3.5 mg/kg. These results indicated that AHSLE improved the sleep behavior in pentobarbital-treated rats, possibly through GABA-gated channel-related mechanisms.

Anthocyanins extracted from Chinese blueberry (Vaccinium uliginosum L.) and its anticancer effects on DLD-1 and COLO205 cells.

BACKGROUND: Vaccinium uliginosum L. is a type of blueberry found in the Chinese Changbai Mountains. We extracted Vaccinium uliginosum Anthocyanins (A(V.uli)) to investigate its bioactivity on suppressing cancer cells. METHODS: A(V.uli) was extracted under different conditions of temperature (10°C - 35°C), pH 1.0 - 3.0, and diatomaceous earth (1.0 g - 3.0 g), followed by a HPLC analysis for the determination of the ingredients. Its anticancer bioactivities on human colon and colorectal cancer cells (DLD-1 and COLO205) were compared with those on Lonicera caerulea Anthocyanins (A(L.cae)) and Vaccinium myrtillus Anthocyanins (A(V.myr)), using cell viability assays, DNA electrophoresis and nuclear morphology assays. RESULTS: The optimum process of A(V.uli) extraction involved conditions of temperature 20°C, pH 2.0, and diatomaceous earth 1.0 g/50 g of fruit weight. A(V.uli) contained 5 main components: delphinidin (40.70 ± 1.72)%, cyanidin (3.40 ± 0.68)%, petunidin (17.70 ± 0.54)%, peonidin (2.90 ± 0.63)% and malvidin (35.50 ± 1.11)%. The malvidin percentage was significantly higher (P < 0.05) than it in A(V.myr). A(V.uli) complied with a dose-dependent repression of cancer cell proliferation with an IC(50) (50% inhibitory concentration) value of 50 µg/ml, and showed greater anticancer efficiency than A(L.cae) and A(V.myr) under the same cell treatment conditions. These observations were further supported by the results of nuclear assays. CONCLUSIONS: The extraction protocol and conditions we used were effective for anthocyanin extraction. A(V.uli) could be a feasible practical research tool and a promising therapeutic source to suppress human colon or colorectal cancers.