Effect of three unsaturated fatty acids on the protein oxidation and structure of myofibrillar proteins from rainbow trout (Oncorhynchus mykiss).

In this study, the impact of three unsaturated fatty acids (Oleic acid: OA, Eicosapentaenoic acid: EPA, Docosahexaenoic acid: DHA) on the oxidation and structure of rainbow trout myofibrillar protein (MP) was explored. The findings revealed a notable increase in carbonyl content (P < 0.05) and a significant decrease in total sulfhydryl content (P < 0.05) of MP with the concentration increase of the three unsaturated fatty acids. Endogenous fluorescence spectroscopy and surface hydrophobicity analyses showed that unsaturated fatty acids can cause unfolding and exposure of hydrophobic groups in MP. In addition, SDS-PAGE showed that disulfide bonds were associated with MP cross-linking and aggregate size induced by unsaturated fatty acids. Overall, three unsaturated fatty acid treatments facilitated the oxidation of myofibrillar proteins, and the extent of protein oxidation was closely associated with the concentration of unsaturated fatty acids.

Mitochondria of Live Mizuhopecten yessoensis Scallops Can Sensitively Respond to Quality Changes during Dry/Reimmersed Storage as Determined by TMT-Labeled Proteomic Analysis.

Dry/reimmersed storage is often used in the transportation of live scallops. In this study, tandem mass tag (TMT)-labeled protein omics were used to quantitatively analyze the protein changes in scallops during dry/reimmersed stress. The results showed that during dry storage, scallops maintained cellular redox homeostasis through the upregulation of SCO1-like protein and thioredoxin domain-containing protein and reduced organic acids from the ATP synthetic process by the downregulation of NADH dehydrogenase, thereby reducing the damage caused during dry storage. During reimmersed storage, mitochondrial proteins underwent very sensitive changes. By upregulating aerobic respiration-related proteins (including proteins involved in glucose phosphate metabolism, glyceraldehyde 3-phosphate metabolism, etc.), the ATP synthesis ability was improved. However, the damage to the mitochondrial structure by dry storage could not be completely recovered, even by reimmersion. This included some apoptosis-related proteins that were obviously upregulated. In summary, compared with ATP-related indexes, mitochondria can respond more sensitively to dry storage stress.

Mitochondrial activity as an indicator of scallop (Mizuhopecten yessoensis) adductor muscle in early cold storage.

The purpose of this study was to reveal the role of mitochondria in indicating a change in the freshness of the adductor muscle of Mizuhopecten yessoensis during cold storage and hardening. The adductor muscle hardens after 96 h of cold storage and reaches the maximum degree of hardening in 6 h. In this study, after hardening (102 h), the muscle fiber structure obviously broke and curled, and the sarcomere structure disappeared at 150 h. After hardening (102 h), the morphology of the mitochondria changed, including swelling, cristea breaking and membrane structure disappearance. The arginine phosphate content decreased gradually from the initial 4.618 to 1.306 µmol/g at 48 h, and there was no further obvious change. The ATP content decreased from 6.02 to 1.07 µmol/g in 120 h. The mitochondrial membrane potential (Δφ) was measured by a fluorescence method (JC-1). The changes in freshness could be divided into three classes according to the Δφ difference between the mitochondria in the adductor muscle after adding ADP and CCCP. Mitochondrial function was complete from 0 to 6 h; mitochondrial function began to decline at 6 to 48 h; and mitochondrial function completely disappeared after 48 h. The results showed that the mitochondrial membrane potential compared with other indicators could more sensitively reflect the changes in freshness during the cold storage hardening process of the adductor muscle. PRACTICAL APPLICATION: It is very important to interprete the post-mortem energy metabolism for controlling the muscle quality of Yesso scallop. The K value which was developed based on ATP-related compounds is widely used in evaluating the freshness of seafood. Mitochondria is the main sites of cellular energy metabolism and the changes of its structure and activity can sensitively reflect the quality changes in muscle cell. It is expected to develop a more sensitive freshness evaluation index.

Inonotus obliquus polysaccharide ameliorates serum profiling in STZ-induced diabetic mice model.

BACKGROUND: Diabetes mellitus is a systemic disease mainly caused by the disorder of metabolism, which has become huge threat to human health. Polysaccharides are the main active substance from Inonotus obliquus (I. obliquus) with hypoglycemic effect. This study aims to evaluate the hypoglycemic activity and investigate the molecular mechanism of I. obliquus polysaccharide (IOP) in streptozotocin (STZ)-induced diabetic mice using metabolomics based on UPLC-Q-Exactive-MS method. RESULTS: The results showed that the oral administration of IOP in high dose (1.2 g/kg) can significantly reduce the blood glucose with 31% reduction comparing with the diabetic model and relieve dyslipidemia in diabetic mice. By UPLC-Q-Exactive-MS method and multivariate statistical analysis, a total of 15 differential metabolites were identified, including 4 up-regulated and 11 down-regulated biomarkers, of which L-tryptophan, L-leucine, uric acid, 12-HETE, arachidonic acid, PC(20:1(11Z)/14:1(9Z)) and SM(d18:0/24:1(15Z)) were exhibited an important variation, as the potential biomarkers in diabetes. Pathway analysis indicated that phenylalanine, tyrosine and tryptophan biosynthesis and arachidonic acid metabolism were prone to interference in diabetes. Moreover, leucine and proline were reversed and phytosphingosine was further reduced in diabetic mice under the intervention of IOP. CONCLUSION: IOP has predominant hyperglycemic effect on STZ-induced diabetic mice via ameliorating serum profiling.

Condition-dependent adenosine monophosphate decomposition pathways in striated adductor muscle from Japanese scallop (Patinopecten yessoensis).

The purpose of this study was to confirm inosine monophosphate (IMP) generation and to clarify the decomposition pathway of adenosine monophosphate (AMP) by investigating the properties of AMP, IMP, and adenosine (AdR) decomposition enzymes in Japanese scallop (Patinopecten yessoensis). The results showed that IMP accumulated due to AMP decomposition via endogenous enzymes in scallops stored at both 4 °C and 20 °C. The AMP decomposition rate was highest in the supernatant of homogenized scallop adductor muscle, followed by the suspended solution and precipitate, while IMP could not be decomposed in scallop. The results indicated that the activity of AdR deaminase was very high, and this enzyme was involved in an intracellular process in scallop. Moreover, 1 min of heating exerted little influence on the AMP and AdR decomposition rates, while 5 min of heating induced enzyme denaturation. The IMP generation rate increased dramatically in scallop crude enzyme solution containing 5 mM ethylenediaminetetraacetic acid (EDTA). This suggests that the major pathway of AMP decomposition might change with variations in metal ion concentrations in Japanese scallop. PRACTICAL APPLICATION: IMP generation in Japanese scallop (Patinopecten yessoensis) caused by endogenous enzymes was confirmed. IMP is very important for the umami taste (a pleasant savory taste) of aquatic products. As IMP accumulation might be achieved by changing the concentration of divalent metal ions and no IMP 5'-nucleotidase activity was detected in scallop, a suitable process to produce good flavor scallops with high IMP contents might be developed.

Effects of thawing methods on the biochemical properties and microstructure of pre-rigor frozen scallop striated adductor muscle.

Postmortem biochemical properties (pH, salt solubility, Ca2+-ATPase activity, ATP-related compounds) and microstructural changes in the striated adductor muscle of pre-rigor frozen Japanese scallops (Patinopecten yessoensis) were studied after thawing and during storage at 4℃. Four thawing methods were used: running water (18℃, R); ice-water (0℃, I); air (4℃, A) and ice-saltwater (-2℃, S). The pH values and salt solubility of R group were lower than the other three thawing groups while I group was highest after thawing. However, no significant difference (P < 0.05) in Ca2+-ATPase activity were detected among 4 groups. The microstructure results indicated that the structure of I group was close to that of fresh scallop. Moreover, ATP decomposition rate was the slowest. Therefore, ice-water thawing is the best method because it induced the least changes in the biochemical properties and microstructures of scallop adductor muscle.

Nutritional and Digestive Properties of Protein Isolates Extracted from the Muscle of the Common Carp Using pH-Shift Processing.

This study details the nutritional and digestive properties of protein isolates that are extracted from carp (Cyprinus Carpio L.) muscle using pH shifting methods. Alkaline (ALPI) and acid (ACPI) protein isolates exhibit higher protein yields (87.6%, 76.3%, respectively). In addition to the high recovery of myofibrillar protein, a portion of the water-soluble proteins is also recovered. The moisture contents of ACPI and ALPI are 85.5% and 88.5%, respectively, and the crude protein contents of these two fractions are 83.20% and 83.0%, respectively, both contents of which are higher than those for fresh muscle. Most part of the ash and fat are removed in the separation process. The protein isolation is also found to be lighter and whiter than the fresh muscle and there is no difference between amino acid content of protein isolation and that of fresh muscle. The maximum solubility of water washed surimi is 73.21%, while solubility of ACPI-2 and ALPI-2 (pH 7.0) are 66.67% and 62.08%, respectively. The digestibility of ALPI and ACPI is improved after being treated with chymotrypsin, which is about 7-8 times as that of fresh muscle. The results indicate that the protein isolates have better nutritional and digestive properties than the fresh muscle does in food processing. PRACTICAL APPLICATIONS: Common carp is a lower additional value fish that exists in large amount in China. This study investigates nutritional and digestive properties of protein from carp extracted by pH shifting methods. According to the obtained data in this study, pH shifting method is a good protein recovery method that can effectively remove bone spurs, skin, fat and other impurities. In addition, sarcoplasmic proteins can also be recovered. The nutritional properties of protein isolates of carp were suitable for supplementing as an ingredient for human consumption. The pH-shift process greatly improves the protein digestibility. Therefore, there are broad application prospects of the protein isolation as protein ingredients in food industry.

Structural and functional characterization of CATH_BRALE, the defense molecule in the ancient salmonoid, Brachymystax lenok.

Thick-lipped lenok, Brachymystax lenok is one of the ancient fish species in China and northeast Asia countries. Due to the overfishing, the population of lenok has been declined significantly. Cathelicidins are innate immune effectors that possess both bactericidal activities and immunomodulatory functions. This report identifies and characterizes the salmonoid cathelicidin (CATH_BRALE) from this ancient fish. It consists of open reading frame (ORF) of 886 bp encoding the putative peptide of 199 amino acids. Sequence alignment with other representative salmonid cathelicidins displayed two distinctive features of current lenok cathelicidin: high level of arginine, resulting in high positive charge and glycine residues, which is significantly different from most acknowledged types of cathelicidins; and the six-aminoacid tandem repeated sequence of RPGGGS detected in a variable number of copies among fish cathelicidins, suggesting the existence of a genetically unstable region similar to that found in some mammalian cathelicidins. Expression of CATH_BRALE is predominantly found in gill, with lower levels in the gastrointestinal tract and spleen. The homology modeled structure of CATH_BRALE exhibits structural features of antiparallel b-sheets flanked by a-helices that are representative of small cationic cathelicidin family peptides. CATH_BRALE possesses much stronger antimicrobial activity against gram-negative bacteria than that of the human ortholog, LL-37. The growth of two typical fish bacterial pathogens, gram-negative bacterium of Aeromonas salmonicida and Aeromonas hydrophila was substantially inhibited by synthetic CATH_BRALE, with both MICs as low as 9.38 mM.