Actin Degradation Research Articles

Effects of Stocking Density and Pre-Slaughter Handling on the Fillet Quality of Largemouth Bass (Micropterus salmoides): Implications for Fish Welfare.

There is currently insufficient acknowledgment of the relationship between fish welfare and ultimate fillet quality. The purpose of this study was to assess the impacts of pre-slaughter handling and stocking density as fish welfare markers on fillet quality of largemouth bass (Micropterus salmoides). Fish from three stocking densities of 35, 50, and 65 kg·m-3 were reared in a recirculating aquaculture system (RAS) for 12 weeks and received commercial feed. Ultimately, the fish were either stunned with percussion on the head (control group) or subjected to air exposure for 3 min (anoxia group) before stunning and subsequent collection of blood and fillet samples. Western blot analysis revealed the degradation of actin in both groups. Additionally, higher oxidation progress and lower hardness and pH were observed in anoxia compared to the control group. We observed higher hardness at 35 kg·m-3 in anoxia compared to 50 and 65 km-3. The initial hardness values at 35, 50, and 65 km-3 were 1073, 841, and 813 (g) respectively in the anoxia group. Furthermore, the anoxia and control groups had rigor mortis after 6 and 10 h, respectively. Cortisol and glucose levels, and oxidative enzymes activity were higher in anoxia than in the control group. In conclusion, oxidation induced by anoxia likely plays a crucial role as a promoter of the quality deterioration of largemouth bass fillets.

The Impact of Oxygen and Ascorbic Acid Treatments on the Physicochemical Attributes of Live Sea Cucumber During Extended Transportation/Storage

ABSTRACT Sea cucumbers were stored live in 25% oxygen or 25 mg/mL of ascorbic acid for 7 days, for comparison purposes. The condition of the individuals was assessed through the sensory scores, bacterial communities, and biochemical indexes. The oxygen treatment extended preservation by 2 days compared to ascorbic acid. After 7 days, the total volatile basic nitrogen and the microbial content in all sea cucumbers were within acceptable limits. The actin degradation rate reached 1.3416 ± 0.03%, and cathepsin B and L increased by 94.6% and 59.1%, respectively, in the Control group. The quality decline of sea cucumbers during storage was caused by autolysis rather than spoilage organisms.

Macrophage derived Exosomal Docetaxel (Exo-DTX) for pro-metastasis suppression: QbD driven formulation development, validation, in-vitro and pharmacokinetic investigation

Exosomes, biogenic nano-vesicles, are renowned for their ability to encapsulate diverse payloads, however the systematic development and validation of exosomal formulation with significant biological implications have been overlooked. Herein, we developed and validated Exo-DTX, a QbD-driven optimized RAW 264.7 cell derived exosomal anti-cancer formulation of docetaxel (DTX) and evaluate its anti-metastatic and apoptotic efficacy in TNBC 4T1 cells. RAW264.7-derived exosomes were having particle size (112.5 ± 21.48 nm) and zeta-potential (−10.268 ± 3.66 mV) with polydispersity (PDI:0.256 ± 0.03). The statistical optimization of exosomes (200 μg) with Exo: DTX ratio 4:1 confirmed encapsulation of 23.60 ± 1.54 ng DTX/ µg exosomes. Exo-DTX (∼189 nm, −11.03 mV) with 100 ng/ml DTX as payload exhibited ∼5 folds’ improvement in IC50 of DTX and distinct cytoskeletal deformation in TNBC 4T1 cells. It also has shown enormous Filamentous actin (F-actin) degradation and triggered apoptosis explained Exo-DTX's effective anti-migratory impact with just 2.6 ± 6.33 % wound closure and 4.56 ± 1.38 % invasion. The western blot confirmed that Exo-DTX downregulated migratory protein EGFR and β1-integrin but raised cleaved caspase 3/caspase 3 (CC3/C3) ratio and BAX/BCL-2 ratio by about 2.70 and 4.04 folds respectively. The naive RAW 264.7 exosomes also contributed positively towards the effect of Exo-DTX formulation by suppressing β1-integrin expression and increasing the CC3/C3 ratio in TNBC 4T1 cells as well. Additionally, significant improvement in PK parameters of Exo-DTX was observed in comparison to Taxotere, 6-folds and 3.04-folds improved t1/2 and Vd, proving the translational value of Exo-DTX formulation. Thus, the Exo-DTX so formulated proved beneficial in controlling the aggressiveness of TNBC wherein, naive exosomes also demonstrated beneficial synergistic anti-proliferative effect in 4T1.

Lactic acid fermentation of kamaboko, a heated Alaska pollock surimi, enhances angiotensin I-converting enzyme inhibitory activity via fish protein hydrolysis.

To enhance the value of surimi, efforts have been made to develop a fermentation method with lactic acid bacteria (LAB) to proteolyze fish protein. However, fermenting unheated surimi poses a spoilage risk due to its high bacterial content. Surimi heat treatment can prevent spoilage, but gel formation induced by heating introduces another technical issue: it hinders uniform fermentation. Thus, this study aims to observe the proteolysis and enhance the functionality of seafood product through lactic acid fermentation of kamaboko, a heated surimi. Upon analyzing the kamaboko fermented with Lactobacillus helveticus JCM1004, we observed that LAB produced protease, resulting in the degradation of myosin heavy chain and actin during fermentation. Lactic acid fermentation significantly augmented the peptide content of kamaboko, subsequently elevating the angiotensin Ⅰ-converting enzyme (ACE) inhibitory activity in 200-fold diluted extract of fermented kamaboko to approximately 70% and higher. Notably, our investigation revealed that proteolysis was confined to the surface of kamaboko, as evidenced by SDS-PAGE analysis. This observation implies that the surface area of kamaboko influences the ACE inhibitory activity. Through a comparative analysis of various bacterial strains, we demonstrated that the increase in ACE inhibitory activity is contingent on the protease generated by LAB. These results suggest that LAB-mediated proteolysis of fish proteins liberates bioactive peptides, thereby manifesting in the ACE inhibitory activity. In summary, this study underscores that the fermentation of kamaboko employing proteolytic LAB holds promise in the development of novel functional seafood products.

Protein Phosphorylation Induced by Pyruvate Kinase M2 Inhibited Myofibrillar Protein Degradation in Post-Mortem Muscle.

Myofibrillar protein degradation is primarily related to meat tenderness through protein phosphorylation regulation. Pyruvate kinase M2 (PKM2), a glycolytic rate-limiting enzyme, is also regarded as a protein kinase to catalyze phosphorylation. The objective of this study was to investigate the relationship between myofibrillar protein degradation and phosphorylation induced by PKM2. Myofibrillar proteins were incubated with PKM2 at 4, 25, and 37 °C. The global phosphorylation level of myofibrillar proteins in the PKM2 group was significantly increased, but it was sensitive to temperature (P < 0.05). Compared with 4 and 25 °C, PKM2 significantly increased the myofibrillar protein phosphorylation level from 0.5 to 6 h at 37 °C (P < 0.05). In addition, the degradation of desmin and actin was inhibited after they were phosphorylated by PKM2 when incubated at 37 °C. These results demonstrate that phosphorylation of myofibrillar proteins catalyzed by PKM2 inhibited protein degradation and provided a possible pathway for meat tenderization through glycolytic enzyme regulation.

Application and Effect of Pediococcus pentosaceus and Lactiplantibacillus plantarum as Starter Cultures on Bacterial Communities and Volatile Flavor Compounds of Zhayu, a Chinese Traditional Fermented Fish Product.

Zhayu is a type of traditional fermented fish product in China that is made through the fermentation of salted fish with a mixture of cereals and spices. Inoculation fermentation was performed using Pediococcus pentosaceus P1, Lactiplantibacillus plantarum L1, and a mixture of two strains, which were isolated from cured fish in Hunan Province. Compared with the natural fermentation, inoculation with lactic acid bacteria (LAB) accelerated the degradation of myosin and actin in Zhayu, increased the trichloroacetic acid (TCA)-soluble peptide content by about 1.3-fold, reduced the colony counts of Enterobacteriaceae and Staphylococcus aureus by about 40%, and inhibited their lipid oxidation. In the texture profile analysis performed, higher levels of hardness and chewiness were observed in the inoculation groups. In this study, the bacterial community and volatile flavor compounds were detected through 16S high-throughput sequencing and headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). Inoculation with L. plantarum L1 reduced around 75% abundance of Klebsiella compared with the natural fermentation group, which was positively correlated with 2,3-Butanediol, resulting in a less pungent alcohol odor in Zhayu products. The abundances of 2-pentylfuran and 2-butyl-3-methylpyrazine were increased over threefold in the L1 group, which may give Zhayu its unique flavor and aroma.

The Role of Vitamin D and Vitamin D Binding Protein in Chronic Liver Diseases.

Vitamin D (calciferol) is a fat-soluble vitamin that has a significant role in phospho-calcium metabolism, maintaining normal calcium levels and bone health development. The most important compounds of vitamin D are cholecalciferol (vitamin D3, or VD3) and ergocalciferol (vitamin D2, or VD2). Besides its major role in maintaining an adequate level of calcium and phosphate concentrations, vitamin D is involved in cell growth and differentiation and immune function. Recently, the association between vitamin D deficiency and the progression of fibrosis in chronic liver disease (CLD) was confirmed, given the hepatic activation process and high prevalence of vitamin D deficiency in these diseases. There are reports of vitamin D deficiency in CLD regardless of the etiology (chronic viral hepatitis, alcoholic cirrhosis, non-alcoholic fatty liver disease, primary biliary cirrhosis, or autoimmune hepatitis). Vitamin D binding protein (VDBP) is synthesized by the liver and has the role of binding and transporting vitamin D and its metabolites to the target organs. VDBP also plays an important role in inflammatory response secondary to tissue damage, being involved in the degradation of actin. As intense research during the last decades revealed the possible role of vitamin D in liver diseases, a deeper understanding of the vitamin D, vitamin D receptors (VDRs), and VDBP involvement in liver inflammation and fibrogenesis could represent the basis for the development of new strategies for diagnosis, prognosis, and treatment of liver diseases. This narrative review presents an overview of the evidence of the role of vitamin D and VDBP in CLD, both at the experimental and clinical levels.

Control on moisture distribution and protein changes of Antarctic krill meat by antifreeze protein during multiple freeze-thaw cycles.

Control on the moisture distribution, protein structure changes, and protein degradation of Antarctic krill meat during freeze-thaw (F-T) cycles by presoaking with antifreeze protein (AFP) was investigated. The results from the thawing loss rate and cooking loss rate indicated that 0.1% was the optimal AFP concentration. Magnetic resonance imaging and low-field nuclear magnetic resonance results showed that AFP inhibited the changes in moisture distribution and maintained the moisture in Antarctic krill meat. The contents of nonprotein nitrogen and trichloroacetic acid-soluble peptides indicated that AFP reduced protein degradation. Further, SDS-PAGE showed that AFP reduced the degradation of actin, troponin T, and myosin light chain. The results of fluorescence spectra, circular dichroism, and chemical bond contents indicated that AFP reduced the damage of the protein tertiary and secondary structures of Antarctic krill meat by holding it in a weak polar environment. This study supplied basic theory for the quality control of Antarctic krill meat. PRACTICAL APPLICATION: Protein degradation, moisture distribution, and protein structure changes occurred to Antarctic krill meat during freeze-thaw cycles due to ice crystal growth and recrystallization, which leads to the decrease in quality. Antifreeze protein has been proven to avoid ice crystals' growth and inhibit ice recrystallization. During freeze-thaw cycles, the moisture distribution of Antarctic krill meat treated with antifreeze protein was more uniform, the degree of protein degradation was lower, and the protein structure was protected. This study demonstrated the potential of antifreeze protein as a water and protein protectant of Antarctic krill meat during freeze-thaw cycles.

Effects of oxidation on the physicochemical properties and degradation of mutton myofibrillar proteins

Tenderness affects mutton quality and price, and the degradation of myofibrillar protein (MP) is critical to improve tenderness. We investigated the oxidative modification of mutton MP by hydroxyl radicals (OH) and the effects of this modification on the proteolysis of MP by µ-calpain. As the H2 O2 concentrations increased, the carbonyl and dityrosine contents and the surface hydrophobicity of MP all display an increasing trend, whereas the total sulfhydryl and intrinsic fluorescence intensity of MP declines significantly. SDS-PAGE electrophoresis indicates that disulfide bonds and other covalent bonds led to protein cross-linking and aggregation. After adding µ-calpain, with increasing oxidation, the degradation percentage of myosin heavy chain (MHC) increases considerably and actin degradation is promoted, while the proteolysis of troponin-T and desmin is inhibited. These data suggest that·OH can change MP physicochemical properties and its susceptibility to µ-calpain. Future investigations will focus on the effect of oxidation on the degradation of MP by other proteases, such as cathepsins and caspase and the effect of oxidation on these enzymes. PRACTICAL APPLICATION: The calpain system, particularly µ-calpain, plays a pivotal role in postmortem tenderization of meat. Protein oxidative modifications influence meat tenderness mainly by regulating proteolysis. An investigation of the effect of oxidation on the proteolytic susceptibility of MP to degradation by µ-calpain allows for the monitoring of the association between protein oxidation and meat tenderness.

Ultrasonic pretreatment improves the gelation properties of low-salt Penaeus vannamei (Litopenaeus vannamei) surimi

The effects of different ultrasonic pretreatments (120–600 W, 20 min; 360 W, 10–30 min) on the gel properties of shrimp surimi were investigated. Gel properties and protein functional properties were analysed to clarify the mechanism of action of ultrasound. The gel strength, water holding capacity and surface hydrophobicity of shrimp surimi gel increased initially and then decreased with the increase in ultrasound power or time, but the change in total sulfhydryl content showed the opposite trend, which indicated that proper ultrasound pretreatment could improve the gel properties of shrimp surimi, expand the protein to a greater extent and expose more SH groups and hydrophobic groups. According to scanning electron microscopy observation, ultrasound made shrimp surimi gel form a denser gel network. Fourier transform infrared analysis indicated that the α-helix content in shrimp surimi gel decreased initially and then increased with the increase of in ultrasound power or time, whereas the change in β-sheet content showed the opposite trend. And the protein was the most stable in 360 W/20 min pretreatment. SDS-PAGE patterns showed that proper ultrasound inhibited the degradation of actin and troponin C. In addition, dynamic rheology illustrated that the G′ values of the ultrasonic pretreatment group were higher than that of the control group, indicating that ultrasound could improve the elasticity and stability of shrimp surimi gel. The results suggested that the shrimp surimi gel pretreated by 360 W/20 min ultrasound showed the best gel properties. Furthermore, the correlation between the indexes affecting the properties of the gel was analyzed. This study provides a new technical means to improve the gel properties of shrimp surimi.

Early postmortem degradation of actin muscle protein in Algerian Sahraoui dromedaries.

The present study aimed to evaluate actin degradation during the early postmortem time in Longissimus Lumborum muscle according to Sahraoui dromedary's age. A sample of eight males, young (2 years old) and adult (8 years old) dromedaries, was used to investigate meat quality traits and actin proteolysis during the conversion of muscle to meat. Results demonstrated higher pH values in young compared to adult with a polyphasic pH drop profile. While, age did not affect drip loss (DL) and the values at 72 h postmortem varied from 5 to 9%. Western blot revealed that actin proteolysis occurred since 1 h postmortem and that it was affected by age and postmortem time. In particular, the 32 and 25 kDa actin fragments could be potential markers of ongoing meat tenderization.

EhRho6-mediated actin degradation in Entamoeba histolytica is associated with compromised pathogenicity.

Entamoeba histolytica causes amoebiasis which is a major health concern in developing countries. E. histolytica pathogenicity has been implicated to a large repertoire of small GTPases which switch between the inactive GDP bound state and the active GTP bound state with the help of guanine nucleotide exchange factors (GEFs) and GTPase activating protein (GAPs). Rho family of small GTPases are well known to modulate the actin cytoskeletal dynamics which plays a major role in E. histolytica pathogenicity. Here, we report an atypical amoebic RhoGEF, and its preferred substrate EhRho6, which, upon overexpression abrogated the pathogenic behavior of the amoeba such as adhesion to host cell, monolayer destruction, erythrophagocytosis, and formation of actin dots. A causative immunoblot analysis revealed actin degradation in the EhRho6 overexpressing trophozoites that could be inhibited by blocking the amoebic proteasomal pathway. A careful analysis of the results from a previously published transcriptomics study, in conjunction with our observations, led to the identification of a clade of Rho GTPases in this pathogenic amoeba which we hypothesize to have implications during the amoebic encystation.

Inhibitory mechanism of tyramine-degrading strains on reducing tyramine accumulation in Harbin dry sausage during fermentation

To reduce tyramine accumulation in Harbin dry sausage, two tyramine-degrading strains, Lactobacillus sakei and Staphylococcus epidermidis, were inoculated in Harbin dry sausage. Changes in pH, aw, protein degradation, microbial growth, and tyramine content during fermentation were determined. The results showed that two strains could increase the abundance of corresponding genera and form population dominance in the early fermentation stage. Particularly, L. sakei could significantly reduce the pH and aw of Harbin dry sausage (P < 0.05) and promote the degradation of myosin heavy chain and actin. The α-amino group content was increased due to L. sakei, which resulted in an increase in tyramine content in the early fermentation stage. After fermentation, the tyramine content in the compound fermentation group decreased to 31.91 mg/kg, which was 55% less than that in the spontaneous fermentation group (70.21 mg/kg). Therefore, co-inoculation with tyramine-degrading strains could reduce tyramine accumulation in Harbin dry sausage.

Inhibition of endogenous protease activity and protection of histomorphical integrity during refrigerated storage of grass carp fillets by treatment with natural edible di‐ and tri‐carboxylic acids

SummaryIn this study, by dipping grass carp fillets with 10 mM di‐ and tri‐carboxylic acid solutions individually before storage at 4 °C for 10 days, the application potential of these acids in mitigating softening of fish was investigated through analysing shear force, pH, endogenous cathepsin activities, myofibrillar proteolysis and histomorphological characteristics. The results suggested that maleic acid, fumaric acid and tartaric acid were significantly effective against softening of grass carp muscle, as shown by shear force 26%–31% higher than that of control after storage. Acid treatments led to a decline in the pH of muscle. Besides, the inhibitory effect of these acids on cathepsins was demonstrated by activity reduction by 20%–90%. Further, sodium dodecyl sulphate–polyacrylamide gel electrophoresis and heat shock protein (HSP) analysis supported the protection of these acids against degradation and dissociation of myosin and actin through suppressing HSP loss. Moreover, histomorphological images with higher intensity of acridine orange fluorescence in muscle revealed the improvement of tissue integrity by acid treatments. The inhibitory potential of these acids in suppressing softening of fish muscle is particularly relevant to their capacity of mitigating cathepsin‐induced proteolysis and improving lysosome stability. Therefore, di‐ and tri‐carboxylic acids could play pleiotropic functions in the quality control of fish fillets.

UBE2L3, a Partner of MuRF1/TRIM63, Is Involved in the Degradation of Myofibrillar Actin and Myosin.

The ubiquitin proteasome system (UPS) is the main player of skeletal muscle wasting, a common characteristic of many diseases (cancer, etc.) that negatively impacts treatment and life prognosis. Within the UPS, the E3 ligase MuRF1/TRIM63 targets for degradation several myofibrillar proteins, including the main contractile proteins alpha-actin and myosin heavy chain (MHC). We previously identified five E2 ubiquitin-conjugating enzymes interacting with MuRF1, including UBE2L3/UbcH7, that exhibited a high affinity for MuRF1 (KD = 50 nM). Here, we report a main effect of UBE2L3 on alpha-actin and MHC degradation in catabolic C2C12 myotubes. Consistently UBE2L3 knockdown in Tibialis anterior induced hypertrophy in dexamethasone (Dex)-treated mice, whereas overexpression worsened the muscle atrophy of Dex-treated mice. Using combined interactomic approaches, we also characterized the interactions between MuRF1 and its substrates alpha-actin and MHC and found that MuRF1 preferentially binds to filamentous F-actin (KD = 46.7 nM) over monomeric G-actin (KD = 450 nM). By contrast with actin that did not alter MuRF1–UBE2L3 affinity, binding of MHC to MuRF1 (KD = 8 nM) impeded UBE2L3 binding, suggesting that differential interactions prevail with MuRF1 depending on both the substrate and the E2. Our data suggest that UBE2L3 regulates contractile proteins levels and skeletal muscle atrophy.

Prediction of top round beef meat tenderness as a function of marinating time based on commonly evaluated parameters and regression equations.

This study investigates the influence of 24‐hr marination (with different plant extracts and vinegar) at refrigerated conditions on commonly evaluated physicochemical and textural attributes, including pH, water‐holding capacity (WHC), collagen solubility, moisture, drip loss, and shear force values of beef meat. The results reflected the appropriate correlation between each pair and indicated the efficiency of the household marination procedure to acquire more palatability and tender beef meat. Therefore, to predict beef meat tenderness by applying the Warner–Bratzler shear force (WBSF), a strong positive correlation with the drip loss (p < .01) and a notable negative correlation with the moisture content (p < .01) emphasized the importance of moisture improvement and shear force reduction in affecting tenderness of baked beef meat. The regression equations and R‐squared values were revealed the favorable correlation between collagen solubility and WHC (y = 0.1035x−0.8431, R 2 = .98) as well as moisture and WBSF (y = −0.3297x + 102.58, R 2 = .99) in marinated beef meat. Electrophoresis patterns of isolated myofibrillar proteins disclosed remarkable degradation of myosin heavy chain (MHC), desmin, actin, and tropomyosin during the first day of aging. The noticeable ultrastructural destruction and connective tissue solubilization were observed by microscopy images. These outcomes were a good tenderness predictor be utilized in retailing and industrial scale.

Effects of pectin/chitosan composite and bi-layer coatings combined with Artemisia dracunculus essential oil on the mackerel’s shelf life

This study investigated the effects of pectin coating incorporated with chitosan (CH) and Tarragon essential oil (Artemisia dracunculus) (TEO) on the microbiological, physicochemical, and sensory properties of Scomberomorus commerson muscle during refrigerated storage. The results showed that pectin coating incorporated with CH and TEO could develop active coatings with excellent antibacterial activity to inhibit bacterial growth (mesophilic, psychrotrophic, and lactic acid bacteria). The initial mesophilic, psychrotrophic, and lactic acid bacteria (log10 CFU/g) in the all samples of fillet were 3.21, 4.03 and 1.23–1.72 log10 CFU/g. Pectin-CH coating on mackerel samples enriched with TEO could retard physicochemical properties and preserve the fish quality during refrigerated storage. There was a statistical difference between composite and bi-layer coatings (CC and BC) incorporating TEO (P < 0.05). The SDS‐PAGE analysis of the samples during storage exhibited higher degradation of proteins (myosin heavy chain (MHC), paramyosin, actin, troponin T, and tropomyosin bands) in the control samples than in the treated samples. According to the quality attributes, the incorporation of essential oils or other biopolymers into edible coatings could decrease the bacterial and physicochemical deterioration of seafood during chilling.

Impacts of a novel live shrimp (Litopenaeus vannamei) water-free transportation strategy on flesh quality: Insights through stress response and oxidation in lipids and proteins

A novel strategy of water-free transportation (WFT) for live shrimp was recently developed. In this study, survival rate, hormone response, free radical content, oxidation in lipids and proteins, and flesh quality were evaluated after WFT. Meanwhile water transportation (WT) was employed as a reference for comparison. Results showed only 5.1% mortality after 10 h of WFT, while crustacean hyperglycemic hormone (CHH), reactive nitrogen species (RNS), free fatty acid content (FFA), peroxide value (PV), carbonyl concentration, intrinsic fluorescence intensity (IFI), and surface hydrophobicity significantly increased. Myosin heavy chain (MHC) and actin degradation were also detected after WFT. Although oxidation occurred in lipids and proteins, no significant changes in texture and water-holding capacity (WHC) were detected after WFT or WT (p > 0.05). The 10 h of WFT showed little influence on flesh quality of shrimp even though lipids and proteins oxidation occurred during the mimicked water-free transportation.

Cathepsin D in prawn reproductive system: its localization and function in actin degradation.

Cathepsin D (CAT-D) is a well-known aspartic protease that serves a function as house-keeping lysosomal enzyme in all somatic cells. Its existence in reproductive tissues is highly variable, even in the somatic derived epithelial cells of reproductive tract. In Macrobrachium rosenbergii, existence of MrCAT-D and its translational product was detected in both somatic cells (Sertoli-like supporting cells) and developing spermatogenic cells as well as along accessory spermatic ducts. Specifically, MrCAT-D was localized onto the sperm surface rather than within the acrosomal matrix, as evident by similar staining pattern of anti-CAT-D on live and aldehyde fixed sperm. MrCAT-D in testicular extracts and sperm isolates showed active enzyme activities towards its specific fluorogenic substrate (MCA-Gly-Lys-Pro-Ile-Leu-Phe-Phe-Arg-Leu-Lys (Dnp)-D-Arg-NH2). MrCAT-D also exerted its function towards hydrolyzing filamentous actin, the meshwork of which is shown to be localized at the junction between germ cells and supporting cells and spermatogonia in M. rosenbergii testicular epithelium. Together, we have localized MrCAT-D transcript and its translational product in both supporting and germ cells of testis and claimed its enzymatic function towards actin degradation, which may be related to sperm release from the epithelial cell interaction.

Effects of natural trypsin inhibitor from soybean on texture deterioration of the bay scallop (Argopecten irradians) during cold storage and its mechanism

SummaryIn this study, the inhibitory effects of natural trypsin inhibitor from soybean (SBTI) on texture deterioration of adductor muscle of scallop (AMS) during cold storage and its mechanism were revealed by determining the changes of structural proteins of AMS in situ and in vitro, respectively. Results indicated that the endogenous proteolysis resulted in the disintegration of connective tissue proteins, the degradation of myosin and actin, as well as the reduction of α‐helix, the increase of β‐sheet and amino acid side chains, and the exposure of tryptophan residues of isolated myofibrillar proteins (MPs). These changes in structural proteins contribute to the texture deterioration of AMS as reflected in a significant decrease in hardness, chewiness, adhesiveness and springiness, but a significant increase in cohesiveness. SBTI significantly inhibited the conformational changes of structural proteins caused by endogenous proteolysis, improved the MP gel‐forming ability and consequently retarded the texture deterioration of AMS.