Protein Hydrolysis as a Way to Valorise Squid-Processing Byproducts: Obtaining and Identification of ACE, DPP-IV and PEP Inhibitory Peptides.

The industrial processing of Argentine shortfin squid to obtain rings generates a significant amount of protein-rich waste, including the skin, which is rich in collagen and attached myofibrillar proteins. This waste is generally discarded. In this study, skin was used as a source of proteins that were hydrolysed using Trypsin, Esperase® or Alcalase®, which released peptides with antioxidant potential and, in particular, antihypertensive (ACE inhibition), hypoglycemic (DPP-IV inhibition) and/or nootropic (PEP inhibition) potential. Among the three enzymes tested, Esperase® and Alcalase produced hydrolysates with potent ACE-, DPP-IV- and PEP-inhibiting properties. These hydrolysates underwent chromatography fractionation, and the composition of the most bioactive fractions was analysed using HPLC-MS-MS. The fractions with the highest bioactivity exhibited very low IC50 values (16 and 66 µg/mL for ACE inhibition, 97 µg/mL for DPP-IV inhibition and 55 µg/mL for PEP inhibition) and were mainly derived from the hydrolysate obtained using Esperase®. The presence of Leu at the C-terminal appeared to be crucial for the ACE inhibitory activity of these fractions. The DPP-IV inhibitory activity of peptides seemed to be determined by the presence of Pro or Ala in the second position from the N-terminus, and Gly and/or Pro in the last C-terminal positions. Similarly, the presence of Pro in the peptides present in the best PEP inhibitory fraction seemed to be important in the inhibitory effect. These results demonstrate that the skin of the Argentine shortfin squid is a valuable source of bioactive peptides, suitable for incorporation into human nutrition as nutraceuticals and food supplements.

Integrated extraction, structural characterization, and activity assessment of squid pen protein hydrolysates and ß-chitin with different protease hydrolysis.

Squid pen (SP) is a valuable source of protein and ß-chitin. However, current research has primarily focused on extracting ß-chitin from SP. This study innovatively extracted both SP protein hydrolysates (SPPHs) and SP ß-chitin (SPC) simultaneously using protease hydrolysis. The effects of different proteases on their structural characteristics and bioactivity were evaluated. The results showed that SP alcalase ß-chitin (SPAC) had the highest degree of deproteinization (DP, 98.19 %) and SP alcalase hydrolysates (SPAH) had a degree of hydrolysis (DH) of 24.47 %. The analysis of amino acid composition suggested that aromatic amino acids accounted for 17.44 % in SPAH. Structural characterization revealed that SP flavourzyme hydrolysates (SPFH) had the sparsest structure. SPC exhibited an excellent crystallinity index (CI, over 60 %) and degree of acetylation (DA, over 70 %). During simulated gastrointestinal digestion (SGD), the hydroxyl radical scavenging activity, ABTS radical scavenging activity, Fe2+ chelating activity, and reducing power of the SPPHs remained stable or increased significantly. Additionally, SPFC exhibited substantial inhibitory effects on Staphylococcus aureus and Escherichia coli (S. aureus and E. coli), with inhibition circle diameters measuring 2.4 cm and 2.1 cm. These findings supported the potential use of SPPHs as natural antioxidant alternatives and suggested that SPC could serve as a potential antibacterial supplement.

Voltage-calibrated, finely tunable protein assembly.

Neuronally triggered phosphorylation drives the calibrated and cyclable assembly of the reflectin signal transducing proteins, resulting in their fine tuning of colours reflected from specialized skin cells in squid for camouflage and communication. In close parallel to this physiological behaviour, we demonstrate for the first time that electrochemical reduction of reflectin A1, used as a surrogate for charge neutralization by phosphorylation, triggers voltage-calibrated, proportional and cyclable control of the size of the protein's assembly. Electrochemically triggered condensation, folding and assembly were simultaneously analysed using in situ dynamic light scattering, circular dichroism and UV absorbance spectroscopies. The correlation of assembly size with applied potential is probably linked to reflectin's mechanism of dynamic arrest, which is controlled by the extent of neuronally triggered charge neutralization and the corresponding fine tuning of colour in the biological system. This work opens a new perspective on electrically controlling and simultaneously observing reflectin assembly and, more broadly, provides access to manipulate, observe and electrokinetically control the formation of intermediates and conformational dynamics of macromolecular systems.

Sensory specializations drive octopus and squid behaviour.

The evolution of new traits enables expansion into new ecological and behavioural niches. Nonetheless, demonstrated connections between divergence in protein structure, function and lineage-specific behaviours remain rare. Here we show that both octopus and squid use cephalopod-specific chemotactile receptors (CRs) to sense their respective marine environments, but structural adaptations in these receptors support the sensation of specific molecules suited to distinct physiological roles. We find that squid express ancient CRs that more closely resemble related nicotinic acetylcholine receptors, whereas octopuses exhibit a more recent expansion in CRs consistent with their elaborated 'taste by touch' sensory system. Using a combination of genetic profiling, physiology and behavioural analyses, we identify the founding member of squid CRs that detects soluble bitter molecules that are relevant in ambush predation. We present the cryo-electron microscopy structure of a squid CR and compare this with octopus CRs1 and nicotinic receptors2. These analyses demonstrate an evolutionary transition from an ancestral aromatic 'cage' that coordinates soluble neurotransmitters or tastants to a more recent octopus CR hydrophobic binding pocket that traps insoluble molecules to mediate contact-dependent chemosensation. Thus, our study provides a foundation for understanding how adaptation of protein structure drives the diversification of organismal traits and behaviour.

Study on the preparation and biological activities of low molecular weight squid ink polysaccharide from Sepiella maindroni.

Sepiella maindroni ink polysaccharide (SIP) from the ink of cuttlefish Sepiella maindroni and its sulfated derivative (SIP-SII) have been demonstrated to possess diverse biological activities. But little is known about low molecular weight squid ink polysaccharides (LMWSIPs). In this study, LMWSIPs were prepared by acidolysis, and the fragments with molecular weight (Mw) distribution in the ranges of 7 kDa to 9 kDa, 5 kDa to 7 kDa and 3 kDa to 5 kDa were grouped and named as LMWSIP-1, LMWSIP-2 and LMWSIP-3, respectively. The structural features of LMWSIPs were elucidated, and their anti-tumor, antioxidant and immunomodulatory activities were also studied. The results showed that with the exception of LMWSIP-3, the main structures of LMWSIP-1 and LMWSIP-2 did not change compared with SIP. Though there were no significant differences in the antioxidant capacity between LMWSIPs and SIP, the anti-tumor and immunomodulatory activities of SIP were enhanced to a certain extent after degradation. It is particularly noteworthy that the activities of LMWSIP-2 in anti-proliferation, promoting apoptosis and inhibiting migration of tumor cells as well as promoting the proliferation of spleen lymphocytes were significantly higher than those of SIP and the other degradation products, which is promising in the anti-tumor pharmaceutical field.

The impact of ocean acidification on the eye, cuttlebone and behaviors of juvenile cuttlefish (Sepiella inermis).

The cuttlefish (Sepiella inermis) is an economically important species in the coastal seas of China. The impacts of ocean acidification on the ability of juvenile cuttlefish to select a suitable habitat, its hunting and swimming behavior, remains unknown. We examined behavior-related responses and the eye and cuttlebone structure of juvenile cuttlefish following short-term exposure to CO2-enriched seawater. The predation success rate decreased with the elevation in CO2 concentration. In the CO2 treatment groups, cuttlefish spent more time in the dark zone and the average swimming speed and total swimming distance significantly decreased. The structure of the retina and cuttlebone was affected by seawater acidification. Moreover, apoptotic cells were significantly increased in the eyes. In the wild, the impairment of the eye and cuttlebone may decrease the predation ability of juvenile cuttlefish and negatively affect their ability to select a suitable habitat, which would be detrimental to its population.

The inhibitory mechanism of 2-amino-3,8-dimethylimidazo [4,5-f] quinoxaline (MeIQx) formation by ultraviolet-gallic acid (UV-GA) during the oil-frying process of squid.

The inhibitory effect of ultraviolet-gallic acid (UV-GA) on carbonyl valence and intermediates and precursors of 2-amino-3,8-dimethylimidazo [4,5-f] quinoxaline (MeIQx) was investigated to futher clarify the inhibitory mechanism for safety control the quality of oil-fried squid. Ultraviolet C-treated gallic acid (UVC-GA) and ultraviolet B-treated gallic acid (UVB-GA) were produced by ultraviolet 225 nm of band C and 300 nm of band B, respectively. The MeIQx contents in oil-fried squid were significantly higher, and UVC-GA and UVB-GA could significantly inhibit the MeIQx formation and the formation rates of carbonyl valence and precursors (threonine (Thr), creatinine, and glucose). The UVB-GA inhibited formaldehyde formation, while UVC-GA significantly reduced the formaldehyde, acetaldehyde, and 2,5-dimethyl pyrazine contents. In conculsion, UV-GA reduced carbonyl produced from the lipid oxidation to further weaken the catalysis of carbonyl, rendering the MeIQx precursor degrading into the intermediates during Strecker degradation. Thus, the MeIQx formation was inhibited.

Functional and physicochemical properties of protein from giant squid (Dosidicus gigas) extracted using foam-aided pH-shift processing.

Jumbo squid (Dosidicus gigas) is a commercially valuable mollusk in Mexico; 80% of its body is edible. Despite the high protein content (∼18%) and low cholesterol content of this species, its high proteolytic activity (microbial and endogenous enzymes) induces protein degradation and consequent reduction in functional properties from a structural viewpoint. Gelation capacity (texture profile of the gels obtained), solubility, water holding capacity, foaming capacity, emulsification capacity, and emulsion stability were evaluated in protein concentrates obtained by foam-aided pH-shift processing: (A) myofibrillar protein extraction with distilled water and no pH-shifting; (B) alkaline solubilization and isoelectric precipitation; (C) acidic solubilization and isoelectric precipitation; and (D) process A and isoelectric precipitation. Process B showed superior gelation capacity, D had high emulsion stability across a wide range of pH values (4.0-8.0) and C lower plate counts of aerobic mesophilic. Therefore, all three alternative extraction processes showed techno-functional advantages. PRACTICAL APPLICATION: Jumbo squid is an abundant protein source in México, most of which is exported. Functional and physicochemical properties of muscle protein were improved by pH-shift processing. The recovered protein showed modifications of technological properties, using one of the methods described can lead to produce a protein extract with the most desirable attributes, such as foaming, emulsifying, or gelling capacities. The functional and physicochemical properties of protein from squid can be enhanced by selecting a certain pH-shift processing, depending on the desirable use. There is a broad perspective on the use of these protein extracts as ingredients or additives.

Squid Skin Cell-Inspired Refractive Index Mapping of Cells, Vesicles, and Nanostructures.

The fascination with the optical properties of naturally occurring systems has been driven in part by nature's ability to produce a diverse palette of vibrant colors from a relatively small number of common structural motifs. Within this context, some cephalopod species have evolved skin cells called iridophores and leucophores whose constituent ultrastructures reflect light in different ways but are composed of the same high refractive index material─a protein called reflectin. Although such natural optical systems have attracted much research interest, measuring the refractive indices of biomaterial-based structures across multiple different environments and establishing theoretical frameworks for accurately describing the obtained refractive index values has proven challenging. Herein, we employ a synergistic combination of experimental and computational methodologies to systematically map the three-dimensional refractive index distributions of model self-assembled reflectin-based structures both in vivo and in vitro. When considered together, our findings may improve understanding of squid skin cell functionality, augment existing methods for characterizing protein-based optical materials, and expand the utility of emerging holotomographic microscopy techniques.

Evaluating the effects of nanoparticles combined ultrasonic-microwave thawing on water holding capacity, oxidation, and protein conformation in jumbo squid (Dosidicus gigas) mantles.

Magnetic nanoparticles (MNPs) have a dual role in acting as magnetic and sonosensitizer agents, which can combine the synergistic effects of microwave and ultrasonic waves. To study the effects of MNPs combined ultrasonic-microwave thawing (NUMT) on the water holding capacity (WHC), oxidation of protein and lipid, and protein conformation, jumbo squid mantles were subjected to cold storage thawing (CST), MNPs combined ultrasonic thawing (NUT), MNPs combined microwave thawing (NMT) and NUMT. Results showed that NUMT treatment had a higher WHC, lower oxidation, effectively reduced myofibrillar protein aggregation and degradation, and stabilized the structure of the protein of the jumbo squid. The muscle fiber structure of NUMT treated jumbo squid mantles was dense, orderly with a smooth surface, and the fiber network gaps were small and uniformly distributed. This study shows that NUMT can ameliorate the thawing qualities of jumbo squid, and is an effectively thawing method.

A Mini-Review on Reflectins, from Biochemical Properties to Bio-Inspired Applications.

Some cephalopods (squids, octopuses, and cuttlefishes) produce dynamic structural colors, for camouflage or communication. The key to this remarkable capability is one group of specialized cells called iridocytes, which contain aligned membrane-enclosed platelets of high-reflective reflectins and work as intracellular Bragg reflectors. These reflectins have unusual amino acid compositions and sequential properties, which endows them with functional characteristics: an extremely high reflective index among natural proteins and the ability to answer various environmental stimuli. Based on their unique material composition and responsive self-organization properties, the material community has developed an impressive array of reflectin- or iridocyte-inspired optical systems with distinct tunable reflectance according to a series of internal and external factors. More recently, scientists have made creative attempts to engineer mammalian cells to explore the function potentials of reflectin proteins as well as their working mechanism in the cellular environment. Progress in wide scientific areas (biophysics, genomics, gene editing, etc.) brings in new opportunities to better understand reflectins and new approaches to fully utilize them. The work introduced the composition features, biochemical properties, the latest developments, future considerations of reflectins, and their inspiration applications to give newcomers a comprehensive understanding and mutually exchanged knowledge from different communities (e.g., biology and material).

Mechanically strong all-chitin filaments: Wet-spinning of ß-chitin nanofibers in aqueous NaOH.

Herein, a facile wet-spinning strategy was used for the fabrication of mechanically strong all-chitin filaments from an aqueous NaOH solution using ß-chitin nanofibers (ß-ChNFs). It is hypothesized that to reach high mechanical performance it is important to preserve the crystalline structure of chitin during fabrication. To explore this possibility, ß-ChNFs were disintegrated from squid pens by a mild procedure and showed a uniform diameter of 10-25 nm, length of a few microns, and a high aspect ratio of more than 200. An interesting finding was that gel-like ß-ChNF filaments were directly formed in aqueous NaOH without using any organic or ionic agents. The gelation of ß-ChNFS under alkali treatments contributed to the construction of strong nanonetworks and thus facilitated the formation of high-strength filaments. The resulting all-chitin filaments showed a high tensile strength and Young's modulus of 251.3 ± 12.45 MPa and 12.1 ± 0.72 GPa, respectively, which were further investigated for utilization as flexible sensors. The advantages of this strategy included the lack of use of any toxic solvents and the achievement of high mechanical performance for the all-chitin filaments. We believe that this wet-spinning approach may promote the functional utilization of chitin to develop high-strength filaments in smart textiles, biosensors, and structural reinforcements.

Tailoring nanohole sizes through the deacetylation process in chitosan powders obtained from squid pens.

An experimental study on the evolution of the physicochemical, thermal and nanostructural properties of chitosan samples obtained from squid pens as the deacetylation treatment proceeds is presented. To this aim, potentiometric titration, capillary viscosimetry, infrared spectroscopy, differential scanning calorimetry and positron annihilation lifetime spectroscopy were used. The results obtained are discussed in terms of the influence of the deacetylation time on the deacetylation degree, average molecular weight, thermal parameters and average free nanohole size of the different samples. A way of preparing chitosan matrices with tailored nanostructural characteristics for specific applications through the deacetylation process is explored.

A New Method for Albuminuria Measurement Using a Specific Reaction between Albumin and the Luciferin of the Firefly Squid Watasenia scintillans.

This study demonstrates that the luciferin of the firefly squid Watasenia scintillans, which generally reacts with Watasenia luciferase, reacted with human albumin to emit light in proportion to the albumin concentration. The luminescence showed a peak wavelength at 540 nm and was eliminated by heat or protease treatment. We used urine samples collected from patients with diabetes to quantify urinary albumin concentration, which is essential for the early diagnosis of diabetic nephropathy. Consequently, we were able to measure urinary albumin concentrations by precipitating urinary proteins with acetone before the reaction with luciferin. A correlation was found with the result of the immunoturbidimetric method; however, the Watasenia luciferin method tended to produce lower albumin concentrations. This may be because the Watasenia luciferin reacts with only intact albumin. Therefore, the quantification method using Watasenia luciferin is a new principle of urinary albumin measurement that differs from already established methods such as immunoturbidimetry and high-performance liquid chromatography.

Molecular Insights into the Self-Assembly of Block Copolymer Suckerin Polypeptides into Nanoconfined ß-Sheets.

Suckerin in squid sucker ring teeth is a block-copolymer peptide comprised of two repeating modules-the alanine and histidine-rich M1 and the glycine-rich M2. Suckerin self-assemblies display excellent thermo-plasticity and pH-responsive properties, along with the high biocompatibility, biodegradability, and sustainability. However, the self-assembly mechanism and the detailed role of each module are still elusive, limiting the capability of applying and manipulating such biomaterials. Here, the self-assembly dynamics of the two modules and two minimalist suckerin-mimetic block-copolymers, M1-M2-M1 and M2-M1-M2, in silico is investigated. The simulation results demonstrate that M2 has a stronger self-association but weaker ß-sheet propensities than M1. The high self-assembly propensity of M2 allows the minimalist block-copolymer peptides to coalesce with microphase separation, enabling the formation of nanoconfined ß-sheets in the matrix formed by M1-M2 contacts. Since these glycine-rich fragments with scatted hydrophobic and aromatic residues are building blocks of many other block-copolymer peptides, the study suggests that these modules function as the "molecular glue" in addition to the flexible linker or spacer to drive the self-assembly and microphase separation. The uncovered molecular insights may help understand the structure and function of suckerin and also aid in the design of functional block-copolymer peptides for nanotechnology and biomedicine applications.

Cephalopod-Mimetic Tunable Photonic Coatings Assembled from Quasi-Monodispersed Reflectin Protein Nanoparticles.

The remarkable dynamic camouflage ability of cephalopods arises from precisely orchestrated structural changes within their chromatophores and iridophores photonic cells. This mesmerizing color display remains unmatched in synthetic coatings and is regulated by swelling/deswelling of reflectin protein nanoparticles, which alters platelet dimensions in iridophores to control photonic patterns according to Bragg's law. Toward mimicking the photonic response of squid's skin, reflectin proteins from Sepioteuthis lessioniana were sequenced, recombinantly expressed, and self-assembled into spherical nanoparticles by conjugating reflectin B1 with a click chemistry ligand. These quasi-monodisperse nanoparticles can be tuned to any desired size in the 170-1000 nm range. Using Langmuir-Schaefer and drop-cast deposition methods, ligand-conjugated reflectin B1 nanoparticles were immobilized onto azide-functionalized substrates via click chemistry to produce monolayer amorphous photonic structures with tunable structural colors based on average particle size, paving the way for the fabrication of eco-friendly, bioinspired color-changing coatings that mimic cephalopods' dynamic camouflage.

Marine cuttlefish derived 2H-benzochromenone: Pharachromenone as a dual inhibitor of pro-inflammatory 5-lipoxygenase and cyclooxygenase-2.

Cephalopod cuttlefish, Sepia pharaonis, has been considered as a commercially important resource, which is widely regarded as nutritious food in the southwest of Indian and Mediterranean coasts. Chemical analysis of the crude extract of S. pharaonis resulted in the isolation of an undescribed 2H-benzochromenone, pharachromenone, which was characterized as methyl-2″-(7-hydroxy-4-(5'-methylpent-5'-en-1'-yl-oxy-methyl)-2-oxo-2H-benzo[h]chromen-5-yl-methyl)-butanoate by mass and nuclear magnetic resonance spectral experiments. Pharachromenone revealed effective biopotency against 5-lipoxygenase (IC50 1.85 mM) and cyclooxygenase-2 (IC50 0.52 mM) than that displayed by nonsteroidal anti-inflammatory drug ibuprofen (IC50 4.36 mM, p < .05). Promising antioxidant property for pharachromenone (IC50 1.42-1.61 mM) compared with those exhibited by antioxidative agents butylated hydroxyl anisole (BHA) and α-tocopherol (IC50 1.40-1.90 mM) could conceivably validate its dual inhibition potential against 5-lipoxygenase and cyclooxygenase-2. Greater electronic parameters, lesser steric bulkiness, along with acceptable lipophilic-hydrophobic balance significantly contributed toward its promising anti-inflammatory activities. Molecular docking studies showing significantly greater inhibition constant (Ki) 8.24 nM and binding energy (-11.03 kcal/mol) of pharachromenone than the standard ibuprofen (Ki 4.65 µM, binding energy -7.27 kcal/mol) at the binding site of 5-lipoxygenase recognized its noncompetitive binding, which could describe the promising anti-inflammatory potential. Pharachromenone could be developed as a functional food component against oxidative stress-related inflammatory disorders. PRACTICAL APPLICATIONS: The cuttlefish Sepia pharaonis (family Sepiidae) comprises a major share in the global fishery sector due to its culinary delicacy and nutritionally valued high-quality meat. Furthermore, cephalopod mollusks are gaining pharmaceutical acceptance as resources to derive bioactive compounds with therapeutic significance. Bioassay-guided chromatographic fractionation of crude extract of S. pharaonis could result in the isolation of a 2H-benzochromenone derivative, pharachromenone exhibiting potent antioxidant and anti-inflammatory properties. This study recognized the therapeutic potential of a marine cuttlefish-originated food constituent against inflammatory conditions, and could be anticipated as a high-value functional food lead to minimize oxidative stress-related inflammatory disorders.

Low molecular weight sulfated chitosan isolation, characterization and anti-tuberculosis activity derived from Sepioteuthis lessoniana.

The research focused on tuberculosis as it is one of the world's most serious health problems. The extracted chitin from the gladius of Sepioteuthis lessoniana converted into Chitosan (CH). The purified and freeze-dried CH was refined as Sulfated Chitosan (SCH). The SCH was converted into low molecular weight of SCH with various doses of Gamma Irradiation (GIR). Fluorescence characteristics of GIR-SCH and elemental analysis were confirmed. The structure and molecular weights of GIR-SCH were determined with FT-IR, NMR and MALDI-TOF/Mass Spectroscopy. 100 Gy of GIR-SCH significantly showed the minimum inhibitory concentration (MIC) against Mycobacterium smegmatis. The MIC against M. smegmatis was not affected by the varied sulfate levels in the identical molecular weight GIR-SCH. However, the lowest molecular weight GIR-SCH displayed a significantly MIC against M. smegmatis. In docking analysis, the 6ZT3 ligand had the lowest binding energy of -1.57 kcal/Mol indicating a superior binding interaction with GIR-SCH. The effect of molecular weight reduction by GIR on the anti-tuberculosis capacity of GIR SCH was investigated in this study which had antimicrobial implications.

The drip loss inhibitory mechanism of nanowarming in jumbo squid (Dosidicus gigas) mantles: protein structure and molecular dynamics simulation.

BACKGROUND: The magnetic nanoparticles plus microwave thawing (MNPMT), a new rewarming technology entitled 'nanowarming', can serve as an effective method to achieve rapid and uniform thawing, thus reducing drip loss. The purpose of this study was to decipher the drip loss inhibitory mechanism of MNPMT in jumbo squid (Dosidicus gigas) from the perspectives of protein structure and ice crystal recrystallization. A number of different techniques such as dynamic rheology, Raman spectra, intrinsic fluorescence measurement, and ultraviolet (UV) absorption spectra were conducted to analyze myofibrillar protein conformation and stability of jumbo squid. Scanning electron microscopy (SEM) and myofibrillar fragmentation index (MFI) were used to observe the growth of ice crystals. The interaction between magnetic nanoparticles (MNPs) and ice crystals was studied by using molecular dynamic (MD) simulation. RESULTS: MNPMT exhibited the highest storage modulus (G') value at 90 °C, suggesting the protein conformation was more stable. The increase in α-helices, fluorescence intensity and characteristic absorption peak of MNPMT illustrated that MNPMT can effectively maintain the secondary and tertiary structure of the protein. Compared with cold storage thawing (CST) and microwave thawing (MT), the MFI value of MNPMT was significantly decreased (P < 0.01). The result of MD simulation showed that MNPs displayed a tendency to gradually approach the surface of ice crystals, and induced a certain degree of damage to the ice crystal surface, thereby markedly inhibiting ice crystal recrystallization. CONCLUSION: MNPMT can reduce the drip loss by keeping the protein conformation stable and inhibiting the recrystallization of ice crystals during the thawing process. © 2022 Society of Chemical Industry.

Effect of vacuum frying and atmospheric frying on the quality and protein oxidation of squid (Loligo chinensis).

Squid are commercial marine species that have high nutritional value. This study aimed to compare the influences of vacuum frying and atmospheric frying on the physicochemical properties and protein oxidation of three main parts (ring, tentacle, and fin) of the squid Loligo chinensis. The results showed that the vacuum-fried (VF) group had lower moisture and total fat contents and looser microstructures than the atmospheric-fried (AF) group. The amino acid contents and molecular weight revealed that the proteins were well preserved during vacuum frying. Carbonyl content in the VF ring, tentacle, and fin samples increased nearly 2.53-, 1.54-, and 2.56-fold, respectively, compared to that in the corresponding fresh group, but these increases were lower than those of the corresponding AF group. In addition, the secondary structures of proteins revealed a slight decrease in the α-helix and ß-turn contents and a significant increase in the ß-sheet content during vacuum frying. Therefore, vacuum frying can be used as an efficient processing method to conserve the high nutritive quality of the product. PRACTICAL APPLICATION: As a developing alternative technology to prepare healthier fried products, vacuum frying has been the focus of recent researches. Vacuum frying produced squid products that had lower TBARS values, carbonyl contents, and Schiff base substances compared to atmospheric frying. And the protein secondary structures of the vacuum-fried group retained better. The study proved that vacuum frying could be an effective method with the advantages of high protein stability and product quality.