Heterologous Expression and Characterization of a pH-Stable Chitinase from Micromonospora aurantiaca with a Potential Application in Chitin Degradation.

To promote the bioconversion of marine chitin waste into value-added products, we expressed a novel pH-stable Micromonospora aurantiaca-derived chitinase, MaChi1, in Escherichia coli and subsequently purified, characterized, and evaluated it for its chitin-converting capacity. Our results indicated that MaChi1 is of the glycoside hydrolase (GH) family 18 with a molecular weight of approximately 57 kDa, consisting of a GH18 catalytic domain and a cellulose-binding domain. We recorded its optimal activity at pH 5.0 and 55 °C. It exhibited excellent stability in a wide pH range of 3.0-10.0. Mg2+ (5 mM), and dithiothreitol (10 mM) significantly promoted MaChi1 activity. MaChi1 exhibited broad substrate specificity and hydrolyzed chitin, chitosan, cellulose, soluble starch, and N-acetyl chitooligosaccharides with polymerization degrees ranging from three to six. Moreover, MaChi1 exhibited an endo-type cleavage pattern, and it could efficiently convert colloidal chitin into N-acetyl-D-glucosamine (GlcNAc) and (GlcNAc)2 with yields of 227.2 and 505.9 mg/g chitin, respectively. Its high chitin-degrading capacity and exceptional pH tolerance makes it a promising tool with potential applications in chitin waste treatment and bioactive oligosaccharide production.

Astaxanthin Esters as Functional Food: A Review of Their Nutrition, Phytochemical Structure, Biological Features, and Food Industry Prospects.

Astaxanthin (AST), mainly found in algae and shrimp, is a liposoluble ketone carotenoid with a wide range of biological activities and is commonly used in healthcare interventions and cosmetics. AST has a long chain of conjugated double bonds with hydroxyl and ketone groups at both ends, enabling it to form astaxanthin esters (AST-Es) through esterification with fatty acids. The fatty acid structure of AST plays a key role in the stability, antioxidant activity, and bioavailability of AST-Es. Antarctic krill (Euphausia superba) and blood-red algae Haematococcus Pluvialis (H pluvialis)-derived AST-Es exhibit strong antioxidant activity and numerous biological activities, such as improving insulin resistance, preventing Parkinson's disease, regulating intestinal flora, and alleviating inflammatory bowel disease. This review discusses the significance of AST-Es as functional food ingredients, highlighting their nutritional value, phytochemical structure, biological activities, and potential applications in the food industry.

Ultrasonic-Assisted Extraction and Gastrointestinal Digestion Characteristics of Polysaccharides Extracted from Mallotus oblongfolius.

The polysaccharides were extracted from the leaves of Mallotus oblongifolius (MO) using an ultrasonic-assisted extraction method in this study. The main variables affecting the yield of polysaccharides extracted from Mallotus appallatus (MOPS) were identified and optimized while concurrently investigating its antioxidant capacity, hypoglycemic activity, and digestive properties. The results indicated that the optimal ultrasound-assisted extraction of MOPS involved an ultrasound power of 200 W, a liquid-to-solid ratio of 25:1 (mL:g), an extraction temperature of 75 °C, and an ultrasound time of 45 min, leading to an extraction yield of (7.36 ± 0.45)% (m/m). The MOPS extract exhibited significant scavenging activity against DPPH and ABTS radicals with IC50 values of (25.65 ± 0.53) µg/mL and (100.38 ± 0.38) µg/mL, respectively. Furthermore, it effectively inhibited the enzymatic activities of α-glucosidase and α-amylase with IC50 values of (2.27 ± 0.07) mg/mL and (0.57 ± 0.04) mg/mL, respectively. The content of MOPS remained relatively stable in the stomach and small intestine; however, their ability to scavenge DPPH radicals and ABTS radicals and exhibit reducing power was attenuated, and the inhibition of α-amylase and α-glucosidase activity was diminished. In conclusion, the ultrasonic extraction of MOPS showed feasibility and revealed antioxidant and hypoglycemic effects. However, the activities were significantly reduced after gastric and small intestinal digestion despite no significant change in the MOPS content.

Effects of Mactra chinenesis Peptides on Alcohol-Induced Acute Liver Injury and Intestinal Flora in Mice.

Food-borne bioactive peptides have shown promise in preventing and mitigating alcohol-induced liver injury. This study was the first to assess the novel properties of Mactra chinenesis peptides (MCPs) in mitigating acute alcoholic liver injury in mice, and further elucidated the underlying mechanisms associated with this effect. The results showed that MCPs can improve lipid metabolism by modulating the AMPK signaling pathway, decreasing fatty acid synthase activity, and increasing carnitine palmitoyltransferase 1a activity. Meanwhile, MCPs ameliorate inflammation by inhibiting the NF-κB activation, leading to reduced levels of pro-inflammatory cytokines (tumor necrosis factor-α and interleukin-1ß). Additionally, a 16S rDNA sequencing analysis revealed that MCPs can restore the balance of gut microbiota and increase the relative abundance of beneficial bacteria. These findings suggest that supplementation of MCPs could attenuate alcohol intake-induced acute liver injury, and, thus, may be utilized as a functional dietary supplement for the successful treatment and prevention of acute liver injury.

Novel hydrocolloids synthesized by polyphenols grafted onto chitosan: A promising coating to inhibit PhIP formation during pan-frying of golden pompano fillets.

Hydrocolloids synthesized by gallic acid (GA) and ferulic acid (FA) grafting onto chitosan (CS) were characterized, and their effects on PhIP formation in pan-fried golden pompano were investigated. Spectrograms including nuclear magnetic resonance, Fourier transform infrared spectroscopy and ultraviolet-visible confirmed that GA and FA were successfully grafted onto CS via covalent bonds, with grafting degree of 97.06 ± 2.56 mg GA/g and 93.56 ± 2.76 mg FA/g, respectively. The CS-g-GA and CS-g-FA exerted better solubility and antioxidant activities than CS. For the 8-min pan-fried golden pompano fillets, CS-g-GA and CS-g-FA (0.5 %, m/v) significantly reduced the PhIP formation by 61.71 % and 81.64 %, respectively. Chemical models revealed that CS-g-GA and CS-g-FA inhibited PhIP formation mainly by decreasing the phenylacetaldehyde contents from Maillard reaction and competing with creatinine to react with phenylacetaldehyde. Therefore, it was suggested that CS-g-phenolic acids emerge as novel coating for aquatic products during processing and inhibit heterocyclic amines generation.

Synergistic effects of tilapia head protein hydrolysate and walnut protein hydrolysate on the amelioration of cognitive impairment in mice.

BACKGROUND: Cognitive impairment (CI) is a significant public health concern, and bioactive peptides have shown potential as therapeutic agents. However, information about their synergistic effects on cognitive function is still limited. Here, we investigated the synergistic effects of tilapia head protein hydrolysate (THPH) and walnut protein hydrolysate (WPH) in mitigating CI induced by scopolamine in mice. RESULTS: The results showed that the combined supplementation of THPH and WPH (mass ratio, 1:1) was superior to either individual supplement in enhancing spatial memory and object recognition abilities in CI mice, and significantly lessened brain injury in CI mice by alleviating neuronal damage, reducing oxidative stress and stabilizing the cholinergic system. In addition, the combined supplementation was found to be more conducive to remodeling the gut microbiota structure in CI mice by not only remarkably reducing the ratio of Firmicutes to Bacteroidota, but also specifically enriching the genus Roseburia. On the other hand, the combined supplementation regulated the disorders of sphingolipid and amino acid metabolism in CI mice, particularly upregulating glutathione and histidine metabolism, and displayed a stronger ability to increase the expression of genes and proteins related to the brain-derived neurotrophic factor (BDNF)/TrkB/CrEB signaling pathway in the brain. CONCLUSION: These findings demonstrate that tilapia head and walnut-derived protein hydrolysates exerted synergistic effects in ameliorating CI, which was achieved through modulation of gut microbiota, serum metabolic pathways and BDNF signaling pathways. © 2024 Society of Chemical Industry.

The effect of crystal structure of MnO2 electrode on DMAC removal: degradation performance, mechanism, and application evaluation.

The crystal structure has a significant impact on the electrochemical properties of electrode material, and thus influences the electrocatalytic activity of the electrode. In this work, α-, ß-, and γ-MnO2 electrodes were fabricated and applied for investigating the effect of crystal structure on electro-oxidation treatment of N,N-dimethylacetamide (DMAC) containing wastewater. The prepared MnO2 electrodes were characterized by scanning electron microscopy and X-ray diffraction, suggesting that different crystal structures of MnO2 electrodes with the same morphology of stacking-needle structure were successfully prepared. The electrochemical performances, including removal efficiencies of DMAC, chemical oxygen demand (COD) and total nitrogen (TN), and energy consumption, were compared between different MnO2 electrodes. Results indicated that ß-MnO2 electrode presented the excellent electrochemical activity, and could remove 93% DMAC, 62% COD, and 78.9% TN, which was much higher than that of α- and γ-MnO2; moreover, energy consumptions of 11.3, 9.7, and 10.5 kWh/m3 were calculated for α-, ß-, and γ-MnO2, respectively. Additionally, the oxidation mechanism of the MnO2 electrodes was presented, indicating that DMAC was mainly oxidized by hydroxyl radical through reactions of hydroxylation, demethylation, and deamination, and electrode characteristics of specific surface area, oxygen evolution potential, and hydroxyl radical production were the key factors for degrading DMAC on MnO2 electrodes. Finally, an actual DMAC containing wastewater was applied for testing the electrochemical performance of the three electrodes, and ß-MnO2 electrode was verified as the suitable electrode for potential application which achieved removal efficiencies of 100%, 64.5%, and 73% for DMAC, COD, and TN, respectively, after system optimization.

Chitosan-based edible film incorporated with wampee (Clausena lansium) seed essential oil: Preparation, characterization and biological activities.

Chitosan (Ch)-based edible composite films were prepared by incorporating blending wampee seed essential oil (WSEO) into a Ch matrix, using the incorporation ratio as a variable. The physical, mechanical properties, structure morphology and rheological properties were determined using tensile strength (TS), elongation at break (EB), water vapor permeability (WVP) tests together with Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) observations and apparent viscosity and shear rate. In addition, the antimicrobial, antioxidant activities were investigated by the DPPH & ABTS radicals scavenging and inhibition zone assays, respectively. Compared with Ch, the incorporation of WSEO significantly decreased (P < 0.05) the TS, EB, and WVP values, especially when the WSEO ratio reached 1.0 % or higher. Meanwhile, the films exhibited greatly improved visible light barrier performance after WSEO incorporation. Both FTIR spectroscopy and SEM observations reflected the crosslinking between WSEO and Ch. Meanwhile, the composite films demonstrated smaller particle size and weaker rheological viscosities, which enhanced the antimicrobial and antioxidant capabilities when compared with those of Ch. Therefore, this study suggested that WSEO incorporated with Ch is an effective ingredient for the preparation of edible films with enhanced physicochemical and biological properties.

Interaction between liposome and myofibrillar protein in surimi: Effect on gel structure and digestive characteristics.

This study investigated the effects of the interaction between liposomes and myofibrillar protein (MP) on tilapia surimi. The strong interaction between liposomes and MP was primarily mediated through hydrogen bonding and hydrophobic interaction. Liposomes caused the unfolding of MP structure, resulting in the decrease of α-helix content and transformation of spatial structure. Notably, the appropriate ratio of liposomes improved the gel properties of tilapia surimi. The water distribution, microstructure, and texture characteristics further confirmed that liposomes strengthened the structure of surimi gel through non-covalent bonds. However, excessive liposomes (1.0 %) weakened gel characteristics and texture. Moreover, the proper ratio of liposomes enhanced the stability of surimi gels during digestion, reducing protein digestibility from 66.0 % to 54.8 %. Curcumin-loaded liposomes in gel matrix notably delayed digestion and improved bioavailability. This delay in digestion was attributed to the ability of liposomes to decrease the interaction between MP and digestive enzymes. This study provides new insight into the application of liposomes in protein-rich food matrixes.

Optimization of ultrasonic-assisted extraction of polyphenol from Areca nut (Areca catechu L.) seeds using response surface methodology and its effects on osteogenic activity.

Areca nut (Areca catechu L.) seeds are rich in polyphenols, while few studies focused on it. This study was designed to obtain the maximum extraction yield of areca nut seed polyphenol (ACP). An ultrasonic-assisted extraction method optimized by response surface methodology (RSM) was established to extract ACP. Under the optimal conditions (ultrasonic power of 87 W, ethanol concentration of 65%, extraction temperature of 62℃, and extraction time of 153 min), the actual extraction yield of ACP was 139.62 mg/g. Then we investigated the effects of ACP on the proliferation, differentiation and mineralization of MC3T3-E1 pre-osteoblasts. Results suggested that ACP notably promoted the proliferation of MC3T3-E1 cells without cytotoxicity, and the contents of collagen type Ⅰ (COL-Ⅰ) and osteocalcin (OCN) were rising. Meanwhile, the alkaline phosphatase (ALP) activity and mineralized nodules were enhanced. These findings demonstrated that ACP could induce the proliferation, differentiation and mineralization of osteoblasts in vitro. This work provided a certain experimental basis for the developing and utilization of polyphenols from Areca nut seeds.

Study of unripe and inferior banana flours pre-gelatinized by four different physical methods.

This study aimed to prepare the pre-gelatinized banana flours and compare the effects of four physical treatment methods (autoclaving, microwave, ultrasound, and heat-moisture) on the digestive and structural characteristics of unripe and inferior banana flours. After the four physical treatments, the resistant starch (RS) content values of unripe and inferior banana flours were decreased from 96.85% (RS2) to 28.99-48.37% (RS2 + RS3), while C∞ and k values were increased from 5.90% and 0.039 min-1 to 56.22-74.58% and 0.040-0.059 min-1, respectively. The gelatinization enthalpy (ΔHg) and I1047/1022 ratio (short-range ordered crystalline structures) were decreased from 15.19 J/g and 1.0139 to 12.01-13.72 J/g, 0.9275-0.9811, respectively. The relative crystallinity decreased from 36.25% to 21.69-26.30%, and the XRD patterns of ultrasound (UT) and heat-moisture (HMT) treatment flours maintained the C-type, but those samples pre-gelatinized by autoclave (AT) and microwave (MT) treatment were changed to C + V-type, and heat-moisture (HMT) treatment was changed to A-type. The surface of pre-gelatinized samples was rough, and MT and HMT showed large amorphous holes. The above changes in structure further confirmed the results of digestibility. According to the experimental results, UT was more suitable for processing unripe and inferior banana flours as UT had a higher RS content and thermal gelatinization temperatures, a lower degree and rate of hydrolysis, and a more crystalline structure. The study can provide a theoretical basis for developing and utilizing unripe and inferior banana flours.

Characterization of coacervation behavior between whey protein isolate and gum Arabic: Effects of heat treatment.

Currently, the effect of heat treatment on the complex coacervation behavior of whey isolate protein (WPI) with gum arabic (GA) is undiscussed. In this work, the complex coacervation behavior of WPI with or without heat treatment and GA in different environments was investigated. The results showed that coacervates were formed at a mass ratio of 2:1 and a pH of 3.5, which was confirmed by the fluorescence spectroscopy results. Heat treatment increased the surface charge of WPI, reduced the saturated adsorption concentration of GA, and enhanced the sensitivity of the complex coacervation reaction to salt ions. Fourier infrared spectroscopy, intermolecular force analysis and molecular docking results confirm that the formation of coacervates is the result of electrostatic interactions. From the scanning electron microscope and differential scanning calorimetry results, it is clear that the whey isolate protein combined with gum arabic forms a gel-like conjugate with higher thermal stability and a dense structure. This study provides more in-depth theoretical guidance for the application of WPI and GA based coacervation and more advanced theoretical data for the study of hWPI.

Glycolipids improve the stability of liposomes: The perspective of bilayer membrane structure.

The storage and thermal stability of liposomes, which are amphiphilic carriers, cause very large challenges. However, glycolipid modification may be a potential method to improve the stability of liposomes. In this study, the mechanism by which tilapia head glycolipids improve the stability of liposomes was studied. The head groups of glycolipids and liposomes have a strong interaction (Ka = 633.650 M-1), mainly due to hydrogen bonds, which promote the formation of microstructure domains between glycolipids and liposomes. In addition, glycolipids caused the bilayer structure of liposomes to rearrange, resulting in an increase in the phase transition temperature, tight arrangement of membrane molecules, and increase in membrane thickness (from 2.4 nm to 3.5 nm). Novelty, the formation of microstructure domains helped prevent the liposomes membrane structure from being disrupted during storage and heat. Therefore, glycolipid modification improved the stability of liposomes. This study can provide new insights into the development of high-stability liposomes.

Preparation of liposomes by glycolipids/phospholipids as wall materials: Studies on stability and digestibility.

Glycolipids may be potential materials to improve the instability of liposomes during storage and consumption. Curcumin-loaded liposomes with high stability were successfully prepared by glycolipids and phospholipids extracted from tilapia. The physicochemical properties analysed showed that glycolipids enhanced the surface charge of liposomes and the encapsulation ability of curcumin. The enhanced affinity of liposomes for curcumin was attributed to the stronger interaction between the head group of glycolipids and curcumin through hydrogen bonding. As predicted, glycolipids improved the storage stability of liposomes, and the thermal stability of curcumin increased from 35.95% to 54.13%. Moreover, glycolipids could resist the degradation of liposomes in the gastrointestinal tract, reducing the encapsulation efficiency changes of curcumin from 60.67% to 43.63%. Simultaneously, the liposomes formed by glycolipids could more effectively protect nerve cells from oxidative damage. Therefore, the substitution of phospholipids with glycolipids is an effective strategy to improve the stability and bioactivity of liposomes.

Emulsion gel stabilized by tilapia myofibrillar protein: Application in lipid-enhanced surimi preparation.

In this study, surimi products rich in lipids were prepared by using myofibril protein (MP) emulsion gel as carriers. The MP emulsion gel (MP concentration, c = 1.5%, oil fraction, ø = 0.68) was prepared by one-step homogenization. The emulsion gel maintained a high elastic modulus (G') after heating and freezing treatment. Confocal laser scanning microscopy revealed that the structure of the emulsion gel was a hybrid network consisting of polymers of cross-linked MP and aggregated protein-stabilized emulsion (W/O/W multiple structures) droplets. The double emulsification of the emulsion gel and MP stabilized the oil droplets in the surimi product, preventing water and oil from leaching out. The microstructure also showed smaller gaps between MPs with increased porosity, while oil droplets were stably embedded in the surimi gel matrix. Moreover, adding MP emulsion gel significantly reduced the surimi gel strength compared to adding oil directly (p < 0.05).

Identification and virtual screening of novel salty peptides from hydrolysate of tilapia by-product by batch molecular docking.

Introduction: Tilapia produces a large number of by-products during processing, which contain potentially flavorful peptides. Methods: The application of PyRx software enabled batch molecular docking andscreening of 16 potential salty peptides from 189 peptides identified in the enzymaticdigestion of tilapia by-products. Results: According to sensory analysis, all 16 peptides werepredominantly salty with a threshold of 0.256 - 0.379 mmol/L with some sournessand astringency, among which HLDDALR had the highest salty intensity, followedby VIEPLDIGDDKVR, FPGIPDHL, and DFKSPDDPSRH. I addition, moleculardocking results showed these four core peptides with high salt intensity bound to thesalt receptor TRPV1 mainly via van der Waals interactions, hydrogen bonds, andhydrophobic forces; Arg491, Tyr487, VAL441, and Asp708 were the key sites for thebinding of salty peptides to TRPV1. Therefore, the application of batch moleculardocking using PyRx is effective and economical for the virtual screening of saltypeptides.

Protective Effect of Polyphenols Purified from Mallotus oblongfolius on Ethanol-Induced Gastric Mucosal Injury by Regulating Nrf2 and MAPKs Pathways.

Mallotus oblongifolius (MO), which is rich in polyphenols, is a characteristic tea resource with medicinal value. In this study, a total of 45 polyphenolic components of MO, including narirutin, isoquercitrin, rutin and digallic acid, were identified by UPLC-Q-TOF/MS analysis. In addition, the gastroprotective effect of Mallotus oblongifolius polyphenols (MOP) on ethanol-induced gastric mucosal injury in rats was investigated. The rats received anhydrous ethanol after continuous gavage of MOP or lansoprazole for one week. In addition, the macro- and micro-damage induced by ethanol in the gastric tissue was significantly reduced after MOP pretreatment for one week. Further analysis showed that MOP prevented ethanol-induced acute gastric mucosal injury by increasing the expression of antioxidant enzymes (SOD, CAT, GSH-Px) and decreasing the expression of reactive oxygen species (ROS), lipid oxidation product (MDA) and myeloperoxidase (MPO). Meanwhile, MOP inhibited the phosphorylation of p38/ERK/JNK and promoted the activation of the Nrf2 pathway. These results suggested that MOP may be a promising therapeutic target for the prevention of ethanol-induced gastric mucosal injury by improving oxidative stress, inhibiting the p38/ERK/JNK signaling pathways and activating Nrf2 expression.

A fucoidan-gelatin wound dressing accelerates wound healing by enhancing antibacterial and anti-inflammatory activities.

Microbial infections and the slow regression of inflammation are major impediments to wound healing. Herein, a tilapia fish skin gelatin-fucose gum-tannic acid (Gel&Fuc-TA) hydrogel wound dressing (Gel&Fuc-TA) was designed to promote wound healing by mixing and reacting tannic acid (TA) with tilapia fish skin gelatin (Gel) and fucoidan (Fuc). Gel&Fuc-TA hydrogel has a good network structure as well as swelling and release properties, and shows excellent antibacterial, antioxidant, cell compatibility, and hemostatic properties. Gel&Fuc-TA hydrogel can promote the expression of vascular endothelial growth factor (VEGF), platelet endothelial cell adhesion molecule-1 (CD-31), and alpha-smooth muscle actin (α-SMA), enhance collagen deposition, and accelerate wound repair. Gel&Fuc-TA hydrogel can change the wound microbiome, reduce wound microbiome colonization, and decrease the expression of microbiome-related proinflammatory factors, such as lipopolysaccharide (LPS), Toll-like receptor 2 (TLR2), and Toll-like receptor 4 (TLR4). Gel&Fuc-TA hydrogel effectively regulates the conversion of wound macrophages to the M2 (anti-inflammatory phenotype) phenotype, decreases the expression of interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and tumor necrosis factor-alpha (TNF-α), and increases the expression of arginase-1 (Arg-1), interleukin-10 (IL-10) and transforming growth factor-beta (TGF-ß), thereby reducing the inflammatory response. In summary, Gel&Fuc-TA hydrogel prepared using a rational green cross-linking reaction can effectively accelerate wound healing.