Characterization of the extracellular proteases from Bacillus inaquosorum strain E1-8 and its application in the preparation of hydrolysates from plant and animal proteins with antioxidant, antifreeze and anti-browning properties.

BACKGROUND: Bacillus inaquosorum strains is widely recognized for their plant-growth-promoting and biocontrol capabilities, yet their roles in protease production remain unclear. The present study aimed to comprehensively assess the protease-producing performance of B. inaquosorum strain E1-8, at the same time as exploring the novel application of agricultural Bacillus proteases in the preparation of protein hydrolysates for fresh-cut fruits preservation. RESULTS: First, genomic sequencing revealed the diversity of E1-8 proteases, indicating 15 putative extracellular proteases. Subsequently, the fermentation conditions for E1-8 protease production were optimized, with sweet potato powder and soybean meal identified as the most suitable carbon and nitrogen sources, respectively, resulting in a maximum protease activity of 321.48 U mL-1. Upon culturing the strain under these optimized conditions, only an S8 family serine protease and an M48 family metalloprotease were revealed by secretomic analysis and protease inhibitor assays. Additionally, the optimal protease conditions for generating protein hydrolysates from soy, pea, fish and porcine proteins were determined. The molecular weight of the hydrolysates primarily ranged from 2000 to 180 Da, with a total of 17 amino acids identified. The application of these hydrolysates demonstrated a 2,2-diphenyl-1-picrylhydrazyl (i.e. DPPH) scavenging activity ranging from 58.64% to 84.12%, significantly reducing of the melting peaks and the freezing points. Furthermore, the browning index of apple slices stored at 4 °C decreased by 14.81% to 22.15% on the second day, and similar effects were observed in fresh-cut banana stored at 4 °C for 7 days. CONCLUSION: The protein hydrolysates obtained exhibit remarkable antioxidant, antifreeze and anti-browning properties for fresh-cut fruits. © 2024 Society of Chemical Industry.

Recent knowledge in phages, phage-encoded endolysin, and phage encapsulation against foodborne pathogens.

The use of antibiotics had reached a plateau due to antibiotic resistance, overuse, and residue. Bacteriophages have recently attracted considerable attention as alternative biocontrol agents. Here, we provide an up-to-date overview of phage applications in the food industry. We reviewed recently reported phages against ten typical foodborne pathogens, studies of competitive phage-encoded endolysins, and the primary outcomes of phage encapsulation in food packaging and pathogen detection. Furthermore, we identified existing barriers that still need to be addressed and proposed potential solutions to overcome these obstacles in the future.

Versatile wheat gluten: functional properties and application in the food-related industry.

Gluten is a key component that allows wheat flour to form a dough, and it is also a byproduct of the production of wheat starch. As a commercial product, wheat gluten is increasingly used in the food-related industry because of its versatile functional properties and wide range of sources. Wheat gluten is manufactured industrially on a large scale through the Martin process and batter process and variants thereof. Gliadin and glutenin impart cohesiveness and elasticity properties, respectively, to wheat gluten. The formation of gluten networks and polymers depends mainly on covalent bonds (disulfide bonds) and noncovalent bonds (ionic bonds, hydrogen bonds, and hydrophobic interactions). The multifunctional properties (viscoelasticity, gelation, foamability, etc.) of wheat gluten are shown by rehydration and other processing techniques. Wheat gluten has been widely used in wheat-based products, food auxiliary agents, food packaging, encapsulation and release of food functional ingredients, food adsorption and heat insulation materials, special purpose foods, and versatile applications. In the future, wheat gluten protein will be used as an important raw material to participate in the development and preparation of various food and degradable materials, and the application potential of wheat gluten in food-related industries will be massive. This review summarizes the main manufacturing processes, composition, and structure of gluten protein, and the various functional properties that support its application in the food and related industries.

The versatile functional properties of wheat gluten are closely related to structureThe application as a binder and substitute for meat is growing in popularityRenewable and degradable gluten-based materials are important exploited fieldsWheat gluten shows vast potential and application value in food-related industry.

Wheat Flour-Based Edible Films: Effect of Gluten on the Rheological Properties, Structure, and Film Characteristics.

This work investigates the structure, rheological properties, and film performance of wheat flour hydrocolloids and their comparison with that of a wheat starch (WS)-gluten blend system. The incorporation of gluten could decrease inter-chain hydrogen bonding of starch, thereby reducing the viscosity and solid-like behavior of the film-forming solution and improving the frequency-dependence, but reducing the surface smoothness, compactness, water vapor barrier performance, and mechanical properties of the films. However, good compatibility between starch and gluten could improve the density of self-similar structure, the processability of the film-forming solution, and film performance. The films based on wheat flours showed a denser film structure, better mechanical properties, and thermal stability that was no worse than that based on WS-gluten blends. The knowledge gained from this study could provide guidance to the development of other flour-based edible packaging materials, thereby promoting energy conservation and environmental protection.

Starch-based nanoparticles: Stimuli responsiveness, toxicity, and interactions with food components.

In recent years, starch-based nanoparticles have attracted great interest due to their small size, good biocompatibility, and environmental friendliness, as well as their potential applications in foods, drug delivery carriers, and biodegradable edible films. Compared with nonstarch polysaccharides, starch can be enzymatically hydrolyzed into glucose in vivo, so it can be used as an enzyme-responsive carrier. The recent research progress of starch-based nanoparticles, including starch nanoparticles, starch nanospheres, starch micelles, starch vesicles, starch nanogels, and starch nanofibers, are reviewed in this paper. The main focus is on their responsiveness, digestibility, toxicity, interactions with other components, and applications. Starch-based nanoparticles are nontoxic and responsive to pH, temperature, light, and other stimuli. It can interact with proteins, antioxidants, and lipids through electrostatic interactions and hydrogen bonding interactions. Starch-based nanoparticles have a wide range of applications, including enhancing the mechanical properties of films and gels, stabilizing emulsions, as a fluorescent indicator, a catalyst, and a nanocarrier to control the release of active ingredients and drugs.

Double Cross-Linked Chitosan Composite Films Developed with Oxidized Tannic Acid and Ferric Ions Exhibit High Strength and Excellent Water Resistance.

There is tremendous scientific interest in developing biodegradable films through facile and versatile strategies. Although extensive studies on the preparation of chitosan films have been conducted, the reported results commonly present low mechanical strength and weak water resistance. In the present study, high strength and significantly water resistance single-cross-linked chitosan-oxidized tannic acid (SC-CS/OTA) composite films and double cross-linked chitosan/oxidized tannic acid/FeIII (DC-CS/OTA/FeIII) composite films were created through a Schiff base reaction and metal coordination. As a result, the optimal tensile strength of SC-CS/OTA composite films and DC-CS/OTA/FeIII composite films was 35.92 and 209 MPa, respectively. Notably, when compared with other chitosan-based films, the tensile strength of DC-CS/OTA/FeIII composite films was approximately three times stronger. Moreover, the water vapor permeability (WVP) values of the films with FeIII(0.66 ± 0.03 × 10-10 g/m·h·Pa) was lower than that of films without FeIII (1.33 ± 0.01 × 10-10 g/m·h·Pa). More importantly, WVP values of the DC-CS/OTA/FeIII composite films were 3-4 orders of magnitude lower than those of chitosan films previously reported. The SC-CS/OTA composite films (96.69%) and DC-CS/OTA/FeIII composite films (99.06%) also presented high DPPH radical scavenging activity. Furthermore, SC-CS/OTA and DC-CS/OTA/FeIII hydrogels were also prepared. This work can be widely applied in the food, biomedical science, and wastewater treatment fields.

Construction and Characterization of Phthalocyanine-Loaded Particles of Curdlan and Their Photosensitivity.

To optimize the physicochemical properties of phthalocyanine (PC), we examined its behavior in particles of triple helix glucan curdlan (CUR). CUR was denatured and renatured in DMSO, in the presence of PC. Infrared spectroscopy and transmission electron microscopy (TEM) showed that PC and CUR formed an inclusion complex, in which PC was trapped inside CUR molecules. This redshifted the absorption peak of PC, which would improve its usefulness as a photosensitizer, because infrared light can penetrate more deeply into human tissues. The conductivity of the solution of CUR-PC was higher than the conductivities of either a CUR solution or a PC dispersion, indicating that CUR-PC is more water soluble than PC. In addition, CUR-PC was highly stable in water. Thus, the use of CUR as a carrier of PC improves several of its physical properties. PC is used as a photosensitizer for killing cancer cells, but its use is hampered by its low solubility. Further, its absorption range limits its use to a depth of 1⁻3 mm in tissues. CUR-PC, with its high solubility and infrared absorption peak, was highly effective as a photosensitizer. It killed 84% of HeLa cells under 15 min of long wavelength radiation and had little cytotoxicity in the absence of light. These results demonstrate that CUR-PC has promise as a photosensitizer, as well as provide theoretical support for a wide range of applications for PC and CUR.

Effect of acid hydrolysis combined with heat moisture treatment on structure and physicochemical properties of corn starch.

Modification of starch led to new products with new desirable properties. Corn starch samples modified by acid hydrolysis combined with heat moisture treatment (AH-HMT) were made by changing pH, moisture content and treated temperature. After modification, swelling power at temperature higher than 75 °C of corn starches decreased while solubility of the starches increased. After AH-HMT, pasting temperature (PT) of all treated starch samples increased. But lower peak viscosity (PKV), trough viscosity (TV) and break down (BD) of most treated starch samples were observed. AH-HMT increased the gel hardness of all treated starches. And the biggest hardness of modified starch gel was 148.419 g, improving 93.471 g compared with native starch gel. The melting temperatures (To, Tp, Tc) of modified starch increased, but the melting range and △H decreased. The X-ray pattern remained practically unchanged with or without AH-HMT. Acid hydrolysis combined with heat moisture treatment (AH-HMT) improved the functional properties of corn starch.

Different effects of pea protein on the properties and structures of starch gel at low and high solid concentrations.

In the context of starch-protein composite gels, the influence of protein on gel formation significantly shapes the textural attributes of starch gels, leading to distinct outcomes. This study aimed to evaluate how different ratios of pea protein (PP) affect the properties and structures of starch-protein composite gels at low (10 wt%) and high (40 wt%) solid concentrations. The addition of PP had opposite effects on the two gels. Compared to the pure starch gel, the low-concentration composite gel (LCG) with 20 % PP experienced a 48.90 ± 0.33 % reduction in hardness, and the storage modulus (G') decreased from 14,100 Pa to 5250 Pa, indicating a softening effect of PP on LCG. Conversely, the hardness of the high-concentration composite gel (HCG) with 20 % PP exhibited a 62.19 ± 0.03 % increase in hardness, and G' increased from 12,100 Pa to 41,700 Pa, highlighting the enhancing effect of PP on HCG. SEM and fluorescence microscopy images showed that PP induced uneven network sizes in LCG, while HCG with a PP content of 20 %, PP, together with starch, formed a three-dimensional network. This study provides valuable insights and guidance for the design and production of protein-enriched starch gel products with different textural properties.

Novel one-step method to construct gellan gum-zein core-shell structured starch beads for regulating starch digestion.

A simple and efficient one-step method combining ion crosslinking and antisolvent exchange has been developed to construct gellan gum/corn starch@zein (GG/CS@Z) core-shell structured beads. This novel approach aims to reduce the digestibility and digestion rate of starch. The GG/CS@Z beads were comprehensively characterized using scanning electron microscopy (SEM), confocal laser scanning microscope (CLSM), differential scanning calorimetry (DSC), swelling power experiments and in vitro simulated digestion tests, respectively. SEM and CLSM analyses unequivocally confirmed the successful construction of the core-shell structure in GG/CS@Z beads. The encapsulation of starch within the core-shell structure effectively restricted its swelling and gelatinization by inhibiting water contact. Notably, compared to native corn starch, the GG/CS@Z5 beads exhibited significantly enhanced contents of slowly digestible starch (SDS) and resistant starch (RS), reaching 34.07 % and 26.86 %, respectively. These findings demonstrate the potential of GG/CS@Z core-shell structured beads as functional food ingredients for individuals with cardiovascular diseases and diabetes.

Welding Characteristics of Medium Titanium Plates with Autogenous Laser Welding and Narrow-Gap Laser Filling Welding Modes.

Titanium and titanium alloys with a medium thickness of 5 to 12 mm are widely used for ocean platforms, military equipment and in other fields because of their light weight, appropriate strength and corrosion resistance. In this study, autogenous laser welding and narrow-gap laser welding processes were researched and compared, and the welding characteristics, weld microstructure and joint strength were analyzed. The results showed that autogenous laser welding had higher efficiency, narrower weld width and higher microstructure uniformity. Autogenous laser welding can achieve the single pass weld penetration at laser keyhole mode. The weld width of narrow-gap laser welded joint was 12.5 mm, which was nearly three times than that of autogenous laser welding. The grain size of autogenous laser welding was obviously smaller and more uniform in depth than that of narrow-gap laser welding. In the weld zone, the coarse columnar α grains grew from the fusion line, while in the heat-affected zone, equiaxed α grains with needle and sawtooth α morphologies were presented. The microhardness of the heat-affected zone was higher than in the weld zone and the base metal due to the denser needle microstructure. The tensile samples all fractured at the base metal, indicating the welded joint strength efficiency was greater than 1.

Water in water emulsion stabilized by liposomes developed from whey protein isolate and xanthan gum: Environmental stability and photoprotection effect for riboflavin.

The purpose of this work is to explore the feasibility of water in water (W/W) emulsion stabilized with liposomes as a water-soluble nutraceutical carrier. A W/W emulsion system composed of xanthan gum (XG) and whey protein isolate (WPI) with different amount (0.2 %, 0.4 %, and 0.6 %) of liposomes as stabilizer was constructed. Fast green staining observation showed that XG was the internal phase and WPI was the continuous phase respectively. Confocal laser scanning microscopy revealed that with the increase of liposomes concentration from 0.4 % to 0.6 %, the interface thickness of the W/W emulsions was approximately twice that of the 0.2 % liposome-stabilized emulsion.The emulsions remained stable under neutral and weakly alkaline conditions. The droplet sizes of the emulsions were little affected by ionic strength. The binding constant (Ka) for XG to riboflavin (12.22) was approximately 5 times that for WPI to riboflavin (2.46), suggesting that riboflavin had a stronger binding affinity for the XG molecule compared to WPI. The fluorescence spectra of riboflavin showed that 0.4 % and 0.6 % liposome stabilized emulsions could effectively retard the photodegradation of riboflavin under ultraviolet irradiation. The successful construction of liposomes stabilized W/W emulsion provides a novel strategy for delivering water-soluble bioactive substances.

Effect of lecithin on the complexation between different botanically sourced starches and lauric acid.

This research investigated the effect of lecithin on the complexation of lauric acid with maize starch, potato starch, waxy maize starch, and high amylose maize starch. Rapid visco analysis showed that lecithin altered the setback pattern of potato starch-lauric acid and maize starch-lauric acid mixtures but not waxy maize starch-lauric acid. Further investigation, including differential scanning calorimetry, complex index, and X-ray diffraction, showed that lecithin enhanced the complexation of maize starch, potato starch, and high amylose maize starch with lauric acid. Fourier transform infrared and Raman spectroscopy revealed increasingly ordered structures formed in maize starch-lauric acid-lecithin, potato starch-lauric acid-lecithin, and high amylose maize starch-lauric acid-lecithin systems compared to corresponding binary systems. These highly ordered complexes of maize starch, potato starch, and high amylose maize starch also demonstrated greater resistance to in vitro enzymatic hydrolysis. Waxy maize starch complexation however remained unaffected by lecithin. The results of this study show that lecithin impacts complexation between fatty acids and native starches containing amylose, with the starch source being critical. Lecithin minimally impacted the complexation of low amylose starch and fatty acids.

A short linear glucan nanocomposite hydrogel formed by in situ self-assembly with highly elastic, fatigue-resistant and self-recovery.

Polyacrylamide (PAM) hydrogels are widely used in wide-ranging applications in biology, medicine, pharmaceuticals and environmental sectors. However, achieving the requisite mechanical properties, fatigue resistance, self-recovery, biocompatibility, and biodegradability remains a challenge. Herein, we present a facile method to construct a nanocomposite hydrogel by integrating short linear glucan (SLG), obtained by debranching waxy corn starch, into a PAM network through self-assembly. The resulting composite hydrogel with 10 % SLG content exhibited satisfactory stretchability (withstanding over 1200 % strain), along with maximum compressive and shear strengths of about 490 kPa and 39 kPa at 90 % deformation, respectively. The hydrogel demonstrated remarkable resilience and could endure repeated compression and stretching. Notably, the nanocomposite hydrogel with 10 % SLG content exhibited full stress recovery at 90 % compression deformation after 20 s, without requiring specific environmental conditions, achieving an energy dissipation recovery rate of 98 %. Meanwhile, these hydrogels exhibited strong adhesion to various soft and hard substrates, including skin, glasses and metals. Furthermore, they maintain solid integrity at both 37 °C and 50 °C after swelling equilibrium, unlike traditional PAM hydrogels, which exhibited softening under similar conditions. We hope that this PAM-SLG hydrogel will open up new avenues for the development of multifunctional electronic devices, offering enhanced performance and versatility.

Preparation of debranched starch with high thermal stability and crystallinity using a novel thermal cycling treatment.

Herein, the effects of temperature cycling (4 °C/50 °C/100 °C) on the recrystallization, physicochemical properties, and digestibility of debranched starch (DBS) were investigated. Temperature cycling involved heating DBS to 100 °C to dissociate weak heat-sensitive crystalline structures and cooling to 4 °C to induce the rapid growth of crystal nuclei, followed by maintaining the temperature at 50 °C to promote orderly crystalline growth. This procedure aimed to increase the degree of crystalline structure in recrystallized DBS, thereby resulting in DBS that was heat- and digestion-resistant. Temperature cycling increased the dissociation temperature of DBS, and temperatures of up to 114.8 °C were attained after five cycling times. With increasing cycles, the crystalline structure of DBS transitioned from B-type to the more robust and compact A-type, and the crystallinity increased to ∼81.9 % (after seven cycles). Raman and Fourier transform infrared (FTIR) spectra indicated that temperature cycling enhanced the short-range ordered structure of DBS. Moreover, in vitro digestion experiments demonstrated that the resistant starch content of DBS increased to ∼61.9 % after eight cycles. To summarize, this study demonstrated a green and effective method for preparing heat-and digestion-resistant recrystallized DBS, which can be used for developing dietary supplements and low gastrointestinal staples.

The Construction of Sodium Alginate/Carboxymethyl Chitosan Microcapsules as the Physical Barrier to Reduce Corn Starch Digestion.

To enhance the resistant starch (RS) content of corn starch, in this work, carboxymethyl chitosan/corn starch/sodium alginate microcapsules (CMCS/CS/SA) with varying concentrations of SA in a citric acid (CA) solution were designed. As the SA concentration increased from 0.5% to 2%, the swelling of the CMCS/CS/SA microcapsule decreased from 15.28 ± 0.21 g/g to 3.76 ± 0.66 g/g at 95 °C. Comparatively, the onset, peak, and conclusion temperatures (To, Tp, and Tc) of CMCS/CS/SA microcapsules were higher than those of unencapsulated CS, indicating that the dense network structure of microcapsules reduced the contact area between starch granules and water, thereby improving thermal stability. With increasing SA concentration, the intact and dense network of CMCS/CS/SA microcapsules remained less damaged after 120 min of digestion, suggesting that the microcapsules with a high SA concentration provided better protection to starch, thereby reducing amylase digestibility. Moreover, as the SA concentration increased from 0.5% to 2%, the RS content of the microcapsules during in vitro digestion rose from 42.37 ± 0.07% to 57.65 ± 0.45%, attributed to the blocking effect of the microcapsule shell on amylase activity. This study offers innovative insights and strategies to develop functional starch with glycemic control properties, holding significant scientific and practical value in preventing diseases associated with abnormal glucose metabolism.

Effect of curdlan addition and thermal sterilization on the structural and properties of rice starch gel.

This study delves into the effects of curdlan integration and thermal sterilization on the rheological properties, structure, and quality attributes of concentrated rice starch gel. Acting as a heat-set polysaccharide, curdlan established a dual-network gel structure with rice starch gel, displaying strong interactions with rice starch, as confirmed by confocal laser scanning microscopy and Fourier-transform infrared spectroscopy. The addition of curdlan expedited the gel formation of rice starch, yielding a denser gel structure. Consequently, this enhanced G', solid-like behavior, textural properties, and cooking quality while reducing frequency-dependence. Given the cooling-induced gelation behavior of pure rice starch, thermal treatment disrupted inter-chain hydrogen bonding, compromising the structural integrity of the gel. This disruption manifested in a softer texture and diminished mechanical properties and cooking quality. Notably, this decline in mechanical properties and cooking quality of rice starch gel was markedly ameliorated with the incorporation of curdlan, particularly at a content of ≥1.0 %. Compared with pure RS, 1.0 % CD inclusion showed a reduction in cooking breakage rate by 30.69 % and an increase in hardness by 38.04 %. This work provides valuable insights for the advancement of fresh starch gel-based foods that exhibit exceptional quality and an extended shelf life.

On the investigation of composite cooling/heating set gel systems based on rice starch and curdlan.

In pursuit of advancing the understanding of composite gel systems, this study delves into the intricate realm of rheology, structural elucidation, and mechanical attributes. Specifically, it scrutinizes the symbiotic interplay between rice starch, a cooling-set gel, and curdlan, a thermo-irreversible heating-set gel. A higher curdlan content enhances the inter-chain hydrogen bonding between rice starch and curdlan, resulting in a denser gel structure and thus increased moduli, solid-like behavior, and mechanical properties, and reduced frequency-dependence, especially at high temperatures (>65 °C). For example, with 50 % curdlan incorporation, G' (90 °C) improved by 252 %. Notably, thermal treatment can compromise the structural integrity of the rice starch gel, reducing strength and softening texture. However, this textural degradation can be effectively mitigated with, for example, 30 % curdlan incorporation, resulting in a 55-fold hardness increase at 85 °C. The knowledge gained from this work offers valuable guidance for tailoring starch-based gel products to specific properties.

Effects of recrystallization degree on structure and digestibility of debranched starch.

This study developed a novel method for the facile, green and efficient fabrication of highly crystalline and heat-resistant starch via recrystallization with high concentrations of debranched starch (DBS), which greatly reduced the complexity and period compared to conventional preparation methods. The structural, thermal, and digestive properties of recrystallized DBS obtained from different concentrations (0-50 %) have been systematically investigated. For instance, the peak melting temperature of recrystallized DBS increased from around 77.8 °C to 114.7 °C with increasing DBS concentration. Moreover, the crystallinity of the recrystallized DBS increased from around 23.5 % to 73.6 % when the DBS concentration was raised. In addition, the resistant starch content of the recrystallized DBS increased from around 30.8 % to 72.1 % as the DBS concentration increased. These results show that the DBS concentration in water during recrystallization plays a critical role in determining the molecular, physicochemical, and digestion properties of DBS, which may be an economical and effective method for large-scale production of highly crystalline starch and provides a new method for preparing heat-resistant type-3 resistant starch, which can be used in low glycemic index foods designed to prevent diabetes and obesity.

Development of pea protein nanoparticle/hydrolyzed rice glutelin fibril emulsion gels for encapsulation of curcumin.

The potential of using emulsion gels stabilized by binary plant protein nanoparticle mixtures for the encapsulation and delivery of lipophilic nutraceuticals was evaluated. The particle characteristics, physical stability, water diffusivity, microrheology, large amplitude oscillating shear (LAOS) properties, and in vitro digestion of emulsion gels prepared by different ratios of hydrolyzed rice glutelin fibrils (HRGFs) and pea protein nanoparticle (PNP) were characterized. The emulsion gel with P/H = 2:1 (0.84 µm) exhibited the best storage stability and freeze-thaw stability, as seen by the smaller oil droplet size (1.02 and 1.42 µm, respectively). Low-field pulsed NMR indicated that the majority of water in samples was highly mobile. All the samples were predominantly elastic-like materials. The P/H 2:1 emulsion gel had the lowest FI value (6.21 × 10-4 Hz), the highest MVI value (5.57 s/nm2), G'/ G″ values and enclosed area, showing that it had denser 3D network structures, higher stiffness values, and a high sensitivity to changes in strain. Additionally, P/H 2:1 emulsion gel had a relatively high lipid digestibility (96.1 %), curcumin bioaccessibility (58.9 %), and curcumin stability (94.2 %). This study showed that emulsion gels stabilized by binary protein nanoparticle mixtures (PNP/HRGF) have potential as edible delivery systems for lipophilic nutraceuticals.