Effect of multi-mode sweep frequency ultrasound pretreatment on properties of the zeins and ACE inhibitory peptides activity of the hydrolysates.

Effects of sweep frequency ultrasound (SFU) pretreatment of a new multifunctional ultrasonic equipment on hydrolysis characteristics of zeins and angiotensin-converting enzyme (ACE) inhibitory activity of zein hydrolysates were investigated. Degree of hydrolysis of zeins reached the highest of 25.93 % and 25.72 % at 40 kHz and 25/40 kHz, respectively. While 25/40 kHz increased solubility, surface hydrophobicity, particle size uniform of zeins and ACE inhibitory activities of the hydrolysates significantly. Endogenous fluorescence indicated that 25/40 kHz promoted unfolding of protein molecules and exposure of hydrophobic residues, thereby facilitating enzymatic hydrolysis. Circular dichroism spectrum and Fourier transform infrared spectrometer illustrated that 25/40 kHz unfolded protein molecules and decreased α-helical contents remarkably. Gel permeation chromatography showed that more small-molecule active peptides were obtained from hydrolysates at 25/40 kHz. In conclusion, SFU pretreatment at 25/40 kHz with the new equipment before proteolysis is an efficient method to improve ACE inhibitory activity of the hydrolysates.

Transcriptome analysis reveals the molecular mechanisms of the novel Lactobacillus pentosus pentocin against Bacillus cereus.

The objective of this study was to investigate the antibacterial effect and mechanism of Lactobacillus pentosus pentocin against Bacillus cereus. The dynamic growth of B. cereus showed that the pentocin had strong antibacterial activity against the strain. The antibacterial mechanism focused on cytomembrane destruction, biofilms formation, DNA replication and protein synthesis of B. cereus. The scanning electron microscopy, transmission electron microscopy and flow cytometry analysis illustrated that the cytomembranes were destroyed, causing the leakage of internal cellular components. Transcriptome sequencing indicated that the genes (KinB, KinC and Spo0B) in two component systems signal pathway were down-regulated, which resulted in the inhibition of the spores and biofilms formation of B. cereus. The phosphorylation and autoinducer-2 import were inhibited by down-regulating the expression levels of LuxS and LsrB genes in quorum sensing signal pathway, which also suppressed biofilms formation of B. cereus. The K+ leakage activated the K+ transport channels by up-relating the genes (KdpA, KdpB and KdpC), promoting the entry of K+ from the extracellular. In addition, the pentocin interfered DNA replication and protein synthesis by regulating the genes associated with DNA replication (dnaX and holB), RNA degradation (cshA, rho, rnj, deaD, rny, dnaK, groEL and hfq) and ribosome function (rpsA, rpsO and rplS). In this article, we provide some novel insights into the molecular mechanism responsible for high antibacterial activity of the L. pentosus pentocin against B. cereus. And the pentocin might be a very promising natural preservative for controlling the B. cereus contaminations in foods.

Complete Replacement of Nitrite With a Lactobacillus fermentum on the Quality and Safety of Chinese Fermented Sausages.

This study investigated the positive effects of complete replacement of nitrite with a Lactobacillus fermentum on the quality and safety of Chinese fermented sausages, and evaluated the risk of this strain. The effects of the strain on pH, color, nitrite, thiobarbituric acid reactive substances (TBARS), total volatile basenitrogen (TVB-N), metmyoglobin (Met-Mb), biological amines, free amino acid content, and sensory index have been studied. The results revealed that the strain reduced the pH of the sausages, which reduced the risk of food-borne pathogens, and accelerated the acidification and gelation process. The inoculation of the strain produced pink color similar to 50 mg/kg nitrite, significantly reducing the residual risk of nitrite in the sausages. In addition, the strain effectively improved quality and nutrition of the sausages through preventing fat oxidation, protein decomposition, and myoglobin oxidation and increasing free amino acid content. The harmful biogenic amines species of the treated sample were reduced, although the tyramine contents were higher than the control, and the contents of the two groups were all far below the specified limit (800 mg/kg). The sensory analysis showed that the strain enhanced the taste, flavor, sourness, and overall acceptability of the sample sausages. Therefore, replacing nitrite completely with the strain L. fermentum could be a potential strategy to produce healthier and safer acceptable sausages through decreasing the risk of nitrite and improving nutrition and quality of the sausages.

Antifungal properties and AFB1 detoxification activity of a new strain of Lactobacillus plantarum.

Aflatoxin B1 is the most toxic mycotoxin and has strong carcinogenicity. In this study, coumarin was employed as the sole carbon source to isolate the microorganisms that had AFB1 detoxification activity. Among106 strains isolated from fermented foods, one potential strain was identified as Lactobacillus plantarum based on 16S rDNA sequence. Removal ratio of AFB1 was 89.5%, inhibitory ratio to A. flavus growth was 42.8% and inhibitory ratio to A. flavus spores growth was as high as 100%. Coumarin utilization indicated that the AFB1 could be decomposed by the strain. The strong antifungal ability against A. flavus growth and spores growth revealed that AFB1 secretion could be highly inhibited by the strain. In addition, High Performance Liquid Chromatography analysis indicated that fermentation supernatant of the strain could degrade AFB1. Scanning Electron Microscopy and Transmission Electron Microscopy indicated that the strain had a strong resistance to AFB1 and had ability to bind AFB1 on the strain surface. Possible detoxification pathway to AFB1 was proposed. Therefore, the strain with high antifungal, antimycotoxigenic abilities might have great potential and immense value in detoxifying AFB1. The use of the strain might be a promising biocontrol strategy to detoxify AFB1.

A composite chitosan derivative nanoparticle to stabilize a W1/O/W2 emulsion: Preparation and characterization.

For preparing stable water-in-oil-in-water emulsion, the role of nanoparticles in stabilizing the interface is very important. In this study, chitosan hydrochloride-carboxymethyl chitosan (CHC-CMC) nanoparticles were prepared considering electrostatic interactions; then the emulsion was prepared and the stability characteristics in presence of NaCl (0-200 mmol/L) and 30 d storage were studied. CHC-CMC nanoparticles (261 nm) were obtained when the CHC: CMC ratio was 1:2. CHC-CMC formation was verified by FT-IR when a new peak appeared at 1580 cm-1; W2 contained 2 wt % CHC-CMC and W1 contained 1 wt % sodium alginate, the creaming index (81.6 %) was higher for the emulsions than Tween 80 (67.4 %) after 30 d. Confocal laser scanning microscopy confirmed the double microstructures, in contrast to the collapse with Tween 80, because the CHC-CMC nanoparticles were densely adsorbing on the oil-water interface. This indicates that CHC-CMC has a stronger ability to stabilize W1/O/W2 emulsion than Tween 80.

Calcium affects glucoraphanin metabolism in broccoli sprouts under ZnSO4 stress.

CaCl2, Ca2+ chelator (EGTA) and Ca2+ channel blocker (verapamil) were used to investigate mechanism of glucoraphanin metabolism in broccoli sprouts under ZnSO4 stress. CaCl2 treatment promoted sprout growth, reduced MDA (malonaldehyde) content and electrolyte leakage in sprouts under ZnSO4 stress. The highest MDA content and electrolyte leakage were obtained in ZnSO4 plus verapamil-treated sprouts. In addition, ZnSO4 plus CaCl2 treatment significantly enhanced glucoraphanin content and sulforaphane formation, while an opposite result was observed after ZnSO4 plus EGTA treatment; which were further supported by expression of glucoraphanin biosynthetic and hydrolytic genes as well as myrosinase (MYR) and epithiospecifier protein (ESP) activities. These results indicated that exogenous and endogenous calcium promoted glucoraphanin biosynthesis and the conversion rate of glucoraphanin into sulforaphane. Verapamil treatment also stimulated glucoraphanin biosynthesis, but exerted an adverse influence on sulforaphane formation from the hydrolysis of glucoraphanin because of much higher ESP expression and ESP activity than ZnSO4 treatment.

The characteristics of gelation of myofibrillar proteins combined with salt soluble Rhodotorula glutinis proteins by enzymatic crosslinking.

Some microbial single-cell proteins are capable of producing synergistic crosslinking interactions with edible proteins by rational regulation. Herein, we reported that salt soluble proteins (RGP) extracted from Rhodotorula glutinis in an alkaline and saline system may combine with myofibrillar proteins (MP) by transglutaminase (TG) polymerization to form improvable irreversible thermal co-gels. The combination of MP, RGP, and TG, namely restructured MP gels, led to significantly enhanced water holding capacity (WHC), up to 90.76 ± 1.88% (% of retained water) and textural properties (hardness, springiness, and adhesiveness) as well as decreases of 'gauche-gauche-gauche' SS bonds and α-helix conformations and increases of 'gauche-gauche-trans' SS bonds and ß-fold conformations, compared to MP and MP-RGP groups. Differential scanning calorimetry analysis validated that thermostability of myosins and actins from MP was reduced after using RGP, TG, and their combination, and unfolding and denaturation of myosin occurred easily during thermal co-gelation by TG and/or RGP.

Antibacterial Activity and Mechanism of Lacidophilin From Lactobacillus pentosus Against Staphylococcus aureus and Escherichia coli.

The main purpose of this study was to explore the antibacterial activity and mechanism of lacidophilin from Lactobacillus pentosus against Staphylococcus aureus and Escherichia coli. The effects of temperature, enzyme, metal ions, and pH on the antibacterial activity of L. pentosus were evaluated. The result showed that lacidophilin had good thermal stability and could be decomposed by trypsin completely. The antibacterial ability was affected by high concentration of metal ions, and the best antibacterial ability was acquired under acidic conditions. The antibacterial mechanism of lacidophilin was explored through studying cytomembrane injury, phosphorus metabolism, protein changes, and oxidative stress response of the indicator bacteria. It was shown that lacidophilin destroyed the cytomembrane of the bacteria and increased the cytomembrane permeability, which resulted in the leak of proteins, nucleic acids, and electrolytes. In addition, it further restrained phosphorus metabolism, caused changes of some protein contents, and increased cytomembrane lipid peroxidation and cell oxidative damage. All these might inhibit the growth of bacteria and even cause their death. This study identified a natural biological preservative with strong antibacterial activity against both Gram-positive and Gram-negative foodborne pathogens. The high antibacterial activity against the two types of bacteria reflected its potential in food preservation used as a natural food preservative.

Beneficial effects of Jerusalem artichoke powder and olive oil as animal fat replacers and natural healthy compound sources in Harbin dry sausages.

This study aimed to improve the nutritional quality of Harbin dry sausages using natural plant-based Jerusalem artichoke powder (JAP) and olive oil as animal fat replacers. Low-fat Harbin dry sausages were manufactured with 2 different formulations containing JAP and olive oil as pork fat replacers. The texture, rheological properties, microstructure, water holding capacity, muscle protein structure, physicochemical indices, microbiological characteristics, and sensory evaluation of the sausages were analyzed. The result showed that Harbin dry sausages with JAP and olive oil were healthier than control sausages based on the lower fat content and improved fatty acid composition. Scanning electron microscopy showed gel network formation in sausages with a high JAP content. Low-field nuclear magnetic resonance illustrated that the water-holding capacity of the modified sausages was improved, suggesting that the replacers enhanced protein gel formation by changes in C-H stretching and bending vibrations, a reduction in α-helixes, and increases in ß-sheets and random coils accompanying the exposure of reactive groups and microenvironment of the tertiary structure. Dynamic rheological and texture tests indicated that the replacers improved the elasticity of sausages. The reduction of fat and addition of replacers significantly enhanced lipid oxidative resistance. Overall, JAP and olive oil improved the fatty acid composition, gel structure, lipid oxidative resistance, and sensory quality of the sausages. These results may contribute to the development of healthy meat products to further reduce animal fat.

Partial replacement of nitrite with a novel probiotic Lactobacillus plantarum on nitrate, color, biogenic amines and gel properties of Chinese fermented sausages.

This article explored the positive effects of partial replacement of sodium nitrite by Lactobacillus plantarum on reducing nitrite and biogenic amine content, improving color and gel structure of Chinese fermented sausages. The results indicated that the strain had nitrate and nitrite reductase activities, which can reduce nitrate and nitrite. And the final reduction products of nitric oxide can react with myoglobin to generate pink nitrosylmyoglobin (Mb(Fe2+)-NO), which can improve sausage color. Dynamic rheological test showed that the strain improved viscoelasticity and gel properties of the samples. Low field nuclear magnetic resonance (LF-NMR) measurement illustrated that the strain enhanced immobilised water in myofibrillar network of the sausages. The protein gels were improved by changes of C-H stretching and bending vibrations, reduction of α-helix and increase of ß-sheet and random coil in secondary structure, changes of microenvironment in tertiary structure. Analysis of biogenic amines showed that the strain reduced risk from biogenic amines by reducing tyramine content. Overall, our findings demonstrated that combination of L. plantarum and low levels of sodium nitrite could be a potential strategy to produce high-quality, healthier sausage by lowering nitrite and biogenic amine levels, improve color and gel properties of Chinese fermented sausages.

Isolation of Antibacterial, Nitrosylmyoglobin Forming Lactic Acid Bacteria and Their Potential Use in Meat Processing.

The use of nitrite as a colorant and preservative in meat processing is associated with health risks. This study aimed to isolate nitrite-substituting lactic acid bacteria for use as natural biological colorants and preservatives. Among the 106 strains isolated from fermented foods, two strains with excellent ability to convert myoglobin and metmyoglobin (Met-Mb) to red nitrosylmyoglobin (Mb-NO) were selected. The superior ability to form Mb-NO was confirmed through UV-visible spectrophotometry, Fourier transform infrared spectrometry, electron spin resonance analysis, nitric oxide synthase activity assay, and Met-Mb reductase activity assay. The potent antibacterial activity was confirmed through biofilm and cytomembrane breakage of the indicator bacteria. Though performing 16S rDNA sequencing, they were identified as two different strains of Lactobacillus plantarum. Based on their favorable characteristics, their applications in the meat industry were further evaluated. This study identified a novel dual-function natural biological colorant and preservative to substitute nitrite in meat products. The application of the two strains would decrease the hazardous of nitrite to health.

Resveratrol-loaded hollow kafirin nanoparticles via gallic acid crosslinking: An evaluation compared with their solid and non-crosslinked counterparts.

The possibility of combining the health benefits of kafirin and polyphenols and improving the bioavailability of resveratrol using hollow kafirin nanoparticles via gallic acid crosslinking was investigated. The size, morphology, charge state, loading efficiency, physicochemical stability, and redispersity after lyophilization of hollow resveratrol-loaded kafirin nanoparticles formed via gallic acid crosslinking were characterized and compared with their solid counterparts and those without crosslinking. The nanoparticles formed were anionic spheres with an average diameter of <100 nm when loading amounts of resveratrol were less than 20%. The hollow nanoparticles were homogenous and still achieved stable colloidal dispersion after lyophilization. The hollow nanoparticles crosslinked with gallic acid displayed stability against pancreatin and delayed release in stimulated digestion. The results suggested that hollow kafirin nanoparticles could be a favorable colloidal delivery system for incorporating resveratrol.

The antibacterial mechanism of ultrasound in combination with sodium hypochlorite in the control of Escherichia coli.

In the present study, the action mechanism of ultrasound (US) combined with sodium hypochlorite (SH) against Escherichia coli was illustrated by different analysis, including reduction, particle size distribution, scanning electron microscopy (SEM), transmission electron microscopy (TEM), K+ leakage, confocal laser scanning microscopy (CLSM) and fluorescence spectroscopy of Escherichia coli. The results showed that ultrasound improved the antimicrobial effect of SH in control of E. coli. No significant difference was obtained in reduction of E. coli, CLSM analysis and K+ leakage between US + SH30 (US + 30 ppm SH) and SH50 (50 ppm SH) treatment. Smaller particle size was recorded in US and US + SH30 treatment. The changes of morphology and intracellular organization of E. coli cells as a result of these treatments were confirmed by SEM and TEM analyses. Fluorescence spectroscopy results indicated SH30, US + SH30 and SH50 treatment caused the burial of tyrosine residues and tryptophan residues as well as increase of hydrophobicity. Therefore, the mechanism of US + SH30 treatment against E. coli involved decreased particle size, damaged membrane and changes of intracellular organization and protein conformation.

Rapid Detection of Enterobacter Sakazakii in milk Powder using amino modified chitosan immunomagnetic beads.

Chitosan immunomagnetic beads (CIBs) were first prepared through converting hydroxyl groups of natural polymer material-chitosan into amino groups using epichlorohydrin and ethylenediamine as modification agent and then coupling with polyclonal antibodies of Enterobacter sakazakii using glutaraldehyde as cross-linking agent. The beads before coupling with antibodies were characterized by magnetic property measurement, FTIR, SEM and XRD technologies. In the assay a natural polysaccharide-chitosan, which has good biological and chemical properties such as non-toxicity, biocompatibility and high chemical reactivity was first used for synthesis of immunomagnetic beads. The detection method first established in this paper that combined the beads with chromogenic medium together to rapid detect E. sakazakii in milk powder could greatly improve the detection specificity and working efficiency. The beads exhibited a maximum capturing capacity of 1×106cfu/g with the detection sensitivity of 4cfu/g. The results demonstrate that the assay is a straightforward, specific and sensitive alternative for rapid detection of E.sakazakii in food matrix. The total analysis time was as little as about 25h, which greatly shorten the detection time. The method can provides new ideas not only to preparation technique of immunomagnetic beads but to imunne detection technique in food safety.

Bioaccumulation of cadmium by growing Zygosaccharomyces rouxii and Saccharomyces cerevisiae.

Bioaccumulation via growing cells is a potential technique for heavy metal removal from food materials. The cadmium bioaccumulation characteristics by growing Zygosaccharomyces rouxii and Saccharomyces cerevisiae were investigated. Z. rouxii displayed powerful cadmium removal ability at low cadmium concentrations, which mainly depended on the intracellular cadmium bioaccumulation. The percentage of intracellular cadmium bioaccumulation of both yeasts obviously decreased with the increase of initial biomass and cadmium concentrations. Low pH and elevated concentrations of zinc and copper significantly decreased the intracellular cadmium bioaccumulation of both yeasts but improved the cadmium tolerance and the cell-surface cadmium bioaccumulation of Z. rouxii. Cadmium removal of Z. rouxii was improved by zinc and copper conditionally. Z. rouxii that possessed more powerful cadmium tolerance and removal ability at low pH and high concentration of competing ions can be developed into a potential cadmium removal agent using in complex food environment in future.