Jabuticaba peel extract and nisin: A promising combination for reducing sodium nitrite in Bologna-type sausages.

This study investigated the effect of a 50% reduction in sodium nitrite and the addition of nisin (200 mg/kg) and different concentrations (0, 0.5%, 0.75%, and 1%) of jabuticaba peel extract (JPE) on the main attributes affected by this chemical additive in Bologna-type sausages. The modified treatments showed approximately 50% lower residual nitrite than the control throughout the storage (60 days at 4 °C). The proposed reformulation did not affect the color (L*, a*, and b*), and the ΔE values (< 2) demonstrated high color stability during storage. Physicochemical (TBARS and volatile compounds) and sensory analyses performed to evaluate oxidative stability indicated that JPE exhibited antioxidant activity comparable to sodium nitrite. The microbiological quality of the reformulated products was similar to the control, but further studies should be conducted to assess the effect of this reformulation strategy on the growth of pathogenic microorganisms impacted by nitrite.

Chemical Synthesis of Proteins Using an o-Nitrobenzyl Group as a Robust Temporary Protective Group for N-Terminal Cysteine Protection.

We report here a robust and practical strategy for chemical protein synthesis using an o-nitrobenzyl group as a temporary protective group for an N-terminal cysteine residue of intermediate hydrazide fragments. By reinvestigating the photoremoval of an o-nitrobenzyl group, we establish a robust and reliable strategy for its quantitative photodeprotection. The o-nitrobenzyl group is completely stable to oxidative NaNO2 treatment and has been applied to the convergent chemical synthesis of programmed death ligand 1 fragment, providing a practical avenue for hydrazide-based native chemical ligation.

Pomegranate peel as a source of antioxidants for the control of lipid and protein oxidation during the ripening of Iberian dry uncured sausages.

This study aimed to evaluate a pomegranate peel extract (PPE), selected for its level of phytochemical compounds and antioxidant activities, as a nitrite substitute in dry sausages, as well as its effect on lipid and protein oxidative changes and instrumental colour during the drying period (28 days). Of the extraction solvents screened, water: acetone 3:7 (v/v) was the most effective extraction solvent, yielding extracts with the highest content of phenolic compounds, flavonoids and condensed tannins and antioxidant activities (ABTS, DPPH and FRAP). Four batches of dry sausages were produced with different incoming amounts of sodium nitrite (NaNO2) and PPE: 1) 150 ppm NaNO2; 2) 0 ppm NaNO2; 3) 0 ppm NaNO2 + 1% PPE (v/w) and 4) 0 ppm NaNO2 + 2% PPE (v/w). Nitrite removal increased lipid oxidation in uncured dry sausages, while nitrite and PPE caused lower TBA-RS values in cured and PPE treated sausages. During drying, both nitrite and PPE addition significantly decreased carbonyl and thiol contents compared to the uncured dry sausages. A dose-response was found for PPE, with lower carbonyl and thiol concentrations, the higher the level of PPE added. PPE significantly modified instrumental colour coordinates L*a*b* producing significant total colour changes compared to cured dry sausages.

Effects of nitrite-rich and pigment-rich substitutes for sodium nitrite on the quality characteristics of emulsion-type pork sausages during cold storage.

This study was conducted to evaluate the effects of nitrite-rich (celery powder; CP) and pigment-rich (purple sweet potato powder, PSP; red beet powder, RB) substitutes for synthetic sodium nitrite (CON) on the quality characteristics of emulsion-type pork sausages during four weeks of cold storage. Natural substitutes decreased the pH, lightness, and textural properties of pork sausages during storage (P < 0.05). Pigment-rich substitutes showed a decreased antioxidant effect after two weeks of storage when compared to the nitrite-rich groups (CON and CP; P < 0.05). Pigment-rich substitutes also accelerated the discoloration of pork sausages by increasing yellowness (RB and PSP) and decreasing redness (PSP) during storage (P < 0.05). However, these two pigment-rich substitutes showed different trends in redness (higher in PSP and lower in RB) and yellowness (higher in RB and lower in PSP) when compared to the nitrite-rich groups (CON and CP). Different types (nitrite-rich and pigment-rich) of natural substitutes for sodium nitrite had different effects on the quality characteristics of emulsion-type pork sausages throughout the four weeks of storage evaluated in this study.

Effects of nitrite concentrations on the quality and protein oxidation of salted meat.

The objective of this study was to investigate the effects of different concentrations of sodium nitrite on the quality and protein oxidation of salted meat during 21 days of curing. The salted meat was treated with sodium nitrite at 50, 100, and 150 mg/kg for curing, and salted meat without sodium nitrite was used as a control. The results showed that in salted meat added with sodium nitrite, the carbonyl group, disulfide bond, dityrosine, surface hydrophobicity, and the transformation rate from α-helix to ß-sheet were all significantly reduced, whereas the sulfhydryl group content of myofibrillar proteins was significantly increased compared to the control. Meanwhile, the total volatile basic nitrogen and aerobic plate content were significantly decreased, while both the pH and a* value were significantly increased with an increase in nitrite concentration compared to the control group. Importantly, this phenomenon was also observed in salted meat treated with low doses of sodium nitrite (50 mg/kg). In conclusion, the quality of salted beef can be improved by adding low-dose sodium nitrite to inhibit protein oxidation during the curing process. PRACTICAL APPLICATION: A low dose of sodium nitrite inhibited the rate of α-helix to ß-sheet transformation of myofibrillar proteins in salted meat, reducing the exposure of hydrophobic groups and decreasing the production of protein oxidation products and TVB-N to improve the quality of salted meat. These results provided a theoretical basis and technical guidance for the application of low-dose sodium nitrite in meat processing enterprises.

Scalable manufacturing platform for the production of methemoglobin as a non-oxygen carrying control material in studies of cell-free hemoglobin solutions.

Methemoglobin (metHb) arises from the oxidation of ferrous hemoglobin (HbFe2+, Hb) to ferric hemoglobin (HbFe3+, metHb), which is unable to bind gaseous ligands such as oxygen (O2) and carbon monoxide (CO), and binds to nitric oxide (NO) significantly slower compared to Hb. Therefore, metHb does not elicit vasoconstriction and systemic hypertension in vivo due to its extremely slow NO scavenging rate in comparison to cell-free Hb, but will induce oxidative tissue injury, demonstrating the potential of using metHb as a control material when studying the toxicity of cell-free Hb. Hence, the goal of this work was to develop a novel manufacturing strategy for production of metHb that is amenable to scale-up. In this study, small scale (e.g. 1 mL reaction volume) screening experiments were initially conducted to determine the optimal molar ratio of Hb to the oxidization agents hydrogen peroxide (H2O2) or sodium nitrite (NaNO2) to achieve the highest conversion of Hb into metHb. A spectral deconvolution program was employed to determine the molar fraction of various species (hemichrome, metHb, oxyHb, metHb-[Formula: see text], and NaNO2) in solution during the oxidation reaction. From this analysis, either a 1:1 or 1:5 molar ratio was identified as optimal molar ratios of Hb:NaNO2 (heme basis) that yielded the highest conversion of Hb into metHb with negligible amounts of side products. Hence in order to reduce the reaction time, a 1:5 molar ratio was chosen for large scale (i.e. 1.5 L reaction volume) synthesis of bovine metHb (metbHb) and human metHb (methHb). The biophysical properties of metHb were then characterized to elucidate the potential of using the synthesized metHb as a non-O2 carrying control material. The haptoglobin binding kinetics of metHb were found to be similar to Hb. Additionally, the synthesized metHb was stable in phosphate buffered saline (PBS, 50 mM, pH 7.4) at 4°C for approximately one week, indicating the high stability of the material.

Synergistic Effect of Mandarin Peels and Hesperidin with Sodium Nitrite against Some Food Pathogen Microbes.

Food preservatives such as NaNO2, which are widely used in human food products, undoubtedly affect, to some extent, human organs and health. For this reason, there is a need to reduce the hazards of these chemical preservatives, by replacing them with safe natural bio-preservatives, or adding them to synthetic ones, which provides synergistic and additive effects. The Citrus genus provides a rich source of such bio-preservatives, in addition to the availability of the genus and the low price of citrus fruit crops. In this study, we identify the most abundant flavonoids in citrus fruits (hesperidin) from the polar extract of mandarin peels (agro-waste) by using spectroscopic techniques, as well as limonene from the non-polar portion using GC techniques. Then, we explore the synergistic and additive effects of hesperidin from total mandarin extract with widely used NaNO2 to create a chemical preservative in food products. The results are promising and show a significant synergistic and additive activity. The combination of mandarin peel extract with NaNO2 had synergistic antibacterial activity against B. cereus, Staph. aureus, E. coli, and P. aeruginosa, while hesperidin showed a synergistic effect against B. cereus and P. aeruginosa and an additive effect against Staph. aureus and E. coli. These results refer to the ability of reducing the concentration of NaNO2 and replacing it with a safe natural bio-preservative such as hesperidin from total mandarin extract. Moreover, this led to gaining benefits from their biological and nutritive values.

Potentially probiotic or postbiotic pre-converted nitrite from celery produced by an axenic culture system with probiotic lacticaseibacilli strain.

The present study evaluated the use of the probiotic Lacticaseibacillus paracasei DTA-83 as a nitrite-reducing agent to produce potentially probiotic or postbiotic pre-converted nitrite from celery. The results obtained were compared to those achieved by direct addition of sodium nitrite for the typical reddish color formation in cooked pork sausages and the inhibitory potential against the growth of target microorganisms, including the clostridia group. Regarding the sausages color, similar findings were observed when comparing the use of pre-converted nitrite from celery produced by L. paracasei DTA-83 and the direct addition of sodium nitrite. Additionally, it presented an inhibitory effect against Salmonella spp., which was not observed with the direct addition of nitrite, revealing a potential strategy to control salmonellosis in the matrix. However, a non-equivalent preservative effect against Clostridium perfringens (INCQS 215) was determined. The results highlight a promising alternative to produce probiotic or postbiotic meat ingredients; however, further studies should be conducted to investigate doses that achieve microbial control.

Effects of a chemical additive on the fermentation, microbial communities, and aerobic stability of corn silage with or without air stress during storage.

We evaluated the effects of a chemical additive on the microbial communities, fermentation profile, and aerobic stability of whole-plant corn silage with or without air stress during storage. Whole-plant corn was either untreated or treated with a chemical additive containing sodium benzoate, potassium sorbate, and sodium nitrite at 2 or 3 liters/t of fresh forage weight. Ten individually treated and replicated silos (7.5 liters) were made for each treatment. Half of the silos remained sealed throughout a 63-d storage period, and the other half was subjected to air stress for 2 h/wk. The composition of the bacterial and fungal communities of fresh forage and silages untreated or treated with 2 liters/t of fresh forage weight was analyzed by Illumina Miseq sequencing. Treated silage had greater (P < 0.05) aerobic stability than untreated, even when subjected to air stress during storage, but the numbers of yeasts culturable on selective agar were not affected. However, the additive reduced the relative abundance (RA) of the lactating-assimilating yeast Candida tropicalis (P < 0.01). In air-stressed silages, untreated silage had a greater (P < 0.05) RA of Pichia kudriavzevii (also a lactate assimilator) than treated silage, whereas treated silage was dominated by Candida humilis, which is usually unable to assimilate lactate or assimilates it slowly. The additive improved the aerobic stability by specifically preventing the dominance of yeast species that can consume lactate and initiate aerobic spoilage. To the best of our knowledge, this is the first work that identifies the specific action of this additive on shifting the microbial communities in corn silage.

Tomato pomace extract and organic peppermint essential oil as effective sodium nitrite replacement in cooked pork sausages.

The effect of supercritical fluid extract of tomato pomace (TP) and essential oil of organic peppermint (PM) on pH, color, residual nitrite content, lipid oxidation (TBARS value) and total plate count (TPC) of cooked pork sausages produced with 50 mg/kg of sodium nitrite was investigated. Five batches were produced: T1: 100 mg/kg of sodium nitrite; T2: 50 mg of sodium nitrite; T3: 50 mg of sodium nitrite and 0.150 µL/g TP; T4: 50 mg of sodium nitrite, 0.075 µL/g TP and 0.075 µL/g PM; T5: 50 mg of sodium nitrite and 0.150 µL/g PM. The lowest residual nitrite content and TBARS value were observed in treatment T4. The inclusion of TP increased redness of cooked pork sausages. TPC was the lowest in treatment T5. The results of this study showed that the addition of TP and PM enhanced quality of cooked sausages produced with reduced level of sodium nitrite.

Using chitosan and radish powder to improve stability of fermented cooked sausages.

Radish powder and chitosan were evaluated in fermented cooked sausages to replace sodium nitrite. Treatments of fermented cooked sausages were prepared and evaluated during the ripening process and storage time: control (150 ppm nitrite, CONT); and two levels of chitosan (0.25%, CHI1) and (0.5%, CHI2) with radish powder (0.5%) and without the addition of nitrite. During storage, pieces sliced or not were also evaluated regarding the physicochemical, microbiological and sensory. Pure chitosan was evaluated regarding determination of MIC and MBC to Enterobacter aerogenes, Listeria innocua and Lactobacillus rhamnosus, and antioxidant capacity. The addition of chitosan increased aw and decreased weight loss of fermented sausages along processing, but pH was not affected during ripening. Except for aroma, sensory attributes were affected by the addition of chitosan. The addition of 0.5% radish powder and 0.25% chitosan showed promising results to development of nitrite-free fermented cooked sausages, mainly regarding their physicochemical and microbiological stability.

Utility of winter mushroom treated by atmospheric non-thermal plasma as an alternative for synthetic nitrite and phosphate in ground ham.

The utility of plasma-treated winter mushroom powder (PWMP) as an alternative to synthetic nitrite and phosphate in ground ham was evaluated. Treatment of atmospheric non-thermal plasma generated nitrite in winter mushroom homogenate, and PWMP contained 4.87 g/kg of nitrite. Canned ground ham samples without nitrite and phosphate (NC), with sodium nitrite and sodium pyrophosphate (PC), and with PWMP (PWM) were manufactured, sterilized (F0 > 4), and stored under accelerated conditions (40 °C) for 30 days. The nitrosyl-hemochrome content and color of PWM were similar to those of PC, without differences in the thiobarbituric acid reactive substances (TBARS) value. The proportion of jelly and melted fat exuded from ground hams in PWM exceeded that in PC but was lower than that in NC (P < .05). Therefore, PWMP can successfully replace synthetic nitrite, but it is an insufficient substitute for phosphate in ground ham.

Screening of Fungi Isolates for C-4 Hydroxylation of R-2-Phenoxypropionic Acid Based on a Novel 96-Well Microplate Assay Method.

R-2-(4-Hydroxyphenoxy)propionic acid (R-HPPA) is a pivotal intermediate for the synthesis of aryloxyphenoxypropionate (APP) herbicide. To rapidly screen microbial isolates with the capacity of hydroxylating R-2-phenoxypropionic acid to R-HPPA from various environmental samples, a convenient and safe 96-well microplate assay method with sodium nitrite (NaNO2) as chromogenic reagent was proposed and optimized. The optimum assay conditions were as follows: the detection wavelength was 420 nm, the concentration of NaNO2 solution was 6.0 g/L, color reaction temperature was 60 °C, the pH of the NaNO2 solution was 2.4, and the reaction time was 40 min. With the aid of this method, screening for microorganisms with C-4-specific hydroxylation activity of R-PPA was conducted. As a result, 23 strains among 3744 single colonies isolated from various samples exhibited the hydroxylation activity. Among these strains, the highest bioconversion rate was achieved by Penicillium oxalicum A5 and Aspergillus versicolor A12, respectively. After 72-h cultivation in shake flask, their conversion rates of R-HPPA from 10 g/L R-PPA reached 21.18% and 40.24%, respectively. The established method was effective in rapid screening of microbes capable of biosynthesizing R-HPPA through hydroxylation of R-PPA, and the obtained two fungi species could be potentially used for R-HPPA production.

Characterization of the microbial community composition in Italian Cinta Senese sausages dry-fermented with natural extracts as alternatives to sodium nitrite.

Nitrite is widely used in meat products as a multifunctional additive, combining flavour and colour properties with antioxidant and antimicrobial effects. However, nitrite may form reaction products (i.e., nitrosamine) that are potentially carcinogenic to humans. The meat industry, in response to consumers' demands for nitrite-free products, is seeking natural alternatives to nitrite, such as plant-based extracts. Three types of dry-fermented sausages were manufactured: NIT, containing 30 ppm of sodium nitrite; GSE, containing grape seed extract and olive pomace hydroxytyrosol; and CHE, containing chestnut extract and olive pomace hydroxytyrosol. Next-generation sequencing (NGS) was used to analyse microbial consortia, which were correlated with physical and chemical parameters. The prokaryotic community composition was similar among treatments, with a high relative abundance of Staphylococcus xylosus and Lactobacillus sakei, collectively accounting for 87% of the total community. However, significant differences were observed in both operational taxonomic unit (OTU) presence/absence and relative abundance. Ten genera varied in abundance between treatments. The increase in Lactobacillaceae in CHE may explain the reduced pH levels detected in these samples. In conclusion, NGS analysis showed that the prokaryotic community composition was similar in GSE and NIT, while CHE varied in both the composition and relative abundance of different taxa.

Microbial biotransformation of N-nitro-, C-nitro-, and C-nitrous-type mutagens by Lactobacillus delbrueckii subsp. bulgaricus in meat products.

Processed meats are classified by the International Agency for Research on Cancer (IARC) as carcinogenic to humans. However, information on the responsible agents and the influence of industrial processing on the increased risk of cancer is still lacking. This study aimed to use cultures of Lactobacillus delbrueckii subsp. bulgaricus LB-UFSC 01 to biodegrade harmful C-nitrous, N-nitro, and C-nitro compounds in processed meat matrix. Firstly, positive results for ethylnitrolic acid (ENA) (>5.00 µg kg-1) and 2-methyl-1,4-dinitro-pyrrole (DNMP) (>12.0 µg kg-1) were obtained in mortadellas produced under different experimental conditions employing preservatives and antioxidants. Mortadellas containing nitrite and sorbate in the ratio of 8:1 (w/w) yielded the highest concentrations of mutagens. However, the treatment with the LB-UFSC 01 culture was able to modulate the harmful compounds in the mortadella samples. Several analytical methods employing liquid chromatography coupled to mass spectrometry and statistical models were employed to identify the metabolites and reaction routes during microbial biotransformation. For the first time, relevant information regarding the formation and degradation of ENA and DNMP in a processed meat model simulating real conditions was presented.

Photochemical reaction of tricresyl phosphate (TCP) in aqueous solution: Influencing factors and photolysis products.

Organophosphate triesters (OPEs) have caused great concern as a class of emerging environmental contaminants due to their widespread use and their toxicity to organisms. However, the phototransformation behavior of OPE is still not fully understood, which is important for understanding their environmental fate. In the present study, the photodegradation of tricresyl phosphate (TCP), one of the most widely detected OPEs in aqueous environments, was investigated including the direct photolysis and in the presence of several natural water factors, NO2-, Fe3+ and humic acid. The degradation process followed the pseudo-first-order kinetics, with rate constant increasing slightly with increasing initial TCP concentration. The presence of NO2- and Fe3+ was observed to promote the photochemical loss of TCP, while humic acid played a negative role on TCP transformation. Electron spin resonance (EPR) analysis showed that carbon-centered radical was produced in the photolysis process of TCP, and hydroxyl radical contributed to the promotion of rate constant for Fe3+ and NO2-. Four photolysis products were tentatively identified by HPLC-LTQ-Orbitrap MS analysis, and the possible degradation pathways of TCP were proposed. These findings provide a meaningful reference for the fate and transformation of OPEs in natural water.

Coriander essential oil as natural food additive improves quality and safety of cooked pork sausages with different nitrite levels.

The effect of coriander essential oil (CEO) at concentrations of 0.075-0.150 µL/g on pH, color, lipid oxidation (TBARS), residual nitrite concentration and microbial growth of cooked pork sausages produced with different levels of sodium nitrite (0, 50 and 100 mg/kg) was investigated. Artificial neural networks modeling and the multi-response optimization were used to determine the optimal combinations of process parameters and storage time. Reduced concentration of sodium nitrite (60 mg/kg) in combination with 0.12 µL/g of CEO resulted in satisfying redness (a* approx. 11.1) and improved oxidative (TBARS approx. 0.12 mg MDA/kg) and microbial stability (total plate count - TPC approx. 2.50 Log CFU/g) of cooked pork sausages during refrigerated storage. Therefore, the results of this paper revealed significant antioxidative and antimicrobial activity of CEO, and consequently its high potential of utilization in processing of cooked pork sausages with enhanced quality and shelf-life.

Quality characteristics of pork loin cured with green nitrite source and some organic acids.

This study was conducted to improve the quality characteristics of cured meat with natural nitrite. Control and treatment were conducted as follows: nitrite free, marinated with sodium nitrite and ascorbic acid, marinated with only fermented spinach (FS), and marinated with FS adding ascorbic acid, malic acid, citric acid, and tartaric acid. Treatments were pickled with regulated brine (8% salt and 0.08% nitrite). Cured meat with FS adding ascorbic acid, malic acid, and citric acid had higher redness values than sodium nitrite with ascorbic acid on cooked meat. There was a positive effect on lipid oxidation except for citric acid. Protein degradation appeared more in malic acid and tartaric acid treatment than others. Residual nitrite level was lower when adding organic acids. Among various organic acid, ascorbic acid had the highest efficient on quality properties of cured meat. Thus, ascorbic acid was a proper ingredient when curing meat product.

Nitrite scavenging impact of fermented soy sauce in vitro and in a pork sausage model.

The objective of this study was to determine nitrite scavenging activity of fermented soy sauce in vitro and in a pork sausage model. In vitro nitrite scavenging activity (pH 6.0) of fermented soy sauce (10 mg/mL) was 13.2%. Within selected concentrations (10-100 mg/mL), in vitro nitrite scavenging activity and total phenol content of fermented soy sauce were highly dose-dependent (P < 0.001). In pork sausage model containing 120 mg/kg of NaNO2, fermented soy sauce resulted in significantly lower residual nitrite content (35.28 mg/kg) compared to only NaNO2 addition (40.12 mg/kg) at initial storage. During 4 weeks of cold storage, however, fermented soy sauce showed little effect on the residual nitrite content. This study indicates that fermented soy sauce could initially contribute to reduce residual nitrite content, and the nitrite scavenging impact of fermented soy sauce was less effective in the pork sausage model than in vitro evaluation.

A novel S-nitrosocaptopril monohydrate for pulmonary arterial hypertension: H2O and -SNO intermolecular stabilization chemistry.

S-nitrosocaptopril (CapNO) possesses dual capacities of both Captopril and an NO donor with enhanced efficacy and reduced side effects. CapNO crystals are difficult to make due to its unstable S-NO bond. Here, we report a novel stable S-nitrosocaptopril monohydrate (CapNO·H2O) that is stabilized by intermolecular five-membered structure, where one H of H2O forms a hydrogen bond with O- of the stable resonance zwitterion Cap-S+=N-O-, and the O in H2O forms the dipole-dipole interaction with S+ through two unpaired electrons. With the chelation and common ion effect, we synthesized and characterized CapNO·H2O that is stable at 4 °C for 180 days and thereafter without significant degradation. Compared to Captopril, CapNO showed direct vasorelaxation and beneficial effect on PAH rats, and could be self-assembled in rat stomach when Captopril and NaNO2 were given separately. This novel CapNO·H2O with low entropy paves an avenue for its clinical trials and commercialization.