Food Technology and Biotechnology最新文献

Research background: Breast cancer is one of the most common cancers and remains a major cause of morbidity and mortality among women worldwide. In developed countries, breast cancer as a multifactorial disease is a major health concern, and its incidence is constantly rising in low and middle-income countries. Numerous studies have demonstrated that phytochemicals such as carotenoids inhibit breast cancer growth and induce apoptosis. We recently enhanced the solubility of capsanthin in water by encapsulating it in diosgenin polyethylene glycol succinate, a novel non-ionic surfactant. Thus, this study aims to evaluate the cytotoxicity of water-soluble capsanthin-loaded micelles in MDA-MB-231 cells in vitro through tetrazolium dye MTT assay.

Experimental approach: In the current study, capsanthin, a hydrophobic carotenoid, is extracted from sweet red pepper (Capsicum annuum). Capsanthin-loaded diosgenin polyethylene glycol succinate 1000 (cap-DPGS-1000) micelles were prepared from capsanthin extract (cap) and diosgenin polyethylene glycol succinate 1000 (DPGS-1000) using the solid dispersion method. The capsanthin extract and cap-DPGS-1000 micelles were characterized by UV-visible spectroscopy, high-performance liquid chromatography (HPLC), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), particle size distribution, polydispersity, and scanning electron microscopy (SEM). The effects of capsanthin extract and cap-DPGS-1000 micelles on a human triple-negative breast cancer cell line (MDA-MB-231) were tested to check the cell viability, proliferation and cytotoxicity of the micelles.

Results and conclusions: The solubility of encapsulated cap-DPGS-1000 micelles in water is greatly enhanced and leads to an increased scope for localized drug delivery, a better delivery option for treating residual cancerous tumours. The encapsulated capsanthin showed a sustained release in simulated intestinal fluid (pH=6.8). Our research proposes a sustained drug delivery system that ensures effective and controlled release to the affected site. The characterization data revealed no change in the structure and functional groups in the encapsulated capsanthin. The IC₅₀ value of the cap-DPGS-1000 micelles against MDA-MB-231 breast cancer cells was (3.10±1.09) μg/mL, which is much lower than of capsanthin extract ((81.1±1.5) μg/mL). Capsanthin extract and capsanthin-loaded micelles are promising drug candidates to induce apoptosis and increase reactive oxygen species (ROS) in cancer cells.

Novelty and scientific contribution: The result shows the cytotoxic effect of capsanthin and capsanthin-loaded micelles on MDA-MB-231 cell line for the first time. Capsanthin from sweet red pepper (Capsicum annuum) showed remarkable cytotoxic effect on the triple-negative MDA-MB-231 cell line.

Research background: This study aims to monitor the growth of the methylotrophic bacteria Methylobacterium organophilum in a culture medium with methanol as a carbon source and to verify the production of unicellular proteins and other biomolecules, such as carotenoids, exopolysaccharides and polyhydroxyalkanoates, making them more attractive as animal feed.

Experimental approach: Bacterial growth was studied in shake flasks using different carbon/nitrogen (C:N) ratios to determine their best ratio for achieving the highest volumetric productivity of cells and substrate consumption rate. This optimal parameter was further used in a fed-batch operating bioreactor system to define the kinetic profile of cell growth. Methanol consumption was measured by HPLC analysis and the extracted pigments were analyzed by liquid chromatography/mass spectrometry. Chemical composition and rheological properties of the produced exopolysaccharides were also determined.

Results and conclusions: The best experimental parameters were verified using an initial methanol concentration of 7 g/L in the culture medium. The same initial substrate concentration was used in the fed-batch operation and after 60 h of cultivation 5 g/L of biomass were obtained. The accumulation of carotenoids associated with cell growth was monitored, reaching a concentration of 1.6 mg/L at the end of the process. These pigments were then analyzed and characterized as a set of xanthophylls (oxidized carotenoids). In addition, two other product types were identified during the fed-batch operation: exopolysaccharides, which reached a concentration of 8.9 g/L at the end of the cultivation, and an intracellular granular structure that was detected by transmission electron microscopy (TEM), suggesting the accumulation of polyhydroxyalkanoate (PHA), most likely polyhydroxybutyrate.

Novelty and scientific contribution: Methylobacterium organophilum demonstrated a unique ability to produce compounds of commercial interest. The distinct metabolic diversity of this bacterium makes room for its use in biorefineries.

Research background: Anthocyanins possess valuable health-promoting activities with significant health benefits for humans. However, their instability is a limiting factor for their usage in functional foods and beverages.

Experimental approach: In this work, a new method to enhance the stability of anthocyanins from mulberry fruit through acylation by using succinic acid as a selected acyl donor was explored. The Box-Behnken design of response surface methodology was applied to determine the optimized conditions for the acylation process.

Results and conclusions: The highest acylation conversion rate was 79.04% at anthocyanins to succinic acid mass ratio 1:8.96, acylation duration 3 h and temperature 50 °C. Structural analysis of acylated anthocyanins revealed that succinic acid introduces a C-O-C bond and a hydroxyl group. The thermostability and light stability of mulberry anthocyanins were significantly improved after acylation, and the antioxidant activity expressed as total reducing power and Fe²⁺-chelating capacity of the acylated anthocyanins was also enhanced.

Novelty and scientific contribution: Succinic acid acylation provides a novel method for stabilizing mulberry anthocyanins, as evidenced by the increased stability and antioxidant ability of anthocyanins, and thus facilitates its use in the food and nutraceutical industries.

Research background: Oral microbiota has become an important factor in obesity, but its association with obesity-related diseases and serum 25-hydroxy vitamin D [25(OH)D] and B complex amounts is still uncertain. The main aim of the paper is to determine the variation in oral microbiota composition as a response to the vitamin status and obesity-related diseases in obese females from Croatia. We hypothesized that the prevalence of probiotic or pathogenic bacteria in the oral cavity of obese women in Croatia depends on the amounts of vitamin B₉ (folic acid), B₁₂ and 25(OH)D in serum and/or hypertension, diabetes and prediabetes diagnosis.

Experimental approach: To test the defined research hypothesis, female individuals with body mass index (BMI)≥30 kg/m² (N=70) were recruited to participate in this study. Obese women were divided into groups according to BMI value, diagnosis of obesity-related diseases and amount of micronutrient in blood. For the quantitative determination of folic acid, vitamin B₁₂ and 25(OH)D in serum, an electrochemiluminescence protein binding assay (ECLIA) was performed. Microorganisms isolated from the saliva of obese women were analyzed by MALDI-TOF mass spectrometer.

Results and conclusions: The presented results do not support the hypothesis that the prevalence of probiotic or pathogenic bacteria in the oral cavity of obese women in Croatia depends on the amount of micronutrients. On the other hand, hypertension and diabetes/prediabetes favour the growth of oral pathogens, specifically increased levels of Candida sp.

Novelty and scientific contribution: To the best of our knowledge, this is the first study showing the relationship between obesity, micronutrient amount, oral microbiota composition, and the incidence of obesity-related disease. We included only obese women from Croatia, so it is regionally specific. Also, we have shown that oral microbiota composition is not connected with micronutrient deficiencies but only with obesity-related diseases.

Research background: Obesity increases mortality and morbidity due to its impact on type 2 diabetes, cardiovascular and gastrointestinal diseases, arthritis and certain cancers. The epidemic of excessive mass and obesity require constant research to improve therapies without undesirable side effects. Therefore, exploring the anti-obesity phytochemicals from food sources is essential. Most pharmacological studies of the anti-obesity potential of Capsicum annuum have been directed towards capsaicin and very few towards capsanthin. However, these studies utilized uncoated capsaicin and capsanthin. This study aims to compare the anti-obesity effects of enteric-coated capsaicin and capsanthin in a high-fat diet-induced obesity in animal model.

Experimental approach: In this study, we investigated the anti-obesity properties of capsanthin-enriched pellets and capsaicin pellets derived from red chili fruit (Capsicum annuum) on high-fat diet (HFD)-induced obesity in C57BL/6J mice. First, the animals received HFD to induce their obesity. Animals were supplemented orally with pellets. The food intake, body mass, obesity and clinical biomarkers were assessed.

Results and conclusions: The mice fed with HFD gained body mass and white adipose tissue mass compared to the mice that consumed a normal diet. The oral administration of capsanthin-enriched pellets and capsaicin pellets significantly reduced the body mass gain. These pellets have a statistically significant (p<0.05) impact on obesity biomarkers by increasing adiponectin and decreasing leptin, free fatty acid and insulin concentrations relative to HFD control. There was no change in the liver mass in all groups, but there was a significant decrease in white adipose tissue amounts. Inguinal adipose tissue amount was reduced by 37.0% and that of epididymal adipose tissue by 43.64% after treatment with capsanthin-enriched pellets. These results suggest that capsanthin-enriched pellets and capsaicin pellets may be useful in combating metabolic diseases, including obesity, without adverse effects.

Novelty and scientific contribution: We increased the content of capsanthin for more than 50% in capsanthin-enriched extract and extended the room temperature stability for more than one year by converting the crystals into capsanthin-enriched pellets. This study breaks new ground by examining the potential of capsanthin >50% in the management of obesity for the first time.

Research background: Haloalkaline proteases are one of the most interesting types of commercial enzymes in various industries due to their high specific activity and stability under extreme conditions. Biochemical characterization of enzymes is an important requirement for determining their potential for application in industrial fields. Most of microbial proteases have been isolated from Bacillus spp. In this study, the purification and characterization of an extracellular haloprotease produced from Bacillus sp. KB111 strain, which was previously isolated from mangrove forest sediments, are investigated for industrial applications.

Experimental approach: The whole genome of KB111 strain was identified by DNA sequencing. Its produced protease was purified by salting out and anion-exchange chromatography, characterized based on protease activity and stability using a peptide substrate, and identified by LC-MS/MS.

Results and conclusions: The strain KB111 was identified as Bacillus licheniformis. The molecular mass of its extracellular protease, termed KB-SP, was estimated to be 70 kDa. The optimal pH and temperature for the activity of this protease were 7 and 50 °C, respectively, while the enzyme exhibited maximal activity in the broad salinity range of 2-4 M NaCl. It was fully stable at an alkaline pH range of 7-11 at 50 °C with a half-life of 90 min. Metal ions such as K⁺, Ca²⁺ and Mg²⁺ could enhance the enzyme activity. Therefore, this protease indicates a high potential for the applications in the food and feed industry, as well as the waste management since it can hydrolyse protein at high alkaline pH and salt concentrations. The amino acid profiles of the purified KB-SP determined by LC-MS/MS analysis showed high score matching with the peptidase S8 of B. licheniformis LMG 17339, corresponding to the mature domain of a minor extracellular protease (Vpr). Amino acid sequence alignment and 3D structure modelling of KB-SP showed a conserved catalytic domain, a protease-associated (PA) domain and a C-terminal domain.

Novelty and scientific contribution: A novel extracellular haloprotease from B. licheniformis was purified, characterized and identified. The purified protease was identified as being a minor extracellular protease (Vpr) and this is the first report on the halotolerance of Vpr. This protease has the ability to work in harsh conditions, with a broad alkaline pH and salinity range. Therefore, it can be useful in various applications in industrial fields.

Research background: Gallic acid is a polyphenol with antioxidant and antitumor activities; however, its use as a nutraceutical or drug is hindered by its low bioavailability. Zein is a natural protein found in corn and has been applied as nanoparticle drug carrier. In this study, zein nanoparticles were obtained and stabilized with polyethylene glycol (PEG) as gallic acid carriers.

Experimental approach: Nanoparticles were obtained by the liquid-liquid method and characterized in terms of mean size, polydispersity index, zeta potential, morphology, solid-state interactions and encapsulation efficiency/drug loading. The stability of nanoparticles was evaluated in simulated gastrointestinal fluids and food simulants, and the antioxidant activity was determined by the scavenging of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical.

Results and conclusions: Zein nanoparticles containing gallic acid were obtained and stabilized only in the presence of PEG. Under optimal conditions, nanoparticles with mean size <200 nm, low polydispersity index (<0.25) and negative zeta potential (-20 mV) were obtained. The gallic acid encapsulation efficiency was about 40%, loading about 5%, and it was encapsulated in an amorphous state. Fourier transform infrared spectroscopy (FTIR) did not identify chemical interactions after gallic acid nanoencapsulation. Zein nanoparticles were more prone to release the gallic acid in gastric than intestinal simulated medium; however, more than 50% of drug content was protected from premature release. In food simulants, the gallic acid release from nanoparticles was prolonged and sustained. Moreover, the nanoencapsulation did not reduce the antioxidant activity of gallic acid.

Novelty and scientific contribution: The results show the importance of PEG in the formation and its effect on the properties of zein nanoparticles obtained by the liquid-liquid dispersion method. This study indicates that PEG-stabilized zein nanoparticles are potential carriers for oral intake of gallic acid, preserving its antioxidant properties and enabling its use in the pharmaceutical and food industries.

Research background: Potato tissue is damaged during fresh-cut production, which makes fresh-cut potato susceptible to the quality loss and microbiological spoilage. At the same time, such products are desirable due to their convenience; however, they are extremely sensitive and have short shelf life. The main challenge of the fresh-cut potato industry is to find possibilities to overcome these drawbacks. UV-C treatment, known for its antibacterial activity, is a promising technique and it shows a potential to improve shelf life of fresh-cut potato products.

Experimental approach: The influence of the UV-C treatment on the safety and quality, as well as sensory traits of fresh-cut potato (Solanum tuberosum L. cv. Birgit) during storage was examined. For this purpose, 0-, 3-, 5- and 10-min UV-C irradiation was applied on vacuum-packed potato slices pretreated with sodium ascorbate solution. During 23 days of storage at (6±1) °C, microbiological, physicochemical and sensory properties of raw samples were monitored, along with sensory properties of boiled and fried fresh-cut potatoes.

Results and conclusions: The 5- and 10-min UV-C treatments significantly reduced microbial growth, increased total solids and lightness (L*), and positively affected odour and firmness of raw potatoes. Cooked UV-C-treated samples were described with more pronounced characteristic potato odour and taste. Overall, UV-C-treated fresh-cut potato retained its good quality and sensory traits up to 15 days at (6±1) °C.

Novelty and scientific contribution: To the best of our knowledge, this is the first scientific article dealing with the effect of UV-C light on durability (safety, quality and sensory traits) of fresh-cut potato cv. Birgit and its suitability for boiling and frying. In general, UV-C treatment is a known antimicrobial technique, but its application on fresh-cut potato is poorly explored. Results confirmed that vacuum-packed fresh-cut potato treated only with UV-C and sodium ascorbate as anti-browning agent, without the addition of chemical preservatives, had twofold longer shelf-life at (6±1) °C than the fresh-cut potato not treated with UV-C. Fresh-cut potato treated with UV-C retained good overall quality and sensory properties either raw, boiled or fried. Results of this study could also be useful for producers in terms of potential UV-C application as a strategy for prolonging the shelf-life of fresh-cut potato.

Research background: Acquisition of migratory potential is pivotal for cancer cells, enabling invasion and metastasis of colorectal carcinoma. Royal jelly and its bioactive component trans-10-hydroxy-2-decenoic acid (10H2DA) showed remarkable antimetastatic potential, but the molecular mechanism underlying this activity is unclear.

Experimental approach: Identification and quantification of 10H2DA in royal jelly originating from Serbia was done by HPLC method. Cytotoxicity of 10H2DA was measured by tetrazolium dye MTT test in concentration range 1-500 μg/mL after 24 and 72 h. Its effect on the collective and single-cell migration was measured by wound healing and transwell migration assays. Invasive potential of cancer cells was evaluated by a transwell method modified with collagen. Immunofluorescence was used for migratory and invasive protein expression, while the gene expression of these markers was evaluated by quantitative real time polymerase chain reaction (qRT-PCR). All assays were applied on human colorectal carcinoma HCT-116 and SW-480 cell lines and, except for MTT, evaluated after 24 h of treatment with two selected concentrations of royal jelly and 10H2DA.

Results and conclusions: According to HPLC, the mass fraction of 10H2DA in royal jelly was 0.92% (m/m). Treatment with 10H2DA showed no cytotoxic effect; however, significant inhibitory potential of royal jelly and 10H2DA on the motility and invasiveness of colorectal cancer cells was observed. More pronounced effect was exerted by 10H2DA, which significantly suppressed collective cell migration and invasiveness of SW-480 cells, as well as single- and collective cell migration and invasive potential of HCT-116 cell line. Treatments increased epithelial markers E-cadherin and cytoplasmic β-catenin in HCT-116 cells, thus stabilizing intercellular connections. In SW-480 cells, 10H2DA increased E-cadherin on protein and gene level, and suppressed epithelial-mesenchymal transition (EMT) markers. In both cell lines, treatments induced significant suppression of promigratory/proinvasive markers: N-cadherin, vimentin and Snail on protein and gene level, which explains decreased migratory and invasive potential of HCT-116 and SW-480 cells.

Novelty and scientific contribution: Our study presents new findings and elucidation of royal jelly and 10H2DA molecular mechanism that underlies their antimigratory/antiinvasive activity on colorectal cancer cells. These findings are shown for the first time indicating that these natural products are a valuable source of anticancer potential and should be reconsidered for further antitumour therapy.

Research background: Baranjski kulen is one of the most popular fermented meat sausages originating from Croatia. It has protected geographical indication, and is traditionally produced in the Baranja region of Croatia. Kulenova seka is a fermented sausage very similar to baranjski kulen, but it has a different calibre and consequently, a shorter time of production. In recent decades, due to the high demand and popularity of these products, industrially produced baranjski kulen and kulenova seka have become available on the market. This work aims to identify specific characteristics of traditional and industrial sausages baranjski kulen and kulenova seka on proteome, peptidome and metabolome level, which could potentially lead to better optimization of the industrial production process in order to obtain an equivalent to the traditional product.

Experimental approach: Protein profiles of baranjski kulen and kulenova seka (traditional and industrial) were analysed using two-dimensional gel electrophoresis followed by differential display analysis and protein identification by mass spectrometry. Peptidomic profile was analysed via liquid chromatography-tandem mass spectrometry. Furthermore, aroma profiles were investigated via headspace solid phase microextraction and gas chromatography-mass spectrometry.

Results and conclusions: The major identified characteristics of each product were: industrial baranjski kulen: specific degradation of myosin-1 and titin, overabundance of stress-related proteins and increased phenylalanine degradation; traditional baranjski kulen: decreased concentration of phenylalanine and overabundance of fructose-bisphosphate aldolase A and carbonic anhydrase 3; industrial kulenova seka - specific myosin-4 and haemoglobin subunit alpha degradation process; traditional kulenova seka - overabundance of dihydropyrimidine dehydrogenase [NADP(+)] and myosin light chain 1/3, skeletal muscle isoform, degradation of albumin and myoglobin, decreased concentrations of almost all free amino acids and increased amounts of smoke-derived volatile compounds. Presented results showed that potential product type-specific quality markers for each sausage could be found.

Novelty and scientific contribution: In this preliminary communication, the first insights into protein degradation processes and generation of peptides, free amino acids and aroma compounds of industrial and traditional baranjski kulen and kulenova seka are presented. Although further research is needed to draw general conclusions, the specific profile of proteins, peptides, amino acids and volatile compounds represents the first step in the industrial production of sausages that meet the characteristics of traditional flavour.