Isolation of an acetyl-CoA synthetase gene (ZbACS2) from Zygosaccharomyces bailii

A gene homologous to Saccharomyces cerevisiae ACS genes, coding for acetyl-CoA synthetase, has been cloned from the yeast Zygosaccharomyces bailii ISA 1307, by using reverse genetic approaches. A probe obtained by PCR amplification from Z. bailii DNA, using primers derived from two conserved regions of yeast ACS proteins, RIGAIHSVVF (ScAcs1p; 210-219) and RVDDVVNVSG (ScAcs1p; 574-583), was used for screening a Z. bailii genomic library. Nine clones with partially overlapping inserts were isolated. The sequenced DNA fragment contains a complete ORF of 2027 bp (ZbACS2) and the deduced polypeptide shares significant homologies with the products of ACS2 genes from S. cerevisiae and Kluyveromyces lactis (81% and 82% identity and 84% and 89% similarity, respectively). Phylogenetic analysis shows that the sequence of Zbacs2 is more closely related to the sequences from Acs2 than to those from Acs1 proteins. Moreover, this analysis revealed that the gene duplication producing Acs1 and Acs2 proteins has occurred in the common ancestor of S. cerevisiae, K. lactis, Candida albicans, C. glabrata and Debaryomyces hansenii lineages. Additionally, the cloned gene allowed growth of S. cerevisiae Scacs2 null mutant, in medium containing glucose as the only carbon and energy source, indicating that it encodes a functional acetyl-CoA synthetase. Also, S. cerevisiae cells expressing ZbACS2 have a shorter lag time, in medium containing glucose (2%, w/v) plus acetic acid (0.1-0.35%, v/v). No differences in cell response to acetic acid stress were detected both by specific growth and death rates. The mode of regulation of ZbACS2 appears to be different from ScACS2 and KlACS2, being subject to repression by a glucose pulse in acetic acid-grown cells.     

Factors affecting the oxidative activity of laccase towards biphenyl derivatives in homogeneous aqueous-organic systems

Catalytic oxidation of biphenyl derivatives was investigated using laccase in a homogeneous aqueous-organic system. A thermostable laccase from Trametes sp. showed the highest catalytic activity for the oxidation of 4-hydroxybiphenyl (4-HB) at a reaction temperature of 60 degrees C when dimethylsulfoxide (DMSO) was employed as a co-solvent. Furthermore, the catalytic performance was successfully enhanced by the incorporation of a laccase mediator system (LMS) into the aqueous-DMSO media. The catalytic performance strongly depended on the type of mediator, and the highest activity was observed with 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as mediator, suggesting the importance of the selection of a suitable mediator. It was verified that this mediator system is applicable to the oxidation of several biphenyl derivatives with hydroxyl groups.     

Enhanced selenium content in buckwheat (<Emphasis Type="Italic">Fagopyrum esculentum</Emphasis> Moench) and pumpkin (<Emphasis Type="Italic">Cucurbita pepo</Emphasis> L.) seeds by foliar fertilisation

The fruit and thin-husked seeds of the pumpkin (Cucurbita pepo L.) and buckwheat grain (Fagopyrum esculentum Moench), both grown in Slovenia, were analysed for selenium (Se) content following foliar application of Se(VI) solution during the period of blooming. Samples were digested by a H₂SO₄-HNO₃-H₂O₂-V₂O₅ mixture and Se determined, based on hydride generation atomic fluorescence spectrometry. The whole procedure from weighing to measuring was carried out in the same Teflon vessel. The detection limit of the method was 0.14 ng g^–1 solution. Buckwheat seeds from untreated plants contained 47 ng g^–1 of Se and 394 ng g^–1 from plants after foliar fertilisation with Se. Pumpkin seeds from untreated plants contained 108 ng g^–1 of Se, and 381 ng g^–1 of Se from Se-treated plants, all per lyophilised sample. Se content in lyophilised pumpkin fruit was 15 ng g^–1 in untreated plants and 20 ng g^–1 in Se-treated pumpkin plants. It is thus feasible to enhance Se content in buckwheat and pumpkin seeds by foliar fertilisation, making them a rich source of dietary Se and useful as a raw material for enriched food products.     

Preparation and Characterization of Nanocomposites from Whey Protein Concentrate Activated with Lycopene

The production and characterization of nanocomposites based on whey protein concentrate (WPC) and montmorilonite (MMT) incorporated with lycopene as a functional substance is presented and discussed as an alternative biomaterial for potential uses in foodstuff applications. A full factorial design with varying levels of MMT (0% and 2% in w/w) and lycopene (0%, 6%, and 12% in w/w) was used. Color, light transmission, film transparency, moisture, density, solubility, water vapor permeability, and antioxidant activity of the resulting materials were evaluated. Results indicated that lycopene and MMT nanoparticles were successfully included in WPC films using the casting/evaporation method. Inclusion of 2% w/w of MMT in the polymeric matrix significantly improved barrier property against water vapor. Lycopene, besides its good red coloring ability, provided to the films antioxidant activity and UV‐vis light protection. These findings open a new perspective for the use of materials for bioactive packaging applications.     

Effect of steaming,freezing, and re-steaming on the texture properties of non-glutinous rice cakes

The effects of steaming time (6, 8, and 10 min), freezing storage period, and re-steaming for thawing on the textural properties of non-glutinous rice cakes (baekseolgi) were investigated. As the steaming time increased, the rice cakes softened. In particular, the sample that was steamed for 10 min showed lower hardness than those steamed for shorter periods. A short period of steaming was insufficient for water bound to the surface of the starch granules to penetrate the granules in the dough. During the re-steaming process of the frozen non-glutinous rice cake samples, the retrogradation of starch and water syneresis contributed to the increased hardness of non-glutinous rice cakes.     

Development of a long‐life vacuum‐packaged ready‐to‐eat meat product based on a traditional Portuguese seasoned meat

Carne do alguidar is a Portuguese traditional pork fried meat, usually manufactured for self‐consumption purposes. This study developed a ready‐to‐eat (RTE) meat product, to meet today's consumers’ convenience, manufactured at the industrial scale evaluating its quality and shelf life, assessing the effect of vacuum packaging and the use of an antioxidant (50 ppm BHT) to enhance oxidative stability. Physicochemical and microbiological parameters were assessed, and a sensory analysis was performed. Interestingly, no significant differences were recorded between control (non‐BHT) and antioxidant (BHT) samples. Microbiological counts remained at low levels throughout the storage period, ensuring the product's required microbiological quality. At later storage stages, higher values of thiobarbituric acid‐reactive substances arose and off flavours and aromas were perceived. Still, overall appreciation was not affected until 12 months of storage and a significant depreciation was perceived only after 15 months. Fibrousness and rising of off flavours were negatively correlated with overall appreciation.     

Effect of Dietary Red Meat on Colorectal Cancer Risk—A Review

Heme iron overload has been implicated as the main cause of the increased risk of cancer due to the consumption of red meat. However, fish and shellfish, teas, and spices contain up to five times more iron than red meat. There is insufficient evidence that iron intake in dietary red meat is the primary causal factor for colorectal cancer. In addition, harmful substances produced during the preparation of red meat, including heterocyclic amines (HCAs), polycyclic aromatic hydrocarbons (PAHs), N‐nitroso compounds, and acrylamide, are extrinsic factors that increase carcinogenicity. HCAs are produced during the cooking of red meat, poultry meat, and fish. PAHs may also be produced during the cooking of diverse food groups, such as dairy products, fruits, vegetables, and cereals. The average daily intake of red meat among Korean individuals is 62 g; the amount of PAHs entering the body via red meat is less than the average amount of PAHs the body is exposed to in the air. Therefore, it is difficult to conclude that dietary red meat is the main cause of colorectal cancer. Rather, there may be an intricate influence of multiple factors, including fruit and vegetable intake, alcohol consumption, smoking, overweight, obesity, and stress.     

Food processing increases casein resistance to simulated infant digestion

The objective of this study was to determine whether processing could modify the resistance of casein (CN) to digestion in infants. A range of different dairy matrices was manufactured from raw milk in a pilot plant and subjected to in vitro digestion using an infant gut model. Digestion products were identified using MS and immunochemical techniques. Results obtained showed that CNs were able to resist digestion, particularly κ‐ and αs₂‐CN. Resistant areas were identified and corresponded to fragments hydrophobic at pH 3.0 (gastric conditions) and/or carrying post‐translational modifications (phosphorylation and glycosylation). Milk processing led to differences in peptide patterns and heat treatment of milk tended to increase the number of peptides found in digested samples. This highlights the likely impact of milk processing on the allergenic potential of CNs.     

Development and validation of an HPLC method with fluorescence detection for the determination of fluorescent whitening agents migrating from plastic beverage cups

An HPLC method with fluorescence detection has been developed and validated for the quantification of six fluorescent whitening agents (FWA) in plastic beverage cups after extraction and in food simulants after migration at 70°C for 2 h. The sensitivity of the method was high with LODs ranging from 0.053 to 0.251 μg kg^?1 and LOQs from 0.107 to 0.504 μg kg^?1. Accuracy and precision were highly acceptable, with recoveries greater than 82% and RSDs (%) below 16%. The expanded combined uncertainty was found to be less than 23% for the measurements of all studied FWAs. In extracting the analytes from food contact materials (FCM), accelerated solvent extraction (ASE) and Soxhlet extraction were applied using ethanol as the extraction solvent. The results obtained for FWA in 10 different food plastic cups, made from different polymers, were compared. The ASE technique proved to be faster, more effective and efficient than Soxhlet extraction. Migration tests with official food simulants from Regulation (EU) No 10/2011 showed that the substances studied could potentially migrate using the selected migration conditions. The most pronounced effect was observed in case of simulant D1 (50% w/v ethanol in water). The analytical method proved to be a simple, fast, sensitive and reliable tool for the simultaneous quantification of six of the most used FWAs in both FCM extracts and food simulants after migration experiments.