Barbara Mickowska

Quality of gluten-free supplemented cakes and biscuits

Gluten-free confectionery products were used as controls for comparison with the products, which included different supplements such as linseed meal, amaranth and/or buckwheat. The latter were expected to increase nutritional values of confectionery products. Cookies were analyzed in terms of volume, selected textural parameters (hardness, cohesiveness), organoleptic quality, shelf-life, and different chemical components. All supplemented gluten-free products received high consumer scores, exceeding in some cases those of control samples. Supplementation of gluten-free confectionery products with linseed meal, amaranth and/or buckwheat flours enhanced their final nutritional quality. A significant rise was observed in the protein content and dietary fiber, and in the case of linseed meal also α-linolenic acid. All of the supplemented gluten-free confectionery products contained more macro-elements and micro-elements (i.e. potassium, phosphorus, magnesium, calcium, iron, manganese, zinc and copper), as compared with the controls. Taking into account the amino-acid composition, amaranth proved a more beneficial supplement of gluten-free products than linseed.

Effect of controlled lactic acid fermentation on selected bioactive and nutritional parameters of tempeh obtained from unhulled common bean (Phaseolus vulgaris) seeds

BACKGROUND Tempeh is a traditional Indonesian food of high nutritional quality obtained by fungal fermentation of dehulled, soaked and cooked legumes. The aim of this research was to study the effect of Lactobacillus plantarum DSM 20174 activity on selected parameters of tempeh made from unhulled seeds of common bean (Phaseolus vulgaris). Lactobacillus plantarum cells were applied during soaking of seeds (submerged fermentation) or during solid state fermentation with Rhizopus microsporus var. chinensis (co-cultivation). RESULTS Tempeh obtained from common beans contained 200 g kg⁻¹ protein of 34% in vitro bioavailability. Fungal fermentation caused decomposition of raffinose, stachyose and verbascose levels in seeds, on average by 93, 84 and 73% respectively. Enhanced antiradical (DPPH•, ABTS•+) capacity was accompanied by increased soluble phenol content. Application of Lactobacillus in the fermentation procedure increased tempeh protein and in vitro protein bioavailability by 18 and 17% respectively. Mixed culture tempeh contained lower levels of stachyose (25%), verbascose (64%) and condensed tannins (20%). Co-cultivation enhanced both DPPH•-scavenging activity and antioxidant capacity. CONCLUSION The application of Lactobacillus in most cases improved the nutritional parameters of tempeh from unhulled common beans. It may also be recommended to obtain products with diverse antioxidant properties as compared with fungal fermentation alone.

The influence of inoculum composition on selected bioactive and nutritional parameters of grass pea tempeh obtained by mixed-culture fermentation with Rhizopus oligosporus and Aspergillus oryzae strains

Tempeh is a popular Indonesian product obtained from legume seeds by solid-state fermentation with Rhizopus sp. The aim of this research was to study the effect of simultaneous mixed-culture fermentation of grass pea seeds on selected parameters of products as compared to traditional tempeh. The inoculum contained different ratios of Rhizopus oligosporus and Aspergillus oryzae spores. The simultaneous fermentation of grass pea seeds with inoculum consisting of 1.2 × 106 R. oligosporus and 0.6 × 106 A. oryzae spores (per 60 g of seeds) resulted in a product of improved quality, as compared with traditionally made tempeh (obtained after inoculation with 1.2 × 106 R. oligosporus spores), at the same fermentation time. This product had radical scavenging ability 70% higher than the one obtained with pure R. oligosporus culture and contained 2.23 g/kg dm of soluble phenols. The thiamin and riboflavin levels were above three (340 µg/g dm) and two (50.50 µg/g dm) folds higher than in traditionally made tempeh, respectively. The product had 65% in vitro bioavailability of proteins and 33% in vitro bioavailability of sugars. It also contained 40% less 3-N-oxalyl-L-2, 3-diaminopropionic acid (0.074 g/kg dm), as compared to traditional tempeh.

Mutants of the Saccharomyces cerevisiae VPS genes CCZ1 and YPT7 are blocked in different stages of sporulation

The CCZ1 gene is a member of the class B VPS (vacuolar protein sorting) genes and it is engaged in the last stage of delivery of multiple kinds of cargo to the yeast vacuole. In the process of fusion of the multivesicular body (MVB) with the vacuole, Ccz1p forms a complex with Ypt7p. Both genes are non-essential for vegetative growth, but their deletions cause a complete block in spore formation. The results of this study indicate that ccz1Δ cells initiate the meiotic program, properly proceed through premeiotic DNA replication and through the pairing of homologous chromosomes, but fail to progress through the first meiotic divisions and arrest in prophase I with a single nucleus. The mutant cells are defective in spindle formation as well as in duplication and/or separation of the SPBs. ypt7Δ cells, on the other hand, cannot execute DNA synthesis. We also show that expression of a mutated variant of the YPT7 gene suppresses the sporulation and autophagy defects of ccz1Δ cells to a quantitatively similar level, suggesting that restoration of autophagy in the ccz1Δ strain is sufficient to enable its sporulation.

Probiotic Lactobacillus acidophilus bacteria or synthetic TLR2 agonist boost the growth of chicken embryo intestinal organoids in cultures comprising epithelial cells and myofibroblasts

The intestinal epithelial cells reside in close proximity to myofibroblasts and microbiota, which are supposed to have an impact on intestinal stem cells fate and to influence processes of tissue maturation and regeneration. Mechanism underlying these phenomena and their diversity among vertebrates can be studied in 3D organoid cultures. We investigated the growth of chicken embryo intestinal epithelial organoids in Matrigel with and without Toll-like receptors (TLRs) stimulation. The organoid cultures contained also some myofibroblasts with potential to promote intestinal stem cell survival. Organoid cells, expressing TLR4, TLR2 type 1 and TLR2 type 2 were incubated with their agonists (lipopolysaccharide - LPS and Pam3CSK4) or co-cultured with Lactobacillus acidophilus bacteria (LA-5). Pam3CSK4 and LA-5 promoted organoid growth, which was demonstrated by comparing the morphological parameters (mean number and area of organoids). The profile of prostaglandins (PG), known to promote intestinal regeneration, in supernatants from organoid and fibroblast cultures were evaluated. Both PGE2 and PGD2 were detected. As compared to unstimulated controls, supernatants from the Pam3CSK4-stimulated organoids contained twice as much of PGE2 and PGD2. The changes in production of prostaglandins and the support of epithelial cell growth by myofibroblasts are factors potentially responsible for stimulatory effect of TLR2 activation.

Immunochemical evaluation of proteolysis of cereal proteins causing celiac disease by microbial proteases

ABSTRACT Celiac disease is one of the most prevalent food intolerances worldwide, with a gluten-free diet as the only effective therapy. Therefore, searching for new alternative approaches is a priority, and one of them is the idea of using proteinases for degradation of gluten proteins. The aim of the study was a trial of enzymatic cleavage of prolamins extracted from wheat, barley, rye, triticale and oat. The proteolysis was performed by microbial proteases from Bacillus stearothermophilus, Bacillus licheniformis, Bacillus thermoproteolyticus and Streptomyces griseus. The reaction was performed up to 60 min, stopped by addition of appropriate synthetic inhibitor and products of limited proteolysis were analyzed by SDS-PAGE. Most of the prolamins were susceptible to proteolysis by the examined microbial enzymes and as the result of hydrolysis the molecules of lower molecular weight and peptides occurred (<30,000 Da). The immunoreactivity of proteolysis products was evaluated using the reaction with polyclonal and monoclonal antibodies by Western blot and ELISA analysis, respectively. Immunochemical analyses confirmed that celiac-active sequences of amino acids in prolamins were at least partially destroyed and have lost their “toxicity”, so application of microbial proteases seems to be an alternative method for decreasing of celiac activity of prolamins.

The Effect of Germination on Antioxidant and Nutritional Parameters of Protein Isolates from Grass Pea (Lathyrus sativus) Seeds

The aim of this research was to study the antioxidant and nutritional (selected objects) properties of protein isolates obtained from grass pea seedlings as compared with soaked and raw seeds. Two percent extract of isolate from 5-day-old seedlings showed the highest total antioxidant activity (25%) and the ability to chelate Fe 2+ (2.35 mg/g d.m.) as compared with other isolates. Protein isolates from grass pea seeds had on average 89% total protein, 87% in vitro protein bioavailability, about 5574 TIU/g (d.m.) (trypsin inhibitors activity) and did not contain ODAP. Germination of seeds for 5 days considerably improved the in vitro bioavailability of isolates, by 12%, and profile of sulfur amino acids by 42%, in comparison with isolates obtained from the raw seeds. Isolates from 5-day-old grass pea seedlings had the best antioxidant properties and improved nutritional parameters (as compared with raw seeds), which makes them worthy of being considered as a potential food additive.

Determination of Biological and Sensory Profiles of Biscuits Enriched with Tea (Camellia sinensis L.) Powder

Abstract The aim of this work was to characterise the biological and sensory profile of biscuits enriched with green (1 and 3%) and black tea (1 and 3%) powders. Biscuits without the addition of tea were used as a control. Phenolic concentration, flavonoid concentration, and antioxidant activity were determined spectrophotometrically. Amino acid composition was determined using automatic amino acid analyser AAA 400 and crude fibre content using an Ancom analyser. Sensory profiles were evaluated by comparison of enriched and control biscuit samples. The enriched biscuits showed higher phenolic and flavonoid concentration and antioxidant activity estimated by DPPH and phospholybdenum method in comparison with levels in the control group. The best results for antioxidant activity estimated by DPPH and phosphomolybdenum methods were achieved in biscuits enriched with black tea powder (3%): 2.25 and of 32.64 mg TEAC·g−1, respectively. Total phenolic concentration was 1.16 mg GAE·g−1, and total flavonoid concentration was 0.13 mg QE·g−1. These biscuits had higher concentration of crude fibre in comparison with the control group and the highest concentration (0.64%) was found in biscuits with addition of 3% green tea powder. The amino acid composition in samples, including in the control sample was balanced, with slightly higher concentration of threonine, serine, and methionine in enriched samples, but this parameter was not statistically significant. Biscuits enriched with green and black tea had higher sensory scores for taste, smell and aftertaste.

APPLICATION OF WESTERN BLOT ANALYSIS FOR DETECTION OF PROLAMIN PROTEINS IN CEREAL GRAINS AND BREAD

Celiac disease is an inflammatory condition of the small intestine in genetically susceptible individuals caused by ingestion of wheat gluten and corresponding proteins from barley and rye. Cereal storage proteins (prolamins) are responsible for immunological response of patients with celiac disease. Prolamins are alcohol soluble fractions, namely gliadins (wheat), hordeins (barley) and secalins (rye). The main triggering factor is wheat fraction with low molecular weight (20-30 kDa) called α-gliadins. Immunochemical detection of celiac active proteins is based on reactivity of gluten-detecting antibodies with prolamins extracted from cereals. In our study, we used Western blot analysis for detection of prolamin complex in cereal grains and processed foods (breads). Western blot was carried out by polyclonal antibody raised against wheat gluten. Reaction was positive for all kind of cereal grains. The samples of wheat and spelt wheat show much more positive affinity to antibody than rye and oat. As well as for cereal grains, all samples of bread showed positive immunological reaction with used antibody. Western blot analysis with gluten polyclonal antibody is suitable method for qualitative detection of prolamin complex in cereal grains and processed foods.