Impact of gluten-friendly™ technology on wheat kernel endosperm and gluten protein structure in seeds by light and electron microscopy.

The main aim of this paper was to assess the impact of Gluten-Friendly™ (GF) technology (Italian priority patent n° 102015000084813 filed on 17th December 2015) on wheat kernel endosperm morphology and gluten protein structure, using SEM, light and immunofluorescent microscopy. Microscopy was combined with immunodetection with specific antibodies for gliadins, γ-gliadins, LMW subunits and antigenic epitopes to gain a better understanding of the technology at a molecular level. The results showed significant changes to gluten proteins after GF treatment; cross-reactivity towards the antibodies recognizing almost the entire range of gluten proteins as well as the antigenic epitopes through the sequences QQSF, QQSY, PEQPFPQGC and QQPFP was significantly reduced. The present study confirms the results from our previous work and shows, for the first time, the mechanism by which a chemical-physical treatment abolishes the antigenic capacity of gluten.

Polymeric protein formation during pasta-making with barley and semolina mixtures, and prediction of its effect on cooking behaviour and acceptability.

Spaghetti were produced in a pilot plant from semolina and semolina blended with increasing amounts of barley flour. According to size exclusion high-performance liquid chromatography (SE-HPLC), barley proteins interact with semolina proteins during pasta making, forming polymers of high molecular weight. Of these, the unextractable polymeric proteins (UPP) were at significantly higher concentrations than in spaghetti made from semolina. The decrease of both S-S bonds and -SH free groups in barley semolina spaghetti, with respect to that made of semolina, suggested that polymerisation among the different classes of proteins involves a new bonding arrangement. Due to ß-glucan hydrophilicity and competition with starch for water, the replacement of increasing amounts of semolina with barley flour was able to increase the optimal cooking time. The sensory properties of composite spaghetti were judged as better than the control because of the higher firmness and the lower bulkiness and stickiness of the former.

Application of capillary electrophoresis to determine the technological properties of wheat flours by a glutenin index.

Capillary electrophoresis was used to characterize glutenin proteins from ancient varieties of Southern Italy common wheat and to determine the technological properties of wheat flours based on a glutenin index. Three zones were identified in the electropherograms, indicated as A, B, and C according to electroelution order. The three zones corresponded to the low molecular weight glutenin subunits and to the y- and x-type high molecular weight subunits, respectively. The ratio B/C was correlated to the alveographic parameter P/L. These results indicated that flours resulting in a B/C ratio lower than 2 produced elastic doughs whereas flours resulting in a B/C ratio higher than 2 produced doughs more resistant to extension. This study showed that capillary electrophoresis is useful for determining the types and quantities of gluten proteins in the evaluation of wheat-flour technological properties of a limited number of noncommercial varieties as evidenced by the x-type content which seems to strongly influence the flour technological parameters.

Changes in phenolic content and antioxidant activity of Italian extra-virgin olive oils during storage.

Phenolic composition and antioxidant activity of extra-virgin olive oils extracted from several Italian varieties were studied at production and during storage. The antioxidant activity was measured according to the following tests: in the aqueous phase, by radical scavenging of the 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation; and in the lipid phase, using the beta-carotene bleaching method. The phenolic content was not correlated to the oxidation indices (peroxide value and spectrophotometric constants). The phenolic contents and profiles of the various cultivars showed remarkable differences. The phenolic content was strongly correlated with the antioxidant activity measured according to the beta-carotene test and weakly correlated with the radical scavenging ability.

Use of natural compounds to improve the microbial stability of Amaranth-based homemade fresh pasta.

A study on the use of natural antimicrobial compounds to improve the microbiological stability of refrigerated amaranth-based homemade fresh pasta is presented in this work. In particular, the antimicrobial activity of thymol, lemon extract, chitosan and grapefruit seed extract (GFSE) has been tested against mesophilic and psychrotrophic bacteria, total coliforms, Staphylococcus spp., yeasts and moulds. A sensory analysis on both fresh and cooked pasta was also run. Results suggest that chitosan and GFSE strongly increase the microbial acceptability limit of the investigated spoilage microorganisms, being the former the most effective. Thymol efficiently reduces the growth of mesophilic bacteria, psychrotrophic bacteria and Staphylococcus spp., whereas it does not affect, substantially, the growth cycle of total coliforms. Lemon extract is the less effective in preventing microbial growth. In fact, it is able to delay only total mesophilic and psychrotrophic bacterial evolution. From a sensorial point of view no significant differences were recorded between the control samples and all the types of loaded amaranth-based pasta.

Endosperm-specific activity of a storage protein gene promoter in transgenic wheat seed.

The characterization of the promoter of a wheat (Triticum aestivum) cv. Cheyenne high molecular weight glutenin subunit (HMW subunit) gene, Glu-1D-1 is reported. The nucleotide sequence of the promoter from position -1191 to -650 with respect to the transcription start site was determined, to add to that already determined. Analysis of this region of the promoter revealed the presence of an additional copy of part of the primary enhancer sequence and sequences related to regulatory elements present in other wheat seed protein genes. A chimaeric gene was constructed comprising the 5' flanking region of the Glu-1D-1 gene from position -1191 to +58, the coding region of the UID:A (Gus) gene, and the nopaline synthase (Nos) gene terminator. This chimaeric gene was introduced into wheat (Triticum durum cv. Ofanto) by particle bombardment of inflorescence explants. Two independent transgenic lines were produced, and both showed expression of the Gus gene specifically in the endosperm during mid-development (first detected 10-12 d after anthesis). Histochemical analysis of homozygous T(2) seed confirmed this pattern of expression, and showed that expression was initiated first in the central lobes of the starchy endosperm, and then spread throughout the endosperm tissue, while no expression was detected in the aleurone layer. Native HMW subunit protein was detectable by Western analysis 12-14 d after anthesis, consistent with concurrent onset of activity of the native and introduced HMW subunit gene promoters.