Browsing by discipline "Elintarviketeknologia (viljateknologia)"

The literature review dealt with celiac-toxic Triticeae prolamins and their enzymatic degradation. Also the immunochemical methods for prolamin analysis were introduced. The gluten-derived immunogenic peptides are proline-rich and thereby remarkably resistant to proteolytic degradation. Most of the triggering prolamins can, however, be degraded by combining endogenous cereal enzyme activity with acidic incubation. Despite of this residual prolamins still exist and their concentration exceeds the threshold considered to be safe for gluten intolerants. The objective of the experimental work was to further hydrolyse the residual prolamins present in malt autolysates of wheat, barley and rye, with a food grade proline endopeptidase from Aspergillus niger (AN-PEP). Size-exclusion chromatography (SEC), free amino nitrogen (FAN) and SDS-PAGE analysis determined the extent of protein hydrolysis. Actual prolamin degradation was observed with immunological methods. Hydrolysis of residual prolamins was extensive in all malt systems – more than 96% of the prolamins were hydrolysed. The SEC and FAN data revealed that continuation of the hydrolysis overnight converted the polypeptides into smaller hydrolysis products. According to enzyme-linked immunosorbent assay analyses, 22 h incubation decreased the prolamin contents of wheat and rye malt hydrolysates below the level of 100 mg/kg. This level was achieved with AN-PEP concentration of 35 ?L/g in relation to freeze-dried autolysate. According to the Codex Alimentarius, food products containing gluten up to 100 mg/kg can be labelled 'very low gluten' and thus included in coeliac diet. AN-PEP treated rye malt ingredient could especially be a promising low-gluten ingredient to enhance the flavour of often poor-quality gluten-free bread. Before commercial applications can be devised the potential as a flavouring agent as well as the clinical safety of the product must be evaluated.

Most barley is used for animal feed or malting. However, barley contains technologically and nutritionally valuable components. The compositions of different barley cultivars as well as the factors affecting the concentrations of different nutrients (beta-glucan, protein, starch and ash) were surveyed in the literature part of this Master’s thesis. Special attention was paid to beta-glucan as it has attained significant interest in the food industry due to its positive health effects. In addition, the effect of grain composition on the pearling, milling and air classification properties of barley were surveyed. The nutrient composition and milling properties of ten different barley varieties were examined in the experimental part of the thesis. The varieties were divided into four subgroups based on their potential end usage: 1) speciality barley, 2) feed barley, and 3) starch barley, and 4) malting barley. The milling properties were analysed by sieve analysis and volumetric particle size distribution from the whole grain barley flours. In addition, coarse fractions were separated from the whole grain flour by air classification. These fractions were also analysed by the abovementioned particle analyses. Beta-glucan, protein and ash concentrations were usually higher in the speciality barley varieties and their coarse fractions. Starch concentrations were lower in these varieties and fractions. Feed and starch barleys had somewhat higher beta-glucan concentration compared to malting barleys, which respectively had the highest starch concentrations. Zero to 25 % of the grain’s outer layers was removed by pearling. Compositional analyses revealed that pearling decreased the amount of ash and increased the concentration of starch and betaglucan until about 15 % of the grain was pearled off. Starch and beta-glucan concentrations did not change significantly after this pearling level. Pearled barley flour was manufactured by pearling off 15–20 % of the grain’s outer layers, and milling the remaining pearled grains with a fine impact mill. Flour was then air classified to give fine and coarse fractions. The coarse fractions contained enriched concentrations of beta-glucan, protein and ash. However, their concentrations and yields were dependent on the speed of the classifier wheel. By contrast, starch was enriched in the fine fractions. The highest beta-glucan concentration was obtained with the beta-glucan-rich speciality variety D, which initially had 9.4 % beta-glucan. The beta-glucan concentration was enriched up to 11.4 % by pearling. Air classification of whole grain flour resulted maximally in 13.5 % and air classification of pearled grain flour in 15.5 % beta-glucan concentration. Based on the results, the beta-glucan concentration of the raw material seems to play the most important role in the enrichment. However, proper milling technology and air classifier settings are of utmost importance.

The literature review focused on the composition of the barley hordeins and the known extraction methods. The metal-catalysed oxidation of proteins and amino acids was also reviewed. The aim of the experimental part was to develop a simple selective extraction method for the B and C hordeins and to study how metal-catalysed oxidation affects these hordeins. Hulles barley cf. Jorma was selected as test material since milling of this cultivar was simple with a Brabender Junior mill. From the milled barley, water and salt-soluble material were removed and the rest was freeze-dried. The freeze-dried sample flour was studied by gel separation, precipitation and extraction with aqueous alcohols. The aqueous alcohol extracted the C hordeins completely although there was some B hordeins present. Two-dimensional electrophoresis showed that the isoelectric point of C hordeins was between the pH 5–7. Aqueous alcohols, extraction temperatures and pH were studied for hordein extraction. None of the studied methods improved the extraction of B and C hordeins. The hordein sample used in further experiments was extracted with 55% 2-propanol at 50 ºC. The metal-catalysed oxidation of the extracted hordeins was studied by using copper or iron as a catalyst and hydrogen peroxide or ascorbic acid as an oxidant. The reactions were analysed with SDS-PAGE and SE-HPLC. The results showed that the most effective reaction was with copper and hydrogen peroxide where the B and C hordeins were degraded efficiently after 24 hours of incubation. The results from SE-HPLC showed aggregated B hordeins in the extracted sample, which were partly degraded after two hours of incubation with hydrogen peroxide and copper. The results of this study indicated that the biggest groups of hordeins, the B and C hordeins, cannot be selectively separated with extraction. The barley hordeins efficiently degraded in the metal-catalysed oxidation with hydrogen peroxide and copper.

The aim of the literature review was to research barley proteins, metal-catalyzed oxidation and subjects related to it, like antioxidants and oxidation reactions in beer. In addition ACE inhibition was looked into. The object of the experimental part was to find out if the proteins of a barley-based industrial side product can be modified by metal-catalyzed oxidation or enzymatic hydrolysis, and how these treatments affect the different proteins in the sample material. In addition, the possible ACE inhibition activity of the reaction products was determined. The sample material was a protein-rich side-product of barley starch production. Two protein fractions were extracted from the material; an alcohol soluble fraction and a reduced fraction. The modification of the proteins in the sample fractions by oxidation and hydrolysis was determined with gel electrophoresis and size exclusion chromatography. The ACE inhibitory activity of the small peptides from these reactions was determined with UV-VIS spectroscopy. Three protein groups were identified from the sample material; polymeric B hordein, monomeric B hordein and C hordein. Contrary to expectations metal-catalyzed oxidation did not break down any of the proteins in the sample; instead it aggregated the proteins into bigger units. The enzyme treatment hydrolyzed the proteins effectively. Small peptides from the enzyme hydrolysis had an ACE inhibition IC50 of 246 µg/ml, which is similar to gluten hydrolysates IC50 of 29 µg/ml. IC50 is the inhibitor concentration where 50% of enzyme activity is inhibited. Instead of breaking down the subject proteins metal-catalyzed oxidation aggregated them, and thus it could not be used to make ACE inhibitory peptides. Enzyme hydrolysis was found to be a valid method of inhibitor peptide production. The peptides produced had an ACE inhibition capacity similar to previously known ACE inhibitory food hydrolysates.