The overarching goal of this research was to investigate the effect of cultivar and environment on the physicochemical and functional properties of pea protein isolates using a structure-function approach. Six pea cultivars (Agassiz, CDC Golden, CDC Dakota, CDC Striker, CDC Tetris, Cooper) were collected from two years (2011, 2012) over two locations in Saskatchewan (Saskatoon and Rosthern) from two field replicates. Pea protein isolates were prepared from defatted flours by alkaline extraction (pH 9.0) followed by isoelectric precipitation (pH 4.5), and then neutralized to pH 7.0 prior to freeze-drying. Samples were evaluated for composition (amino acid profile, legumin/vicilin ratio), surface characteristics (zeta potential, surface hydrophobicity), and functional properties (nitrogen solubility, oil holding capacity, foaming capacity, foam stability, emulsion stability). In addition, samples were assessed for seed weight and colour, and compared against the functional characteristics of six commercially produced protein isolates (whey, wheat, egg, pea, and two soy ingredients). The extracted pea protein isolates had protein contents of ~91% (d.b.), as well as isolate and protein yields of ~18% and ~72%, respectively. Although cultivars exhibited a range of legumin/vicilin ratios from 0.36 (Agassiz) to 0.79 (CDC Golden), such differences were not reflected in their amino acid profiles. Differences amongst cultivars, as well as significant cultivar × environment interactions, were found for only surface hydrophobicity (195-267 a.u.), solubility (63-75%), and foaming capacity (167-244%). No differences in either cultivar or environment were observed in other surface (zeta potential = ~-24 mV) or functional (oil holding capacity = ~3.2 g/g; foam stability = ~75%; emulsion stability = ~96%) properties. All functional properties were significantly correlated with legumin/vicilin ratio and/or surface hydrophobicity. However, such relationships were weak (r = -0.19 to -0.20, and r = 0.17 to 0.32). The strongest correlation was observed between the legumin/vicilin ratio and surface hydrophobicity at r = 0.63 for the pea protein isolates. Meanwhile, zeta potential did not display a significant correlation to any property tested. In comparison to commercial protein isolates, the pea protein isolates behaved most similarly to soy except for solubility. Whey and egg were superior in solubility and the foaming properties, whereas wheat and the commercial pea protein product underperformed in almost all functionality tests. These findings suggest that while inherent protein material source may be important to functional behaviours, the method of extraction could pose even greater effects. This was observed between the laboratory- and commercially-prepared pea protein isolates, which at minimum differed in processing (defatting) and method of drying (freeze- vs. spray-dried). Coupled with the weak correlations between physicochemical and functional properties, findings overall indicate that method of protein isolate production play a more significant role in protein functional characteristics than cultivar, environment, or composition. Findings also suggest that secondary processors may not need to specify either cultivar or environment of their raw materials, thus creating advantages in their feedstock sourcing.

The overarching goal of this research was to investigate the differences in the physicochemical and functional properties of protein isolates produced from seven different faba bean genotypes (CDC Fatima, Taboar, SSNS-1, FB9-4, FB18-20, Snowbird and CDC Snowdrop) grown at different locations in Canada (Saskatchewan, Alberta and Manitoba) in 2011 and 2012. The protein isolates were prepared by alkaline extraction (pH 9.5) followed by isoelectric precipitation at pH 4.5. The isolates had an average protein content of ~94% and average protein and isolate yields of ~77% and ~25%, respectively. The physicochemical properties assessed in this study included surface charge/zeta potential (ZP), surface hydrophobicity (SH), and surface and interfacial tension (ST and IT). The functional properties tested included foaming capacity (FC) and foam stability (FS), emulsion capacity (EC) and creaming stability (CS), emulsion activity index (EAI) and emulsion stability index (ESI), oil holding capacity (OHC), and protein solubility. The findings indicated that all physicochemical properties for all isolates were independent of genotype. Overall, an average ZP of + 22.1 mV, SH of 47.2 arbitrary units, and ST and IT of 65.0 mN/m and 10.7 mN/m, respectively, were observed. However, with the exception of ZP considerable differences were observed due to the effect of environment. The ratio of the major globulin protein fractions [legumin:vicilin (L/V)] was found to shift during processing, from 3.8 (range: 3.4-4.6) in the flour to 4.5 (range 4.0-4.9) in the isolates. The L/V ratio for faba bean flour and isolate samples was also found to be independent of genotype. For all genotypes, with the exception of the zero-tannin varieties (Snowbird and CDC Snowdrop), the L/V ratio was affected by the environment. Similar to the physicochemical properties, all functional attributes were found to be independent of genotype. However, environmental effects were observed for all functional properties with the exception of EAI and ESI. Average values for FC of 162.0%, for FS of 65.0%, for EC of 184.0 g/g, for CS of 94.0%, for OHC of 5.7 g/g, for EAI of 13.0 m2/g, for ESI of 10.7 min and for solubility of 81.0% were reported. Zeta potential was observed to be positively correlated with CS (r = 0.46; p<0.05) and FS (r = 0.54; p<0.01), whereas SH and L/V ratio were not. The L/V ratio in the isolate, however, was correlated positively with SH (r = 0.40; p<0.05) and negatively with ZP (r = -0.39; p<0.05). Moreover, the solubility of faba bean isolates was found to be positively correlated with ZP (r = 0.44; p<0.05) and negatively correlated with both IT (r = -0.38; p<0.05) and OHC (r = -0.38; p<0.05). The functional properties of some commercial protein isolates (soy, pea, whey, egg and wheat) were evaluated for comparative purposes. The OHC of the faba bean isolate was found to be higher than that of any of the commercial isolates. With the exception of CS (soy and pea) and FC (egg), all of the emulsifying (EC, EAI and ESI) and foaming (FC and FS) properties of the faba bean protein isolates were comparable to those of soy, pea and egg isolates. In contrast, values for most of the other functional properties were greater for faba bean isolates than for the pea and wheat isolates, but lower than for the whey isolate. For example, the solubility of the protein isolates was observed to decrease in the following order: whey (89.0%) = egg (88.1%) > faba bean (81.0%) > soybean (30.5%) > pea (20.1%) > wheat (10.7%).

Dry bean utilization by the food industry can be increased by developing value-added processing applications. The goals of this research were to evaluate (1) the effect of milling method on the physical, chemical and functional properties of whole black bean flour and its fractions and (2) the effect of removing soluble phenolic compounds on the functional and rheological properties of black bean protein isolates. Black bean was milled with five laboratory mills [cyclone mill, hammer mill, stone mill (fine, medium, coarse), disc mill (fine, coarse), and centrifugal mill (10,000 or 12,000 rpm and 250, 500, 1000 μm aperture screen)] and the resulting flours were evaluated for their physical, chemical and flow properties of bulk samples and particle size fractions. Whole black bean flour and cotyledon flour were subjected to phenolic extraction and protein isolation, resulting in protein isolates with and without soluble phenolics. Solubility, wettability, dispersibility, water binding capacity, foam capacity and stability, emulsification capacity, and gelation properties of protein isolates were evaluated. Variation in milling method produced flours with significantly different flour characteristics. Geometric mean size of whole bean flour was negatively correlated with starch damage (r = -0.92), L* (r = -0.94), angle of repose (r = -0.94), and angle of slide (r = -0.80 to -0.90) and positively correlated with moisture (r = 0.72), and loose bulk density (r = 0.72). Milling method and particle size interaction was significant on characteristics of black bean flour fractions. Particle circularity of flour fractions had a negative correlation of r = -0.93, r = -0.81, r ≈ -0.95, and r = -0.94 with L*, angle of repose, angle of slide and compact density, respectively. Particle circularity had a positive correlation of r = 0.93 and r = 0.89 with average minimum particle size and loose bulk density, respectively. The removal of soluble phenolic compounds improved the brightness, solubility, wettability, dispersibility, foaming capacity, foaming stability, emulsion capacity, emulsion stability and gelling properties of protein isolates. These findings will help food manufacturers to process black bean ingredients using different mill settings to achieve different functionalities depending on the consumer requirements.

This diploma thesis deals with the use of various types of bran as a by-product in the milling of cereals. Mills create a huge amount of this material per year. The most common way of processing bran is mostly incineration and to a lesser extent it is used as feed for livestock. Depending on the type of cereal, bran contains 10-20% of protein, which disappears from the food chain due to combustion. Within the framework of sustainability and valorisation of waste, which has recently been largely discussed, great emphasis is placed on waste minimization whether in the field of its production or further processing. Due to the relatively high protein content, bran appears to be a suitable starting material to produce protein supplements. Proteins can be extracted from bran based on their different solubility at different pH. In the alkaline method, the proteins are first dissolved in an alkaline pH and then precipitated in an acidic medium. Lyophilization is followed by characterization of the extract in terms of yield, protein content, moisture, amino acid profile and digestibility. The highest yield was obtained with the oat bran isolate (13,5 ± 0,6 g of isolate per 100 g of bran). In terms of protein content, the best protein isolate was also obtained from oat bran (95,2 ± 0,4% protein in the isolate). Another determination was the analysis of the amino acid profile, in which a high content of arginine was found in all analyzed protein isolates from bran. Determination of digestibility showed very good digestibility of all produced protein extracts from bran.

Orientador: Flavia Maria Netto / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-07T16:15:22Z (GMT). No. of bitstreams: 1 Souza_AparecidaSoniade_D.pdf: 1116607 bytes, checksum: 498527d5fa8ce5e4faecf37aa8409d46 (MD5) Previous issue date: 2006 / Resumo: O presente trabalho teve como objetivo estudar os efeitos da irradiação e do tempo de armazenamento na composição química de grãos de soja e funcionais de seus isolados protéicos. Os grãos de soja foram divididos em quatro lotes, sendo que dois foram irradiados em fonte de 60Cobalto, com doses de 2,0 ou 5,0 kGy, um lote foi irradiado em acelerador de elétrons com dose de 2,0 kGy e um lote foi utilizado como controle (não irradiado). Os lotes foram armazenados por um período de 12 meses. A cada quatro meses, uma porção de cada lote foi retirada para produção laboratorial de farinhas desengorduradas (FDSs) e isolados protéicos (IPSs). A composição centesimal nos grãos, FDSs e IPSs foi determinada. Nos grãos foram analisados a atividade de água, teor de isoflavonas e atividade antioxidante (IA) e nas FDSs, os teores de fitato e inibidores de tripsina (IT). O teor de acidez foi avaliado nos óleos obtidos do desengorduramento das farinhas. As características estruturais das proteínas dos IPSs obtidos foram avaliadas por análises de eletroforese em gel de poliacrilamida na presença de dodecil sulfato de sódio (SDS-PAGE) em condições não redutoras, das proteínas solúveis em diferentes sistemas, calorimetria diferencial de varredura (CDV), grupos sulfidrila livres (SH), fluorescência intrínseca (FI), cromatografia liqüida de alta eficiência de fase reversa (CLAE-FR) e cromatografia liqüida de alta eficiência de exclusão molecular (CLAE-EM). As propriedades funcionais avaliadas foram: solubilidade, gelificação, capacidade de retenção de água (CRA), capacidade emulsificante (CE), índice de atividade emulsificante (IAE) e estabilidade de emulsão (EE). O teor das isoflavonas totais e o índice de antioxidação, analisados nos grãos, não diferiu significativamente (p>0,05) e permaneceu estável ao longo do armazenamento, independente do tipo ou dose de radiação. No entanto, houve aumento das concentrações relativas dos b-glicosídeos e diminuição dos malonil glicosídeos ao longo do armazenamento. O teor de acidez aumentou em até 81,6% no óleo de grãos irradiados com dose de 5 kGy e armazenados por doze meses. As FDSs de grãos irradiados apresentaram os menores valores dos inibidores de tripsina e fitato. As análises de calorimetria diferencial de varredura, fluorescência intrínseca e cromatografia liqüida de alta eficiência de exclusão molecular indicaram não ter ocorrido alterações estruturais importantes nos isolados protéicos devido à irradiação ou ao armazenamento. Entretanto, os grupos sulfidrila livres aumentaram nos IPSs de grãos irradiados, mas sofreram pouca alteração ao longo dos 12 meses de armazenamento. Houve aumento da proporção relativa de frações hidrofóbicas observadas por CLAE-FR em todos os isolados após quatro meses de armazenamento dos grãos. Os IPSs de grãos irradiados apresentaram maior solubilidade protéica e capacidade emulsificante, enquanto que a estabilidade de emulsão e o índice de atividade emulsificante não diferiram significativamente (p>0,05). Os valores de dureza e capacidade de retenção de água dos géis dos IPSs não diferiram significativamente (p>0,05) em função do tratamento empregado. No entanto, os géis dos IPSs de grãos armazenados tiveram maiores valores de dureza. Os valores coesividade e elasticidade dos géis dos isolados, independente da dose e tipo de radiação, não diferiram significativamente (p>0,05). Nas condições estudadas, as radiações gama com doses de até 5 kGy e de feixe de elétrons com dose de 2 kGy podem ser utilizadas sem prejuízo as propriedades químicas de grãos e funcionais de seus isolados protéicos, durante o armazenamento / Abstract: The aim of this work was to study the effects of both irradiation and storage on the chemical composition of soybean grains and on the physicochemical and functional properties of their protein isolates. The soy grains were divided into four lots, two of them were irradiated with of 60Cobalt source in doses of 2.0 or 5.0 kGy, a third one was irradiated with an electron beam at 2.0 kGy and another was used as control (non-irradiated). All the lots were stored for a maximum of 12 months. Every four months, a portion of each lot was removed to produce deffated soy flour (DFS) and soy protein isolate (SPI). The proximal compositions of the grains, FDSs and SPIs were determined. In the grains, the water activity, total isoflavone content and antioxidant index (AI) were measured and in DFSs, the phytate content and trypsin inhibitor activity (TIA). The acidity content was measured in the oil obtained from the whole flours. The structural characteristics of SPIs were evaluated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate (SDSPAGE) under non reducing conditions of the soluble proteins using different systems. Additionally, differential scanning calorimetry (DSC), free sulfhydryl groups (SH), intrinsic fluorescence (IF), reversed-phase high performance liquid chromatography (RP-HPLC) and of size exclusion high performance liquid chromatography (SE-HPLC) were also used to evaluate the effects. The analyses of the functional properties in SPIs were: solubility, gelation, water holding capacity (WHC), emulsifying capacity (EC), emulsifying activity index (EAI) and emulsifying stability (ES). The total isoflavone content and antioxidant index analyzed in the grains did not differ significantly (p>0.05) and remained stable throughout storage, as a function of the dose or type of irradiation. However, there was an increase in the b-glycosides and decrease in the mean percentage of malonyl glycosides. The acidity content increased in the percentage of 81.6% in the oil grains irradiated with 5 kGy and stored for 12 months. DFSs of irradiated grains exhibited smaller values of the trypsin inhibitor and phytate. The analyses of differential scanning calorimetry, intrinsic fluorescence and size exclusion high performance liquid chromatography showed that no important structural alterations occurred in the protein isolates due irradiation or storage. However, the free sulfhydryl groups increased in SPIs of irradiated grains, without further significant changes along the storage. There was an increase of the proportion of the hydrophobic fractions, showed by reversed-phase high performance liquid chromatography profile, for all the isolated after four months of storage of the grains. SPIs of irradiated grains presented high solubility, emulsifying capacity while emulsifying stability and emulsifying activity index did not differ significantly (p>0.05). The values of hardness and water holding capacity of the SPIs gels did not differ significantly (p>0.05) for all treatment. However, SPIs gels of stored grains had high values of hardness. The values cohesiveness and elasticity of the SPIs gels did not differ significantly (p>0.05), independent of the dose and type of irradiation. Under the studied conditions, gamma irradiation at doses of 5 kGy and of electron beam of 2 kGy can be used without damage to the grain composition or storage property or to the physicochemical properties of the protein isolates / Doutorado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Doutor em Alimentos e Nutrição

Nach den Ergebnissen der zweiten Nationalen Verzehrsstudie sind in Deutschland bereits 66 % der Männer und 51 % der Frauen übergewichtig (BMI > 25) oder adipös (BMI > 30) [BMELV, 2008]. Bisher auf dem Markt befindliche „Light-Lebensmittel“ mit Fettaustausch- bzw. Fettersatzstoffen weisen jedoch häufig sensorische Mängel auf. Im Kooperationsprojekt „Pflanzliche Fettaustauschstoffe aus sphärischen Proteinmizellen“ (Universität Leipzig: Institut für Lebensmittelhygiene; Freising: Fraunhofer IVV) wurde ein Lupinenproteinisolat entwickelt, welches micellare Strukturen mit hydrophober Oberfläche ausbilden kann und sich aufgrund seiner fettähnlichen Eigenschaften als neuer proteinbasierter Fettaustauschstoff in Lebensmitteln eignet. Aufgrund der geringen mikrobiologischen Stabilität und einer hohen Belastung mit sporenbildenden Bakterien, z. T. Bacillus cereus, waren jedoch Maßnahmen zur Entkeimung der Rohstoffe sowie des Proteinisolats notwendig. Die Arbeit stellt diese Maßnahmen und deren Einfluss auf die mikrobiologische Beschaffenheit sowie sensorische, technofunktionelle und ausgewählte ernährungsphysiologische Eigenschaften dar. In der vorliegenden Arbeit wurde eine physikalische Methode der Saatgutentkeimung etabliert (130 °C/60 min), welche die mikrobielle Stabilisierung des lupinenproteinbasierten Fettaustauschstoffes sicherstellte, wobei die sensorische Qualität (Geschmack, Cremigkeit, Farbe) nur minimal, die ernährungsphysiologische (in-vitro-Verdaubarkeit, Maillard-Produkte, Polyphenolgehalt) jedoch nicht beeinflusst wurde. Starke Veränderungen der technofunktionellen Eigenschaften (z. B. Gelbildung, Wasserbindung, Emulgierbarkeit, Schaumbildung etc.) konnten sowohl im positiven als auch im negativen Sinne nicht beschrieben werden. Lichtmikroskopische Aufnahmen und Untersuchungen der Proteine mittels SDS-PAGE und DSC bestätigten eine nur geringfügige Beeinflussung der micellaren Struktur und Proteinzusammensetzung. Die Anwendung als Fettaustauschstoff in Lebensmitteln würde somit nicht beeinträchtigt. Der Einfluss der Saatgutbehandlung auf das Protein war wesentlich geringer als eine direkte thermische Behandlung des Proteinisolats. Im Hinblick auf den Gesamtprozess sollte eine Pasteurisierung der feuchten Proteinisolate im nichtproteinschädigenden Temperaturbereich (75 °C/5 min) dennoch durchgeführt werden, um während des Prozesses eingetragene Mikroorganismen zu inaktivieren.

Cette étude a été menée dans le but d’explorer les nouvelles sources de protéines à valeur nutraceutique. Les graines de Ricinodendron heudelotii (Bail. Pierre ex Pax) et de Tetracarpidium conophorum (Müll. Arg) ont d’abord été cuites dans de l’eau bouillante pendant 90 et 30 min qui sont respectivement leurs temps optimums de cuisson. Après délipidation et tamisage des tourteaux, la fraction 400 - 500 µm s’est révélée la plus représentative avec plus de 70 % et riche en azote protéique (6–7% MS). Les concentrats et les isolats protéiques ont été préparés à partir des tourteaux respectivement dans l’eau distillée à pH 4,5 et dans une solution de NaOH à 0,2 % (R. heudelotii), une solution de NaCl 0,6 M (T. conophorum) à pH 11. Ces concentrats (65 – 75 % MS de protéines) et ces isolats protéiques (81 – 92 % MS de protéines) ont une composition physico-chimique différente (P < 0,05) de celle des tourteaux. Pour les deux Euphorbiacées, les capacités de rétention d’eau (367 – 467 g / 100 g d’échantillon), de rétention d’huile (256 – 410 g / 100 g d’échantillon) et moussante (68 – 71 %) sont maximales dans les isolats protéiques tandis que les capacités gélifiante (6 – 14 %) et émulsifiante (63 – 87 %) le sont dans les concentrats protéiques. Les teneurs en acides aminés essentiels des tourteaux de R. heudelotii et de T. conophorum sont comparables à celle de la protéine de référence. L’étude de la digestibilité enzymatique in vitro a montré que l’azote libéré après 6 h est supérieur à 90 % dans les concentrats et les isolats protéiques. La digestibilité protéique in vivo indique que le gain de poids des rats mâles âgés de 21 ± 3 jours durant 15 jours d’expérimentation ainsi que les paramètres de rétention azotée sont plus importants avec les régimes à base de l’aliment de référence (caséine) et du tourteau de T. conophorum. Les valeurs corrigées des paramètres de digestibilité par l’indice chimique des acides aminés laissent apparaître que le tourteau de T. conophorum renferme les protéines de très bonne qualité nutritionnelle, autant que la caséine / This study was conducted in order to look for alternative sources of proteins having nutraceutic value. The grains of Ricinodendron heudelotii (Bail.) and Tetracarpidium conophorum (Müll. Arg) were first cooked in boiled water at their optimal cooking time for 90 and 30 min respectively. After defating, sieving of the defatted cakes showed that samples with a granulometry of 400 - 500 µm were most representative (more than 70%), containing more proteic nitrogen (6–7 %). Protein concentrates and protein isolates were prepared from defatted cakes respectively in distilled water at pH 4.5 and in NaOH 0.2% (R. heudelotii) and NaCl 0.6M (T. conophorum) at pH 11. Physico-chemical properties of protein concentrates (65 – 75 % of proteins) and protein isolates (81–92 % of proteins) were different from those of the defatted cakes. Water holding (367 – 467 g / 100 g of sample), oil holding (256 – 410 g / 100 g of sample) and foaming capacities (68 – 71 %) were highest with protein isolates whereas gelling (6 – 14 %) and emulsion capacities (63 – 87 %) were highest with concentrates. The amounts of essential amino acids in both defatted flours were comparable to the value in FAO / WHO (2007) scoring pattern. Nitrogen liberated after 6 h of enzymatic digestibility was more than 90 % both in the proteins concentrates and isolates. In vivo studies carried out on 21 ± 3 days old Sprague Dawley male rats for 15 days showed that gain of weight and nitrogen retention parameters were higher for rats that consumed casein and T. conophorum defatted cake. Corrected values of nitrogen digestibility of the analysed samples showed that T. conophorum defatted cake contains protein source with good nutritional quality

Nach den Ergebnissen der zweiten Nationalen Verzehrsstudie sind in Deutschland bereits 66 % der Männer und 51 % der Frauen übergewichtig (BMI > 25) oder adipös (BMI > 30) [BMELV, 2008]. Bisher auf dem Markt befindliche „Light-Lebensmittel“ mit Fettaustausch- bzw. Fettersatzstoffen weisen jedoch häufig sensorische Mängel auf. Im Kooperationsprojekt „Pflanzliche Fettaustauschstoffe aus sphärischen Proteinmizellen“ (Universität Leipzig: Institut für Lebensmittelhygiene; Freising: Fraunhofer IVV) wurde ein Lupinenproteinisolat entwickelt, welches micellare Strukturen mit hydrophober Oberfläche ausbilden kann und sich aufgrund seiner fettähnlichen Eigenschaften als neuer proteinbasierter Fettaustauschstoff in Lebensmitteln eignet. Aufgrund der geringen mikrobiologischen Stabilität und einer hohen Belastung mit sporenbildenden Bakterien, z. T. Bacillus cereus, waren jedoch Maßnahmen zur Entkeimung der Rohstoffe sowie des Proteinisolats notwendig. Die Arbeit stellt diese Maßnahmen und deren Einfluss auf die mikrobiologische Beschaffenheit sowie sensorische, technofunktionelle und ausgewählte ernährungsphysiologische Eigenschaften dar. In der vorliegenden Arbeit wurde eine physikalische Methode der Saatgutentkeimung etabliert (130 °C/60 min), welche die mikrobielle Stabilisierung des lupinenproteinbasierten Fettaustauschstoffes sicherstellte, wobei die sensorische Qualität (Geschmack, Cremigkeit, Farbe) nur minimal, die ernährungsphysiologische (in-vitro-Verdaubarkeit, Maillard-Produkte, Polyphenolgehalt) jedoch nicht beeinflusst wurde. Starke Veränderungen der technofunktionellen Eigenschaften (z. B. Gelbildung, Wasserbindung, Emulgierbarkeit, Schaumbildung etc.) konnten sowohl im positiven als auch im negativen Sinne nicht beschrieben werden. Lichtmikroskopische Aufnahmen und Untersuchungen der Proteine mittels SDS-PAGE und DSC bestätigten eine nur geringfügige Beeinflussung der micellaren Struktur und Proteinzusammensetzung. Die Anwendung als Fettaustauschstoff in Lebensmitteln würde somit nicht beeinträchtigt. Der Einfluss der Saatgutbehandlung auf das Protein war wesentlich geringer als eine direkte thermische Behandlung des Proteinisolats. Im Hinblick auf den Gesamtprozess sollte eine Pasteurisierung der feuchten Proteinisolate im nichtproteinschädigenden Temperaturbereich (75 °C/5 min) dennoch durchgeführt werden, um während des Prozesses eingetragene Mikroorganismen zu inaktivieren.:1 Einleitung und Zielstellung 1 2 Stand des Wissens 4 2.1 Die Lupine 4 2.1.1 Anbau und Verbreitung 4 2.1.2 Einsatz von Lupinenprodukten und -proteinen in der Humanernährung 5 2.1.3 Inhaltsstoffe und deren Verteilung 5 2.1.4 Lupinenproteine 10 2.1.4.1 Einteilung und Struktur der Lupinenproteine 10 2.1.4.2 Lupinenproteine und Allergenität 12 2.1.5 Eigenschaften der verschiedenen Lupinenproteinfraktionen 13 2.1.5.1 Ernährungsphysiologische Eigenschaften 13 2.1.5.2 Funktionelle Eigenschaften 15 2.1.5.3 Modifikation der Proteinstruktur 15 2.1.5.4 Herstellung verschiedener Lupinenproteinpräparate 16 2.1.5.5 Micellare Proteine 17 2.2 Möglichkeiten der Fettreduktion in Lebensmitteln 18 2.2.1 Fettaustauschstoffe 18 2.2.1.1 Fettaustauschstoffe auf Proteinbasis (Mikropartikulierte Proteine) 18 2.2.1.2 Fettaustauschstoffe auf Kohlenhydratbasis 19 2.2.1.3 Quellstoffe 19 2.2.2 Fettersatzstoffe 19 2.2.2.1 Spezielle Triglyceride 20 2.2.2.2 Kohlenhydratpolyester 20 2.2.2.3 Retrofette 20 2.3 Herstellung des lupinenproteinbasierten Fettaustauschstoffes 20 2.4 Saatgutbehandlung 21 2.4.1 Methoden der Lebensmittelkonservierung 22 2.5 Proteinfunktionalität 25 2.5.1 Definition und Zusammenhang zu Proteinen 25 2.5.2 Ausgewählte funktionelle Eigenschaften 26 2.5.2.1 Wasserbindevermögen 26 2.5.2.2 Ölbindevermögen 26 2.5.2.3 Löslichkeit 27 2.5.2.4 Emulgiervermögen 27 2.5.2.5 Schaumbildungsvermögen 28 2.5.2.6 Gelbildungsvermögen 29 2.5.2.7 Oberflächenhydrophobität 30 2.5.2.8 Bedeutung für die Lebensmittelentwicklung 30 3 Material und Methoden 32 3.1 Material 32 3.1.1 Saatgut 32 3.1.2 Geräte, Chemikalien, Verbrauchsmaterial, Software 32 3.1.3 Pufferlösungen 39 3.1.4 Herstellung Bradford-Reagenz, 5-fach 39 3.1.5 Auswahl der Vergleichssubstanzen 39 3.2 Methoden 40 3.2.1 Herstellung der Proteinisolate 40 3.2.2 Mikrobiologische Analysen 41 3.2.3 Bestimmung der Trockenmasse 41 3.2.4 Bestimmung des Proteingehalts 42 3.2.5 Thermische Behandlungsmethoden im Prozess 42 3.2.5.1 UHT-Erhitzung des Extraktes 42 3.2.5.2 Pasteurisierung des Isolats 44 3.2.6 Saatgutentkeimung 44 3.2.6.1 UVC-Bestrahlung 44 3.2.6.2 Trockene Erhitzung 45 3.2.6.3 Autoklavieren 46 3.2.7 Sensorische Untersuchungen 46 3.2.8 Proteinfunktionalität 47 3.2.8.1 Ölbindevermögen 47 3.2.8.2 Wasserbindevermögen 47 3.2.8.3 Gelbildungsvermögen 47 3.2.8.4 Emulgiereigenschaften 47 3.2.8.5 Schaumbildungsvermögen 48 3.2.8.6 Proteinlöslichkeit 48 3.2.8.7 Oberflächenhydrophobität 49 3.2.9 Ernährungsphysiologische Eigenschaften 50 3.2.9.1 in-vitro-Verdaubarkeit 50 3.2.9.2 Maillard-Produkte 50 3.2.9.3 Nachweis reduzierender Zucker .50 3.2.9.4 Nachweis von Glykoproteinen 50 3.2.9.5 Polyphenolgehalt der Lupinenflocken und Proteinisolate 51 3.2.10 Proteincharakterisierung 51 3.2.10.1 Lichtmikroskopie 51 3.2.10.2 Dynamische Differenzkalorimetrie 51 3.2.10.3 Natriumdodecylsulfat-Polyacrylamidgelelektrophorese 52 4 Ergebnisse und Diskussion 54 4.1 Thermische Behandlungsmethoden im Prozess 54 4.1.1 UHT-Erhitzung des Extraktes: Einfluss auf Mikrobiologie und Proteinausbeute 54 4.1.2 Pasteurisierungsversuche: Einfluss auf Mikrobiologie und Proteinqualität 55 4.2 Saatgutentkeimung - Mikrobiologie und Proteinausbeute 56 4.2.1 Versuchsreihe I 56 4.2.2 Versuchsreihe II 61 4.3 Sensorische Untersuchungen 63 4.3.1 Verkostungen 64 4.3.2 Farbmessung der Proteinisolate und Flocken 65 4.4 Proteinfunktionalität 69 4.4.1 Wasser- und Ölbindevermögen 69 4.4.2 Gelbildungsvermögen 72 4.4.3 Emulgiereigenschaften 74 4.4.4 Schaumbildungsvermögen 78 4.4.5 Proteinlöslichkeit 81 4.4.6 Oberflächenhydrophobität 83 4.5 Ernährungsphysiologische Eigenschaften 86 4.5.1 Maillard-Produkte 86 4.5.2 Nachweis reduzierender Zucker 87 4.5.3 Nachweis von Glykoproteinen 87 4.5.4 Verdaubarkeit 88 4.5.5 Polyphenolgehalte 89 4.6 Proteincharakterisierung 91 4.6.1 Lichtmikroskopie 91 4.6.2 Dynamische Differenzkalorimetrie 95 4.6.3 Natriumdodecylsulfat-Polyacrylamidgelelektrophorese 98 5 Zusammenfassung 105 Anhang 109