Tiger nuts (Cyperus esculentus L) are recognized as a high potential, alternative source of food nutrients. However, there is limited scientific literature on the technological possibilities for developing value-added foods, such as fermented products from tiger nut milk. Therefore, strategies for producing and improving the properties of fermented tiger nut milk were investigated for generating lactose-free, nutritious yogurt-like products with acceptable sensory properties and a prolonged shelf life quality. A wet-milling procedure was standardized for extracting tiger nut milk from tiger nuts, and the effects of the extraction process on nutrient distribution, colour properties and colloidal stability of the milk were analyzed. Next, tiger nut milk was enriched with proteins and/or hydrocolloids and the impact of the additives on the physical properties of the milk were determined. Enriched tiger nut milk was fermented by using classical yogurt cultures and the obtained products were analyzed for the microbiological, physico-chemical and sensory characteristics. Additionally, effects of enriching tiger nut milk with microbial transglutaminase cross-linked proteins on the microbiological and physico-chemical properties were evaluated. Higher wet-milling intensity improved the nutrient composition, colloidal stability and colour of the milk. Enrichment of tiger nut milk with milk proteins and xanthan gum enhanced the viscosity and stability, and after fermentation, led to homogenous gel-like products with superior microbiological, physico-chemical and different sensory properties compared to the fermented plain tiger nut milk. Microbial transglutaminase cross-linked proteins improved the physical characteristics of the fermented product, especially during storage. This product would be relevant in many developing countries with high prevalence of lactose intolerance, limited access to nutritious food but show a high distribution of tiger nut vegetation. / Erdmandeln (Cyperus esculentus L) haben ein hohes Potential als alternative Quelle Lebensmittelinhaltsstoffen. Allerdings gibt es nur in begrenztem Ausmaß Literatur über technologische Möglichkeiten zur Entwicklung von Mehrwert-Lebensmitteln wie fermentierter Erdmandelmilch. Daher wurden Strategien zur Herstellung und Verbesserung der Eigenschaften von fermentierter Erdmandelmilch zur Erzeugung laktosefreier joghurtähnlicher Produkte mit akzeptablen sensorischen Eigenschaften untersucht. Für die Extraktion der Erdmandelmilch wurde ein Nassmahlverfahren standardisiert und der Einfluss des Verfahrens auf die Nährstoffverteilung, die Farbeigenschaften und die kolloidale Stabilität der Milch analysiert. Als nächstes wurde Erdmandelmilch mit Proteinen und/oder Hydrokolloiden angereichert, und der Einfluss der Additive auf die physikalischen Eigenschaften des Extrakts bestimmt. Angereicherte Erdmandelmilch wurde mit klassischen Joghurtkulturen fermentiert, und die mikrobiologischen, physikalisch-chemischen und sensorischen Eigenschaften der Produkte wurden untersucht. Zusätzlich wurden Effekte der Anreicherung von Erdmandelmilch mit enzymatisch vernetzten Proteinen auf die mikrobiologischen und physikalisch-chemischen Eigenschaften bewertet. Eine höhere Nassmahlintensität verbesserte die Nährstoffzusammensetzung, die kolloidale Stabilität und die Farbe der Milch. Die Anreicherung erhöhte die Viskosität und Stabilität und führte nach der Fermentation zu homogenen gelartigen Produkten mit verbesserten mikrobiologischen, physikalisch-chemischen und sensorischen Eigenschaften im Vergleich zur fermentierten Erdmandelmilch. Mikrobielle Transglutaminase-vernetzte Proteine verbesserten die physikalischen Eigenschaften des fermentierten Produkts, insbesondere während der Lagerung. Dieses Produkt wäre in vielen Entwicklungsländern mit hoher Prävalenz von Laktoseintoleranz und begrenztem Zugang zu nahrhaften Lebensmitteln als Alternative von Interesse.

Tiger nuts (Cyperus esculentus L) are recognized as a high potential, alternative source of food nutrients. However, there is limited scientific literature on the technological possibilities for developing value-added foods, such as fermented products from tiger nut milk. Therefore, strategies for producing and improving the properties of fermented tiger nut milk were investigated for generating lactose-free, nutritious yogurt-like products with acceptable sensory properties and a prolonged shelf life quality. A wet-milling procedure was standardized for extracting tiger nut milk from tiger nuts, and the effects of the extraction process on nutrient distribution, colour properties and colloidal stability of the milk were analyzed. Next, tiger nut milk was enriched with proteins and/or hydrocolloids and the impact of the additives on the physical properties of the milk were determined. Enriched tiger nut milk was fermented by using classical yogurt cultures and the obtained products were analyzed for the microbiological, physico-chemical and sensory characteristics. Additionally, effects of enriching tiger nut milk with microbial transglutaminase cross-linked proteins on the microbiological and physico-chemical properties were evaluated. Higher wet-milling intensity improved the nutrient composition, colloidal stability and colour of the milk. Enrichment of tiger nut milk with milk proteins and xanthan gum enhanced the viscosity and stability, and after fermentation, led to homogenous gel-like products with superior microbiological, physico-chemical and different sensory properties compared to the fermented plain tiger nut milk. Microbial transglutaminase cross-linked proteins improved the physical characteristics of the fermented product, especially during storage. This product would be relevant in many developing countries with high prevalence of lactose intolerance, limited access to nutritious food but show a high distribution of tiger nut vegetation.:1. Introduction and aim 1 2. Literature review 4 2.1 Tiger nut, origin, nutritional value and food use 4 2.2 Tiger nut milk, preparation and nutrient composition 7 2.3 Colloidal characteristics of tiger nut milk 9 2.4 Factors accounting for the dispersion stability of tiger nut milk 10 2.5 Enhancing tiger nut milk stability 12 2.6 Properties of fermented tiger nut milk 17 2.7 Microbial transglutaminase and properties of fermented tiger nut milk 18 3. Methodology 21 3.1 Extraction and characterisation of tiger nut milk 21 3.1.1 Sample collection and preparation 21 3.1.2 Tiger nut milk extraction 21 3.1.3 Nutrient analysis of tiger nuts 22 3.1.4 Analysis of tiger nut products 23 3.1.5 Particle size distribution 24 3.1.6 Colloidal stability 25 3.1.7 Colour measurement 25 3.2 Stabilisation of tiger nut milk dispersion 26 3.2.1 Tiger nut milk preparation 26 3.2.2 Preparation of tiger nut milk enrichments 26 3.2.3 Gravitational stability of enriched tiger nut milk 27 3.2.4 Accelerated gravitational stability of enriched tiger nut milk 28 3.2.5 Viscosity of TNM mixtures 29 3.3 Extraction and characterisation of globular tiger nut proteins 29 3.3.1 Protein extraction and fractionation 29 3.3.2 Molecular mass of globular tiger nut proteins 31 3.3.3 Denaturation temperature of globular tiger nut proteins 32 3.3.4 Isoelectric point of globular tiger nut protein 33 3.4 Properties of fermented tiger nut milk enriched with proteins 34 3.4.1 Materials and Reagents 34 3.4.2 Preparation of plain and enriched tiger nut milk 34 3.4.3 Fermentation of plain and enriched tiger nut milk 35 3.4.4 Viable counts of starter cultures in fermented tiger nut milk systems 36 3.4.5 Chemical analysis of unfermented and fermented tiger nut milk 36 3.4.6 Physical analysis of fermented tiger nut milk products 37 3.4.7 Sensory analysis of fermented tiger nut milk products 38 3.5 Microbial transglutaminase and fermented tiger nut milk property 38 3.5.1 Preparation of plain and enriched tiger nut milk 38 3.5.2 Fermentation of plain and enriched tiger nut milk 39 3.5.3 Analysis of the enzymatically cross-linked proteins 39 3.5.4 Viable counts 40 3.5.5 pH and titratable acidity 40 3.5.6 Syneresis and viscosity 41 3.5.7 Colour of fermented tiger nut products 41 3.6 Statistical analysis 41 4. Results and discussion 43 4.1 Extraction and characteristics of tiger nut milk 43 4.1.1 Material recovery, mass transfer and yield of tiger nut solids 43 4.1.2 Nutrient composition of tiger nut products 45 4.1.3 Physical properties of tiger nut milk 48 4.1.3.1 Particle size distribution of extracted tiger nut milk 48 4.1.3.2 Colloidal stability of tiger nut milk 49 4.1.3.3 Colour stability of tiger nut milk 51 4.2 Stabilisation of tiger nut milk 53 4.2.1 Effects of enrichments on the stability of tiger nut milk 53 4.2.2 Effects of pH and temperature on the stability of enriched TNM 56 4.2.3 Effects of enrichments on the rheology of tiger nut milk 58 4.3 Tiger nut protein extraction and characterisation 60 4.3.1 Protein extraction and fractionation 60 4.3.2 Molecular mass of tiger nut protein 62 4.3.3 Thermal denaturation of tiger nut protein 63 4.3.4 Isoelectric point of tiger nut proteins 66 4.4 Properties of fermented tiger nut milk enriched with proteins 67 4.4.1 Acidification and gel formation during fermentation 67 4.4.2 Microbiological properties of fermented enriched tiger nut milk 70 4.4.3 Physico-chemical properties of fermented enriched tiger nut milk 71 4.4.4 Sensory properties of fermented tiger nut milk products 76 4.5 Microbial transglutaminase and fermented tiger nut milk property 77 4.5.1 Effects on tiger nut milk fermentation 77 4.5.2 Microbiological properties during storage of fermented product 81 4.5.3 Physico-chemical properties during storage of fermented product 83 4.5.4 Effects on colour of fermented tiger nut product 86 5. Conclusions and outlook 88 Bibliography 90 List of figures 111 List of tables 115 List of Publications 116 Poster and presentations 116 / Erdmandeln (Cyperus esculentus L) haben ein hohes Potential als alternative Quelle Lebensmittelinhaltsstoffen. Allerdings gibt es nur in begrenztem Ausmaß Literatur über technologische Möglichkeiten zur Entwicklung von Mehrwert-Lebensmitteln wie fermentierter Erdmandelmilch. Daher wurden Strategien zur Herstellung und Verbesserung der Eigenschaften von fermentierter Erdmandelmilch zur Erzeugung laktosefreier joghurtähnlicher Produkte mit akzeptablen sensorischen Eigenschaften untersucht. Für die Extraktion der Erdmandelmilch wurde ein Nassmahlverfahren standardisiert und der Einfluss des Verfahrens auf die Nährstoffverteilung, die Farbeigenschaften und die kolloidale Stabilität der Milch analysiert. Als nächstes wurde Erdmandelmilch mit Proteinen und/oder Hydrokolloiden angereichert, und der Einfluss der Additive auf die physikalischen Eigenschaften des Extrakts bestimmt. Angereicherte Erdmandelmilch wurde mit klassischen Joghurtkulturen fermentiert, und die mikrobiologischen, physikalisch-chemischen und sensorischen Eigenschaften der Produkte wurden untersucht. Zusätzlich wurden Effekte der Anreicherung von Erdmandelmilch mit enzymatisch vernetzten Proteinen auf die mikrobiologischen und physikalisch-chemischen Eigenschaften bewertet. Eine höhere Nassmahlintensität verbesserte die Nährstoffzusammensetzung, die kolloidale Stabilität und die Farbe der Milch. Die Anreicherung erhöhte die Viskosität und Stabilität und führte nach der Fermentation zu homogenen gelartigen Produkten mit verbesserten mikrobiologischen, physikalisch-chemischen und sensorischen Eigenschaften im Vergleich zur fermentierten Erdmandelmilch. Mikrobielle Transglutaminase-vernetzte Proteine verbesserten die physikalischen Eigenschaften des fermentierten Produkts, insbesondere während der Lagerung. Dieses Produkt wäre in vielen Entwicklungsländern mit hoher Prävalenz von Laktoseintoleranz und begrenztem Zugang zu nahrhaften Lebensmitteln als Alternative von Interesse.:1. Introduction and aim 1 2. Literature review 4 2.1 Tiger nut, origin, nutritional value and food use 4 2.2 Tiger nut milk, preparation and nutrient composition 7 2.3 Colloidal characteristics of tiger nut milk 9 2.4 Factors accounting for the dispersion stability of tiger nut milk 10 2.5 Enhancing tiger nut milk stability 12 2.6 Properties of fermented tiger nut milk 17 2.7 Microbial transglutaminase and properties of fermented tiger nut milk 18 3. Methodology 21 3.1 Extraction and characterisation of tiger nut milk 21 3.1.1 Sample collection and preparation 21 3.1.2 Tiger nut milk extraction 21 3.1.3 Nutrient analysis of tiger nuts 22 3.1.4 Analysis of tiger nut products 23 3.1.5 Particle size distribution 24 3.1.6 Colloidal stability 25 3.1.7 Colour measurement 25 3.2 Stabilisation of tiger nut milk dispersion 26 3.2.1 Tiger nut milk preparation 26 3.2.2 Preparation of tiger nut milk enrichments 26 3.2.3 Gravitational stability of enriched tiger nut milk 27 3.2.4 Accelerated gravitational stability of enriched tiger nut milk 28 3.2.5 Viscosity of TNM mixtures 29 3.3 Extraction and characterisation of globular tiger nut proteins 29 3.3.1 Protein extraction and fractionation 29 3.3.2 Molecular mass of globular tiger nut proteins 31 3.3.3 Denaturation temperature of globular tiger nut proteins 32 3.3.4 Isoelectric point of globular tiger nut protein 33 3.4 Properties of fermented tiger nut milk enriched with proteins 34 3.4.1 Materials and Reagents 34 3.4.2 Preparation of plain and enriched tiger nut milk 34 3.4.3 Fermentation of plain and enriched tiger nut milk 35 3.4.4 Viable counts of starter cultures in fermented tiger nut milk systems 36 3.4.5 Chemical analysis of unfermented and fermented tiger nut milk 36 3.4.6 Physical analysis of fermented tiger nut milk products 37 3.4.7 Sensory analysis of fermented tiger nut milk products 38 3.5 Microbial transglutaminase and fermented tiger nut milk property 38 3.5.1 Preparation of plain and enriched tiger nut milk 38 3.5.2 Fermentation of plain and enriched tiger nut milk 39 3.5.3 Analysis of the enzymatically cross-linked proteins 39 3.5.4 Viable counts 40 3.5.5 pH and titratable acidity 40 3.5.6 Syneresis and viscosity 41 3.5.7 Colour of fermented tiger nut products 41 3.6 Statistical analysis 41 4. Results and discussion 43 4.1 Extraction and characteristics of tiger nut milk 43 4.1.1 Material recovery, mass transfer and yield of tiger nut solids 43 4.1.2 Nutrient composition of tiger nut products 45 4.1.3 Physical properties of tiger nut milk 48 4.1.3.1 Particle size distribution of extracted tiger nut milk 48 4.1.3.2 Colloidal stability of tiger nut milk 49 4.1.3.3 Colour stability of tiger nut milk 51 4.2 Stabilisation of tiger nut milk 53 4.2.1 Effects of enrichments on the stability of tiger nut milk 53 4.2.2 Effects of pH and temperature on the stability of enriched TNM 56 4.2.3 Effects of enrichments on the rheology of tiger nut milk 58 4.3 Tiger nut protein extraction and characterisation 60 4.3.1 Protein extraction and fractionation 60 4.3.2 Molecular mass of tiger nut protein 62 4.3.3 Thermal denaturation of tiger nut protein 63 4.3.4 Isoelectric point of tiger nut proteins 66 4.4 Properties of fermented tiger nut milk enriched with proteins 67 4.4.1 Acidification and gel formation during fermentation 67 4.4.2 Microbiological properties of fermented enriched tiger nut milk 70 4.4.3 Physico-chemical properties of fermented enriched tiger nut milk 71 4.4.4 Sensory properties of fermented tiger nut milk products 76 4.5 Microbial transglutaminase and fermented tiger nut milk property 77 4.5.1 Effects on tiger nut milk fermentation 77 4.5.2 Microbiological properties during storage of fermented product 81 4.5.3 Physico-chemical properties during storage of fermented product 83 4.5.4 Effects on colour of fermented tiger nut product 86 5. Conclusions and outlook 88 Bibliography 90 List of figures 111 List of tables 115 List of Publications 116 Poster and presentations 116

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