The aim of this study was to explore the characteristics of a non-gelling exopolysaccharide (EPS) obtained from Lactobacillus delbrueckii subsp. bulgaricus NCFB 2483 and a gelling EPS obtained from Sphingomonas elodea ATCC 31461 (31461). The EPSs were isolated from the two bacterial strains grown in milk permeate-based media. They were purified and then characterised using light scattering and viscometric techniques. A greater emphasis of this research was placed on 2483 EPS since its physical characteristics have not been reported to date. In the case of 31461 EPS. a model for gelation of the sodium gellan was proposed based on rheological and light scattering measurements. The rheological properties of the two EPSs were also compared with several commercial polysaccharides. Microscopy examination of 2483 EPS was carried out using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). In CLSM, the lectin SBA (from Glycine max) Alexa Fluor 488 conjugate was used to stain the EPS since it has affinity for galactopyranosyl residues present in 2483 EPS. The CLSM micrographs showed a random distribution of EPS aggregates in the culture medium. At high magnification, the SEM micrographs showed web-like EPS structures. These structures were formed during the critical point drying process, when the EPS, which filled the interstices and channels of the protein aggregates, dehydrated. The 2483 EPS aggregates were found to be stable at neutral or low pH (~3.9) but were disrupted at high pH (pH 8-10). Procedures commonly used to quantify EPS from culture medium were found to be unreliable. In the development of an improved EPS assay, each of the processing steps was examined. Key improvements included the use of Flavourzyme for protein hydrolysis; optimising ethanol concentration to prevent lactose crystallisation yet allowing complete EPS precipitation; and a suitable centrifugation regime to minimise EPS loss. The improved EPS assay gave reproducible results (5% coefficient of variation). The isolation of 2483 EPS from milk media proved to be a difficult task because of interference from non-EPS components. An effective and simple approach allowing maximum EPS recovery involved the use of a hydrolysed milk medium which was ultrafiltered (UF) to remove molecular species larger than 2.5 x 105 Da. The UF permeate was suitable for the growth of 2483 with an EPS yield of ~400mg/L. Two EPS fractions (namely a soluble and an insoluble fraction) were isolated by ethanol precipitation and the soluble 'ropy' fraction was further purified to achieve ~98% purity. The elemental analysis of the purified fraction revealed the presence of nitrogen (~2.7% w/w). This could be due to the interaction of some peptides (from the growth medium) with the EPS. The polysaccharide composition of the soluble EPS fraction comprised of galactose, glucose, rhamnose and mannose residues (5:1:0.6:0.5). Traces of glucosamine were also found in the fraction. The purified fraction of 2483 EPS was characterised. Using a capillary viscometer, an intrinsic viscosity of ~2013mL/g was determined. The flow curves of the 2483 EPS solutions obtained using a rotational viscometer showed shear-thinning behaviour and an exponent value of ~0.76 (based on the Cross-type model) is typical of random coil polymers. The concentration dependence of the viscosity plot produced gradients of ~1.1 in the dilute domain and ~3.3 in the semi-dilute to concentrated domain. The coil overlap parameters at three concentration domains (c*[ŋ],ccr[ŋ] and c**[ŋ]) were 0.55, 2.86 and 5.67 respectively. The molecular parameters of the 2483 EPS were found via static light scattering measurements to have a weight-average molar mass (Mw.) of ~2 x 106 Da, a z-average root-mean-square radius ((r2g)z1/2) of ~165nm and a low polydispersity index (Mw/Mn ~1.15). The plot of Mw versus (r2g)z1/2 gave a gradient of approximately 0.5, which also suggested that the EPS polymer adopted a random coil conformation. The second part of the research involved gellan gum. Two gellan samples were studied. The first gellan sample was obtained from the fermentation of Sphingomonas elodea ATCC 31461 using milk permeate-based medium (31461). The second sample was a commercial high acyl gellan (LT100). Both gellan samples were converted to their sodium forms (Na-31461 and Na-LT100 gellan) using cation exchange resin and purified. The Na-gellan samples were highly sensitive to changes in Na+ concentrations. From oscillatory measurements, it was found that the complex moduli of the two Na-gellan samples superimposed closely at a specific Na+ concentration. The model for the conformational changes of Na-gellan molecules from a solution to a gel was proposed based on rheological and light scattering data. At very low Na+ concentrations (<19mM, in the case of Na-LT100). Na-gellan molecules were single-stranded (Mw ~2.5 x 10 5 Da) and adopted random coil conformation (exponent value based on the Cross-type model of ~0.76). At a slightly higher Na+ concentration (~19-24mM), Na-gellan molecules formed double-helices which led to a two-fold increase in molecular weight (M w ~5.2 x 105 Da). The double-stranded molecule appeared to be stiffer (exponent value of the Cross-type model ~0.82) and the mechanical spectra (G',G”) demonstrated 'weak ge' characteristics. A further increase in the Na+ concentration (>24mM) resulted in the formation of a gel network. The study also found that at low Na+concentration, both single-stranded and double-stranded Na-gellan molecules had a tendency to form aggregates under zero-shear conditions. The interactions involved in these aggregates were considered weak and transient, according to the Cox-Merz plot and light scattering data.

The technique of micro-oxygenation involves the deliberate addition of continuous, metered amounts of oxygen into a vessel of bulk wine during the maturation period (between the end of fermentation and bottling). The aim of the process is to improve the sensory properties of red wine, particularly the mouthfeel characteristics associated with the various polyphenol constituents. The success of the process appears to depend strongly on the ability to control the rate of oxygen dosage. The effect of dosage rate on the chemical and corresponding sensory changes of a red wine is the central theme of this thesis. A method of dosing oxygen (at typical micro-oxygenation rates) into small volumes of wine (<100 litres) was developed using a dense polymer membrane diffuser. It was clearly demonstrated that wine could be reliably oxygenated at very low rates using a coiled length of FEP as the diffuser material. Oxygen dosage was regulated by adjusting the oxygen pressure inside the tube. The advantage with a dense polymer diffuser is that no bubbles are generated and the oxygenation efficiency is 100%. The diffuser was fully modeled and characterised for use in the laboratory scale trials detailed in Chapters Four and Six. The small scale oxygenation equipment was used to conduct a fully replicated experiment to investigate the evolution of a Cabernet Sauvignon wine under four oxygenation treatments at dosage rates of 0, 10, 23 and 36 mg/L/mth. The total period of the trial was 105 days. HPLC analysis indicated that the rate change of low molecular weight polyphenols is directly related to the oxygen dosage rate. The concentration of the majority of the identifiable monomers, most notably the anthocyanins decreased throughout the course of the trial. The rate of decrease was directly related to oxygen dosage rate. Thiolysis results showed an increase in mDP for all treatments over the course of the trial until day 77 when they were observed to decrease for all treatments. The decrease in mDP coincided with an addition of SO2 which was investigated in a subsequent trial. Spectrophotometric results indicated that the rate of formation of non-bleachable pigments was directly related to the rate of oxygen dosage with significant differences between the high rates (23 and 36 mg/L/mth) and the low rates (0 and 10 mg/L/mth). The trend for all treatments was for increased levels of stable pigments. The sensory results show that the measured organoleptic temporal development exhibits a similar oscillatory behaviour compared to the anecdotally derived curve presented in figure 1-2. The distinction between the respective phases described in section 1.1.1 was, however less clear. The most significant factor in the model weighting was mouthfeel and astringency which correlates with the observed changes occurring in the wine polypenols during maturation. Overall the laboratory scale trial showed that the chemical polyphenol development was directly related to the oxygen dosage rate. The sensory evolution also appeared to be accelerated with higher oxygen dosage rates, although the oscillatory nature of the sensory response given a single linear input indicates a complex underlying mechanism driving the changes. The effect of SO2 on the development of wine polyphenols with and without oxygen was also investigated. The presence of SO2 was found to have a significant effect on both mDP and the concentration of non-bleachable pigments. mDP was observed to decrease over the six week trial period irrespective of whether oxygen had been added or not. The mDP for the treatments without SO2 increased steadily over the course of the trial. Similarly the formation of non-bleachable pigments was suppressed and even retarded with SO2 present whereas for the treatments without SO2 a steady increase was observed. The implication of these results is that SO2 may have a much larger effect on tannin development than oxygen. The use of electrochemical micro-oxidation (or ELMOX) was examined ostensibly to determine proof of concept and also compare the performance of glassy carbon and titanium as electrode materials against traditional micro-oxygenation. Notable transformations occurred with titanium showing higher levels of ethanal than the other treatments both chemically and by sensory measure. A greater rate of stable pigment formation was also observed for the titanium compared to the other treatments. The respective dosage rates for the glassy carbon ELMOX and traditional micro-oxygenation treatments were too low to be able to discriminate any significant differences compared to the control wine. / AGMARDT Doctoral Scholarship

The overall aim of this thesis is to determine if the interactions between normal and waxy rice starch and milk proteins from four milk protein ingredients, namely skim milk powder (SMP), milk protein concentrate (MPC), sodium caseinate (NaCAS) and whey protein isolate (WPI) do occur, and to identify the mechanisms underlying these interactions. Different milk protein ingredients at various concentrations (0 to 10%, w/w) affected markedly and differently the pasting behaviour of 10% (w/w) rice starches. SMP delayed the pasting of both rice starches by increasing the onset temperature (Tonset) and the peak viscosity temperature (Tpeak) of pasting. This was mainly due to the presence of lactose and ions, which was further supported by the investigation of the effects of UFSMP (a solution of salts and lactose present in SMP at their proper concentration) and lactose. The addition of NaCAS also delayed the pasting of rice starch; Tpeak in the case of both starches was increased. For normal rice starch paste, MPC and WPI decreased the Tpeak. MPC had no affect on Tpeak of waxy rice starch paste. The qualitative viscoelastic behaviour of rice starch/milk protein ingredient gels obtained from the above pastes was dominated by the continuous phase made of the starch molecules. There was evidence, as indicated by confocal microscopy, of phase separation between the milk proteins of SMP and MPC and the two starches. The phase separation was not observed in the addition of either NaCAS or WPI. Studies on the thermal behaviour of rice starch/milk protein ingredient mixtures by differential scanning calorimetry (DSC) showed that SMP, similarly to UFSMP, delayed the gelatinization of both starches. NaCAS also delayed the gelatinisation of both starches but had a greater effect on waxy than normal rice starch. The addition of NaCAS did not affect Tonset but increased Tpeak for normal rice starch, whereas the gelatinisation temperature of waxy rice starch was highly affected by the addition of NaCAS with both Tonset and Tpeak shifted to higher temperatures. MPC had no affect on the gelatinization temperature of normal rice starch, whereas the gelatinization temperature of waxy rice starch was increased by the addition of MPC. The addition of WPI to both rice starches showed two thermal transitions. The first of these was due to the gelatinisation of the starches and the second to the denaturation of ß-lactoglobulin (ß-lg). The addition of WPI to normal rice starch showed that the thermal behaviour of normal starch and protein were independent from each other. In contrast, the thermal behaviour of waxy rice starch was modified by the addition of WPI; both Tonset and Tpeak were increased. SMP decreased the Tonset of swelling, swelling ratio and the amount of starch leaching from both starches. These observed changes were due to the presence of lactose and ions in SMP. NaCAS slightly increased Tonset of swelling but the amount of starch leaching was reduced for both rice starches. The rigidity of both starches tended to increase in the presence of NaCAS. MPC and WPI affected the swelling behaviour of normal and waxy rice starch differently. A dramatic increase in the swelling of normal rice starch/MPC or WPI mixtures was observed, whereas this trend was not evident for waxy rice starch/ MPC or WPI mixtures. The difference in the water holding ability and gelatinization peak temperatures of the two starches over the temperature range at which whey proteins denature and form gels are believed to be responsible for the observed differences. The results from confocal microscopy showed that milk proteins, such as a-casein, ß- casein, ß-lg and a-lactalbumin (a-la), were adsorbed onto the granule surface of both normal and waxy rice starch. The mechanism for this adsorption is the hydrophilic interactions; hydrogen bonds between hydroxyl group from terminated glucan molecule that protrude around starch granule surface-hydroxyl; amino, or other electron-donation or electron-accepting groups of the added proteins. Using sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) it was found that for SMP and MPC the adsorbed as- to ß-casein ratio on both starches was similar to the as-casein to ß- casein ratio in the casein micelle at low SMP and MPC concentrations. But at high concentrations of SMP or MPC, this ratio decreased indicating that more ß-casein was adsorbed preferentially to as-casein. In the case of NaCAS, as-casein was adsorbed preferentially to ß-casein. Moreover, there was evidence of multilayer adsorption of ascasein into the surface of rice starch granules. Compared to the other milk protein ingredients, very small amounts of the ß-lg and a-la from WPI were adsorbed onto starch granules. However, the adsorbed amounts of ß-lg and a-la from WPI continuously increased with increasing WPI concentration, suggesting that these two proteins, particularly ß-lg, adsorbed in multilayers too.

A shelf life trial using a fully balanced factorial experimental design was used to analyse the effects of acidity and light on broccoli juice made on a semi commercial scale over an eight week period in simulated retail refrigerated storage conditions. The research focused on making broccoli juice on a pilot scale, and what happens to the colour, composition and flavour during storage. A pilot scale production of pasteurised broccoli juice was conducted and the juice satisfied microbiological safety limits for the eight week shelf life trial in retail storage conditions. The stability of the green colour of fresh broccoli through processing and storage was assessed. Neutral broccoli juice remained green for four weeks before the colour became more yellow. The acidified juice became yellow on acidification and did not change significantly during storage. Dietary fibre and pectin levels did not change during storage. Chlorophyll and carotenoids levels decreased during storage and were directly influencing the colour changes in the juices. Ascorbic acid levels decreased significantly during processing resulting in low ascorbic acid levels (12 - 15 mg /100ml of juice) at the start of the shelf life trial and dropped further to 2-6 mg /100ml of juice after eight weeks. Acidification and storage in the dark had a protective effect on the degradation of ascorbic acid with only a 58% reduction in ascorbic acid levels compared to an 84% reduction in neutral light stored broccoli juice. The effect of processing and storage on the flavour of the beverage was assessed using a trained sensory panel providing descriptive analysis. The sensory profiles for neutral and acidified juices were extremely different with the unbalanced acidity suppressing the perception of the basic tastes, sweet, salty and bitter. The neutral juice sensory profile only changed slightly in aroma attributes during storage for seven weeks. The astringent aftertaste of the acidified juice increased while the broccoli smell decreased during storage. The results from this research indicate that the production of a broccoli juice with a yellow green colour and some retained nutritional components is achievable with a refrigerated (4 °C) shelf life of 30 days in light excluding glass packaging. The neutral juice is recommended as it was greener and had a broccoli flavour.A shelf life trial using a fully balanced factorial experimental design was used to analyse the effects of acidity and light on broccoli juice made on a semi commercial scale over an eight week period in simulated retail refrigerated storage conditions. The research focused on making broccoli juice on a pilot scale, and what happens to the colour, composition and flavour during storage. A pilot scale production of pasteurised broccoli juice was conducted and the juice satisfied microbiological safety limits for the eight week shelf life trial in retail storage conditions. The stability of the green colour of fresh broccoli through processing and storage was assessed. Neutral broccoli juice remained green for four weeks before the colour became more yellow. The acidified juice became yellow on acidification and did not change significantly during storage. Dietary fibre and pectin levels did not change during storage. Chlorophyll and carotenoids levels decreased during storage and were directly influencing the colour changes in the juices. Ascorbic acid levels decreased significantly during processing resulting in low ascorbic acid levels (12 - 15 mg /100ml of juice) at the start of the shelf life trial and dropped further to 2-6 mg /100ml of juice after eight weeks. Acidification and storage in the dark had a protective effect on the degradation of ascorbic acid with only a 58% reduction in ascorbic acid levels compared to an 84% reduction in neutral light stored broccoli juice. The effect of processing and storage on the flavour of the beverage was assessed using a trained sensory panel providing descriptive analysis. The sensory profiles for neutral and acidified juices were extremely different with the unbalanced acidity suppressing the perception of the basic tastes, sweet, salty and bitter. The neutral juice sensory profile only changed slightly in aroma attributes during storage for seven weeks. The astringent aftertaste of the acidified juice increased while the broccoli smell decreased during storage. The results from this research indicate that the production of a broccoli juice with a yellow green colour and some retained nutritional components is achievable with a refrigerated (4 °C) shelf life of 30 days in light excluding glass packaging. The neutral juice is recommended as it was greener and had a broccoli flavour.

Emulsions containing three different types of milk fat fractions (MF13, MF27 and MF42) and anhydrous milk fat (AMF) were prepared at oil to water (O/W) ratios of 1:9, 3:7, 5:5 and 7:3 using rhamnolipids as emulsifiers. The prepared emulsions were analyzed for their storage stability and properties (colour, particle size, zeta potential and rheology). The effects of various factors (freezing/thawing, heating, pH, salts and ionic strength) on the stability of emulsions were also investigated. All emulsions prepared with an O/W ratio of 7:3, regardless of the type of milk fat, rendered a highly condensed, semi solid and cream-like substance whereas other emulsions containing less oil were in a liquid form. Among the four different O/W ratios tested, the highest emulsion stability during the storage of 12 weeks was observed from the emulsions containing 1:9 O/W ratios, due to a combine effect of smaller emulsion particle size and lower collision frequency between droplets. Interestingly, the emulsions with 7:3 O/W ratios were found to be more stable than the ones with 5:5 O/W ratios. This might be due to the limited movements of closely-packed emulsion droplets induced by the high oil concentration of 7:3 O/W ratios. The emulsion stability was significantly affected by low pH, especially at lower than pH 4, due to the loss of electrostatic repulsions between droplets leading to droplet coalescence and also possibly due to hydrolysis of rhamnolipid molecules. The presence of salts (NaCl, KCl and CaCl2) also rendered the emulsion unstable. The degree of instability was gradually increased with increasing salt concentrations. CaCl2 had the most significant effect even at a very low concentration. The viscosity of emulsions increased with increasing oil concentration but was not affected by the types of milk fats. Emulsions with 3:7, 5:5 and 7:3 O/W ratios exhibited non-Newtonian and shear thinning flow behaviour. At 7:3 O/W ratios, MF13 exhibited gel-like properties whereas both MF42 and AMF emulsions became more solid-like at higher frequency.

The overall aim of this thesis is to determine if the interactions between normal and waxy rice starch and milk proteins from four milk protein ingredients, namely skim milk powder (SMP), milk protein concentrate (MPC), sodium caseinate (NaCAS) and whey protein isolate (WPI) do occur, and to identify the mechanisms underlying these interactions. Different milk protein ingredients at various concentrations (0 to 10%, w/w) affected markedly and differently the pasting behaviour of 10% (w/w) rice starches. SMP delayed the pasting of both rice starches by increasing the onset temperature (Tonset) and the peak viscosity temperature (Tpeak) of pasting. This was mainly due to the presence of lactose and ions, which was further supported by the investigation of the effects of UFSMP (a solution of salts and lactose present in SMP at their proper concentration) and lactose. The addition of NaCAS also delayed the pasting of rice starch; Tpeak in the case of both starches was increased. For normal rice starch paste, MPC and WPI decreased the Tpeak. MPC had no affect on Tpeak of waxy rice starch paste. The qualitative viscoelastic behaviour of rice starch/milk protein ingredient gels obtained from the above pastes was dominated by the continuous phase made of the starch molecules. There was evidence, as indicated by confocal microscopy, of phase separation between the milk proteins of SMP and MPC and the two starches. The phase separation was not observed in the addition of either NaCAS or WPI. Studies on the thermal behaviour of rice starch/milk protein ingredient mixtures by differential scanning calorimetry (DSC) showed that SMP, similarly to UFSMP, delayed the gelatinization of both starches. NaCAS also delayed the gelatinisation of both starches but had a greater effect on waxy than normal rice starch. The addition of NaCAS did not affect Tonset but increased Tpeak for normal rice starch, whereas the gelatinisation temperature of waxy rice starch was highly affected by the addition of NaCAS with both Tonset and Tpeak shifted to higher temperatures. MPC had no affect on the gelatinization temperature of normal rice starch, whereas the gelatinization temperature of waxy rice starch was increased by the addition of MPC. The addition of WPI to both rice starches showed two thermal transitions. The first of these was due to the gelatinisation of the starches and the second to the denaturation of ß-lactoglobulin (ß-lg). The addition of WPI to normal rice starch showed that the thermal behaviour of normal starch and protein were independent from each other. In contrast, the thermal behaviour of waxy rice starch was modified by the addition of WPI; both Tonset and Tpeak were increased. SMP decreased the Tonset of swelling, swelling ratio and the amount of starch leaching from both starches. These observed changes were due to the presence of lactose and ions in SMP. NaCAS slightly increased Tonset of swelling but the amount of starch leaching was reduced for both rice starches. The rigidity of both starches tended to increase in the presence of NaCAS. MPC and WPI affected the swelling behaviour of normal and waxy rice starch differently. A dramatic increase in the swelling of normal rice starch/MPC or WPI mixtures was observed, whereas this trend was not evident for waxy rice starch/ MPC or WPI mixtures. The difference in the water holding ability and gelatinization peak temperatures of the two starches over the temperature range at which whey proteins denature and form gels are believed to be responsible for the observed differences. The results from confocal microscopy showed that milk proteins, such as a-casein, ß- casein, ß-lg and a-lactalbumin (a-la), were adsorbed onto the granule surface of both normal and waxy rice starch. The mechanism for this adsorption is the hydrophilic interactions; hydrogen bonds between hydroxyl group from terminated glucan molecule that protrude around starch granule surface-hydroxyl; amino, or other electron-donation or electron-accepting groups of the added proteins. Using sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) it was found that for SMP and MPC the adsorbed as- to ß-casein ratio on both starches was similar to the as-casein to ß- casein ratio in the casein micelle at low SMP and MPC concentrations. But at high concentrations of SMP or MPC, this ratio decreased indicating that more ß-casein was adsorbed preferentially to as-casein. In the case of NaCAS, as-casein was adsorbed preferentially to ß-casein. Moreover, there was evidence of multilayer adsorption of ascasein into the surface of rice starch granules. Compared to the other milk protein ingredients, very small amounts of the ß-lg and a-la from WPI were adsorbed onto starch granules. However, the adsorbed amounts of ß-lg and a-la from WPI continuously increased with increasing WPI concentration, suggesting that these two proteins, particularly ß-lg, adsorbed in multilayers too.

The overall aim of this thesis is to determine if the interactions between normal and waxy rice starch and milk proteins from four milk protein ingredients, namely skim milk powder (SMP), milk protein concentrate (MPC), sodium caseinate (NaCAS) and whey protein isolate (WPI) do occur, and to identify the mechanisms underlying these interactions. Different milk protein ingredients at various concentrations (0 to 10%, w/w) affected markedly and differently the pasting behaviour of 10% (w/w) rice starches. SMP delayed the pasting of both rice starches by increasing the onset temperature (Tonset) and the peak viscosity temperature (Tpeak) of pasting. This was mainly due to the presence of lactose and ions, which was further supported by the investigation of the effects of UFSMP (a solution of salts and lactose present in SMP at their proper concentration) and lactose. The addition of NaCAS also delayed the pasting of rice starch; Tpeak in the case of both starches was increased. For normal rice starch paste, MPC and WPI decreased the Tpeak. MPC had no affect on Tpeak of waxy rice starch paste. The qualitative viscoelastic behaviour of rice starch/milk protein ingredient gels obtained from the above pastes was dominated by the continuous phase made of the starch molecules. There was evidence, as indicated by confocal microscopy, of phase separation between the milk proteins of SMP and MPC and the two starches. The phase separation was not observed in the addition of either NaCAS or WPI. Studies on the thermal behaviour of rice starch/milk protein ingredient mixtures by differential scanning calorimetry (DSC) showed that SMP, similarly to UFSMP, delayed the gelatinization of both starches. NaCAS also delayed the gelatinisation of both starches but had a greater effect on waxy than normal rice starch. The addition of NaCAS did not affect Tonset but increased Tpeak for normal rice starch, whereas the gelatinisation temperature of waxy rice starch was highly affected by the addition of NaCAS with both Tonset and Tpeak shifted to higher temperatures. MPC had no affect on the gelatinization temperature of normal rice starch, whereas the gelatinization temperature of waxy rice starch was increased by the addition of MPC. The addition of WPI to both rice starches showed two thermal transitions. The first of these was due to the gelatinisation of the starches and the second to the denaturation of ß-lactoglobulin (ß-lg). The addition of WPI to normal rice starch showed that the thermal behaviour of normal starch and protein were independent from each other. In contrast, the thermal behaviour of waxy rice starch was modified by the addition of WPI; both Tonset and Tpeak were increased. SMP decreased the Tonset of swelling, swelling ratio and the amount of starch leaching from both starches. These observed changes were due to the presence of lactose and ions in SMP. NaCAS slightly increased Tonset of swelling but the amount of starch leaching was reduced for both rice starches. The rigidity of both starches tended to increase in the presence of NaCAS. MPC and WPI affected the swelling behaviour of normal and waxy rice starch differently. A dramatic increase in the swelling of normal rice starch/MPC or WPI mixtures was observed, whereas this trend was not evident for waxy rice starch/ MPC or WPI mixtures. The difference in the water holding ability and gelatinization peak temperatures of the two starches over the temperature range at which whey proteins denature and form gels are believed to be responsible for the observed differences. The results from confocal microscopy showed that milk proteins, such as a-casein, ß- casein, ß-lg and a-lactalbumin (a-la), were adsorbed onto the granule surface of both normal and waxy rice starch. The mechanism for this adsorption is the hydrophilic interactions; hydrogen bonds between hydroxyl group from terminated glucan molecule that protrude around starch granule surface-hydroxyl; amino, or other electron-donation or electron-accepting groups of the added proteins. Using sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) it was found that for SMP and MPC the adsorbed as- to ß-casein ratio on both starches was similar to the as-casein to ß- casein ratio in the casein micelle at low SMP and MPC concentrations. But at high concentrations of SMP or MPC, this ratio decreased indicating that more ß-casein was adsorbed preferentially to as-casein. In the case of NaCAS, as-casein was adsorbed preferentially to ß-casein. Moreover, there was evidence of multilayer adsorption of ascasein into the surface of rice starch granules. Compared to the other milk protein ingredients, very small amounts of the ß-lg and a-la from WPI were adsorbed onto starch granules. However, the adsorbed amounts of ß-lg and a-la from WPI continuously increased with increasing WPI concentration, suggesting that these two proteins, particularly ß-lg, adsorbed in multilayers too.

The overall aim of this thesis is to determine if the interactions between normal and waxy rice starch and milk proteins from four milk protein ingredients, namely skim milk powder (SMP), milk protein concentrate (MPC), sodium caseinate (NaCAS) and whey protein isolate (WPI) do occur, and to identify the mechanisms underlying these interactions. Different milk protein ingredients at various concentrations (0 to 10%, w/w) affected markedly and differently the pasting behaviour of 10% (w/w) rice starches. SMP delayed the pasting of both rice starches by increasing the onset temperature (Tonset) and the peak viscosity temperature (Tpeak) of pasting. This was mainly due to the presence of lactose and ions, which was further supported by the investigation of the effects of UFSMP (a solution of salts and lactose present in SMP at their proper concentration) and lactose. The addition of NaCAS also delayed the pasting of rice starch; Tpeak in the case of both starches was increased. For normal rice starch paste, MPC and WPI decreased the Tpeak. MPC had no affect on Tpeak of waxy rice starch paste. The qualitative viscoelastic behaviour of rice starch/milk protein ingredient gels obtained from the above pastes was dominated by the continuous phase made of the starch molecules. There was evidence, as indicated by confocal microscopy, of phase separation between the milk proteins of SMP and MPC and the two starches. The phase separation was not observed in the addition of either NaCAS or WPI. Studies on the thermal behaviour of rice starch/milk protein ingredient mixtures by differential scanning calorimetry (DSC) showed that SMP, similarly to UFSMP, delayed the gelatinization of both starches. NaCAS also delayed the gelatinisation of both starches but had a greater effect on waxy than normal rice starch. The addition of NaCAS did not affect Tonset but increased Tpeak for normal rice starch, whereas the gelatinisation temperature of waxy rice starch was highly affected by the addition of NaCAS with both Tonset and Tpeak shifted to higher temperatures. MPC had no affect on the gelatinization temperature of normal rice starch, whereas the gelatinization temperature of waxy rice starch was increased by the addition of MPC. The addition of WPI to both rice starches showed two thermal transitions. The first of these was due to the gelatinisation of the starches and the second to the denaturation of ß-lactoglobulin (ß-lg). The addition of WPI to normal rice starch showed that the thermal behaviour of normal starch and protein were independent from each other. In contrast, the thermal behaviour of waxy rice starch was modified by the addition of WPI; both Tonset and Tpeak were increased. SMP decreased the Tonset of swelling, swelling ratio and the amount of starch leaching from both starches. These observed changes were due to the presence of lactose and ions in SMP. NaCAS slightly increased Tonset of swelling but the amount of starch leaching was reduced for both rice starches. The rigidity of both starches tended to increase in the presence of NaCAS. MPC and WPI affected the swelling behaviour of normal and waxy rice starch differently. A dramatic increase in the swelling of normal rice starch/MPC or WPI mixtures was observed, whereas this trend was not evident for waxy rice starch/ MPC or WPI mixtures. The difference in the water holding ability and gelatinization peak temperatures of the two starches over the temperature range at which whey proteins denature and form gels are believed to be responsible for the observed differences. The results from confocal microscopy showed that milk proteins, such as a-casein, ß- casein, ß-lg and a-lactalbumin (a-la), were adsorbed onto the granule surface of both normal and waxy rice starch. The mechanism for this adsorption is the hydrophilic interactions; hydrogen bonds between hydroxyl group from terminated glucan molecule that protrude around starch granule surface-hydroxyl; amino, or other electron-donation or electron-accepting groups of the added proteins. Using sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) it was found that for SMP and MPC the adsorbed as- to ß-casein ratio on both starches was similar to the as-casein to ß- casein ratio in the casein micelle at low SMP and MPC concentrations. But at high concentrations of SMP or MPC, this ratio decreased indicating that more ß-casein was adsorbed preferentially to as-casein. In the case of NaCAS, as-casein was adsorbed preferentially to ß-casein. Moreover, there was evidence of multilayer adsorption of ascasein into the surface of rice starch granules. Compared to the other milk protein ingredients, very small amounts of the ß-lg and a-la from WPI were adsorbed onto starch granules. However, the adsorbed amounts of ß-lg and a-la from WPI continuously increased with increasing WPI concentration, suggesting that these two proteins, particularly ß-lg, adsorbed in multilayers too.

The overall aim of this thesis is to determine if the interactions between normal and waxy rice starch and milk proteins from four milk protein ingredients, namely skim milk powder (SMP), milk protein concentrate (MPC), sodium caseinate (NaCAS) and whey protein isolate (WPI) do occur, and to identify the mechanisms underlying these interactions. Different milk protein ingredients at various concentrations (0 to 10%, w/w) affected markedly and differently the pasting behaviour of 10% (w/w) rice starches. SMP delayed the pasting of both rice starches by increasing the onset temperature (Tonset) and the peak viscosity temperature (Tpeak) of pasting. This was mainly due to the presence of lactose and ions, which was further supported by the investigation of the effects of UFSMP (a solution of salts and lactose present in SMP at their proper concentration) and lactose. The addition of NaCAS also delayed the pasting of rice starch; Tpeak in the case of both starches was increased. For normal rice starch paste, MPC and WPI decreased the Tpeak. MPC had no affect on Tpeak of waxy rice starch paste. The qualitative viscoelastic behaviour of rice starch/milk protein ingredient gels obtained from the above pastes was dominated by the continuous phase made of the starch molecules. There was evidence, as indicated by confocal microscopy, of phase separation between the milk proteins of SMP and MPC and the two starches. The phase separation was not observed in the addition of either NaCAS or WPI. Studies on the thermal behaviour of rice starch/milk protein ingredient mixtures by differential scanning calorimetry (DSC) showed that SMP, similarly to UFSMP, delayed the gelatinization of both starches. NaCAS also delayed the gelatinisation of both starches but had a greater effect on waxy than normal rice starch. The addition of NaCAS did not affect Tonset but increased Tpeak for normal rice starch, whereas the gelatinisation temperature of waxy rice starch was highly affected by the addition of NaCAS with both Tonset and Tpeak shifted to higher temperatures. MPC had no affect on the gelatinization temperature of normal rice starch, whereas the gelatinization temperature of waxy rice starch was increased by the addition of MPC. The addition of WPI to both rice starches showed two thermal transitions. The first of these was due to the gelatinisation of the starches and the second to the denaturation of ß-lactoglobulin (ß-lg). The addition of WPI to normal rice starch showed that the thermal behaviour of normal starch and protein were independent from each other. In contrast, the thermal behaviour of waxy rice starch was modified by the addition of WPI; both Tonset and Tpeak were increased. SMP decreased the Tonset of swelling, swelling ratio and the amount of starch leaching from both starches. These observed changes were due to the presence of lactose and ions in SMP. NaCAS slightly increased Tonset of swelling but the amount of starch leaching was reduced for both rice starches. The rigidity of both starches tended to increase in the presence of NaCAS. MPC and WPI affected the swelling behaviour of normal and waxy rice starch differently. A dramatic increase in the swelling of normal rice starch/MPC or WPI mixtures was observed, whereas this trend was not evident for waxy rice starch/ MPC or WPI mixtures. The difference in the water holding ability and gelatinization peak temperatures of the two starches over the temperature range at which whey proteins denature and form gels are believed to be responsible for the observed differences. The results from confocal microscopy showed that milk proteins, such as a-casein, ß- casein, ß-lg and a-lactalbumin (a-la), were adsorbed onto the granule surface of both normal and waxy rice starch. The mechanism for this adsorption is the hydrophilic interactions; hydrogen bonds between hydroxyl group from terminated glucan molecule that protrude around starch granule surface-hydroxyl; amino, or other electron-donation or electron-accepting groups of the added proteins. Using sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) it was found that for SMP and MPC the adsorbed as- to ß-casein ratio on both starches was similar to the as-casein to ß- casein ratio in the casein micelle at low SMP and MPC concentrations. But at high concentrations of SMP or MPC, this ratio decreased indicating that more ß-casein was adsorbed preferentially to as-casein. In the case of NaCAS, as-casein was adsorbed preferentially to ß-casein. Moreover, there was evidence of multilayer adsorption of ascasein into the surface of rice starch granules. Compared to the other milk protein ingredients, very small amounts of the ß-lg and a-la from WPI were adsorbed onto starch granules. However, the adsorbed amounts of ß-lg and a-la from WPI continuously increased with increasing WPI concentration, suggesting that these two proteins, particularly ß-lg, adsorbed in multilayers too.

The overall aim of this thesis is to determine if the interactions between normal and waxy rice starch and milk proteins from four milk protein ingredients, namely skim milk powder (SMP), milk protein concentrate (MPC), sodium caseinate (NaCAS) and whey protein isolate (WPI) do occur, and to identify the mechanisms underlying these interactions. Different milk protein ingredients at various concentrations (0 to 10%, w/w) affected markedly and differently the pasting behaviour of 10% (w/w) rice starches. SMP delayed the pasting of both rice starches by increasing the onset temperature (Tonset) and the peak viscosity temperature (Tpeak) of pasting. This was mainly due to the presence of lactose and ions, which was further supported by the investigation of the effects of UFSMP (a solution of salts and lactose present in SMP at their proper concentration) and lactose. The addition of NaCAS also delayed the pasting of rice starch; Tpeak in the case of both starches was increased. For normal rice starch paste, MPC and WPI decreased the Tpeak. MPC had no affect on Tpeak of waxy rice starch paste. The qualitative viscoelastic behaviour of rice starch/milk protein ingredient gels obtained from the above pastes was dominated by the continuous phase made of the starch molecules. There was evidence, as indicated by confocal microscopy, of phase separation between the milk proteins of SMP and MPC and the two starches. The phase separation was not observed in the addition of either NaCAS or WPI. Studies on the thermal behaviour of rice starch/milk protein ingredient mixtures by differential scanning calorimetry (DSC) showed that SMP, similarly to UFSMP, delayed the gelatinization of both starches. NaCAS also delayed the gelatinisation of both starches but had a greater effect on waxy than normal rice starch. The addition of NaCAS did not affect Tonset but increased Tpeak for normal rice starch, whereas the gelatinisation temperature of waxy rice starch was highly affected by the addition of NaCAS with both Tonset and Tpeak shifted to higher temperatures. MPC had no affect on the gelatinization temperature of normal rice starch, whereas the gelatinization temperature of waxy rice starch was increased by the addition of MPC. The addition of WPI to both rice starches showed two thermal transitions. The first of these was due to the gelatinisation of the starches and the second to the denaturation of ß-lactoglobulin (ß-lg). The addition of WPI to normal rice starch showed that the thermal behaviour of normal starch and protein were independent from each other. In contrast, the thermal behaviour of waxy rice starch was modified by the addition of WPI; both Tonset and Tpeak were increased. SMP decreased the Tonset of swelling, swelling ratio and the amount of starch leaching from both starches. These observed changes were due to the presence of lactose and ions in SMP. NaCAS slightly increased Tonset of swelling but the amount of starch leaching was reduced for both rice starches. The rigidity of both starches tended to increase in the presence of NaCAS. MPC and WPI affected the swelling behaviour of normal and waxy rice starch differently. A dramatic increase in the swelling of normal rice starch/MPC or WPI mixtures was observed, whereas this trend was not evident for waxy rice starch/ MPC or WPI mixtures. The difference in the water holding ability and gelatinization peak temperatures of the two starches over the temperature range at which whey proteins denature and form gels are believed to be responsible for the observed differences. The results from confocal microscopy showed that milk proteins, such as a-casein, ß- casein, ß-lg and a-lactalbumin (a-la), were adsorbed onto the granule surface of both normal and waxy rice starch. The mechanism for this adsorption is the hydrophilic interactions; hydrogen bonds between hydroxyl group from terminated glucan molecule that protrude around starch granule surface-hydroxyl; amino, or other electron-donation or electron-accepting groups of the added proteins. Using sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) it was found that for SMP and MPC the adsorbed as- to ß-casein ratio on both starches was similar to the as-casein to ß- casein ratio in the casein micelle at low SMP and MPC concentrations. But at high concentrations of SMP or MPC, this ratio decreased indicating that more ß-casein was adsorbed preferentially to as-casein. In the case of NaCAS, as-casein was adsorbed preferentially to ß-casein. Moreover, there was evidence of multilayer adsorption of ascasein into the surface of rice starch granules. Compared to the other milk protein ingredients, very small amounts of the ß-lg and a-la from WPI were adsorbed onto starch granules. However, the adsorbed amounts of ß-lg and a-la from WPI continuously increased with increasing WPI concentration, suggesting that these two proteins, particularly ß-lg, adsorbed in multilayers too.