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京都大学 / 新制・課程博士 / 博士(農学) / 甲第23237号 / 農博第2444号 / 新制||農||1083(附属図書館) / 学位論文||R3||N5327(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 植田 充美, 教授 小川 順, 教授 栗原 達夫 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Canada is a major global producer of pulse products including pulse starch, which notably contributes to a healthy diet. Strategically, Canada is taking steps to research methods of adding greater value to these crop products, and functional foods like resistant starch are particularly interesting. The primary objectives of this study were to develop an effective reactive extrusion process to produce gelatinized lipid complexed pea starches with enhanced enzyme resistance and examine the effects of bulk lipid complexing conditions on physicochemical and functional properties of extruded pea starches. One type of commercially available pea starch, Nutri-Pea, was chosen as the research subject in this study. A number of methods including; Englyst digestion method, differential scanning calorimetry (DSC), infrared spectroscopy (FTIR), contact angle, titrations, residence time distribution (RTD) and rapid visco analysis (RVA) were used to characterize the properties of extruded pea starches. The effects of feed formulation and extrusion conditions on lipid complexing and Englyst digestion profiles were systematically examined on two mixing devices. An extensive kinetics study was conducted on a lab scale twin-screw compounder, DSM-Xplore. The process was then scaled up to produce bulk lipid complexed pea starch on a Leistritz twin-screw extruder. The results showed that lipid complexing and digestion profiles were highly dependent on feed moisture and induced screw shear. Reactive extrusion of pea starches under optimized conditions achieved a significant but moderate increase in either resistant starch (RS) content (from 13.3% to 20.2%) or slowly digestible starch (SDS) content (from 7.85% to 23.3%) compared to their native counterparts. However, RS and SDS content could not be improved simultaneously based on the pea starch and extrusion process in this study. Increased degree of substitution (DS) was found for myristic acid complexed pea starches (nominal DS= ~0.8) when compared to palmitic acid complexed pea starch (nominal DS= ~0.5). Contact angle measurements, FTIR and DSC thermograms confirmed the presence of lipids. Lipid complexed starch films showed increasing hydrophobicity with increasing lipid content. As an alternative product compared to functional foods, the modified starch was considered as a biodegradable film for industrial applications. The material was produced at the highest moisture content for extruded native starch and two concentrations of lipid complexed starch using an intensive screw design. Preliminary results show that increasing lipid content and adding 1% glycerol to samples decreases the force per film thickness required to puncture films. However further investigation is required to determine effect of heat and moisture deformation. / Thesis / Master of Applied Science (MASc) / Incorporation of pulses into food products has been a major area of Canadian research for its potential to create new avenues of enzyme resistant food starches. Extrusion cooking is commonly used in industry for producing various food products such as snacks and cereals but little research has been reported on using an extruder to rapidly produce resistant pulse starches as a functional ingredient on a large scale; resistant starch is a functional food beneficial to humans in the same manner as insoluble fiber but exhibits improved textural properties. This study aimed to develop an effective reactive extrusion process to produce lipid complexed pea starches with enhanced enzyme resistance (i.e. increased slowly digestible starch (SDS) and resistant starch (RS) content) by an examination of the effects of reaction conditions on the properties of extrusion products. The lipid complexed pea starches under optimized conditions achieved a significant but moderate increase in either RS content or SDS content depending on the sample formulation compared to their native counterparts. However, RS and SDS content could not simultaneously be improved in this study.

Research studies show that an average American consumes 3,400 to 4,500 mg/day of sodium in their diet against the dietary recommendation of 1,500 to 2,300 mg/day. The majority of this sodium comes from processed foods. Excess sodium in the diet can potentially cause health issues such as hypertension, heart attack, kidney failure, and bone problems. The objective of this study is to understand sodium diffusion in a protein model using turkey breast meat and determine the opportunities of its reduction by changing the process conditions and combining table salt with other sodium salts. Since proteins are a complicated system and there are very few research studies available on meat in this area, it is necessary to understand the trend of sodium diffusion in the meat while cooking in presence of salt at different temperatures followed by analyzing the change in the diffusion in presence of different sodium salts with anions larger than sodium chloride. Since protein denaturation can have impact on the diffusion process, the denaturation profile for turkey breast meat protein was analyzed by Differential Scanning Colorimetry (DSC). The DSC results were visually confirmed by SEM analysis. After obtaining results from DSC and SEM analysis, the processing temperature of 4oC, 23oC, 50oC, 70oC and 90oC was implemented to obtain the diffusion trend before, during, and after protein denaturation. 1-inch meat cubes were cooked in 5% (0.86M/L) salt solution at different time and temperature combinations. Sodium analysis was performed on Ion Selective Electrode (ISE) and the results were confirmed by ICP-OES. This study showed that the sodium content of the processed samples was not proportional to the treatment temperature. The Study was also conducted to analyze the effects of sodium salts with larger anions than sodium chloride on sodium diffusion during thermal applications. Sodium Phosphate, Sodium Tripolyphosphate, Sodium Sulfate and Monosodium Glutamate were incorporated in the study. Sodium content in the samples processed in this salts showed 32-46% reduction compared to those processed in sodium chloride. Since sodium phosphate is widely used in the meat industry, it may have the most potential to partially replace salt. Texture analysis was performed on the samples cooked in sodium chloride and sodium phosphate solutions in order to determine changes in textural properties. A brief sensory test was also conducted with 10 participants to investigate the preference of sodium phosphate incorporation in the meat and to identify any after taste.

This thesis investigates a newly developed and patented technology for its ability to inactivate spore- forming bacteria and non-spore-forming microorganisms. This new technology “Pressure Assisted Thermal Sterilization©” (PATS) is based on the theory of the thermal expansion of liquids. The efficiency of inactivating spore-forming and non-spore-forming microorganisms by PATS was compared with the thermal treatment alone. A combination treatment consisting of high pressure processing and gaseous carbon dioxide was also investigated for its ability to inactivate bacterial spores in model and real food matrices. The structural damage caused by treatments to the spores and non-spore-forming bacteria was assessed by scanning electron microscopy. Geobacillus stearothermophilus spores suspended in Milli-Q water, UHT milk and pumpkin soup, treated by PATS were found to have significantly lower decimal reduction times (D values) compared with the thermal treatment alone. Spores suspended in UHT milk were more heat resistant compared with those in Milli-Q water and pumpkin soup. Bacillus cereus spores suspended in Milli-Q water and pumpkin soup treated with PATS were more effectively inactivated compared with spores treated by the thermal treatment alone. Clostridium botulinum spores in saline buffer subjected to PATS treatment were inactivated more effectively compared with the thermal treatment alone. Overall, the results show that PATS was a better processing technique for inactivation of bacterial spores compared with thermal treatment alone. However, PATS had no added benefit in inactivating the non-spore-forming bacteria Escherichia coli and Saccharomyces cerevisiae cells compared with the thermal treatment. A shelf life study showed that B. cereus spores in pumpkin soup retained a low spore count (<5 LogCFU/mL) for approximately 40 days in 30oC storage after treatment with PATS. No additional degradation of colour pigments of pumpkin soup and model pumpkin juice was observed following PATS compared with the thermal treatment. Spore-forming microorganisms can be resistant to pressure treatment alone, which limits the application of high pressure processing (HPP). Therefore, a combination approach was investigated. The mechanism of inactivating spores by combining HPP with other treatments is that the pressure assists in spore germination. Then a secondary treatment (thermal or CO2 gas) can be used to inactivate the germinated spores. A combined application of HPP and a consecutive CO2 treatment was investigated for the efficiency of spore inactivation. Results showed that HPP (200 MPa for 30 min) followed by a CO2 treatment inactivated Bacillus subtilis 168 in nutrient broth, tomato juice and liquid whole egg by 2.5, 1.0 and 1.5 LogCFU/mL respectively. These results indicated that this technique is inadequate for practical use. Scanning electron micrographs showed that pressure processing of B. subtilis 168 and B. subtilis natto spores resulted in deformation of the spore structure. This structural deformation of spores may have been due to water absorption during HPP and subsequent release upon decompression. PATS treated G. stearothermophilus and B. cereus spores were more severely damaged compared with the same spores which underwent thermal treatment alone. However, the extent to which E. coli and S. cerevisiae cells were damaged by both PATS and thermal treatment was similar.

This thesis investigates a newly developed and patented technology for its ability to inactivate spore- forming bacteria and non-spore-forming microorganisms. This new technology “Pressure Assisted Thermal Sterilization©” (PATS) is based on the theory of the thermal expansion of liquids. The efficiency of inactivating spore-forming and non-spore-forming microorganisms by PATS was compared with the thermal treatment alone. A combination treatment consisting of high pressure processing and gaseous carbon dioxide was also investigated for its ability to inactivate bacterial spores in model and real food matrices. The structural damage caused by treatments to the spores and non-spore-forming bacteria was assessed by scanning electron microscopy. Geobacillus stearothermophilus spores suspended in Milli-Q water, UHT milk and pumpkin soup, treated by PATS were found to have significantly lower decimal reduction times (D values) compared with the thermal treatment alone. Spores suspended in UHT milk were more heat resistant compared with those in Milli-Q water and pumpkin soup. Bacillus cereus spores suspended in Milli-Q water and pumpkin soup treated with PATS were more effectively inactivated compared with spores treated by the thermal treatment alone. Clostridium botulinum spores in saline buffer subjected to PATS treatment were inactivated more effectively compared with the thermal treatment alone. Overall, the results show that PATS was a better processing technique for inactivation of bacterial spores compared with thermal treatment alone. However, PATS had no added benefit in inactivating the non-spore-forming bacteria Escherichia coli and Saccharomyces cerevisiae cells compared with the thermal treatment. A shelf life study showed that B. cereus spores in pumpkin soup retained a low spore count (<5 LogCFU/mL) for approximately 40 days in 30oC storage after treatment with PATS. No additional degradation of colour pigments of pumpkin soup and model pumpkin juice was observed following PATS compared with the thermal treatment. Spore-forming microorganisms can be resistant to pressure treatment alone, which limits the application of high pressure processing (HPP). Therefore, a combination approach was investigated. The mechanism of inactivating spores by combining HPP with other treatments is that the pressure assists in spore germination. Then a secondary treatment (thermal or CO2 gas) can be used to inactivate the germinated spores. A combined application of HPP and a consecutive CO2 treatment was investigated for the efficiency of spore inactivation. Results showed that HPP (200 MPa for 30 min) followed by a CO2 treatment inactivated Bacillus subtilis 168 in nutrient broth, tomato juice and liquid whole egg by 2.5, 1.0 and 1.5 LogCFU/mL respectively. These results indicated that this technique is inadequate for practical use. Scanning electron micrographs showed that pressure processing of B. subtilis 168 and B. subtilis natto spores resulted in deformation of the spore structure. This structural deformation of spores may have been due to water absorption during HPP and subsequent release upon decompression. PATS treated G. stearothermophilus and B. cereus spores were more severely damaged compared with the same spores which underwent thermal treatment alone. However, the extent to which E. coli and S. cerevisiae cells were damaged by both PATS and thermal treatment was similar.

This thesis investigates a newly developed and patented technology for its ability to inactivate spore- forming bacteria and non-spore-forming microorganisms. This new technology “Pressure Assisted Thermal Sterilization©” (PATS) is based on the theory of the thermal expansion of liquids. The efficiency of inactivating spore-forming and non-spore-forming microorganisms by PATS was compared with the thermal treatment alone. A combination treatment consisting of high pressure processing and gaseous carbon dioxide was also investigated for its ability to inactivate bacterial spores in model and real food matrices. The structural damage caused by treatments to the spores and non-spore-forming bacteria was assessed by scanning electron microscopy. Geobacillus stearothermophilus spores suspended in Milli-Q water, UHT milk and pumpkin soup, treated by PATS were found to have significantly lower decimal reduction times (D values) compared with the thermal treatment alone. Spores suspended in UHT milk were more heat resistant compared with those in Milli-Q water and pumpkin soup. Bacillus cereus spores suspended in Milli-Q water and pumpkin soup treated with PATS were more effectively inactivated compared with spores treated by the thermal treatment alone. Clostridium botulinum spores in saline buffer subjected to PATS treatment were inactivated more effectively compared with the thermal treatment alone. Overall, the results show that PATS was a better processing technique for inactivation of bacterial spores compared with thermal treatment alone. However, PATS had no added benefit in inactivating the non-spore-forming bacteria Escherichia coli and Saccharomyces cerevisiae cells compared with the thermal treatment. A shelf life study showed that B. cereus spores in pumpkin soup retained a low spore count (<5 LogCFU/mL) for approximately 40 days in 30oC storage after treatment with PATS. No additional degradation of colour pigments of pumpkin soup and model pumpkin juice was observed following PATS compared with the thermal treatment. Spore-forming microorganisms can be resistant to pressure treatment alone, which limits the application of high pressure processing (HPP). Therefore, a combination approach was investigated. The mechanism of inactivating spores by combining HPP with other treatments is that the pressure assists in spore germination. Then a secondary treatment (thermal or CO2 gas) can be used to inactivate the germinated spores. A combined application of HPP and a consecutive CO2 treatment was investigated for the efficiency of spore inactivation. Results showed that HPP (200 MPa for 30 min) followed by a CO2 treatment inactivated Bacillus subtilis 168 in nutrient broth, tomato juice and liquid whole egg by 2.5, 1.0 and 1.5 LogCFU/mL respectively. These results indicated that this technique is inadequate for practical use. Scanning electron micrographs showed that pressure processing of B. subtilis 168 and B. subtilis natto spores resulted in deformation of the spore structure. This structural deformation of spores may have been due to water absorption during HPP and subsequent release upon decompression. PATS treated G. stearothermophilus and B. cereus spores were more severely damaged compared with the same spores which underwent thermal treatment alone. However, the extent to which E. coli and S. cerevisiae cells were damaged by both PATS and thermal treatment was similar.

This thesis investigates a newly developed and patented technology for its ability to inactivate spore- forming bacteria and non-spore-forming microorganisms. This new technology “Pressure Assisted Thermal Sterilization©” (PATS) is based on the theory of the thermal expansion of liquids. The efficiency of inactivating spore-forming and non-spore-forming microorganisms by PATS was compared with the thermal treatment alone. A combination treatment consisting of high pressure processing and gaseous carbon dioxide was also investigated for its ability to inactivate bacterial spores in model and real food matrices. The structural damage caused by treatments to the spores and non-spore-forming bacteria was assessed by scanning electron microscopy. Geobacillus stearothermophilus spores suspended in Milli-Q water, UHT milk and pumpkin soup, treated by PATS were found to have significantly lower decimal reduction times (D values) compared with the thermal treatment alone. Spores suspended in UHT milk were more heat resistant compared with those in Milli-Q water and pumpkin soup. Bacillus cereus spores suspended in Milli-Q water and pumpkin soup treated with PATS were more effectively inactivated compared with spores treated by the thermal treatment alone. Clostridium botulinum spores in saline buffer subjected to PATS treatment were inactivated more effectively compared with the thermal treatment alone. Overall, the results show that PATS was a better processing technique for inactivation of bacterial spores compared with thermal treatment alone. However, PATS had no added benefit in inactivating the non-spore-forming bacteria Escherichia coli and Saccharomyces cerevisiae cells compared with the thermal treatment. A shelf life study showed that B. cereus spores in pumpkin soup retained a low spore count (<5 LogCFU/mL) for approximately 40 days in 30oC storage after treatment with PATS. No additional degradation of colour pigments of pumpkin soup and model pumpkin juice was observed following PATS compared with the thermal treatment. Spore-forming microorganisms can be resistant to pressure treatment alone, which limits the application of high pressure processing (HPP). Therefore, a combination approach was investigated. The mechanism of inactivating spores by combining HPP with other treatments is that the pressure assists in spore germination. Then a secondary treatment (thermal or CO2 gas) can be used to inactivate the germinated spores. A combined application of HPP and a consecutive CO2 treatment was investigated for the efficiency of spore inactivation. Results showed that HPP (200 MPa for 30 min) followed by a CO2 treatment inactivated Bacillus subtilis 168 in nutrient broth, tomato juice and liquid whole egg by 2.5, 1.0 and 1.5 LogCFU/mL respectively. These results indicated that this technique is inadequate for practical use. Scanning electron micrographs showed that pressure processing of B. subtilis 168 and B. subtilis natto spores resulted in deformation of the spore structure. This structural deformation of spores may have been due to water absorption during HPP and subsequent release upon decompression. PATS treated G. stearothermophilus and B. cereus spores were more severely damaged compared with the same spores which underwent thermal treatment alone. However, the extent to which E. coli and S. cerevisiae cells were damaged by both PATS and thermal treatment was similar.