In this study, we isolated a koji mold from urushi (Rhus javanica) leaves and used it to produce soybean miso. The general component quantification of the produced soybean miso indicated moisture, protein, lipid, ash, and carbohydrate contents of 45.2, 16.0, 8.6, 19.0, and 11.2g/100g, respectively. Judging from the components and color, this soybean miso was sufficiently matured. Moreover, despite the very short maturation period of 3 months, the components did not significantly differ from those of general miso, and the decomposition of the raw materials was good. We confirmed that koji mold decomposes the raw materials well and that miso could be produced using urushi leaves. Based on these results, we inferred that the koji mold isolated from Rhus javanica display a high protease activity, as the protein content was high despite the short maturation period of 3 months.

As part of a study on the processing and use of beans as functional foods, this study attempted to develop a new type of miso using kidney beans, red kidney beans, pinto beans, and large white kidney beans as ingredients. This study has been the first to attempt producing miso using kidney beans given the absence of related studies. Our results showed that miso made using kidney beans, pinto beans, and large white kidney beans all had higher peptide and amino acid contents, which are umami components, than did rice miso and were strong-tasting and highly nutritious. Furthermore, we were able to produce functional miso with antioxidant effects, high gamma-aminobutyric acid content, and high polyphenol content, which was not possible with rice miso. These miso were highly evaluated for its color, aroma, and taste. Based on our findings, new products using kidney beans can be expected in the future.

Soybean whey (SW) is a by-product generated from the production of soybean protein isolates (SPI) from defatted soybean. SW is not commercially utilized and is discarded as industrial waste in Japan. Here, we examined whether SW could improve the physical properties of salmon, a protein-rich food. Salmon fillets were soaked in water (WT), citric acid-NaOH solution (CN), or SW before baking. The effects of presoaking with each solution on the qualities of baked salmon fillets were compared. We confirmed that SW contained numerous soluble proteins and carbohydrates in descending order of content, namely, sucrose, stachyose, and raffinose. Additionally, the mineral component ratios of SW closely resembled those of soymilk rather than SPI. Presoaking in SW caused a significantly higher yield rate and lower hardness of salmon fillets than presoaking in WT and CN. Although the salmon fillet moisture content only tended to be increased by SW presoaking when compared to CN presoaking, it was significantly increased when compared to WT presoaking. The results of sensory test showed that salmon fillets presoaked in SW were significantly softer with higher moisture and had a deeper orange color than those presoaked in WT and CN. However, they did not differ in smell. In all evaluations, except of smell, the salmon fillets presoaked in SW were significantly preferred as compared to those presoaked in WT and CN. These results suggested that SW can protect salmon against heat-induced quality reduction. In conclusion, SW could be used for improving the physical properties of protein-rich foods.

We investigated the factors determining color stability and discoloration in camu-camu fruit juice. Heat treatment deepened the red color of the juice, although it faded subsequently. Fading of the red color was also observed when juices were not heated. These findings suggest that the stability of red color is affected by the high concentrations of ascorbic acid in camu-camu juice. Heat treatment at high temperatures and for long durations turned the juice dark yellow. Under conditions of accelerated heating at 50℃, camu-camu juices varied from light pink to yellow and from yellow to brown. Polymerized polyphenols were more abundant in the browning juice than in light pink juice. Polyphenol oxidase activity was not detected, indicating that the browning was non-enzymatic. Regarding preservation conditions using model juices prepared from camu-camu juice containing polyphenol (PP), ascorbic acid (AA), and polyphenol and ascorbic mix (PP+AA), the b＊ value increased in the order PP < AA < PP+AA. These findings suggest that high concentrations of ascorbic acid greatly influence non-enzymatic browning. Thus, it was clarified that red fading and browning are mainly caused by the presence of ascorbic acid in camu-camu juice.

We made a fish sauce using mackerels from Kozushima, Japan. We then evaluated the nutritional components and taste of the sauce and proposed its applications in cooking. The nutritional components of the fish sauce were water (65.5%), protein (10.4%), carbohydrates (1.1%), ash (23%), lipid (0%), and sodium (8000mg/100g, salt equivalent 20.3%). The fish sauce had 1.3 times more magnesium and 8 times more copper than soy sauce. It had a clear light golden color, with a higher lightness level (L＊) than soy sauce. We compared the fish sauce with commercially available fish sauces using an electronic taste sensor. Our fish sauce had relatively low umami taste, but there were no contaminating flavors, resulting in an outstanding umami taste. This fish sauce is a suitable seasoning for seafood and carbohydrates and also enhances the taste of vegetables. In addition to its excellent flavor, Kozushima mackerel fish sauce has high protein and mineral content. This fish sauce has the potential to be marketed as a specialty product of Kozushima in the future.

We thought of using black soybean in foods and tried making miso with it. Black soybean miso had a strong taste and functionality. The total nitrogen content of black soybean miso was 2% and that of yellow soybean miso was 1.83%. Protein content was 11.42% in black soybean miso and 10.45% in the yellow soybean miso, showing that black soybean miso had more total nitrogen and protein content. It was inferred that formol nitrogen and free amino acids were higher in black soybean miso, and black soybean miso contained more peptides and amino acids. The total sugar content and the amount of direct reducing sugar were slightly higher in yellow soybean miso. The acidities Ⅰ and Ⅱ and the content of organic acids and basic amino acids was higher in black soybean miso. Yellow soybean miso had more of calcium and magnesium, while black soybean miso contained more potassium, phosphorus, iron, and zinc. Black soybean miso had somewhat higher amount of γ-aminobutyric acid. Also, it had a slightly higher amount of polyphenol. Antioxidant activity was high in both the miso.

お麩の製造の際に副生される小麦のデンプンは，約2年ものあいだ屋外発酵槽で発酵を経て，発酵小麦デンプン製品となり，関東でいう久寿餅の原料となる。著者らは，この発酵槽中の微生物変化や発酵前後のデンプンの性質の変化について，はじめて明らかにした。個人的には，身近な和菓子である久寿餅に発酵の力が使われていたのは知らなかったので大きな驚きである。興味深い小麦デンプンの発酵について解説していただいた。

Fermented wheat starch (FWS) is produced from starch as a byproduct of gluten manufacture. FWS is used in the traditional Japanese sweet, kuzu-mochi, which originates from the Edo period. The elastic but non-sticky texture of kuzu-mochi is attributable to the properties of FWS. To elucidate the characteristic properties of FWS, the comparison morphology, and physicochemical and pasting properties of starch with/without fermentation treatment were examined. Fermentation of wheat starch (WS) led to a decrease in the abundance of small starch granules, as small starch granules formed large aggregates. The degrees of damaged starch and amylose content in FWS were lower than those in unfermented WS. The λmax of the iodine absorption spectrum of FWS was blue-shifted relative to that of WS. The maximum and final viscosities of FWS, and its consistency in the rapid visco analyzer viscogram were lower than those of WS. These results revealed that fermentation endows starch with low viscosity and the ability to retard retrogradation.

Factors that deteriorate the quality of the fish paste products containing threadfin bream and the ability to suppress these effects were investigated. The myosin heavy chain (MHC) in the gel of threadfin bream surimi was degraded by heating from 40 to 60℃. The gel strength of threadfin bream was softer than that of Alaska pollack. The protease activity in threadfin bream surimi was 799 mU/g, which was approximately 6-fold higher the activity of Alaska pollack surimi. One trypsin-like serine type protease and two metalloproteases were detected using threadfin bream surimi. MHC degradation in the gel was suppressed by the addition of protease inhibitor. These results indicate that the decrease in the gel-forming ability in threadfin bream surimi was caused by internal protease that decomposed MHC. The addition of the trypsin inhibitor (TI) and soybean whey containing TI was effective for suppressing these effects.

Kuzu-mochi made from fermentation treated wheat starch is a traditional Japanese sweet from the Edo period. Kuzu-mochi has a unique texture, which is not found in wheat starch gels. In this study, to clarify the texture characteristics of kuzu-mochi, the physical properties of kuzu-mochi were compared with that of various gelatinous foods such as dango, warabi-mochi, and yokan. Kuzu-mochi was softer than yokan, but it was as hard as dango. The chewiness and the stickiness of kuzu-mochi were lower than that of dango. Kuzu-mochi had an elastic texture and not a sticky one. Many starch granules were observed within the gel matrix of kuzu-mochi. The gelatinous state of starch in kuzu-mochi is suggested to contribute to the texture and the internal structure of kuzu-mochi. The firmness of the fermented wheat starch gels was lower than that of kuzu-mochi. After storing for 72h at 4℃，the firmness of the fermented wheat starch gel underwent few changes. The results of this study suggest that the texture of kuzu-mochi is characterized by fermented wheat starch.

製パン性の改善に小麦タンパク質が用いられ，グルテンのみでなくグリアジン添加の有効性が明らかになっている。製パン改良材であるグリアAに対しPDI-ERO1を作用させS-S結合を増加させることによる性状変化と製パン性改善効果を解析した。未処理のグリアAの添加によって小麦粉生地の形成時間は短くなり，生地からのガス漏洩率は低下したが，パンの比容積には有意な影響を及ぼさなかった。TaPDIおよびTaERO1によってS-S結合を増加させたグリアAは，グリアAと比較して生地のガス漏洩率が大きく減少し，パンの比容積が有意に増加した。TaPDIおよびTaERO1処理によってグリアジンの分子内S-S結合が増加し，グリアAの表面疎水性が低くなった。以上の結果から，グリアジンの生地形成および製パン性に与える影響を確認する共に，グリジアジンの表面疎水性が製パン性に大きな影響を及ぼすことが示唆された。

製パンにおいて小麦生地中でのSS結合形成は重要な要因である。本報告では小麦生地中におけるSS結合に対するリコンビナント小麦プロテインジスルフィドイソメラーゼ（TaPDI）およびエンドプラズミックレティキュラムオキシドレダクターゼ1（TaERO1）の影響について検討した。まず，タンパク質へSS結合を形成することで還元化したPDIのERO1酸化再生能を解析した。その結果，小麦遺伝情報から取得したERO1はPDIに対し酸化再生能を有していた。また，還元変性オボムコイドへのSS結合形成能は，PDIのみではなくERO1およびFADが共存することで約3倍に増加した。強力粉から調製した小麦酸可溶性タンパク質にPDIおよびERO1, FADを作用させ，対角線電気泳動にて解析したところ，PDIはグリアジンに対し分子内にSS結合を形成することが示唆された。さらに，製パン適性の劣る中力粉を用い製パン試験を行ったところ，比容積は無添加で4.69cm3/gであったのが，PDIおよびERO1，FADを添加することで，5.34cm3/gとなり強力粉から調製するパンと同等となった。本結果より，製パン性が劣る小麦粉に対するPDIの製パン改良効果を確認することができた。

Kuzu-mochi, which is a traditional Japanese sweet, is made from fermented wheat starch. In this study, we examined changes in wheat starch granules and the microflora during the fermentation of wheat starch. Large, broken granules of starch and gram-positive bacilli were observed during the fermentation process. The pH ranged from 3 to 4 throughout the process. Lactic acid and acetic acid were detected in each fermentation period, with lactic acid being the predominant acid. This result indicates that lactic acid bacteria (LAB) are heterolactic fermenters. Viable cell count was determined at the same pH as that of the fermentation medium, and microorganisms that can grow in acidic conditions were isolated. The microflora present during the fermentation process included 2—3 species of aerobic bacteria, fungi and yeast, and approximately 10 species of LAB. PCR-denaturing-gradient-gel-electrophoresis profiles indicated that the microflora remained stable from the pre-fermentation period to about 6 months post-fermentation. Furthermore, the supernatant collected during the pre-fermentation period showed starch hydrolysis activity, and wheat starch was broken by its enzyme. These results suggest that the bacterial community present at the pre-fermentation period can serve as a starter culture for the traditional processing of fermented wheat starch.

Spoilage of cooked rice occurred as a result of heat insulation. The cooked rice had an off-flavor that was caused by spoilage. This phenomenon is thought to be caused by microorganisms. Therefore, in this study, we attempted to isolate the contaminating microorganisms from cooked rice and identify them by using polymerase chain reaction (PCR) and PCR-denaturing gradient gel electrophoresis (DGGE). No microorganisms were detected in normal cooked rice; however, microorganisms were detected at 4.6×106 cfu/g in rice spoiled by 72h of heat insulation. The isolates were able to grow at 60～65℃ on the cooked rice, and they produced an off-flavor. Based on phylogenetic analysis, the isolates were identified as members of the genus Geobacillus, and were closely related to Geobacillus thermoleovorans. Contaminating microorganisms in the cooked rice were identified by PCR using bacterial universal primers and G. thermoleovorans was only detected using PCR-DGGE. These results suggest that G. thermoleovorans was the contaminating microorganism in the spoiled cooked rice and produced the off-flavor.

We constructed a gramineous plant-specific primer set-matK 792F1GC and matK 979R1, targeting the maturase K (matK) gene, for detection of gramineous raw materials in processed food. For this, we used polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The primer set was evaluated using agarose gel electrophoresis and DGGE. Our results confirmed that the primer set was specific for gramineous plants. From our analysis, we confirmed that up to 6 types of gramineous plants can be simultaneously detected using PCR-DGGE. Results of a model test showed that by using this method, it was possible to detect wheat from a sample, if present more than 0.1% (w/w). Thus, our method of PCR-DGGE with matK 792F1GC and matK 979R1 can be used to detect the presence of gramineous raw materials in processed food.

In this study, α-glucosidase from rice bran was purified by chromatography and electrophoresis. This enzyme has a molecular weight of 310 kDa and consists of 4 subunits with a molecular weight of 76 kDa or 36 kDa. The N-terminal amino acid sequence of the 76-kDa protein was found to be similar to that previously reported for a rice glycosidase. However, the molecular weight of the enzyme differed from that reported for any rice glycosidase. The enzymatic properties of this enzyme were almost similar to those of most rice α-glucosidases. However, 2 properties, that is, the pH activity and thermostability, were a little different from those reported for polished rice. This enzyme was stable in ethanol up to a concentration of 10% ethanol. In addition, its activity in ethanol did not decrease up to a concentration of 20% ethanol. Ethyl-α-D-glucoside was optimally produced at pH4.0 and 40°C.

The effect of ovomucin, a glycoprotein, on the viscosity and foaming properties of emu egg white was determined. Compared to Gallus gallusdomesticus (hen) egg white, emu egg white was more viscose and had better foaming properties and stability. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis followed by periodic acid-Schiff staining were performed for emu egg white, and carbohydrate chain bands of size 52, 78, and 207 kDa were observed. The N-terminal amino acid sequence of the 207-kDa protein showed 62.5% homology with the N-terminal region of α2-ovomucin obtained from hen. The ovomucin content in emu egg white was approximately 6.3%, over one from hen egg white. In addition, the ovomucin solution prepared from emu egg white was more viscose than that prepared from hen egg white. The viscosity of ovomucin obtained from each egg white was closely related to the viscosity of the egg white. Therefore, these results suggest that ovomucin affects the viscosity and foaming properties of emu egg white.

Ero 1 encodes a flavoprotein oxidase that promotes oxidative protein folding through protein disulfide isomerase and generates reactive oxygen species (hydrogen peroxide) as byproducts. Ero 1 was cloned and sequenced from wheat (Triticum aestivum cv. Haruyutaka). Three homologous Ero 1 genes were obtained, which showed high sequence conservation. These Ero 1 genes were 455 amino acids in length with a conserved catalytic Cys-X-X-Cys-X-X-Cys motif (Cys370, Cys373, Cys376) and a transmembrane signal in the N-terminal domain. Wheat Ero 1 without the signal sequence was expressed in Escherichia coli as a His-tagged fusion protein. Recombinant Ero 1 was purified by affinity chromatography using a TALON resin column. The activity of purified recombinant Ero 1 was measured by both the catalytic reduction of FAD and an oxygen consumption assay. FAD was reduced in an Ero 1 concentration-dependent manner. Oxygen consumption at 0.1 μM wheat Ero 1 was slow; however, the rate increased with increased Ero 1 concentration. These results indicated that recombinant Ero 1 from wheat has FAD-reducing ability, and utilizes oxygen as a terminal electron accepter in the PDI-Ero 1 system.

Two-dimensional polyacrylamide gel electrophoresis was used for analyzing emu and hen egg white proteins. The 78 kDa protein spots of emu and hen egg whites had varied isoelectric points, 9.0 and 6.0, respectively. The protein spot of emu egg white showed 75% homology with the N-terminal region of ovotransferrin from Gallus gallus domesticus (hen). The differential scanning calorimetry (DSC) pattern of emu egg white was different from that of hen egg white. In addition, on DSC analysis using purified ovotransferrin from emu egg white, the 2 endothermic peaks were observed at 60℃ and 80℃. This might suggest that emu and hen egg whites contain distinct heat shock (HS) proteins. Moreover, these results suggest the effect of HS proteins on the thermal properties of emu egg white.