Characterization of peptides generated in proteolytic digest of steamed rice grains by sake koji enzymes

High-molecular-weight peptides (approximately 10-30 kDa) generated in a digest of steamed rice grains by sake koji enzymes were characterized. Among 13 major spots resolved by 2-D gel electrophoresis, 12 contained peptides having N-termini of rice glutelin as determined by mass fingerprinting analysis and/or MS/MS. The source of these peptides was presumed to be the acidic subunit of rice glutelin. An addition of up to 25% glucose in the digestion of an isolated rice protein body induced the accumulation of these peptides. The level of accumulation of these peptides in the digest of 70% polished rice samples correlated well with the crude protein content of the rice grains. The degree of accumulation of these peptides in Yamadanishiki and low-polish-rate rice was low, whereas that observed in 90% polished rice samples was extremely low.     

Brewer's yeast cell wall affects microbiota composition and decreases Bacteroides fragilis populations in an anaerobic gut intestinal model

Brewer's yeast cell wall (BYC) has been reported to have prebiotic activity that improves the microbiotal composition of the human gut. To understand the precise effect of BYC on gut microbiota and its metabolism, we used a three-stage continuous-flow reactor system that mimicked the environment of the large intestine. The reactor system was able to maintain the bacterial community stably for a week. The Bacteroides fragilis population decreased drastically after the addition of BYC into this system while the number of Lactobacillus was stably maintained. In addition, propionate and acetate levels increased drastically. This metabolic change correlated with an increase in a number of specific operational taxonomic units annotated to the genus Veillonella and Megasphaella. These results suggest that BYC affects the composition of microbiota in an in vitro model system.     

Determination of bovine lactoferrin in lactoferrin-supplemented dairy products and raw milk by an automated latex assay

Latex immune agglutination method with a multipurpose auto-analyser (the automated latex assay) was validated for determination of bovine lactoferrin (BLF) in various dairy products. Reproducibility-within-laboratory (intermediate precision) due to day for infant formula, UHT milk and yogurt supplemented with BLF at 50 mg/100 g for infant formula and UHT milk, and at 100 mg/100 g for yogurt were 1.62 to 3.10 mg/100 g. Reproducibility-within-laboratory due to analysis (morning, noon, and evening) for raw milk was 1.59 mg/100 g. Trueness, accuracy in determining known amounts added for BLF-supplemented dairy products was -4.7 to -2.0 mg/100 g. BLF concentration in raw milks was 20.3 to 21.8 mg/100 g. Although interference by the matrixes took place in infant formula and raw milk, BLF assays were accurately carried out by 2000-fold dilution or standard-addition method. Automated latex assay for BLF is simple, rapid, precise and accurate enough for a routine method in various dairy products containing BLF.     

Identification of 2,4,6-trichloroanisole (TCA) causing a musty/muddy off-flavor in sake and its production in rice koji and moromi mash

2,4,6-Trichloroanisole (TCA), which has been identified as the main component responsible for the cork taint in wine, was detected in sake samples having a musty/muddy off-flavor by stir bar sorptive extraction (SBSE). We confirmed that TCA is one of the components causing this off-flavor in sake, as in other alcoholic beverages, from a sensory analysis showing the correlation between TCA concentration and the intensity of the musty/muddy off-flavor. We investigated the route of TCA production in the rice koji preparation process and in the moromi mash process for sake brewing. We found that TCA is produced mainly by the biomethylation of 2,4,6-trichlorophenol (TCP) by rice koji in brewing and that TCP originates from the wooden tools used in preparing rice koji.     

Characterisation and threshold measurement of aroma compounds contributing to the quality of Honkaku shochu and Awamori

Honkaku shochu and Awamori are traditional Japanese spirits. The detection and thresholds of 16 compounds in these spirits were determined so as to understand the compounds contributing to the aroma characteristics of Honkaku shochu and Awamori. The odour activity values calculated using detection thresholds suggested that 14 compounds affect the quality of Honkaku shochu and Awamori. The odour activity values of β-damascenone and rose oxide were much higher in sweet potato shochu and these two compounds are considered to contribute to the characteristics of sweet potato shochu. Similarly, the odour activity value of 1-octen-3-ol was higher in Awamori than in the other categories, suggesting that 1-octen-3-ol produces the characteristic aroma of Awamori. In addition, the sensory attribute terms of 16 compounds are summarised. © 2019 The Institute of Brewing & Distilling     

pDUAL, a multipurpose, multicopy vector capable of chromosomal integration in fission yeast

A novel series of plasmid vectors named pDUAL have been developed. These vectors enable one to introduce not only multicopies of genes with episomal maintenance but also a single copy with chromosomal integration into the fission yeast, Schizosaccharomyces pombe. The multicopy plasmids can be easily converted to fragments for chromosomal integration by digestion of the plasmids with a certain restriction endonuclease before transformation of the yeast cells. The resultant fragments, lacking the autonomously replicating sequence, are designed for targeting into the chromosomal leu1 locus by homologous recombination. Whether the transformants are the results of episomal maintenance of the plasmid or homologous gene targeting can be readily checked by their requirement for uracil or leucine, or by the PCR diagnostic analysis. Furthermore, we propose the use of pDUAL derivatives for PCR-based chromosomal tagging of a gene to introduce several tags into 5'-terminus of a gene, employing a set of primers. Using these all-in-one vectors, a suitable mode of expression of a cloned gene can be selected for individual analysis without any complicated subcloning processes.     

DL-alpha-tocopherol acetate mitigates maternal hyperthermia-induced pre-implantation embryonic death accompanied by a reduction of physiological oxidative stress in mice

Maternal hyperthermia induces pre-implantation embryo death, which is accompanied by enhanced physiological oxidative stress. We evaluated whether the administration of DL-alpha-tocopherol acetate (TA) to hyperthermic mothers mitigated pre-implantation embryo death. Mice were exposed to heat stress (35 degrees C, 60% relative humidity) for 12 h or not heated (25 degrees C) on the day of mating. Twelve hours before the beginning of temperature treatment, TA was injected intraperitoneally at a dose of 1 g/kg body weight. After the treatment, zygotes were recovered and the developmental abilities and intracellular glutathione (GSH) levels were evaluated. Another set of mice, with or without TA treatment, was exposed to heat stress for 12, 24 and 36 h, and the urinary levels of the oxidative stress marker 8-hydroxy-2'-deoxyguanosine (8-OHdG) were measured. Heat stress significantly decreased the blastocyst development rate and the GSH content in zygotes, as compared with the non-heat-stressed embryos, while TA administration significantly mitigated the deleterious effects of heat stress with regard to both parameters. Moreover, although the urinary levels of 8-OHdG gradually increased according to the duration of heat exposure, with or without TA administration, the levels were lower in the TA-administered group than in the placebo-injected mice. These results suggest that heat stress enhances physiological oxidative stress, and that TA administration alleviates the hyperthermia-induced death of pre-implantation embryos by reducing physiological oxidative stress.     

Resuspension of bottom sediment by an internal wave in Lake Biwa

An internal wave‐induced resuspension of bottom sediment and resultant changes in water chemistry were investigated in the sublittoral area of Lake Biwa in the late stage of the stagnant period. The thermocline was measured at between depths of 15 and 20 m at the beginning of the observation and rapidly dropped by 5 m within 3 h. The downward movement of the thermocline resulted in markedly increased turbidity at the top of the thermocline (20 m in water depth). Concentrations of dissolved methane synchronistically increased with turbidity, especially in the top of the thermocline where there was the highest turbidity. Total dissolved phosphorus and particulate phosphorus also increased in the thermocline. In contrast, the soluble reactive phosphate concentration decreased with time. An increase in the concentrations of particulate metals (aluminium, iron and titanium), which probably originated from mineral particles, was more marked in the water sample collected at 1 m above the bottom than at the peak of turbidity. The results indicated that an internal wave caused resuspension of the bottom sediment and also affected the water chemistry.     

Synthesis of Various Water-Soluble C60 Derivatives and Their Superoxide-Quenching Activity

In order to examine the utility of fullerene as a medicinal application, we evaluated the reaction between fullerene derivatives and active oxygen species in vitro. This paper describes the synthesis of various water-soluble C₆₀ derivatives (cationic and anionic) and evaluation of their superoxide (O₂^-) quenching activity. Cationic C₆₀ derivatives showed fairly high efficiency.