ABSTRACT
Sake is made from steamed rice, malted rice, and water. Sake production begins with the preparation of a small-scale starter (moto); the quality of moto significantly influences the flavor and richness of sake. In the traditional starter, yamahai-moto, the growth of naturally occurring lactic acid bacteria represses the putrefactive micro-organisms, whereas in the modern starter, sokujo-moto, this is achieved by adding lactic acid. In this study, the successive change in bacterial flora of yamahai-moto was analyzed by pyrosequencing 16S ribosomal RNA genes. Lactobacillus was dominant throughout the process (93-98%). Nitrate-reducing bacteria that have been generally assumed to be the first colonizers of yamahai-moto were scarcely found in the early stage, but Lactobacillus acidipiscis dominated. Lactobacillus sakei drastically increased in the middle stage. This is the first report, though one case study, to show how the early stage microbiota in Japanese yamahai-moto is varyingly controlled without nitrate-reducing bacteria using next-generation sequencing.
Subject(s)
Alcoholic Beverages/microbiology , Food Microbiology , Lactobacillaceae/genetics , Microbiota/genetics , Oryza/metabolism , Phylogeny , Alcoholic Beverages/analysis , Bacterial Load , Ethanol/metabolism , Fermentation , High-Throughput Nucleotide Sequencing , Lactobacillaceae/classification , Lactobacillaceae/metabolism , RNA, Ribosomal, 16S/geneticsABSTRACT
The processing of archetypal Japanese sushi involves microbial fermentation. The traditional sushi kaburazushi, introduced in the middle ages, is made by fermenting salted yellow tail, salted turnip, and malted rice, and is distinguished from the ancient sushi narezushi, made from fish and boiled rice. In this study, we examined changes in the microbial population during kaburazushi fermentation by pyrosequencing the 16S ribosomal RNA genes (rDNA) of the organisms in the fermentation medium. Ribosomal Database Project Classifier analysis identified 31 genera, among which Lactobacillus drastically increased during fermentation (150-fold increment over 8 d), while the relative populations of the other gram-positive bacteria (Staphylococcus and Bacillus) decreased. Basic Local Alignment Search Tool analysis revealed the dominant species to be L. sakei. This organism constituted approximately 90% of Lactobacillus and 79% of total microbiota. The taxonomic diversity and species richness (assayed by Shannon-Weiner Index and Chao 1, respectively) were not significantly different between middle-ages kaburazushi and ancient narezushi. Both types were characterized by the preferential growth of Lactobacillales.
Subject(s)
Bacteria/genetics , Fishes/microbiology , Food Microbiology , High-Throughput Nucleotide Sequencing , Sequence Analysis , Animals , Bacteria/isolation & purification , DNA, Bacterial/genetics , DNA, Ribosomal/genetics , Fermentation , Japan , MicrobiotaABSTRACT
Fish sauce prepared from squid organs contains cadmium (Cd), which may be present at hazardous concentrations. In this study, we report a new, inexpensive, and acceptable method for removing Cd from fish sauce using tannin, which is an approved food additive in Japan. Decreases in Cd concentrations of 13-fold were observed (0.39-0.03 mg/100 mL) by incorporating the soluble Cd into a precipitate generated by tannin treatment. The total nitrogen content, free amino acid content, 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, and angiotensin I-converting enzyme inhibitory activity of the treated fish sauce were the same as those of the untreated fish sauce.
Subject(s)
Cadmium/analysis , Fish Products/analysis , Food Technology/methods , Animals , Fishes , Food Contamination , Food Technology/instrumentation , Japan , Tannins/analysisABSTRACT
Modern Japanese sushi is derived from an archetype, narezushi, which is made by the fermentation of salted fish with rice. Several studies have demonstrated that lactic acid bacteria are dominantly present in narezushi, but no studies have addressed how microbial composition changes during fermentation. In this study, we examined changes in the microbial population in aji (horse mackerel)-narezushi during fermentation by pyrosequencing the 16S ribosomal RNA gene (rDNA). Ribosomal Database Project Classifier analysis revealed that among the 53 genera present, the Lactobacillus population drastically increased during fermentation, while the populations of other bacteria remained unchanged. Basic Local Alignment Search Tool analysis revealed that L. plantarum and L. brevis were the major species. Comparison with other fermented food microbiota indicated high product-dependency of the bacterial composition, which might have been due to the starter-free fermentation process.
Subject(s)
Bacteria/genetics , Fermentation , Food Handling , Metagenome/genetics , Sequence Analysis , Animals , Bacteria/isolation & purification , Bacteria/metabolism , Fishes , Food Microbiology , Lactic Acid/metabolism , OryzaABSTRACT
AmyR is a Zn(II)(2)Cys(6) transcriptional activator that regulates expression of the amylolytic genes in Aspergillus species. Subcellular localization studies of GFP-fused AmyR in A. nidulans revealed that the fusion protein preferentially localized to the nucleus in response to isomaltose, the physiological inducer of the amylolytic genes. The C-terminal domains of AmyR, designated MH3 (residues 419-496) and MH4 (residues 516-542), were essential for sensing the inducing stimulus and regulating the subcellular localization. The MH2 domain (residues 234-375) located in the middle of AmyR was required for transcriptional activation of the target genes, and the nuclear localization signals were identified within the N-terminal Zn(II)(2)Cys(6) DNA binding motif.
