Kuratsuki bacteria and sake making.

Kuratsuki bacteria enter during the sake-making process and interact with sake yeast until their growth is attenuated by the ethanol produced by sake yeast. Due to the interaction between kuratsuki bacteria and sake yeast, the metabolism of sake yeast changes, affecting the composition of esters and organic acids and subsequently the flavor and taste of sake. We cultivated kuratsuki bacteria and sake yeast, and performed test making at sake breweries to clarify the interaction among microorganisms in the sake-making process.

Effect of kuratsuki Kocuria on sake's taste varies depending on the sake yeast strain used in sake brewing.

Although sake yeast mainly produces the taste of sake, sake brewery-inhabiting (kuratsuki) bacteria affect the taste of sake. Thus, kuratsuki bacteria may alter the metabolism of sake yeast through interactions between kuratsuki bacteria and sake yeast. This study aimed to confirm the effects of the combination of kuratsuki Kocuria TGY1127_2 and different sake yeast strains, AK25, K901, and K1801 on the taste of sake.

Approaches for introducing large DNA molecules into bacterial cells.

Engineering of the bacterial genome plays a key role in systems biology and synthetic biology. Genetic engineering of the bacterial genome involves the design and synthesis of large DNA molecules. However, functional studies of the designed and synthesized large DNA molecules are lagging.

Effect of kuratsuki Kocuria on sake brewing in different koji conditions.

Koji is made using steamed rice and a koji mold, which plays an essential role in sake brewing. We challenge to build a new sake brewing method using the kuratsuki bacteria that have inhabited each sake brewery. In this paper, effects of the kuratsuki Kocuria strain TGY1127_2 were estimated on sake brewing in different koji conditions.

Effects of heterologous genome microinjection on the enlargement of protoplasts.

The lactic acid bacterium genomic DNA and seven phylogenetically distant bacterial genomic DNAs were microinjected into 126 enlarged protoplasts of . After the microinjection, a time-lapse observation was performed on how the cells enlarged. Most cells did not stop enlarging.

Diversity of Isolates from the Sake Brewing Process at a Sake Brewery.

We collected 92 isolates belonging to the genus from the sake brewing process at Shiraki Tsunesuke Sake Brewery in Gifu, Japan to determine whether there is strain specificity at individual sake breweries. After distributing the isolates into seven groups, we observed that at least two groups (68 isolates) were bacteria at Shiraki Tsunesuke Sake Brewery. The   isolates were collected from different samples at the early and late stages of sake brewing in 2021 and 2019, respectively.

Co-cultivation of sake yeast and Kocuria isolates from the sake brewing process.

Kocuria isolates collected from the sake brewing process have inhabited the Narimasa Sake Brewery in Toyama, Japan. To investigate the effect of these actinobacterial isolates on the growth and metabolism of sake yeast, co-cultivation of sake yeast and Kocuria isolates was performed in a medium containing tryptone, glucose and yeast extract (TGY), and a solution containing koji (steamed rice covered with Aspergillus oryzae) and glucose. In the TGY medium, the ethanol concentration and the number of living cells of each microorganism were measured.

Genomic information of isolates from sake brewing process.

Bacteria belonging to the genus were identified as bacteria peculiar to a sake brewery in Toyama, Japan. Comparison of the 16S rRNA gene sequences revealed two groups of isolates. Among known species, one group was similar to ( type), and the other, ( type).

Novobiocin inhibits membrane synthesis and vacuole formation of protoplasts.

We demonstrate that plasma membrane biosynthesis and vacuole formation require DNA replication in protoplasts. The replication inhibitor novobiocin inhibited not only DNA replication but also cell enlargement (plasma membrane biosynthesis) and vacuole formation during the enlargement of the protoplasts. After novobiocin treatment prior to vacuole formation, the cell size of protoplasts was limited to 6 μm in diameter and the cells lacked vacuoles.

Factors That Affect the Enlargement of Bacterial Protoplasts and Spheroplasts.

Cell enlargement is essential for the microinjection of various substances into bacterial cells. The cell wall (peptidoglycan) inhibits cell enlargement. Thus, bacterial protoplasts/spheroplasts are used for enlargement because they lack cell wall.

Species-dependent protoplast enlargement involves different types of vacuole generation in bacteria.

Vacuole generation occurs frequently during the enlargement of bacterial protoplasts and spheroplasts. Gram-positive Enterococcus faecalis protoplasts and gram-negative Lelliottia amnigena spheroplasts had large and small vacuoles inside the cytoplasm, respectively. Although no vacuoles were found at the early stage of cell enlargement, all enlarged cells used in the microinjection procedures had vacuoles.

DNA replication and cell enlargement of protoplasts.

Protoplasts of did not divide but enlarged in Difco Marine Broth containing penicillin. Our previous studies have demonstrated that transcription and translation were essential for bacterial cell enlargement. However, it was uncertain whether replication was also essential.

Sensitivity of deletion mutant to calcium ions results in enhanced spheroplast size.

RodZ is a cytoskeletal protein associated with bacterial cell shape. It is a transmembrane protein located on the plasma membrane, and it binds to another cytoskeletal protein MreB. contains a homolog.

Chemical and Bacterial Components in Sake and Sake Production Process.

Together with the worldwide Washoku (traditional Japanese foods and drinks) boom, interest in sake, a traditional Japanese alcoholic drink, is increasing around the world. There are few scientific analyses and studies on the production of sake or the final product itself. We show the diversity of bacterial contaminants during sake production and investigated the effects of different ingredients on sake (for example, amino acids).

Sugar enhances outer membrane fusion in Deinococcus grandis spheroplasts to generate calcium ion-dependent extra-huge cells.

In our previous study, we showed that cell fusion occurred in spheroplasts of Deinococcus grandis at 200 mM calcium chloride in the incubation medium. Extra-huge cells (> 0.1 mm in diameter) were observed at this concentration with a low frequency of appearance.

Calcium ion induces outer membrane fusion of Deinococcus grandis spheroplasts to generate giant spheroplasts with multiple cytoplasms.

Generally, enlarged spheroplasts of the Gram-negative bacterium Deinococcus grandis contain a single cytoplasm and a large periplasmic space. Enlargement of D. grandis spheroplasts requires the presence of divalent cation Ca2+ or Mg2+.

Enlargement of Deinococcus grandis spheroplasts requires Mg or Ca.

While the cell wall strictly controls cell size and morphology in bacteria, spheroplasts lack cell walls and can become enlarged in growth medium under optimal conditions. Optimal conditions depend on the bacterial species. We frequently observed extreme enlargement of spheroplasts of the radiation-resistant bacterium Deinococcus grandis in Difco Marine Broth 2216, but not in TGY broth (a commonly used growth medium for Deinococcus).

The enigmatic Mixia osmundae revisited: a systematic review including new distributional data and recent advances in its phylogeny and phylogenomics.

The enigmatic basidiomycete genus Mixia includes intracellular parasites of Osmunda and Osmundastrum ferns. Here, the authors review the systematic and phylogenetic history of M. osmundae, originally known as Taphrina osmundae, and provide new data from investigations of specimens of Osmunda japonica collected in Yunnan Province, China, which we determine to be conspecific with M.

Comparison of gene expression levels of appA, ppsR, and EL368 in Erythrobacter litoralis spheroplasts under aerobic and anaerobic conditions, and under blue light, red light, and dark conditions.

We compared the gene expression levels of the blue-light-responsive genes, appA (encoding photosynthesis promoting protein AppA), ppsR (encoding photosynthesis suppressing protein PpsR), and EL368 (encoding a blue-light-activated histidine kinase with a light, oxygen, or voltage domain) between aerobic and anaerobic conditions in spheroplasts of the aerobic photosynthetic bacterium Erythrobacter litoralis. The spheroplasts conducted photosynthesis under red light but not under blue light. All three blue-light-responsive genes showed higher expression under aerobic conditions than under anaerobic conditions under blue light.

Detection of Bacterial DNA During the Process of Sake Production Using Sokujo-Moto.

There are two types of starter cultures used in Japanese rice wine (sake) fermentation, namely, sokujo-moto and yamahai-moto. Analyses of microbiota changes during sake production using yamahai-moto have already been reported. In this study, we analyzed microbiota changes during sake production using sokujo-moto.

Bacterial DNA Detected in Japanese Rice Wines and the Fermentation Starters.

As Japanese rice wine (sake) brewing is not done aseptically, bacterial contamination is conceivable during the process of sake production. There are two types of the fermentation starter, sokujo-moto and yamahai-moto (kimoto). We identified bacterial DNA found in various sakes, the sokujo-moto and the yamahai-moto making just after sake yeast addition.

Changes in nucleosome formation at gene promoters in the archiascomycetous yeast .

We measured the degree of nucleosome formation at the gene promoters in trichostatin A-treated (1, 2, and 3 µg/mL) cells of the archiascomycete and those in enlarged cells after zymolyase treatment. TSA-treated and enlarged cells showed similar changes in nucleosome occupancy in five out of six positions in the gene promoters. These results suggest that changes in nucleosome formation at the gene promoters could serve as stress response mechanisms elicited in response to spheroplast (zymolyase treatment) and TSA treatment.