• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.175-179
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• 1986

In order to develope a protoplast fusion of the genus Cellulomonas having high assimilibility of cellulose, the optimum conditions for the protoplast formation of Cellulomonas flavigena NCIB 12901 was investigated and observed by means of Scanning Electron Microscope. The results suggested that the susceptibility of the cell wall by lysozyme treatment on protoplast formation was considerably depend on the cultural periods of the cells. Cells of C. flavigena at mid exponential phase could more efficiently convert to protoplast cells than those at late exponential phase did. The rate of the protoplast formation was 95%, even though the rate was over 99.9% on counting by indirect method after osmotic shock treatment, when cells of the organism at mid exponential phase were treated with lysozyme (400$\mu\textrm{g}$/$m{\ell}$) for 6 hours and observed by SEM. In the evaluation of protoplast formation of the genus Cellulomonas, direct method of the observation under Scanning Electron Microscope was much more reliable than the counting method of protplasts after osmotic shock treatment. Because defferences between the number of spheroplast and protoplast were not able to be figured out on counting the number of protoplast after osmotic shock treatment.

• Food Science of Animal Resources
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• v.37 no.1
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• pp.134-138
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• 2017

Salmonella enterica infects a broad range of host animals, and zoonostic infection threatens both public health and the livestock and meat processing industries. Many antimicrobials have been developed to target Salmonella envelope that performs essential bacterial functions; however, there are very few analytical methods that can be used to validate the efficacy of these antimicrobials. In this study, to develop a potential biosensor for Salmonella envelope stress, we examined the transcription of the S. enterica serovar typhimurium spy gene, the ortholog of which in Escherichia coli encodes Spy (${\underline{s}}pheroplast$ ${\underline{p}}rotein$ ${\underline{y}}$). Spy is a chaperone protein expressed and localized in the periplasm of E. coli during spheroplast formation, or by exposure to protein denaturing conditions. spy expression in S. typhimurium was examined by constructing a spy-gfp transcriptional fusion. S. typhimurium spy transcription was strongly induced during spheroplast formation, and also when exposed to membrane-disrupting agents, including ethanol and the antimicrobial peptide polymyxin B. Moreover, spy induction required the activity of regulator proteins BaeR and CpxR, which are part of the major envelope stress response systems BaeS/BaeR and CpxA/CpxR, respectively. Results suggest that monitoring spy transcription may be useful to determine whether a molecule particularly cause envelope stress in Salmonella.

• Applied Biological Chemistry
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• v.31 no.1
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• pp.26-32
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• 1988

This study was performed to construct killer wine yeasts which might suppress the growth of wild yeasts, reduce the consumption of starter and condense the fermentation period. Saccharomyces cerevisiae M524, a commercial wine yeast, was treated with N-methyl-N'-nitro-N-nitrosoguanidine to induce auxotrophic mutants, i.e., CHM $2(thr^-)$, CHM 3 $(asp^-)$ and CHM 6 $(tyr^-)$. These auxotrophs were fused successfully with a killer yeast, S. cerevisiae $1368R({\alpha}\;his\;4\;kar\;1-1(kil-k)\;(k_0)$, respiratory deficient) using sphoroplast techniques and the fusants were designated as CHF 21$(th^-\;kil^+)$, CHF 22$(thr^-\;kil^+)$, CHF 31$(asp^-\;kil^+)$ and GHF 61$(tyr^-\;kil^+)$. Combined cultivation of CHF 31 with 1368R or S. cerevisiae $5{\times}47$ (killer sensitive) proved out that CHF 31 had the characteristic of killing and produced the same amount of ethanol as the prototroph, M524.