Kazuhito Watabe

Protein synthesis in the isolated forespores from sporulating cells of Bacillus subtilis.

Developing forespores were isolated from Bacillus subtilis at different stages of sporulation and protein synthesis in the forespore compartment was examined. Pulse‐labeling experiments indicated that [14C]phenylalanine was continuously incorporated into the sporangium throughout sporulation, and at t5 (early stage V of sporulation) 58% of the radioactivity was located in the forespore compartment. Significantly high incorporation of [14C]phenylalanine was observed when the isolated forespores at t5 were incubated with the corresponding mother‐cell cytoplasmic fraction or an amino acid mixture. About 73% of the radioactivity incorporated into the isolated forespore at t5 was found in the cytoplasmic fraction and 26% in the membranous fraction. Analysis by sodium dodecyl sulfate‐gel electrophoresis showed that the 14C‐labeled cytoplasmic protein had a molecular weight of about 20,000, and that a protein having the same molecular weight was present in the t5 forespore as a slight protein band and also in the mature spore as a clear protein band. Gel electrophoresis also revealed that the 14C‐labeled membranous‐soluble protein (prepared by solubilization with detergents) had broad peaks with molecular weights of about 74,000, 33,000, 20,000, and 12,000.

Studies on the indicator for heavy metal contamination in environments (1) Heavy metal contents of hair, nail and moustache

Abstract Heavy metal (Hg, Pb, Cd, Cu, Ni, Fe, Zn, and Mn) contents of the samples from 10 males were followed throughout a year. The values were characteristic of the individuals, and some correlationships between the contents of different metals were shown.

Characterization of forespores isolated from Bacillus subtilis at each stage of sporulation.

Bacterial sporulation proceeds in a sequence of biochemical and morphogenic changes in the cell (13) and has been viewed as a model system of cellular differentiation. During the early stage of sporulation, the forespore compartment is engulfed by the rapid growth of the folded mother cell cytoplasmic membrane and consequently becomes a discrete cell within the mother cell cytoplasm. To study the gene expression and biochemical events taking place in both the mother cell and forespore compartment, separation of these fractions is required. Recently a procedure for isolating the forespores from bacilli has been developed (1, 4, 6). This paper describes briefly some biochemical and functional properties of forespore and mother cell cytoplasm isolated from Bacillus subtilis at each stage of sporulation. The transformable Marburg strain of B. subtilis 60015 (try-, met-) originally obtained from Ernst Freese (National Institutes of Health, Md., USA) was used. The cells were cultivated in a nutrient sporulation medium as described previously (14). The growth stage was identified by tn, where n is the number of hours after the end of exponential growth. All subsequent stages of sporulation followed synchronously. Mother cell cytoplasm, forespores, and forespore cytoplasm were

Purification and Biosynthesis of a Spore Coat Protein in Bacillus megaterium QM B1551

Summary: A spore coat protein of Bacillus megaterium QM B1551 was purified from an SDS-DTT extract of lysozyme-digested spores using Sephadex, DEAE-cellulose chromatography and polyacrylamide disc gel electrophoresis. The molecular weight of the protein was approximately 12000. It constituted about 13% of spore dry weight and was rich in lysine, aspartic acid, glycine and alanine residues. The sum of these four amino acids constituted 53.9% of the total amino acids. Small amounts of phosphorus and sugar were detected, but no amino sugar. The protein was synthesized in the cytosol of sporulating mother cells at t 3, and accumulated on the forespores.

Studies on automated survey system of environmental chemicals by gas chromatography-mass spectrometry-computer. II: Three dimensional mass chromatography

Abstract A three dimensional mass chromatography (TDMC), which is the extended method of mass chromatography, has been developed for the analysis of environmental chemicals. A considerable performance was suggested by the combination of TDMC to gas chromatography-mass spectrometry-computer system.