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T Beppu

Publications and source records attributed to T Beppu.

402 records · Page 23Linked to original sources

Induction by mercuric ion of extensive degradation of cellular ribonucleic acid in Escherichia coli.

Low concentrations of HgCl(2) were found to induce extensive degradation of ribonucleic acid (RNA) in exponentially growing Escherichia coli cells but not in stationary-phase cells. Whereas 80% of cellular RNA was degraded during 90 min of incubation with 10(-5)m HgCl(2) at 37 C, HgCl(2) caused only slight degradation in stationary cells, even when present at concentrations higher than 5 x 10(-5)m. Inhibition of RNA synthesis occurred at almost the same concentration of HgCl(2) as degradation, and the ability of stationary-phase cells to synthesize RNA was also resistant to HgCl(2). The transition of cells from complete sensitivity to HgCl(2) to a fully insensitive state took place simultaneously with the cessation of growth. p-Chloromercuribenzoate was also found to induce remarkable degradation of RNA. In E. coli Q13, a mutant deficient for ribonuclease I, no degradation of RNA was evident, even in the exponential growth phase. 3'-Mononucleotides but not 5'-mononucleotides were found among the degradation products of cellular RNA. 2',3'-Cyclic mononucleotides were produced when RNA was degraded by the cell-free extracts of the Hg treated cells. Almost complete unmasking of the latent ribonuclease occurred in the particle fraction containing subribosomal particles of the Hg-treated cells. These data suggest that the incubation of exponentially growing E. coli cells with HgCl(2) led to the unmasking of ribonuclease I, which resulted in the extensive degradation of cellular RNA. The activation of ribonuclease by HgCl(2) in the isolated particulate fraction of E. coli K-12 which occurred in vitro suggested the presence of an Hg-sensitive inhibitor for ribonuclease I.

Bacterial Proteins↗

Genetic organization of Acetobacter for acetic acid fermentation.

Plasmid vectors for the acetic acid-producing strains of Acetobacter and Gluconobacter were constructed from their cryptic plasmids and the efficient transformation conditions were established. The systems allowed to reveal the genetic background of the strains used in the acetic acid fermentation. Genes encoding indispensable components in the acetic acid fermentation, such as alcohol dehydrogenase, aldehyde dehydrogenase and terminal oxidase, were cloned and characterized. Spontaneous mutations at high frequencies in the acetic acid bacteria to cause the deficiency in ethanol oxidation were analyzed. A new insertion sequence element, IS1380, was identified as a major factor of the genetic instability, which causes insertional inactivation of the gene encoding cytochrome c, an essential component of the functional alcohol dehydrogenase complex. Several genes including the citrate synthase gene of A. aceti were identified to confer acetic acid resistance, and the histidinolphosphate aminotransferase gene was cloned as a multicopy suppressor of an ethanol sensitive mutant. Improvement of the acetic acid productivity of an A. aceti strain was achieved through amplification of the aldehyde dehydrogenase gene with a multicopy vector. In addition, spheroplast fusion of the Acetobacter strains was developed and applied to improve their properties.

Acetates↗

A-factor and streptomycin biosynthesis in Streptomyces griseus.

Accumulating data have shown that the metabolites with a gamma-butyrolactone ring functions as an autoregulatory factor or a microbial hormone for the expression of various phenotypes not only in a variety of Streptomyces spp. but also in the distantly related bacteria. A-factor, as a representative of this type of autoregulators, triggers streptomycin biosynthesis and cellular differentiation in Streptomyces griseus. A model for the A-factor regulatory cascade on the basis of recent work is as follows. At an early step in the A-factor regulatory relay, the positive A-factor signal is first received by an A-factor receptor protein that is comparable in every aspect to eukaryotic hormone receptors, and then, via one or more regulatory steps, transmitted to an A-factor-responsive protein that binds to the upstream activation sequence of the strR gene, a regulatory gene in the streptomycin biosynthetic gene cluster. The StrR protein thus induced appears to activate the other streptomycin biosynthetic genes. This review summarizes the characteristics of A-factor as a microbial hormone and the A-factor regulatory relay leading to streptomycin production.

4-Butyrolactone↗

Expression of cloned calf prochymosin cDNA under control of the tryptophan promoter.

To increase yields of calf prochymosin (PC) produced in Escherichia coli, PC cDNA was cloned in a plasmid vector under control of the trp promoter. The hybrid plasmid pCR501 constructed for this purpose contains cDNA coding for PC (from the 5th Arg to the C-terminal Ile) fused to the N-terminal fragment of the trpE gene preceded by the trp promoter and attenuator region. E. coli C600 harboring this plasmid produces approx. 300 000 molecules of PC per cell. This is about a tenfold increase above the amount obtained using lacUV5 promoter [Nishimori et al., Gene 19 (1982) 337-344]. A similar plasmid, pCR601, which contains the same coding sequence fused to the trp promoter and N-terminal fragment of the trpL gene, directs the production of PC at the same rate as pCR501. In pCR601 the trp attenuator is deleted. Another plasmid, pCR701, in which construction of a sequence coding for fMet-PC cDNA that was aided by chemical synthesis, was placed under direct control of the trp promoter, produced PC at a much lower rate. Extracts prepared from all these bacterial transformants in the presence of urea showed distinct milk-clotting activity after renaturation and processing.

Amino Acid Sequence↗

Evaluation of a new lipophilic prodrug 3', 5'-dioctanoyl-5-bromodeoxyuridine (BrdU-C8) suspended in Lipiodol as a radiosensitizer for the treatment of AH136B tumor.

The effect of anti-cancer treatment on the AH136B tumor was studied using external beam irradiation in combination with a new oil-soluble agent, 3', 5'-dioctanoyl-5-bromodeoxyuridine (BrdU-C8), a lipophilic prodrug of BrdU. BrdU-C8 was dissolved in an oily lymphographic agent, Lipiodol (BrdU-C8/Lipiodol). BrdU-C8/Lipiodol is selectively accumulated in the neovasculature of the tumor and gradually releases BrdU. The AH136B tumor cell, a transplantable rat ascites hepatoma cell line, was implanted in the dorsal foot of rats. In vivo labeling index (L.I.) of the tumor cells after the intraarterial infusion of BrdU-C8/Lipiodol was significantly increased compared to the L.I. observed after intraarterial or intravenous infusion of BrdU. In addition, X-ray irradiation in combination with intraarterial infusion of BrdU-C8/Lipiodol significantly inhibited the tumor growth, indicating the increased radiosensitizing effect.

Animals↗