• Korean Journal of Microbiology
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• v.29 no.3
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• pp.160-166
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• 1991

The regulatory mechanism of tubercidin biosynthesis in Streptomyces tubercidicus was studied. In a wild type strain, addition of adenine and histidine into the medium decreased the tubercidin production by 60-65% and 40%, respectively. The effects of adenine and histidine were alleviated by the addition of inosine monophosphate and 5-aminoimidazole-4-carboxamide ribotide. The production of tubercidin in S. tubercidicus K115 strain ($ade^{-}$ ) was nearly shut off by histidine. In contrast with K115 strain, adenine inhibited the tubercidin biosynthesis in S. tubercidicus K412 strain ($his^{-}$. In S. tubercidicus F667 strain ($ade^{-}$ , $his^{-}$ ), tubercidin production was increased by adenine and histidine. From the effects of adenine and histidine on tubercidin biosynthesis in S. tubercidicus wild type and mutant strains, it became known that feedback control by adenine and histidine of biosynthetic pathwat for purine ribonucleotide and histidine are involved in the regulation of tubercidin biosynthesis.

• Korean Journal of Microbiology
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• v.29 no.2
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• pp.97-103
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• 1991

Glutamine phosphoribosylpyrophosphate amidotransferase of Streptomyces tubercidicus was purified and characterized. Molecular weight of the isolated enzyme was determined to be approximately 230,000 and was composed foru identical subunits having a molecular weight of 58,000. This enzyme was strongly inhibited by AMP while considerably inhibited by ATP and GTP. Inhibition effect of enzyme activity by AMP was antagonized by increased concentration of substrate, PRPP, and metal ion (especially, $Mg^{++}$) was essential in both catalytic activity and nucleotide inhibition of this enzyme. Therefore, it was confirmed that end product inhibition of glutamine phosphoribosylpyrophosphate amidotransferase by adenine participated in the regulation of tubercidin biosynthesis from Streptomyces tubercidicus.s.

• Korean Journal of Microbiology
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• v.24 no.4
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• pp.364-369
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• 1986

A procedure for the preparation, regeneration and fusion of protoplasts of Streptomyces tubercidicus was confirmed. Also, protoplast releasingprocesses from mycelia were observed by scanning electron microscope. Three types of protoplasts releasing processes-from the hyphal tip, hyphal end regions and lateral regions of the hyphae-were observed. More than 17% regeneration efficiency was obtained by regeneration medium that is composed of tryptone-yeast extract-sodium acetate-$MgCl_2-CaCl_2$-sucrose. Optimal concentrations of $Ca^{++},\;Mg^{++}$ and sucrose in the regeneration medium were 50mM, 0.4-0.5M respectively. Above 30% of fusion frequency of the protoplasts derived from two auxotrophic strains of S. tubercidicus was induced by polyethylene glycol 4000(60% w/v).

• Microbiology and Biotechnology Letters
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• v.24 no.1
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• pp.82-86
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• 1996

Three thousand microbial strains collected from different sources were screened for herbicidal activity. A strain of ME-9189 showed herbicidal activity against Digitaria sanguinalis and Portulaca oleracea was isolated from a mountainy soil. Based on taxonomic studies, the strain was identified as Streptomyces tubercidicus. The active compound of ME-9189 was purified from the culture broth by charcoal, silica gel, sephadex LH-20 column chromatography and crystalization, consecutively. The ME-9189 compound was identified as tubercidin by spectroscopic methods of UV, $^{1}H$ and $^{13}C$-NMR, and EIMS. In the bioassay, growth of radish shoot and root was inhibited by 50% with tubercidin treatment of 10 ppm, showing 2 times higher activity than that of herbicidin A and similar to that of toyocamycin.

• Korean Journal of Microbiology
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• v.25 no.3
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• pp.180-183
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• 1987

Stu I, type II restriction endonuclease, has been purified to homogeneity from Streptomyces tubercidicus (ATCC 25502), and its catalytic properties have been studied. For the purification of Stu I endonuclease free of nonspecific nucleases, DEAE-Sephadex (A-50), QAE-Sephadex (A-50) and Heparin-agarose column chromatography have been performed after ammonium sulfate fractionation of the crude extract. The enzyme was further purified by gel filtration using Sephadex G-100 column to obtain homogeneous form of protein. The single polypeptide species of Stu I endonuclease has a subunit molecular weight of 34,000 $\pm$ 1,000 daltons as judged by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Stu I endonuclease requires $Mg^{2+}$ ion for its activity and is maximally active at neutral pH (7.0-8.0) in the absence of NaCl.

• Archives of Pharmacal Research
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• v.21 no.6
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• pp.692-697
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• 1998

GTP cyclohydrolase I catalyzing the first reaction in the biosynthesis of pterin moiety of folic acid in bacteria, was purified from Streptomyces tubercidicus by at least 203-fold with a yield of 32% to apparent homogeneity, using ammonium sulfate fractionation, DEAE-cellulose, Sepharose CL-6B, and hydroxylapatite column chromatography. The molecular weight of the native enzyme was estimated to be 230,000 daltons by gel permeation chromatography. The purified enzyme gave a single band on sodium dodesyl sulfate-polyacrylamide gel electrophoresis and its molecular weight was apparently 58,000 daltons. These results indicate that the enzyme consists of four subunits with the same molecular weight. The $K_m$ and $V_{max}$ values for GTP of the purified enzyme were determined to be 80${\mu}$M and 90nmol/min (mg protein), respectively. The optimum pH and temperature for the enzyme reaction were pH 7.5-8.5 and $40-42^{\circ}C$, respectively. Coenzyme or metal ion was not required for the enzyme activity. The enzyme activity was inhibited by most divalent cations, while it was slightly activated by potassium ion. In case of nucleotides, CTP, GMP, GDP, and UTP inhibited enzyme activity, among which GDP exhibited the strongest inhibitory effect.