• Korean Journal of Microbiology
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• v.41 no.1
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• pp.87-92
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• 2005

Pronase, a protease produced for commercial purpose by Streptomyces griseus, was composed of serine protease, alkaline protease, aminopeptidase and carboxypeptidase complex, and it has been widely used as anti-inflammatory drugs for human therapy. In this study, we developed a new integration vector, pHJ101 derived from pSET152, containing strong promoter, ermE, to overexpress a certain protease gene. Specific PCR primers for cloning of sprA (a gene for S. griseus protease A) and sprB (a gene for S. griseus protease B) genes were designed from the basis of nucleotide sequence in databases and amplified by PCR. Plasmid pHJ201 and pHJ202 were constructed by inserting of amplified each gene in a vector pHJ101. S. griseus HA and S. griseus HB were respectively obtained by conjugal process of a parent strain, S. griseus IFO 13350 with the recombinant Escherichia coli harboring plasmid pHJ201 or pHJ202. When protease activity was measured in flask cultivation, produced protease levels of S. griseus HA and S. griseus HB increased about 5.3 times and 5 times, respectively, more than that of parent strain. And, the constructed integrating plasmid pHJ101 was applicable for overexpression of a certain gene in Streptomyces sp.

• Journal of Microbiology and Biotechnology
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• v.6 no.4
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• pp.265-269
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• 1996

An isolate of Streptomyces rochei synonym was found to produce antibiotics with narrow anti-microbial spectrum against Streptococcus and Xanthomonas sp. Among the antibiotic complex produced by the strain, the main active compound was isolated, and its physico-chemical properties and biological activities were investigated. Molecular weight of the compound was determined to be ${[M+H]}^+$ 797 (FAB-MS). UV, $^1H \;and\;^{13}C$ NMR, and IR spectra suggested that the compound is a kirromycin-like aurodox group antibiotic. However, the anti-microbial spectrum of the main compound was slightly different from that of kirromycin. In addition, it was newly found that kirromycin showed a selective anti-microbial activity against Streptococcus pyogenes and phytopathogenic Xanthomonas sp.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1978.10a
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• pp.207.5-208
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• 1978

Effects of the plant growth regulator (P. G. R.)on the reaction of proteinase, $\gamma-amylase$ and acid phosphatase were investigated, and also were the conditions of production of P. G. R. by Stroptomyces sp. 445. The P. G. R. had no effect on the act ivities of such enzymes in mung bean seedling. But in germinating seed previously treated with P. G. R. it effected the activity of protease in cotyledon. In the conditions of production of P. G. R., the maxim, activity was appeared in shaking cutlure at $30^{\circ}C$ for 5 days, and by the addition of peptone or casein hydrolysate as nitrogen source, soluble starch as carbon source, and sulfur as metal ion.

• Microbiology and Biotechnology Letters
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• v.27 no.4
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• pp.339-343
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• 1999

Pasteurella haemolytica is well known to cause severe pneumonia, consolidation and oedema of the lung, and fibrinous pleurisy under the stress and infection of virus in the cattle. In the course of our screening for antimicrobial agents against P.haemloytica, compound 51086 has been isolated from the fermentation broth of Streptimyces sp. 51086. The compound 51086 was purified by SiO2, Sephadex LH-20 and ODS column chromatographies and HPLC, subsequently. The structure of compound 51086 was determined as hygromycin A by combination of 1H NMR, 13C NMR, HMBC, and ESI-MS. This compound showed significant antibacterial activity against P.haemolytica and P.multocida.

• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.96-105
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• 2005

For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

• Journal of Microbiology and Biotechnology
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• v.8 no.5
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• pp.455-460
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• 1998

In the course of screening for new antitumor substances, a novel cytotoxic agent inducing apoptotic cell death was isolated from the culture broth of marine bacterial strain SCRC A-20. Strain SCRC A-20 was separated from a mollusk and was chemotaxonomically identified as a Streptomyces sp. The cytotoxic substance was purified by organic solvent extraction followed by silica gel column chromatography and preparative TLC. HRFAB-MS determined its molecular formula to be $C_{30}H_{40}N_2O_5$ (MW 508). The 1D and 2D NMR spectral data demonstrated that the substance has a novel lactam structure of a 20-membered macrocycle coupled with a unique acyl tetramine and bicyclo［3.3.0］ octane, which includes three methyl groups, six olefinic protons, five carbonyl groups, a conjugated diene and a dienone. The substance, named aburatubolactam C, appeared to be cytotoxic for various continuously proliferating tumor cells of human and murine origins. The $IC_{50}$ values determined by MIT assay were in the range of 0.3 to $5.8\mug/ml$. When Jurkat T cells were treated with $3\mug/ml$. of aburatubolactam C, the apoptotic DNA fragmentation was detectable within 3 h, indicating that the cytotoxic effect of aburatubolactam C on tumor cells is attributable to the induced apoptosis.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.895-898
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• 2005

In the course of searching for potent chitin synthase 1 inhibitors from natural resources, Streptomyces sp. A6705 was found to exhibit potent inhibitory activity against the chitin synthase 1 from C. albicans (CaCHS1p). As a result, the inhibitor was isolated and identified using a series of chromatographies. Through chemical analyses with UV spectrophotometry, MS spectrometry, and various NMR techniques, the inhibitor was identified as N,N-bis(2-phenylethyl)urea. The compound exhibited strong inhibitory activity against the chitin synthase 1 from C. albicans with an $IC_{50}$ of 14 ${\mu}g$/ml, representing a similar inhibitory activity to that of the well-known chitin synthase inhibitor, polyoxin D ($IC_{50}$ 15 ${\mu}g$/ml). However, the compound showed no inhibitory activity against the chitin synthase 2 of Saccharomyces cerevisiae up to 280 ${\mu}g$/ml, which is structurally and functionally analogous to CaCHS 1 p. In addition, the compound exhibited weak antifungal activities against Cryptococcus neoformans and Rhizoctonis solani.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.909-912
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• 2005

In the course of screening for selective growth inhibitors against the mycelial form of Candida albicans, we isolated a Streptomyces sp. A6792 from soils. The inhibitor was isolated from the above bacterium and identified through several spectral analyses with UV and mass spectrophotometries, and various NMR. The compound was determined to be a macrocyclic dilactone antibiotic, IKD-8344 (molecular weight: 844, molecular formula: $C_{48}H_{76}O_{12}$). The compound selectively inhibited the growth of mycelial form of C. albicans with an MIC of 6.25 ${\mu}g/ml$. It also exhibited strong inhibitory effect preferentially on the mycelial form of various Candida spp. including C. krusei, C. tropicalis, and C. lusitaniae, with MICs ranging from 1.56 to 25 ${\mu}g$/ml. Furthermore, the compound showed no significant toxicity against SPF ICR mice up to 60 mg/kg. These results suggest that IKD-8344 is a useful lead compound for the development of novel antifungal agents, based on the preferential growth inhibition against Candida spp.

• BMB Reports
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• v.35 no.6
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• pp.568-575
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• 2002

An $\alpha$-amylase (EC 3.2.1.1) was purified that catalyses the production of a high level of maltose from starch without the attendant production of glucose. The enzyme was produced extracellularly by thermophilic Streptomyces sp. that was isolated from Thailand's soil. Purification was achieved by alcohol precipiation, DEAE-Cellulose, and Gel filtration chromatographies. The purified enzyme exhibited maximum activity at pH 6-7 and $60^{\circ}C$. It had a relative molecular mass of 45 kDa, as determined by SDS-PAGE. The hydrolysis products from starch had $\alpha$-anomeric forms, as determined by $^1H$-NMR. This maltose-forming $\alpha$-amylase completely hydrolyzed the soluble starch to produce a high level of maltose, representing up to 90%. It hydrolyzed maltotetrose and maltotriose to primarily produce maltose (82% and 62%, repectively) without the attendant production of glucose. The high maltose level as a final end-product from starch and maltooligosaccharides, and the unique action pattern of this enzyme, indicate an unusual maltose-forming system. After the addition of the enzyme in the bread-baking process, the bread's volume increased and kept its softness longer than when the bread had no enzyme.

• Microbiology and Biotechnology Letters
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• v.6 no.3
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• pp.121-127
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• 1978

This paper deals with investigation on the various effects of phosphodiesterase production of Streptomyces sp. and several properties of the enzyme. The results were as follows. 1) As a carbon source, sucrose was most effective PDase production when it was added. to the basal medium at 3％ concentration. 2) These enzymes were remarkably activated by $Ca^{2+}$, Co$^{2+}$ and Mn$^{2+}$ but inhibited by Cu$^{2+}$. It was observed that concentration of metal ions, 0.1％ of $Ca^{2+}$, 0.01％ of Co$^{2+}$ and 0.04％ of Mn$^{2+}$ were effective on the production of phosphodiesterase and phosphomonoesterase. 3) In case of the effect of aeration volume, 25 ml was very effective, that is, the more sufficient aeration, the better enzyme activity. Enzyme activity was to be found effective at 3％ of inoculation volume, and comparatively more effective at 2％, 4％ of inoculation volume. 4) Initial pH was 8. The enzyme activity reached to the maximum at 48 hours of cultivation time. 5) The optimum pH of phosphodiesterase was about 8 and that of phosphomonoesterase was about 9. The optimum temperature of phosphodiesterase and phosphomonoesterase was 6$0^{\circ}C$ and 5$0^{\circ}C$ respectively. respectively.