• Microbiology and Biotechnology Letters
• /
• v.22 no.5
• /
• pp.459-466
• /
• 1994

Virginiae butanolide C(VB-C) is one of the butyrolactone autoregulators, which triggers the production of virginiamycin in Streptomyces virginiae. Streptomyces longwoodensis was selected as a test strain to investigate new VB-C functions. When 100 ng/ml of the synthetic VB-C was added into the culture at 5 hour and 0 hour, the initial production time of antibiotics and a dark blue pigment were shortened by 4~6 hours and 2~4 hours, respectively. HPLC analysis revealed the production of several new antibiotics by VB-C addition. In the SDS-PAGE analysis of the total protein from mycelium several new protein bands showed up and the amounts of certain protein bands increased in the presense of VB-C. The existence of specific VB-C binding protein was confirmed from S. longwoodensis in relation to VB-C signal transduction. These results suggest that the VB-C might have an ability to induce the production of secondary metabolites in Streptomy- ces longwoodensis.

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

• Korean Journal of Microbiology
• /
• v.46 no.4
• /
• pp.359-365
• /
• 2010

Fifty Actinobacteria strains were isolated from rhizosphere soil of Sasa borealis. In the course of screening for antibacterial activity against bacterial leaf spot of pepper (Xanthomonas axonopodis pv. vesicatoria) of isolates, 12 isolates showed strong antibiotic activity. Basis on the 16S rRNA gene sequence, they were belonging to Streptomyces cluster II. Strain JR-24 exhibited strong antibiotic activity against X. axonopodis pv. vesicatoria, had a minimum inhibitory concentration of 10 ${\mu}l$/disc. The strain JR-24 was most closely related to Streptomyces galbus $DSM40089^T$ (98.1%), Streptomyces longwoodensis $LMG20096^T$ (98%) and Streptomyces capoamus $JCM4734^T$ (97.8%). When assayed with the API 20NE and 50 CHE kit, it is positive for utilization of L-arabinose, D-fructose, D-glucose, D-galactose and hydrolysis of gelatin, protein, starch. The strains contained iso-$C_{14:0}$ (25.93%), iso-$C_{15:0}$ (10.13%), anteiso-$C_{15:0}$ (19.29%) and iso-$C_{16:0}$ (20.35%) as major fatty acids and MK-9 (H4), MK-9 (H6), and MK-9 (H8) as the isoprenoid quinone. Strain JR-24 was suggested new species of genus Streptomyces by nearest neighbors of genotypic relationships and phenotypic characterization. This study was important to microbial resources investigation for environment-friendly agriculture.