• Korean Journal of Microbiology
• /
• v.53 no.2
• /
• pp.118-122
• /
• 2017

Bacterial growth inhibition by lead, zinc and cadmium was measured by using modified Tris minimal medium. The toxicity against Escherichia coli strain was in the order of zinc> cadmium> lead, and the Escherichia coli strain overexpressing iron-containing superoxide dismutase 2 of Streptomyces coelicolor A3(2) was found to have resistance to heavy metals.

• Journal of Microbiology
• /
• v.42 no.1
• /
• pp.64-67
• /
• 2004

The pellets from a culture of Streptomyces coelicolor A3(2) that were submerged shaken were disintegrated into numerous hyphal fragments by DNase treatment. The pellets were increasingly dispersed by hyaluronidase treatment, and mycelial fragments were easily detached from the pellets. The submerged mycelium grew by forming complexes with calcium phosphate precipitates or kaolin, a soil particle. Therefore, the pellet formation of Streptomyces coelicolor A3(2) can be considered a biofilm formation, including the participation of adhesive extracellular polymers and the insoluble substrates.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.3
• /
• pp.480-484
• /
• 2010

AfsR2 is a global regulatory protein that stimulates antibiotic biosynthesis in both Streptomyces lividans and S. coelicolor. Previously, various afsR2-dependent genes including a putative abaA-like regulatory gene, SCO4677, were identified through comparative DNA microarray analysis. To further identify the putative SCO4677-dependent proteins, the comparative proteomics-driven approach was applied to the SCO4677-overexpressing strains of S. lividans and S. coelicolor along with the wild-type strains. The 2D gel electrophoresis gave approximately 277 protein spots for S. lividans and 207 protein spots for S. coelicolor, showing different protein expression patterns between the SCO4677-overexpressing strains and the wild-type strains. Further MALDI-TOF analysis revealed that only 18 proteins exhibited similar expression patterns in both S. lividans and S. coelicolor, suggesting that the SCO4677 could encode an abaA-like regulator that controls a few cross-species common proteins as well as many species-specific proteins in Streptomyces species.

• Korean Journal of Microbiology
• /
• v.33 no.4
• /
• pp.237-241
• /
• 1997

Lipase (EC 3.1.1.3) in the culture filtrate of Streptomyces coelicolor A3(2) was active on ${\alpha}$-naphthyl-butyrate as well as on various triacylglycerols with different lengths of acyl chains. The extracellular lipase was purified 15-fold by ammonium sulfate fractionation, Sephadex G-100, DEAE-Cellulose and Phenyl-Sepharose CL4B column chromatography with overall yield of 16%. It showed an molecular weight of 34.7 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme activity with tributyrin as substrate was optimal at pH 8.0~9.0 and at $37^{\circ}C$. The enzyme activity decreased when the chain length of acyl group of triacyglycerol increased. A-factor, a hormone-like regulator of Streptomyces differentiation inhibited the lipase activity, which might corelate with the low enzyme activity in early exponential growth phase.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.5
• /
• pp.799-803
• /
• 2006

It was reported that an accumulation of Sadenosyl-L-methionine increases production of actinorhodin in Streptomyces lividans and induces antibiotic biosynthetic genes. We also obtained the same result in Streptomyces coelicolor A3(2). Therefore, in order to identify proteins changed by the addition of S-adenosyl-L-methionine in S. coelicolor A3(2), LC/MS/MS analyses were carried out. Thirteen proteins that were not observed in the control were found.

• Korean Journal of Microbiology
• /
• v.39 no.2
• /
• pp.83-88
• /
• 2003

We cloned a gene encoding acetyl xylan esterase(axeA) of Streptomyces coelicolor A3(2) and studied its expression pattern in Escherichia coli. The full sequence of axeA was amplified by PCR. Sequence analysis of the PCR product revealed an open reading frame of 1,008 nucleotides encoding a protein consisted of 335 amino acid residues, with a calculated molecular mass of about 38 kDa. The base sequence showed 98% homology to the same gene of Streptomyces lividans. Two different kinds of acetyl xylan esterases were produced in Escherichia coli(pLacI) by IPTG induction; their molecular weights were 38 kDa and 34 kDa, respectively. Of these, 38 kDa protein seemed to be a total protein holding N-terminal signal peptide region, whereas 34 kDa protein seemed to be a matured protein without signal peptide which was produced by peptide bond cleavage between two amino acid residues of alanine 41 and alanine 42.

• Korean Journal of Microbiology
• /
• v.32 no.4
• /
• pp.264-270
• /
• 1994

A gene coding for a novel putative $\sigma$ factor of RNA polymerase has been identified from Streptomyces coelicolor A3(2) using Escherichia coli rpoS gene fragment as a probe. The 486 bp rpoS gene fragment was amplified from E. coli genomic DNA by PCR with two synthetic oligonucleotides, the sequences of which were deduced from the amino acid sequences in the regions 2.3 and 4.2 conserved among various bacterial factors. When E. coli genomic DNA fragments were hybridized with cloned rpoS probe, only one band corresponding to rpoS gene (3.2 kb PvuII fragment or 2.3 kb KpnI fragment) was detected. In S. coelicolor, however, two bands were detected both in PvuII digested DNA and SalI digested DNA. 3.5 kb PvuII fragment which binds the rpoS gene probe was cloned (pMS1) from the sublibrary, and the nucleotide sequences of 1.0 kb BamH'/HincII subclone (pBH2) was partially determined. The nucleotide sequences revealed extensive similarity to other $\sigma$ factor genes of S. coelicolor (hrdA, hrdB, hrdC, hrdD), S. aureofaciens (hrdA, hrdB, hrdC, hrdD), Synechococcus species, Pseudomonas aeruginosa, Stigmatella aurantiaca, and Anabaena species. The nucleotide sequences in regions 1.2 and 4 were compared with the corresponding regions of 5 known ${\sigma}$ factor genes of S. coelicolor by multiple alignment. It turned out that the cloned gene is most closely related to hrdA showing 88% amino acid similarity in region 1.2 and 75% in region 4.

• KSBB Journal
• /
• v.19 no.6 s.89
• /
• pp.462-466
• /
• 2004

Mycothiol (MSH), a low molecular antioxidant thiol compound, was purified and analyzed from Streptomyns coelicolor A[3]2 by the monobromobimane fluorescence detection method modified by this lab. Through HPLC chromatpgram, MSH fraction was obtained following the elution time of standard MSH (donated by Dr. Robert C. Fahey). That MSH showed the highest concentration among the thiol compounds contained in the cell indicated that MSH was the key thiol compound having antioxidant activity. To understand the role of gene of inositol monophosphatase (I-1-Pase) involved in the MSH biosynthesis, it was isolated from S. coelicolor A(3)2 and cloned and overexpressed in the Escherichia coli. The expressed I-1-Pase was purified through Ni-NTA column. The soluble protein consisted of 281 amino acids, and the molecular weight was 32 kDa. I-1-Pase of S. coelicolor A(3)2 had the sequence homology with those of human and E. coli by 24 and $25\%$, respectively, and had two conserved domains (mofif A and motif B) which were typical of I-1-Pase.

• Korean Journal of Microbiology
• /
• v.40 no.3
• /
• pp.232-236
• /
• 2004

We investigated hydrogen peroxide resistance of Escherichia coli possessing acetyl xylan esterase(AxeA) of Streptomyces coelicolor A3(2). The induction of AxeA production by isopropyl-$\beta$-thiogalactoside was confirmed by SDS-polyacrylamide gel electrophoresis. The differences in growth between induced and non-induced E. coli were determined by the changes in optical density of cultures after hydrogen peroxide treatment The lethal effect of hydrogen peroxide was observed for non-induced cultures at all concentrations tested in this study (lmM, 2.5mM and 5mM). However, cultures induced for AxeA production resisted the lethal effect, except at 5mM where cells were killed irrespective of the AxeA production. The axeA induction increased survival against 1.5mM hydrogen peroxide from 59% to 74%. In addition, AxeA producing E. coli showed increased survival at $45^{\circ}C$, near maximum growth temperature. Therefore, it was concluded that AxeA conferred a cross-resistance upon the bacterium against both oxidative- and heat stress.

• Genomics & Informatics
• /
• v.6 no.1
• /
• pp.44-49
• /
• 2008

Protein kinase C (PKC) is a family of kinases involved in the transduction of cellular signals that promote lipid hydrolysis. PKC plays a pivotal role in mediating cellular responses to extracellular stimuli involved in proliferation, differentiation and apoptosis. Comparative analysis of the PKC-${\alpha},{\beta},{\varepsilon}$ isozymes of 200 recently sequenced microbial genomes was carried out using variety of bioinformatics tools. Diversity and evolution of PKC was determined by sequence alignment. The ser/thr protein kinases of Streptomyces coelicolor A3 (2), is the only bacteria to show sequence alignment score greater than 30% with all the three PKC isotypes in the sequence alignment. S.coelicolor is the subject of our interest because it is notable for the production of pharmaceutically useful compounds including anti-tumor agents, immunosupressants and over two-thirds of all natural antibiotics currently available. The comparative analysis of three human isotypes of PKC and Serine/threonine protein kinase of S.coelicolor was carried out and possible mechanism of action of PKC was derived. Our analysis indicates that Serine/ threonine protein kinase from S. coelicolor can be a good candidate for potent anti-tumor agent. The presence of three representative isotypes of the PKC super family in this organism helps us to understand the mechanism of PKC from evolutionary perspective.