• YAKHAK HOEJI
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• v.38 no.6
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• pp.696-702
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• 1994

An interspecific fusant strain, MS1, was obtained by protoplast fusion between S. peucetius subsp. caesius and S. platensis. We studied on the microbiological and cultural characteristics of the fusant MS1. In liquid culture, the viscosity of culture broth of S. peucetius increased during incubation However the fusant MS1 formed pellet like S. platensis without viscosity change. On agar medium the colony morphology of MSI resembled S. platensis but the color was similar to S. peucetius. The fermentation products of the fusant MS1 was identical with S. peucetius.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.66-71
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• 2005

Doxorubicin is a clinically important anticancer polyketide compound that is typically produced by Streptomyces peucetius var. caesius. To improve doxorubicin productivity by S. peucetius, a doxorubicin pathway-specific regulatory gene, dnrI, was cloned into a high-copy-number plasmid containing a catechol promoter system. The S. peucetius containing the recombinant plasmid exhibited approximately 9.5-fold higher doxorubicin productivity compared with the wild-type S. peucetius. The doxorubicin productivity by this recombinant S. peucetius strain was further improved through the optimization of culture media composition. Based on the Fractional Factorial Design (FFD), cornstarch, $K_2HPO_4$, and $MgSO_4$ were identified to be the key factors influencing doxorubicin productivity. The Response Surface Method (RSM) results based on 20 independent culture conditions with varying amounts of key factors predicted the highest theoretical doxorubicin productivity of 11.1 mg/l with corn starch of 46.33 g/l, $K_2HPO_4$ of 4.63 g/l, and $MgSO_4$ of 9.26 g/l. The doxorubicin productivity of the recombinant S. peucetius strain with the RSM-based optimized culture condition was experimentally verified to be 11.46 mg/l, which was approximately 30.8-fold higher productivity compared with the wild-type S. peucetius without culture media optimization.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.10-16
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• 2012

The Streptomyces peucetius OIM (Overproducing Industrial Mutant) strain is a recursively-mutated and optimally-screened strain used for the industrial production of polyketide antibiotics, such as doxorubicin (DXR). Using the S. peucetius OIM mutant strain as a surrogate host, a model minimal polyketide pathway for aloesaponarin II, an actinorhodin shunt product, was cloned in a high-copy conjugative plasmid, followed by functional pathway expression and quantitative metabolite analysis. The level of aloesaponarin II production was noted as being significantly higher in the OIM strain than in the wild-type S. peucetius, as well as in the regulatory network-stimulated S. coelicolor mutant strain. Moreover, the aloesaponarin II production level was seen to be even higher in a down-regulator $wblA_{spe}$-deleted S. peucetius OIM strain, implying that the rationally-engineered S. peucetius OIM mutant strain could be used as an efficient surrogate host for the high expression of foreign polyketide pathways.

• YAKHAK HOEJI
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• v.38 no.6
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• pp.749-755
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• 1994

An interspecific fusant strain, Streptomyces MS1 was obtained by protoplast fusion between S. peucetius subsp. caesius and S. platensis. We studied on the fermentation characteristics of the fusant strain. The fermentation products of the fusant MS1 was identical with S. peucetius, but its production of anthracycline was more stable than S. peucetius under various fermentation conditions in regard to acidogenesis of fermentation broth. The optimal medium composition for anthracycline production by fusant MS1 as follows: sucrose 2.0%, glucose 1.0%, soytone 0.7%, $CaCO_3$ 0.2%, $KH_2PO_4$ 0.013%, casamino acids 0.01%, $K_2SO_4$ 0.025%, $MaCl_2\;6H_2O$ 1.024%, 5M $CaCl_2\;5H_2O$ 0.4%, 1N NaOH 0.7%, 20% L-proline 1.5%. In this condition, the productivity of anthracycline was $80{\sim}100\;{\mu}g/ml$.

• Journal of Industrial Technology
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• v.2
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• pp.21-25
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• 1982

This study was to investigate the basic research about Adriamycin production from Streptomyces peucetius var. caesius. Streptococcus pyogenes(YUFE 2204) was sensitive against Adriamycin and its MIC value was $3.125{\mu}g/ml$. Several mutants were isolated by UV-light. Among 325 mutants, Streptomyces peucetius var. caesius YS-107 was showed highest productivity of Adriamycin.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.895-898
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• 2001

DNA sequence analysis of doxA from Streptomyces peucetius subsp. caesius ATCC 27952 revealed a $95\%$ amino acid identity with that of Streptomyces strain C5. DoxA from S. peucetius subsp. caesius ATCC 27952 encodes a peptide with both conserved heme-binding and dioxygen-binding motifs. Expression of this gene in S. lividans 1326 resulted in bioconversion of daunorubicin to doxorubicin.

• KSBB Journal
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• v.23 no.1
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• pp.44-47
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• 2008

Anthracycline antibiotics doxorubicin (DXR) is clinically important cancer therapeutic agent produced by Streptomyces peucetius. DXR result by further metabolism of rhodomycin D (RHOD) and require a deoxy-sugar component for their biological activity. In this study, production of TDP-L-daunosamine and its attachment to ${\varepsilon}$-rhodomycinone (RHO) to generate RHOD has been achieved by bioconversion in Streptomyces venezuelae that bears eleven genes. S. peucetius seven genes (dnmUTJVZQS) were transformed by plasmid and S. venezuelae two genes desIII, IV and two more S. peucetius drrA, B genes were integrated into chromosomal DNA. To generate the feeding substrate RHO, 6L S. peucetius grown on agar plate was harvested, extracted with organic solvent and then purified using preparative HPLC. Recombinant S. venezuelae grown on agar plate containing RHO was harvested and its n-butanol soluble components were extracted. The glycosylated product of aromatic polyketide RHO using heterologous host S. venezuelae presents the minimal information for TDP-L-daunosamine biosynthesis and its attachment onto aglycone. Moreover, the structure of auxiliary protein, DnrQ, was predicted by fold recognition and homology modeling in this study. This is a general approach to further expand of new glycosides of antitumor anthracycline antibiotics.

• Molecules and Cells
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• v.26 no.4
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• pp.362-367
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• 2008

We identified a 1,134-bp putative type III polyketide synthase from the sequence analysis of Streptomyces peucetius ATCC 27952, named Sp-RppA, which is characterized as 1,3,6,8-tetrahydroxynaphthalene synthase and shares 33% identity with SCO1206 from S. coelicolor A3(2) and 32% identity with RppA from S. griseus. The 1,3,6,8-tetrahydroxynaphthalene synthase is known to catalyze the sequential decarboxylative condensation, intramolecular cyclization, and aromatization of an oligoketide derived from five units of malonyl-CoA to give 1,3,6,8-tetrahydroxynaphthalene, which spontaneously oxidizes to form 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin). In this study, we report the in vivo expression and in vitro synthesis of flaviolin from purified gene product (Sp-RppA).

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1689-1695
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• 2010

Pantothenate kinase (PanK) catalyzes the first step in the biosynthesis of the essential and ubiquitous cofactor coenzyme A (CoA) in all organisms. Here, we report the identification, cloning, and characterization of panK-sp from Streptomyces peucetius ATCC 27952. The gene encoded a protein of 332 amino acids with a calculated molecular mass of 36.8 kDa and high homology with PanK from S. avermitilis and S. coelicolor A3(2). To elucidate the putative function of PanK-sp, it was cloned into pET32a(+) to construct pPKSP32, and the PanK-sp was then expressed in E. coli BL21(DE3) as a His-tag fusion protein and purified by immobilized metal affinity chromatography. The enzyme assay of PanK-sp was carried out as a coupling assay. The gradual decrease in NADH concentration with time clearly indicated the phosphorylating activity of PanK-sp. Furthermore, the ca. 1.4-fold increase of DXR and the ca. 1.5-fold increase of actinorhodin by in vivo overexpression of panK-sp, constructed in pIBR25 under the control of a strong $ermE^*$ promoter, established its positive role in secondary metabolite production from S. peucetius and S. coelicolor, respectively.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.658-661
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• 2015

Genes encoding enzymes with sequence similarity to hopanoids biosynthetic enzymes of other organisms were cloned from the hopanoid (hop) gene cluster of Streptomyces peucetius ATCC 27952 and transformed into Streptomyces venezuelae YJ028. The cloned fragments contained four genes, all transcribed in one direction. These genes encode polypeptides that resemble polyprenyl diphosphate synthase (hopD), squalene-phytoene synthases (hopAB), and squalene-hopene cyclase (hopE). These enzymes are sufficient for the formation of the pentacyclic triterpenoid lipid, hopene. The formation of hopene was verified by gas chromatography/mass spectrometry.