• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.135-137
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• 2008

The metabolic engineering of epothilones, as secondary metabolites, was investigated using Sorangium cellulosum to achieve the selective production of epothilone B, a potent anticancer agent. Thus, the propionyl-CoA synthetase gene (prpE) from Ralstonia solanacearum was heterologously expressed in S. cellulosum to increase the production of epothilone B. Propionyl-CoA synthetase converts propionate into propionyl-CoA, a potent precursor of epothilone B. The recombinant S. celluloslim containing the prpE gene exhibited a significant increase in the resolution of epothilones B/A, with an epothilone B to A ratio of 127 to 1, which was 100 times higher than that of the wild-type cells, demonstrating its potential use for the selective production of epothilone B.

• KSBB Journal
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• v.24 no.1
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• pp.65-69
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• 2009

The biological production of a potent anticancer agent, epothilone, by Sorangium cellulosum was carried out using flask and fermentor cultures. Soluble starch was selected as the main carbon source and the concentrations of lactose and yeast extract were optimized at 4 and 0 g/L, respectively, when using the flask cultures. In the fermentor cultures, the cells were cultivated at a high DO level of more than 80% of air saturation in the growth stage and then the DO level was controlled at about 50, 20 or 1-2% when the carbon source was exhausted. The epothilone production increased with decreasing DO level after the exhaustion of the carbon source, and the maximum concentration of epothilone was 5.4 mg/L. It was found that the DO level had significant regulation effects on the epothilone production.

• Journal of Microbiology and Biotechnology
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• v.18 no.8
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• pp.1416-1422
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• 2008

Epothilone and its analogs are a potent new class of anticancer compounds produced by myxobacteria. Thus, in an effort to identify new myxobacterial strains producing epothilone and its analogs, cellulose-degrading myxobacteria were isolated from Korean soils, and 13 strains carrying epothilone biosynthetic gene homologs were screened using a polymerase chain reaction. A migration assay revealed that Sorangium cellulosum KYC3013, 3016, 3017, and 3018 all produced microtubule-stabilizing compounds, and an LC-MS/MS analysis showed that S. cellulosum KYC3013 synthesized epothilone A.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1653-1659
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• 2015

Epothilone, which is produced by the myxobacterium Sorangium cellulosum, contributes significant value in medicinal development. However, under submerged culture conditions, S. cellulosum will accumulate to form bacterial clumps, which hinder nutrient and metabolite transportation. Therefore, the production of epothilone by liquid fermentation is limited. In this study, diatomite-based porous ceramics were made from diatomite, paraffin, and poremaking agent (saw dust). Appropriate methods to modify the porous ceramics were also identified. After optimizing the preparation and modification conditions, we determined the optimal prescription to prepare high-performance porous ceramics. The structure of porous ceramics can provide a solid surface area where S. cellulosum can grow and metabolize to prevent the formation of bacterial clumps. S. cellulosum cells that do not form clumps will change their erratic metabolic behavior under submerged culture conditions. As a result, the unstable production of epothilone by this strain can be changed in the fermentation process, and the purpose of increasing epothilone production can be achieved. After 8 days of fermentation under optimized conditions, the epothilone yield reached 90.2 mg/l, which was increased four times compared with the fermentation without porous ceramics.

• Korean Journal of Microbiology
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• v.47 no.2
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• pp.170-173
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• 2011

The epothilone biosynthetic gene cluster (epoA~F, epoK) of Sorangium cellulosum KYC3013, an epothilone producing myxobacterium isolated in Korea, was cloned. When the amino acid sequences of the encoded proteins were compared with those from S. cellulosum SMP44, S. cellulosum So ce90, and S. cellulosum So0157-2, which were isolated in other continents or country, the proteins from different strains were 97.4-99.8% identical each other. This suggested that the epothilone-biosynthetic gene clusters are well conserved in S. cellulosum strains.

• Journal of Microbiology and Biotechnology
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• v.16 no.4
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• pp.519-523
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• 2006

To improve epothilones production and the ratio of epothilone B/A, carbon sources were investigated in flask culture of Sorangium cellulosum. Depending on the initial concentration, starch significantly enhanced cell growth, but the maximum epothilones productivity and the maximum epothilones production (0.3 mg/l day and 2.6 mg/l at 15 g/l starch, respectively) were relatively low compared with cell growth. On the other hand, addition of glycerol did not stimulate cell growth, but epothilone production was increased from 2.81 mg/l to 7.59 mg/l. Addition of glycerol to culture medium resulted in more significant enhancement of the production of epothilone A, whereas epothilone B levels were relatively constant. Furthermore, when sodium propionate was added as a precursor of methylmalonyl-CoA, it resulted in increase of both total epothilones production and epothilone B/A resolution. Maximum epothilone A and B concentrations reached 10.9 mg/l and 8.58 mg/l, respectively, at 5 mM sodium propionate.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1208-1212
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• 2007

Production of extracellular epothilone B, one of the potent anticancer agents, by free and immobilized Sorangium cellulosum was studied using the repeated batch culture process. The concentration of alginate used in immobilization was directly related to the mass transfer rate of nutrients, mechanical stability, and the epothilone B production yield. With the optimized 3% (w/v) calcium alginate carrier, a prolonged repeated batch culture was investigated for the 5 repeated batches for 24 days. The maximum productivity of epothilone B obtained from the alginate-immobilized cells was 5.03 mg/l/day, which is 3 times higher than that of free cells (1.68 mg/l/day).

• Research in Plant Disease
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• v.24 no.3
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• pp.213-220
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• 2018

We classified the previously reported antagonistic strains of Sorangium cellulosum into 5 subspecies (A-E). Four strains were antagonistic to Botrytis cinerea (AB group) and two strains were antagonistic to Colletotrichum acutatum (AC group). According to the genetic and sequential analyses with standard genes, xynB1, bglA2, groEL1 for grouping, all strains of AB group were belonged to subspecies C and all strains of AC group were belonged to subspecies D. In addition, high pressure liquid chromatography with the culture filtrates confirmed the genetic results, because AB group had peaks with retention time at 20-22.5 minutes, whereas AC group had no peak. There was positive relationship ($R^2=0.9652$) between the control values of infecting B. cinerea on cherry tomatoes and the main peak areas of chromatograms among the four isolates of AB group. From the subspecies results of AB group, the main peak of KYC 3270 was expected to be epothilone D. However the retention times of the standard of commercial epothilone D and the main peak of KYC 3270 culture filtrate were different as 9.9 and 11.581 min., respectively. Finally, the antagonistic metabolite of AB group was inferred as 7-ketone epothilone D.

• Bulletin of the Korean Chemical Society
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• v.34 no.7
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• pp.1972-1984
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• 2013

Microtubules play an important role in intracellular transport, mobility, and particularly mitosis. Paclitaxel (Taxol$^{TM}$) and paclitaxel-like compounds have been shown to be anti-tumor agents useful for various human tumors. Paclitaxel-like compounds operate by stabilizing microtubules through interface binding at the interface between two ${\beta}$-tubulin monomers in adjacent protofilaments. In this paper we present the elucidation of the structural features of paclitaxel and paclitaxel-like compounds (e.g., epothilones) with microtubule stabilizing activities, and relate their activities to spatial and chemical features of the molecules. CATALYST program was used to generate three-dimensional quantitative structure activity relationships (3D-QSARs) resulting in 3D pharmacophore models of epothilone- and paclitaxel-derivatives. Pharmacophore models were generated from diverse conformers of these compounds resulting in a high correlation between experimental and predicted biological activities (r = 0.83 and 0.91 for epothilone and paclitaxel derivatives, respectively). On the basis of biological activities of the training sets, five- and four-feature pharmacophore hypotheses were generated in the epothilone and paclitaxel series. The validation of generated hypotheses was achieved by using twelve epothilones and ten paclitaxels, respectively, which are not in the training sets. The clustering (grouping) and merging techniques were used in order to supplement spatial restrictions of each of hypothesis and to develop more comprehensive models. This approach may be of use in developing novel inhibitor candidates as well as contributing a better understanding of structural characters of many compounds useful as anticancer agents targeting microtubules.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.15
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• pp.6535-6539
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• 2015

This study was designed to assess, whether a new chemotherapeutic microtubule inhibitor, Epothilone B (EpoB, Patupilone), can induce DNA damage in normal ovarian cells (MM14.Ov), and to evaluate if such damage could be repaired. The changes were compared with the effect of paclitaxel (PTX) commonly employed in the clinic. The alkaline comet assay technique and TUNEL assay were used. The kinetics of DNA damage formation and the level of apoptotic cells were determined after treatment with IC50 concentrations of EpoB and PTX. It was observed that PTX generated significantly higher apoptotic and genotoxic changes than EpoB. The peak was observed after 48 h of treatment when the DNA damage had a maximal level. The DNA damage induced by both tested drugs was almost completely repaired. As EpoB in normal cells causes less damage to DNA it might be a promising anticancer drug with potential for the treatment of ovarian tumors.