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埃坡霉素高产菌株的选育 总被引:1,自引:1,他引:0
采用紫外-亚硝酸盐复合诱变法对纤维堆囊菌(Sorangium cellulosum)ATCC 15384进行诱变,结合红霉素抗性筛选,筛得一株遗传稳定的埃坡霉素高产菌株纤维堆囊菌UNl6H127,埃坡霉素A(Epo A)和埃坡霉素B(Epo B)的产量为289.9和25.14μg/L,总产量(EpoA+Epo B,315.04μg/L)是出发菌株的23.8倍. 相似文献
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目的 用基因重组技术选育埃博霉素高产菌株.方法 本研究首先从不同生境中分离筛选到4株能产生埃博霉素B的纤维堆囊菌,以这些野生菌株作为Genome shuffling递归原生质体融合出发菌株.结果 通过五轮基因组重组成功选育到了3株遗传稳定的高产埃博霉素B重组菌株,其中一株重组菌株So F5-09的埃博霉素B产量达到了15.8mg/L,比原始出发菌株So 07-9埃博霉素B产量提高了35.1倍.结论 本研究证明使用野生菌株作为Genome shuffling递归原生质体融合出发菌株也能有效提高目标代谢产物的产量. 相似文献
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目的 通过对埃莎霉素Ⅰ产生菌WSJ-IA进行诱变选育研究,以期获得埃莎霉素Ⅰ高产菌株。方法 使用多功能等离子体诱变系统(multifunctional plasma mutagenesis system, MPMS)对出发菌株的孢子进行等离子体和紫外复合诱变,设定不同的诱变时间处理孢子悬液,通过致死率确定合适的诱变条件,利用突变株摇瓶发酵效价筛选出正突变菌株。结果 在MPMS射频功率为100W,处理距离5mm,气体流量12.5SLM,等离子体-紫外辐射时间为50s时,菌株致死率为96.08%。在此诱变条件下,以突变株的初筛效价为指标的突变率、正突变率分别达到63.96%和22.52%,复筛效价是出发菌株1.5倍以上的有5株,占复筛菌株的9%。最终筛选出一株发酵单位比出发菌株提高221%、埃莎霉素Ⅰ组分含量提高192%的正突变株IA-425。42L自动发酵罐发酵结果表明,该菌株埃莎霉素Ⅰ产量达到(2000±200)μg/mL左右。结论 新型等离子体复合紫外诱变方式,可有效提高菌株的埃莎霉素Ⅰ发酵产量和组分含量。这为埃莎霉素Ⅰ的大规模发酵和临床前研究奠定了良好基础。 相似文献
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以A40926 B产生菌野野村放线菌SIPI-D-30为出发菌株,通过UV诱变及乙酸钠和甘氨酸抗性平板筛选,获得A40926 B产量提高的突变株SIPI-U-157。经优化,确定摇瓶发酵培养基(g/L)为:葡萄糖5.0、麦芽糊精30.0、可溶性淀粉10.0、玉米淀粉30.0、大豆蛋白胨5.0、肉蛋白胨10.0、酵母浸粉5.0、冷榨黄豆饼粉10.0、棉籽饼粉5.0、L-缬氨酸1.0,发酵培养7 d,突变株的A40926 B发酵单位为893 mg/L。该突变株在5 L发酵罐中培养180 h,A40926 B的产量最高为583 mg/L。 相似文献
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微波诱变及添加氯化镧筛选阿扎霉素B高产菌株 总被引:1,自引:0,他引:1
目的 拟通过微波诱变和添加氯化镧对阿扎霉素B产生菌N98-1634进行选育,获得高产菌株.方法 对菌株进行40~240s的微波诱变,采用金黄色葡萄球菌抑菌圈法获得初筛通过株,并进行HPLC复筛获得高产菌株;在高产菌株发酵培养基中加入5~30mg/L的氯化镧考查其对阿扎霉素B合成的影响.结果 通过微波诱变选出l株阿扎霉素B产量达到945.0μg/mL的突变菌株MW-638,较出发菌株N98-1634的752.4μg/mL提高了25.6%,随后经过添加15mg/L氯化镧阿扎霉素B单位最终达到1413.8μg/mL.结论 微波诱变对提高菌株N98-1634阿扎霉素B发酵单位效果明显,诱变菌株添加适量氯化镧可以显著提高其产量. 相似文献
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目的 通过对达托霉素产生菌进行诱变选育,以及前体物补料发酵方式提高达托霉素的产量。方法 采用常压室温等离子体(atmospheric and room temperature plasma, ARTP)技术对玫瑰孢链霉菌进行诱变,以癸酸铵和甘氨酸耐受作为选择压力进行菌株筛选,在摇瓶和100L发酵罐上进行癸酸铵流加补料试验确定最佳的发酵工艺。结果 经诱变选育获得1株突变株FIM-D1568摇瓶效价达到380mg/L,发酵单位较出发菌株提高了35.7%;通过优化100L发酵罐流加补料癸酸铵溶液工艺,使达托霉素发酵效价达到2276mg/L。结论 以ARTP为诱变源,甘氨酸及癸酸铵耐受性为选择性压力,可以快速筛选获得达托霉素高产菌;高产突变菌株在流加补料发酵工艺上优良性状得以发挥,发酵效价大幅提高。 相似文献
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The biosynthetic origin of the epothilone skeleton was studied by the incorporation of 13C and radioactively labeled precursors by Sorangium cellulosum So ce90. The carbon atoms are derived from acetate, propionate, the methyl group of S-adenosyl-methionine, and cysteine which also introduces the sulfur and nitrogen atoms. Epothilone biosynthesis starts with the formation of the thiazole part from acetate and cysteine. The incorporation of acetate or propionate units results in the formation of epothilones A and B, respectively. To introduce the epoxide function of epothilones A and B molecular oxygen is used. 相似文献
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Discovery and development of the epothilones : a novel class of antineoplastic drugs 总被引:1,自引:0,他引:1
The epothilones are a novel class of antineoplastic agents possessing antitubulin activity. The compounds were originally identified as secondary metabolites produced by the soil-dwelling myxobacterium Sorangium cellulosum. Two major compounds, epothilone A and epothilone B, were purified from the S. cellulosum strain So ce90 and their structures were identified as 16-member macrolides. Initial screening with these compounds revealed a very narrow and selective antifungal activity against the zygomycete, Mucor hiemalis. In addition, strong cytotoxic activity against eukaryotic cells, mouse L929 fibroblasts and human T-24 bladder carcinoma cells was observed. Subsequent studies revealed that epothilones induce tubulin polymerization and enhance microtubule stability. Epothilone-induced stabilisation of microtubules was shown to cause arrest at the G2/M transition of the cell cycle and apoptosis. The compounds are active against cancer cells that have developed resistance to taxanes as a result of acquisition of beta-tubulin overexpression or mutations and against multidrug-resistant cells that overexpress P-glycoprotein or multidrug resistance-associated protein. Thus, epothilones represent a new class of antimicrotubule agents with low susceptibility to key tumour resistance mechanisms.More recently, a range of synthetic and semisynthetic epothilone analogues have been produced to further improve the adverse effect profile (or therapeutic window) and to maximize pharmacokinetic and antitumour properties. Various epothilone analogues have demonstrated activity against many tumour types in preclinical studies and several compounds have been and still are being evaluated in clinical trials. This article reviews the identification and early molecular characterization of the epothilones, which has provided insight into the mode of action of these novel antitumour agents in vivo. 相似文献
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采用均匀设计(U_(16)~3)方案考察树脂提取条件对纤维堆囊菌UNH127产生埃博霉素的影响.结果埃博霉素树脂原位提取的最佳工艺条件为:树脂加入时间12.9h,加入量6.4%,培养时间约8d.上述条件下,埃博霉素产量为2.105 2mg/L,与预测值(2.127 2 mg/L)的误差为1.03%. 相似文献
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The epothilones are a novel class of non-taxane microtubule-stabilizing agents obtained from the fermentation of the cellulose degrading myxobacteria, Sorangium cellulosum. Preclinical studies have shown that the epothilones are more potent than the taxanes and active in some taxane-resistant models. Similar to paclitaxel and other taxanes, the epothilones block cells in mitosis, resulting in cell death. The chief components of the fermentation process are epothilones A and B, with epothilones C and D found in smaller amounts. Trace amounts of other epothilones have also been detected. Pre-clinical studies have shown that epothilone B is the most active form, exhibiting significantly higher antitumor activity than paclitaxel and docetaxel. Several phase I and phase II clinical trials are ongoing with epothilone B and BMS 247550, an epothilone B analog. Preliminary reports indicate these agents are active against human cancers in heavily pre-treated patients. The epothilones appear to be well tolerated, with a side effect profile that is similar to that reported with the taxanes. This article will review some basic aspects of epothilone chemistry and biology, and pre-clinical and preliminary clinical experience with epothilone B and its analog, BMS 247550. 相似文献
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《Expert opinion on therapeutic patents》2013,23(6):951-968
Epothilones A and B are naturally occurring microtubule depolymerisation inhibitors, which inhibit the growth of human cancer cells in vitro at nanomolar or even sub-nanomolar concentrations. In contrast to paclitaxel (Taxol®, Bristol-Myers Squibb) epothilones are also active against multi-drug resistant cancer cell lines and epothilone B exhibits potent in vivo antitumour activity against multidrug-resistant tumours. In addition, epothilones A and B have been shown to be active in vitro against cell lines whose paclitaxel-resistance is derived from specific tubulin mutations. Their attractive preclinical profile has made epothilones important lead structures in the search for improved cytotoxic anticancer drugs and hundreds of analogues and derivatives of epothilones have been prepared and biologically characterised over the past four years. While chemical modifications have been reported for almost every position of the epothilone structural framework, the major focus has been on modifications of the epoxide moiety at C-12/C-13, the C-6- position, the ester linkage and the unsaturated heterocyclic side-chain. Several of the compounds thus produced exhibit low nM IC50 values for the inhibition of human cancer cell proliferation and may represent potential development candidates. Currently, two compounds, natural epothilone B and BMS247550, the lactam analogue of epothilone B, are undergoing clinical trials. An additional analogue, epothilone D, also known as deoxyepothilone B, appears to be in late stage preclinical development and may enter clinical trials in the near future. 相似文献
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Altmann KH 《Current pharmaceutical design》2005,11(13):1595-1613
Epothilones A and B are naturally occurring microtubule-stabilizers, which inhibit the growth of human cancer cells in vitro at nM or even sub-nM concentrations. In contrast to paclitaxel (Taxol) epothilones are also active against different types of multidrug-resistant cancer cell lines in vitro and against multidrug-resistant tumors in vivo (epothilone B). Their attractive preclinical profile has made epothilones important lead structures in the search for improved cytotoxic anticancer drugs and epothilone B is currently undergoing phase II clinical trials. Numerous synthetic and semi-synthetic analogs have been prepared since the absolute stereochemistry of epothilone B was first disclosed in mid-1996 and their in vitro biological activity has been determined. Apart from generating a wealth of SAR information, these efforts have led to the identification of at least four compounds (in addition to epothilone B), which are currently at various stages of clinical evaluation in humans. This review is first intended to provide a summary of the basic features of the in vitro biological profile of epothilone B, with particular emphasis on recent developments in this area. A second part will outline the most relevant aspects of the epothilone SAR with regard to effects on tubulin polymerization, in vitro antiproliferative activity, and in vivo antitumor activity. This will include a brief discussion of research directed at the determination of the bioactive conformation of epothilones. In a final section, the preclinical profile of those epothilone analogs currently in clinical development will be discussed in greater detail. 相似文献
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Altmann KH 《Mini reviews in medicinal chemistry》2003,3(2):149-158
Epothilones are naturally occurring 16-membered macrolides with the ability to promote tubulin polymerization in vitro and to stabilize preformed microtubules against Ca(2+)- or cold-induced depolymerization. In contrast to paclitaxel (Taxol((R))) epothilones are also active in vitro against multidrug-resistant cancer cell lines as well as cell lines whose paclitaxel-resistance is derived from specific beta-tubulin mutations. Based on their attractive in vitro biological profile epothilones have turned into important lead structures in anticancer drug discovery and hundreds of analogs and derivatives of epothilone A and B have been prepared and biologically characterized over the past four years. A number of compounds, including natural epothilone B, deoxyepothilone B, and epothilone B lactam (BMS-247550) have also been reported to exhibit profound in vivo antitumor activity in animal models. Apart from providing a brief summary of the SAR that has emerged from the above in vitro studies, this minireview will largely focus on the biology and chemistry of those analogs for which in vivo antitumor activity has been reported in the literature. Two of these compounds, natural epothilone B and epothilone B lactam (BMS-247550) have advanced to clinical studies in humans. 相似文献
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Epothilones are naturally occurring 16-membered macrolides with the ability to promote tubulin polymerization in vitro and to stabilize preformed microtubules against Ca(2+)- or cold-induced depolymerization. At the cellular level, interference with microtubule functionality results in potent inhibition of cancer cell proliferation at nM to even sub-nM concentrations. Most significantly, epothilones, unlike paclitaxel (Taxol), are equally active against drug-sensitive and multidrug-resistant cell lines in vitro and epothilone B has also shown potent in vivo antitumor activity in Taxol-resistant human tumor models. Epothilone B is currently undergoing Novartis-sponsored Phase II clinical trials. In addition to naturally occurring epothilones, numerous synthetic and semi-synthetic analogs have been prepared since the absolute stereochemistry of epothilone B was first disclosed in mid-1996 and their in vitro biological activity has been determined. These studies have generated a wealth of SAR data in a remarkably short period of time, given the complexity of the synthetic targets pursued. One of these analogs, BMS-247550, is presently in Phase II clinical trials by Bristol-Myers Squibb. In a first part this review is intended to provide a summary of the basic features of the in vitro biological profile of epothilones A and B, including emerging data on potential cellular epothilone resistance mechanisms. The second and third part will feature a comprehensive discussion of the epothilone SAR as it has emerged from the work of various (industrial and academic) laboratories across the world, including our own, with regard to effects on tubulin polymerization, in vitro antiproliferative activity, and in vivo antitumor activity. 相似文献