Isolation and characterization of spliceostatin B, a new analogue of FR901464, from Pseudomonas sp. No. 2663.
Isolation and characterization of spliceostatin B, a new analogue of FR901464, from Pseudomonas sp. No. 2663.
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DOI：
10.1038/ja.2013.38
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发表时间：
2013-09
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FR 901464（图1）是一种从假单胞菌2663号培养液中分离的原型前mRNA剪接抑制剂，是一种强效但不稳定的细胞毒性化合物。1-4剪接抑制素A（图1）是FR 901464的甲基化和更稳定的衍生物，显示与哺乳动物剪接体的U2小核核糖核蛋白颗粒的剪接因子3b亚复合物非共价结合，从而抑制前体mRNA剪接并导致前体mRNA渗漏至细胞质。5、6最近，我们从Burkholderia thailandensis MSMB 43的培养液中发现了作为FR 901464的三种新的且显著更稳定的天然类似物的thailanstatins A、B和C。7为了评价thailanstatins的稳定性、前mRNA剪接抑制活性和细胞毒性，我们使用FR 901464作为参比化合物，其是从假单胞菌2663发酵中新鲜纯化的。在此纯化优化过程中，发现了FR 901464的另一个类似化合物，命名为spliceostatin B（1）。本文报道了从日本工业技术厅生命科学与人类技术研究所获得的产FR 901464菌株假单胞菌2663 - 2的发酵、分离、化学性质和细胞毒性。在Luria-Bertani（LB）琼脂上，在30 ℃下将其从甘油储备液中常规活化2天，作为起始平板。将来自平板的几个菌落接种到含有250 ml LB培养基的烧瓶中，并在旋转振荡器（150 r. pm）以制备种子培养物。对于发酵，将种子培养物以2%（v/v）接种到两个发酵罐中的每一个中（BioFlo IV，20升总体积，新玩法科学公司，恩菲尔德，康涅狄格州，美国）;每种含有12升生产培养基，由可溶性淀粉1%、甘油1%、脱脂豆粕1%、葡萄糖0.5%、玉米浆0.5%、（NH 4）2SO 4 0.2%、CaCO 3 0.2%组成，MgSO 4·7 H2O 0.006%和消泡剂-204 0. 01%（pH 7.0），从
FR901464 (Figure 1), a prototype pre-mRNA splicing inhibitor isolated from the culture broth of Pseudomonas sp. No. 2663, is a potent but unstable cytotoxic compound. 1–4 Spliceostatin A (Figure 1) is a methylated and more stable derivative of FR901464, shown to bind noncovalently to the splicing factor 3b subcomplex of the U2 small nuclear ribonucleoprotein particle of mammalian spliceosome, thus inhibiting pre-mRNA splicing and causing premRNA leakage to the cytoplasm. 5, 6 Recently we discovered thailanstatins A, B and C as three new and significantly more stable natural analogs of FR901464 from the culture broth of Burkholderia thailandensis MSMB43. 7 For evaluations of the stability, pre-mRNA splicing inhibitory activity and cytotoxicity of thailanstatins, we used FR901464 as a reference compound, which was freshly purified from the Pseudomonas sp. No. 2663 fermentation. During this purification optimization process, another analogous compound of FR901464, named spliceostatin B (1), was discovered. Herein we report the fermentation, isolation, chemical characterization and cytotoxicity of this new natural product.The FR901464-producing strain Pseudomonas sp. No. 2663 2 was purchased from the National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology, Japan. It was routinely activated from a glycerol stock on Luria-Bertani (LB) agar at 301C for 2 days as a starting plate. Several colonies from the plate were inoculated into a flask containing 250ml of LB medium and incubated at 301C for 24h on a rotary shaker (150r. pm) to prepare a seed culture. For fermentation, the seed culture was inoculated at 2%(v/v) to each of the two fermentors (BioFlo IV, 20l total volume, New Brunswick Scientific, Enfield, CT, USA); each contains 12l of production medium consisting of soluble starch 1%, glycerin 1%, defatted soybean meal 1%, glucose 0.5%, corn steep liquor 0.5%,(NH4) 2SO4 0.2%, CaCO3 0.2%, MgSO4 Á 7H2O 0.006% and antifoam-204 0.01%(pH 7.0), slightly modified from the
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影响因子：
4.3
作者：
Osman, Sami;Albert, Brian J.;Wang, Yanping;Li, Miaosheng;Czaicki, Nancy L.;Koide, Kazunori
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影响因子：
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MOSMANN, T
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MOSMANN, T