Genomics-guided discovery of a new and significantly better source of anticancer natural drug FK228.
Genomics-guided discovery of a new and significantly better source of anticancer natural drug FK228.
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基因组学引导发现一种新的且明显更好的抗癌天然药物 FK228 来源
DOI：
10.1016/j.synbio.2018.10.011
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发表时间：
2018-12
影响因子：
4.8
通讯作者：
Cheng YQ
中科院分区：
文献类型：
--
作者：
Liu X;Xie F;Doughty LB;Wang Q;Zhang L;Liu X;Cheng YQ
FK 228是FDA批准的抗癌药物，由紫色色杆菌968号在中试工业规模分批发酵中天然产生，浓度高达19 mg/L。在这里，我们报告了一个基因组学指导的发现伯克霍尔德菌泰国MSMB 43作为一个新的和显着更好的源FK 228。B的基因组。结果表明，泰国棉株MSMB 43含有一个功能性生物合成基因簇，该基因簇与C. violaceum No.968，并且该细菌确实产生真实的FK 228。通过在摇瓶中在优选的M8培养基中简单发酵，B.在20 L发酵罐中，以M8培养基为发酵培养基，B. Thailandensis MSMB 43的FK 228产量高达115.9 mg/L，是C. violaceum No.968在相同的实验室发酵条件下。RT-PCR分析表明，FK 228的高产菌株B. thailandensis MSMB 43的突变是由于在发酵过程中以Bth_depA为代表的生物合成基因的高表达。进一步的遗传操作产生了重组菌株B。thailandensis MSMB 43/pBMTL 3-tdpR，其具有表达泰国地辛生物合成途径调控基因tdpR的宽宿主范围载体。该工程菌株在20 L发酵罐中在M8培养基中分批补料发酵中产生高达168.5 mg/L的FK 228。因此，野生型B. thailandensis MSMB 43或其工程衍生物可能是工业方法的良好起点，以改善FK 228的生产，从而扩大其在治疗中的用途。
FK228 is an FDA-approved anticancer drug naturally produced by Chromobacterium violaceum No. 968 up to 19 mg/L in a pilot industry-scale batch fermentation. Here we report a genomics-guided discovery of Burkholderia thailandensis MSMB43 as a new and significantly better source of FK228. The genome of B. thailandensis MSMB43 was found to contain a functional biosynthetic gene cluster highly homologous to that of FK228 in C. violaceum No. 968, and the bacterium indeed produces authentic FK228. By simple fermentation in shaking flasks in a preferred M8 medium, B. thailandensis MSMB43 produced FK228 up to 67.7 mg/L; by fed-batch fermentation in a 20-L fermentor in M8 medium, B. thailandensis MSMB43 produced FK228 up to 115.9 mg/L, which is 95 fold higher than that of C. violaceum No. 968 under the same laboratory fermentation conditions. RT-PCR analysis indicated that the high FK228 yield of B. thailandensis MSMB43 was due to high expression of biosynthetic genes, represented by Bth_depA, during the fermentation process. Further genetic manipulation resulted in a recombinant strain, B. thailandensis MSMB43/pBMTL3-tdpR, which harbors a broad host-range vector expressing the thailandepsin biosynthetic pathway regulatory gene tdpR. This engineered strain produced up to 168.5 mg/L of FK228 in fed-batch fermentation in a 20-L fermentor in M8 medium. Therefore, the wild-type B. thailandensis MSMB43 or its engineered derivative could potentially be a good starting point for an industrial process to improve FK228 production for its expanding use in therapy.
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影响因子：
5.1
作者：
Klausmeyer P;Shipley SM;Zuck KM;McCloud TG
通讯作者：
McCloud TG
影响因子：
14.9
作者：
Ansari, MZ;Yadav, G;Mohanty, D
通讯作者：
Mohanty, D
基因组引导的发现来自Burkholderia sp。
影响因子：
3.4
作者：
Liu, Xiangyang;Cheng, Yi-Qiang
通讯作者：
Cheng, Yi-Qiang
影响因子：
2
作者：
Habibi H;Atashi A;Abroun S;Noruzinia M
通讯作者：
Noruzinia M
影响因子：
15
作者：
Li, KW;Wu, J;Simon, JA
通讯作者：
Simon, JA