Identification of a cluster-situated activator of oxytetracycline biosynthesis and manipulation of its expression for improved oxytetracycline production in Streptomyces rimosus.
Identification of a cluster-situated activator of oxytetracycline biosynthesis and manipulation of its expression for improved oxytetracycline production in Streptomyces rimosus.
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土霉素生物合成簇状激活剂的鉴定及其表达的操纵以提高裂缘链霉菌中土霉素的产量
DOI：
10.1186/s12934-015-0231-7
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发表时间：
2015-04-02
影响因子：
6.4
通讯作者：
Yang K
中科院分区：
文献类型：
--
作者：
Yin S;Wang W;Wang X;Zhu Y;Jia X;Li S;Yuan F;Zhang Y;Yang K
背景土霉素（Oxytetracycline，OTC）是由龟裂链霉菌（Streptomycesrimosus）生产的一种广谱抗生素。尽管它的重要性，鲜为人知的是有关OTC生物合成的调节，这阻碍了任何努力，以提高OTC生产通过工程regulatory genes.ResultsA基因编码链霉菌抗生素调节蛋白（SARP）被发现紧邻theotrB基因ofoxycluster在S. rimosus和指定dotcR。缺失和互补otcR分别抑制或恢复OTC的产生，表明otcR编码OTC生物合成的重要激活因子。然后，从oxycluster的启动子区域中提取预测的SARP结合共有序列。在异源GFP报告系统中的转录分析表明，OtcR直接激活E.在大肠杆菌中，进一步对foxyI启动子的SARP结合序列进行突变分析，证明OtcR直接与共有重复序列相互作用。因此，选择otcR作为工程靶点，通过在SF 14强启动子控制下过表达tcRas串联拷贝，OTC产量显着增加。结论在S. rimosus;它显示直接激活来自moxycluster的五个启动子。过量表达OtcRat可使土霉素产量提高6.49倍，表明调控OtcR对提高土霉素产量具有巨大潜力。
BackgroundOxytetracycline (OTC) is a broad-spectrum antibiotic commercially produced byStreptomyces rimosus. Despite its importance, little is known about the regulation of OTC biosynthesis, which hampered any effort to improve OTC production via engineering regulatory genes.ResultsA gene encoding aStreptomycesantibiotic regulatory protein (SARP) was discovered immediately adjacent to theotrBgene ofoxycluster inS. rimosusand designatedotcR. Deletion and complementation ofotcRabolished or restored OTC production, respectively, indicating thatotcRencodes an essential activator of OTC biosynthesis. Then, the predicted consensus SARP-binding sequences were extracted from the promoter regions ofoxycluster. Transcriptional analysis in a heterologous GFP reporter system demonstrated that OtcR directly activated the transcription of fiveoxypromoters inE. coli, further mutational analysis of a SARP-binding sequence ofoxyIpromoter proved that OtcR directly interacted with the consensus repeats. Therefore,otcRwas chosen as an engineering target, OTC production was significantly increased by overexpression ofotcRas tandem copies each under the control of strong SF14 promoter.ConclusionsA SARP activator, OtcR, was identified inoxycluster ofS. rimosus; it was shown to directly activate five promoters fromoxycluster. Overexpression ofotcRat an appropriate level dramatically increased OTC production by 6.49 times compared to the parental strain, thus demonstrating the great potential of manipulating OtcR to improve the yield of OTC production.
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影响因子：
3.2
作者：
Chen, Yihua;Wendt-Pienkowski, Evelyn;Shen, Ben
通讯作者：
Shen, Ben
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
影响因子：
3.6
作者：
Li, Rui;Liu, Gang;Tan, Huarong
通讯作者：
Tan, Huarong
模因套件：用于发现和搜索的工具。
影响因子：
14.9
作者：
Bailey TL;Boden M;Buske FA;Frith M;Grant CE;Clementi L;Ren J;Li WW;Noble WS
通讯作者：
Noble WS
DOI：
10.1111/j.1749-6632.2011.06354.x
发表时间：
2011-01-01
期刊：
ANTIMICROBIAL THERAPEUTICS REVIEWS: ANTIBIOTICS THAT TARGET THE RIBOSOME
影响因子：
--
作者：
Nelson, Mark L.;Levy, Stuart B.
通讯作者：
Levy, Stuart B.