Novel expression hosts for complex secondary metabolite megasynthetases: Production of myxochromide in the thermopilic isolate Corallococcus macrosporus GT-2.
Novel expression hosts for complex secondary metabolite megasynthetases: Production of myxochromide in the thermopilic isolate Corallococcus macrosporus GT-2.
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DOI：
10.1186/1475-2859-8-1
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发表时间：
2009-01-06
影响因子：
6.4
通讯作者：
Mueller, Rolf
中科院分区：
文献类型：
--
作者：
Perlova, Olena;Gerth, Klaus;Kuhlmann, Silvia;Zhang, Youming;Mueller, Rolf
虽然已经从粘细菌中分离出许多具有不同生物活性的次级代谢产物，但这些生物技术上重要的滑行原核生物的大多数菌株仍然难以进行遗传处理。在这项研究中，我们描述了新的快速增长的粘细菌嗜热分离GT-2作为异源宿主的天然产物的生物合成途径的表达分离自其他粘细菌。根据16 S rDNA序列分析结果，该菌株与大孢珊瑚球菌（Corallococcus macrosporus）亲缘关系较近，因此将其命名为大孢珊瑚球菌（Corallococcus macrosporus）。大孢GT-2。中度嗜热菌株的快速生长导致较短的发酵和世代时间，这些方面对于分子生物学工作以及次级代谢产物的生产具有重要意义。遗传操作系统的开发允许将位于转座片段上的完整粘色素生物合成基因簇引入GT-2的染色体中。通过启动子交换的生物合成基因簇的基因工程导致在异源宿主C.与原始生产者橙黄柱头菌和先前描述的异源宿主恶臭假单胞菌相比，大孢子菌GT-2的浓度（分别为600 mg/L对8 mg/L和40 mg/L）。
Although many secondary metabolites with diverse biological activities have been isolated from myxobacteria, most strains of these biotechnologically important gliding prokaryotes remain difficult to handle genetically. In this study we describe the new fast growing myxobacterial thermophilic isolate GT-2 as a heterologous host for the expression of natural product biosynthetic pathways isolated from other myxobacteria. According to the results of sequence analysis of the 16S rDNA, this moderately thermophilic isolate is closely related to Corallococcus macrosporus and was therefore named C. macrosporus GT-2. Fast growth of moderately thermophilic strains results in shorter fermentation and generation times, aspects which are of significant interest for molecular biological work as well as production of secondary metabolites. Development of a genetic manipulation system allowed the introduction of the complete myxochromide biosynthetic gene cluster, located on a transposable fragment, into the chromosome of GT-2. Genetic engineering of the biosynthetic gene cluster by promoter exchange leads to much higher production of myxochromides in the heterologous host C. macrosporus GT-2 in comparison to the original producer Stigmatella aurantiaca and to the previously described heterologous host Pseudomonas putida (600 mg/L versus 8 mg/L and 40 mg/L, respectively).
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DOI：
10.1016/s0167-4781(99)00041-x
发表时间：
1999-05-14
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION
影响因子：
--
作者：
Beyer, S;Kunze, B;Müller, R
通讯作者：
Müller, R
影响因子：
3.2
作者：
Khalil, MW;Sasse, F;Reichenbach, H
通讯作者：
Reichenbach, H
影响因子：
4.4
作者：
Fieseler, Lars;Hentschel, Ute;Piel, Joern
通讯作者：
Piel, Joern
影响因子：
--
作者：
Gross, Frank;Ring, Michael W.;Mueller, Rolf
通讯作者：
Mueller, Rolf
影响因子：
2.7
作者：
Karlyshev, AV;Pallen, MJ;Wren, BW
通讯作者：
Wren, BW