Heterologous ectoine production in Escherichia coli: by-passing the metabolic bottle-neck.
Heterologous ectoine production in Escherichia coli: by-passing the metabolic bottle-neck.
复制标题
大肠杆菌中异源异源产生：通过代谢瓶颈子。
DOI：
10.1186/1746-1448-4-12
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发表时间：
2008-08-29
期刊：
影响因子：
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通讯作者：
Galinski, Erwin A
中科院分区：
文献类型：
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作者：
Bestvater, Thorsten;Louis, Petra;Galinski, Erwin A
嗜盐海洋球菌（Marinococcus halophilus）四氢嘧啶合成基因ectA、ectB和ectC在重组大肠杆菌DH 5 α中的转录可能起始于每个基因上游的三个σ70/σ A依赖性启动子序列。因此，通过北方印迹分析可以分别检测含有单个基因和基因ectA和ectB或ectB和ectC的组合的mRNA片段。在其自身的调控启动子区（ectUp）的控制下，观察到似乎是受调控的四氢嘧啶生产。此外，天冬氨酸激酶被鉴定为重组E. coli DH 5 α中表达。四氢嘧啶生物合成基因和谷氨酸棒杆菌MH 20 - 22 B的反馈抗性天冬氨酸激酶基因（lysC）的共表达导致E. coli DH 5 α，导致细胞质四氢嘧啶浓度与通过培养基中四氢嘧啶积累达到的浓度相当。
Transcription of the ectoine biosynthesis genes ectA, ectB and ectC from Marinococcus halophilus in recombinant Escherichia coli DH5α is probably initiated from three individual σ70/σA-dependent promoter sequences, upstream of each gene. Consequently, mRNA-fragments containing the single genes and combinations of the genes ectA and ectB or ectB and ectC, respectively, could be detected by Northern blot analysis. Under the control of its own regulatory promoter region (ectUp) a seemingly osmoregulated ectoine production was observed. In addition, aspartate kinases were identified as the main limiting factor for ectoine production in recombinant E. coli DH5α. Co-expression of the ectoine biosynthesis genes and of the gene of the feedback-resistant aspartate kinase from Corynebacterium glutamicum MH20-22B (lysC) led to markedly increased production of ectoine in E. coli DH5α, resulting in cytoplasmic ectoine concentrations comparable to those reached via ectoine accumulation from the medium.