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大肠杆菌中异源依克多因的生产：绕过代谢瓶颈。

Heterologous ectoine production in Escherichia coli: by-passing the metabolic bottle-neck.

作者信息

Bestvater Thorsten, Louis Petra, Galinski Erwin A

机构信息

Institute of Biochemistry, Westfälische Wilhelms-Universität Münster, Münster, Germany.

出版信息

Saline Syst. 2008 Aug 29;4:12. doi: 10.1186/1746-1448-4-12.

DOI:10.1186/1746-1448-4-12
PMID:18759971
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2562377/
Abstract

Transcription of the ectoine biosynthesis genes ectA, ectB and ectC from Marinococcus halophilus in recombinant Escherichia coli DH5alpha is probably initiated from three individual sigma70/sigmaA-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 DH5alpha. 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 DH5alpha, resulting in cytoplasmic ectoine concentrations comparable to those reached via ectoine accumulation from the medium.

摘要

嗜盐海洋球菌ectoine生物合成基因ectA、ectB和ectC在重组大肠杆菌DH5α中的转录可能起始于每个基因上游的三个独立的依赖σ70/σA的启动子序列。因此，通过Northern印迹分析可以检测到分别包含单个基因以及ectA和ectB或ectB和ectC基因组合的mRNA片段。在其自身调控启动子区域（ectUp）的控制下，观察到ectoine的产生似乎受到渗透压调节。此外，天冬氨酸激酶被确定为重组大肠杆菌DH5α中ectoine产生的主要限制因素。ectoine生物合成基因与来自谷氨酸棒杆菌MH20-22B的抗反馈天冬氨酸激酶基因（lysC）的共表达导致大肠杆菌DH5α中ectoine的产量显著增加，使得细胞质ectoine浓度与通过从培养基中积累ectoine所达到的浓度相当。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4705/2562377/b10f29ff8ebb/1746-1448-4-12-8.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4705/2562377/107d47c3143a/1746-1448-4-12-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4705/2562377/b10f29ff8ebb/1746-1448-4-12-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4705/2562377/f3dbc7e49d2d/1746-1448-4-12-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4705/2562377/61d0233f4729/1746-1448-4-12-2.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4705/2562377/9cebafb5edd1/1746-1448-4-12-6.jpg
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