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

Thorsten Bestaver; Petra Louis; Erwin A. Galinski

doi:10.1186/1746-1448-4-12

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

Thorsten Bestaver, Petra Louis, Erwin A. Galinski

The Rowett Institute of Nutrition and Health

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Abstract

Transcription of the ectoine biosynthesis genes ectA, ectB and ectC from Marinococcus halophilus in recombinant Escherichia coli DH5a is probably initiated from three individual s70/sA-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 DH5a. 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 DH5a, resulting in cytoplasmic ectoine concentrations comparable to those reached via ectoine accumulation from the medium.

Original language	English
Article number	12
Journal	Saline Systems
Volume	4
DOIs	https://doi.org/10.1186/1746-1448-4-12
Publication status	Published - 29 Aug 2008

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Bestvater et al Saline Systems 2008
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title = "Heterologous ectoine production in Escherichia coli: By-passing the metabolic bottle-neck",

abstract = "Transcription of the ectoine biosynthesis genes ectA, ectB and ectC from Marinococcus halophilus in recombinant Escherichia coli DH5a is probably initiated from three individual s70/sA-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 DH5a. 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 DH5a, resulting in cytoplasmic ectoine concentrations comparable to those reached via ectoine accumulation from the medium.",

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T2 - By-passing the metabolic bottle-neck

AU - Bestaver, Thorsten

AU - Louis, Petra

AU - Galinski, Erwin A.

PY - 2008/8/29

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N2 - Transcription of the ectoine biosynthesis genes ectA, ectB and ectC from Marinococcus halophilus in recombinant Escherichia coli DH5a is probably initiated from three individual s70/sA-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 DH5a. 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 DH5a, resulting in cytoplasmic ectoine concentrations comparable to those reached via ectoine accumulation from the medium.

AB - Transcription of the ectoine biosynthesis genes ectA, ectB and ectC from Marinococcus halophilus in recombinant Escherichia coli DH5a is probably initiated from three individual s70/sA-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 DH5a. 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 DH5a, resulting in cytoplasmic ectoine concentrations comparable to those reached via ectoine accumulation from the medium.

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JO - Saline Systems

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

Abstract

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