Heterologous ectoine production in Escherichia coli

By-passing the metabolic bottle-neck

Thorsten Bestaver, Petra Louis, Erwin A. Galinski

Research output: Contribution to journalArticle

34 Citations (Scopus)
3 Downloads (Pure)

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 languageEnglish
Article number12
JournalSaline Systems
Volume4
DOIs
Publication statusPublished - 29 Aug 2008

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bottles
neck
Neck
Escherichia coli
gene
aspartate kinase
Aspartate Kinase
genes
Genes
Marinococcus halophilus
promoter regions
biosynthesis
Corynebacterium glutamicum
production economics
Nucleic Acid Regulatory Sequences
Northern blotting
limiting factor
ectoine
Genetic Promoter Regions
Northern Blotting

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Heterologous ectoine production in Escherichia coli : By-passing the metabolic bottle-neck. / Bestaver, Thorsten; Louis, Petra; Galinski, Erwin A.

In: Saline Systems, Vol. 4, 12, 29.08.2008.

Research output: Contribution to journalArticle

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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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