Abstract:
BACKGROUND:L-tryptophan is an aromatic amino acid widely used in the food, chemical and pharmaceutical industries. In Escherichia coli, L-tryptophan is synthesized from phosphoenolpyruvate and erythrose 4-phosphate by enzymes in the shikimate pathway and L-tryptophan branch pathway, while L-serine and phosphoribosylpyrophosphate are also involved in L-tryptophan synthesis. In order to construct a microbial strain for efficient L-tryptophan production from glucose, we developed a one step tryptophan attenuator inactivation and promoter swapping strategy for metabolic flux optimization after a base strain was obtained by overexpressing the tktA, mutated trpE and aroG genes and inactivating a series of competitive steps. RESULTS:The engineered E. coli GPT1002 with tryptophan attenuator inactivation and tryptophan operon promoter substitution exhibited 1.67 ~ 9.29 times higher transcription of tryptophan operon genes than the control GPT1001. In addition, this strain accumulated 1.70 g l(-1) L-tryptophan after 36 h batch cultivation in 300-mL shake flask. Bioreactor fermentation experiments showed that GPT1002 could produce 10.15 g l(-1) L-tryptophan in 48 h. CONCLUSIONS:The one step inactivating and promoter swapping is an efficient method for metabolic engineering. This method can also be applied in other bacteria.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Gu P,Yang F,Kang J,Wang Q,Qi Qdoi
10.1186/1475-2859-11-30subject
Has Abstractpub_date
2012-03-02 00:00:00pages
30issn
1475-2859pii
1475-2859-11-30journal_volume
11pub_type
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