Recombinant Escherichia coli for producing l-tyrosine and application thereof
AssigneeJiangnan University
InventorsJingwen Zhou, Jian Chen, Jurong Ping, Weizhu Zeng
Life of this patent
Estimating the term…
Abstract
Disclosed is recombinant Escherichia coli for producing L-tyrosine and application thereof, and belongs to the technical fields of genetic engineering and bioengineering. According to the present disclosure, genes aroP and tyrP are knocked out, expresses the endogenous gene yddG of E. coli, then heterologously expresses fpk from Bifidobacterium adolescentis, expresses the endogenous genes ppsA and tktA of E. coli, and then expresses aroGfbr and tyrAfbr. Knocking out tyrR, trpE and pheA, so that the synthesis flux of L-tyrosine is increased. Finally, an endogenous gene poxB is knocked out to realize stable fermentation performance at high glucose concentration.
Claims10
A recombinant Escherichia coli for synthesis of L-tyrosine,
- wherein the recombinant E. coli possesses a genome in which several first genes are knocked out, and comprises several added second genes that are expressed or overexpressed,
- wherein the first genes knocked out consist of:
- pheA encoding a fusion of chorismate mutase/prephenate dehydratase,
- trpE encoding an anthranilate synthetase subunit TrpE,
- tyrR encoding a DNA binding transcription double regulator TyrR,
- poxB encoding pyruvate oxidase,
- aroP encoding permease of an aromatic amino acid transporter AroP, and
- tyrP encoding a tyrosine and H (+) symporter,
- wherein the second genes added consist of:
- aroGfbr encoding 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase,
- tyrAfbr encoding chorismate mutase and prephenate dehydrogenase,
- fpk encoding phosphoketolase, and
- yddG encoding an amino acid exportin YddG;
- and
- wherein:
- the pheA gene has the nucleotide sequence set forth in SEQ ID NO:1,
- the trpE gene has the nucleotide sequence set forth in SEQ ID NO:2,
- the aroGfbr gene has the nucleotide sequence set forth in SEQ ID NO:3,
- the tyrAfbr gene has the nucleotide sequence set forth in SEQ ID NO:4,
- the tyrR gene has the nucleotide sequence set forth in SEQ ID NO:5,
- the fpk gene has the nucleotide sequence set forth in SEQ ID NO:8,
- the poxB gene has the nucleotide sequence set forth in SEQ ID NO:9,
- the aroP gene has the nucleotide sequence set forth in SEQ ID NO:10,
- the tyrP gene has the nucleotide sequence set forth in SEQ ID NO:11, and
- the yddG gene has the nucleotide sequence set forth in SEQ ID NO:
- 12;
- wherein the recombinant E. coli genome additionally comprises integrated therein two genes consisting of:
- ppsA expressing phosphoenolpyruvate synthetase, and
- tktA encoding transketolase 1;
- and
- wherein the ppsA gene has the nucleotide sequence set forth in SEQ ID NO:6,
- wherein the tktA gene has the nucleotide sequence set forth in SEQ ID NO:7;
- wherein the ppsA gene is integrated into the genome at a site ykgh-betA, and
- wherein the tktA gene is integrated into the genome at a site dadx-cvra.
Dependencies are inferred from the claim text.