Microorganism for methionine production with enhanced glucose import
View Patent ↗The present invention is related to a recombinant microorganism for improved methionine production comprising modifications to produce methionine from glucose as main carbon source by fermentation, and modifications to improve glucose import, wherein the glucose import is improved by modifying the expression of at least one gene selected from ptsG, sgrT sgrS and dgsA. The invention is also related to a method for the fermentative production of methionine or methionine derivatives comprising the steps of: culturing the recombinant microorganism as described above in an appropriate culture medium comprising a fermentable source of carbon containing glucose and a source of sulphur, and recovering methionine or methionine derivatives from the culture medium.
1. A recombinant microorganism for improved methionine production comprising:
a) at least one modification to produce methionine from glucose as main carbon source by fermentation, and
b) at least one modification to improve glucose import, wherein glucose import is increased compared to the microorganism without the specified modifications by overexpression of the gene ptsG encoding the PTS enzyme IICB Glc and deletion of the gene dgsA encoding a transcriptional regulator,
wherein expression of at least one of the following genes is attenuated: metJ, pykA, pykF, purU, yncA, or udhA, and wherein said microorganism is Escherichia coli.
2. The microorganism of claim 1 , wherein gene ptsG is overexpressed under control of an inducible and/or a constitutive promoter.
3. The microorganism of claim 1 , wherein the gene ptsG does not contain a sequence of a binding site for a small RNA sgrS, which is SEQ ID NO: 19.
4. The microorganism of claim 1 , wherein expression of gene sgrS and/or sgrT is attenuated.
5. The microorganism of claim 4 , wherein the gene sgrS and/or sgrT is deleted.
6. The microorganism of claim 1 , wherein expression of at least one of the following genes is enhanced: pyc, pntAB, cysP, cysU, cysW, cysA, cysMs, cysJ, cysl, cysH, gcvT, gcvH, gcvP, Ipd, serA, serB, serC, cysE, metF, metH, or thrA.
7. The microorganism of claim 6 , wherein at least one gene is under control of an inducible promoter.
8. The microorganism of claim 1 , wherein:
a) gene pstG does not contain an sRNA sgrS binding site and/or gene sgrS is deleted and/or gene sgrT is deleted;
b) expression of genes metH, cysPUWAM, cysJIH, gcvTHP, metF, serA, serB, serC, cysE, and pyc are enhanced; and
c) expression of the genes metJ, pykA, pykF, purU and yncA are attenuated.
9. A method for fermentative production of methionine and/or a methionine derivative comprising:
a) culturing the recombinant microorganism of claim 1 in an appropriate culture medium comprising a fermentable source of carbon comprising glucose and a source of sulphur, and
b) recovering methionine and/or a methionine derivative from the culture medium.
10. The method of claim 9 , wherein the gene ptsG does not contain a sequence of a binding site for a small RNA sgrS, which is SEQ ID NO: 19.
11. The method of claim 9 , wherein expression of gene sgrS and/or sgrT is attenuated.
12. The method of claim 9 , wherein gene sgrS and/or sgrT is deleted.
13. The method of claim 9 , wherein expression of at least one of the following genes is enhanced: pyc, pntAB, cysP, cysU, cysW, cysA, cysts, cyst, cysI, cysH, gcvT, gcvH, gcvP, Ipd, serA, serB, serC, cysE, metF, metH, or thrA.
14. The method of claim 13 , wherein at least one gene is under control of an inducible promoter.
15. The method of claim 9 , wherein expression of at least one of the following genes is attenuated: metJ, pykA, pykF, purU, yncA, or udhA.
16. The method of claim 9 , wherein:
d) gene pstG does not contain a sRNA sgrS binding site, which is SEQ ID NO: 19, and/or gene sgrS is deleted and/or gene sgrT is deleted;
e) expression of genes metH, cysPUWAM, cysJIH, gcvTHP, metF, serA, sere, serC, cysE, and pyc are enhanced; and
f) expression of genes metJ, pykA, pykF, purU and yncA are attenuated.