Polypeptide having ferredoxin-NADP+ reductase activity, polynucleotide encoding the same and uses thereof
The present invention relates to a novel polypeptide having the enzymatic activity of reduction of NADP+ using electrons from reduced ferredoxin (Ferredoxin-NADP+ reductase activity), a polynucleotide having a nucleotide sequence encoding such polypeptide and uses thereof. The invention relates to the modulation of the Ferredoxin NADP+ reductase activity in a microorganism by varying the expression level of the polynucleotide coding for such polypeptide. The invention also relates to the production of commodity chemicals, especially ethanol, n-butanol, 1,3-propanediol, 1,2-propanediol, isopropanol and acetone by fermenting microorganisms wherein their Ferredoxin NADP+ reductase activity is modulated.
1. A modified microorganism having Ferredoxin NADP+reductase enzymatic activity, wherein a polynucleotide coding for the polypeptide having at least 85% identity with the sequence of SEQ ID NO: 1 is overexpressed and having Ferredoxin NADP+ reductase activity as expressed,
wherein the modified microorganism contains at least one modification selected in the group consisting of:
a modification to produce ethanol,
a modification to produce n-butanol,
a modification to produce 1,3-propanediol, and
a modification to produce 1,2-propanediol,
wherein, when the modified microorganism contains at least one modification to produce ethanol, the at least one modification to produce ethanol comprises:
enhancement of the expression of at least one of the following genes: pfor, fdx and udhA,
and
attenuation of the expression of at least one of the following genes: aceE, aceF, lpd, pflA, pflB, frdABCD, ldhA, mgsA, ackA, pta, poxB and iscR,
wherein, when the modified microorganism contains at least one modification to produce n-butanol, the at least one modification to produce n-butanol comprises:
enhancement of the expression of atoB, crt, adhE2 and hbd genes and of at least one of the following genes: pfor, fdx, bcd, etfA, etfB and udhA,
and
attenuation of the expression of at least one of the following genes: aceE, aceF, lpd, pflA, pflB, frdABCD, adhE, ldhA, mgsA, ackA, pta, poxB and iscR,
wherein, when the modified microorganism contains at least one modification to produce 1,3-propanediol, the at least one modification to produce 1,3-propanediol comprises:
enhancement of the expression of pntAB, dhaB1, dhaB2 and yqhD genes and of at least one of the following genes: pfor and,
and
attenuation of the expression of at least one of the following genes: aceE, aceF, lpd, aldA, aldB, ldhA, pflA, pflB, adhE, iscR, glpA, poxB and glpD,
wherein, when the modified microorganism contains at least one modification to produce 1,2-propanediol, the at least one modification to produce 1,2-propanediol comprises:
attenuation of the expression of at least one of the following genes: ptsG, ptsH, ptsl, crr, edd, eda, gloA, aceE, aceF, lpd, aldA, aldB, ldhA, pflA, pflB, adhE, tpiA, gapA, pykA, pykF, ackA, pta, poxB, arcA and ndh,
and
enhancement of the expression of pntAB, gldA and yqhD genes and at least one of the following genes: pfor, fdx, gapN, galP, glk, ppsA, mgsA yafB, ydhF, ycdW, yqhE, yeaE, yghZ, yajO, tas, ydjG, ydbC, and fucO.
2. A method for preparing ethanol comprising culturing a microorganism having a modification to produce ethanol according to claim 1 in culture medium that includes a source of carbon and recovering the ethanol.
3. A method for preparing n-butanol comprising culturing a microorganism having a modification to produce n-butanol according to claim 1 in culture medium that includes a source of carbon and recovering the n-butanol.
4. A method for preparing 1,3-propanediol comprising culturing a microorganism having a modification to produce 1,3-propanediol according to claim 1 in a culture medium that includes a source of carbon and recovering the 1,3-propanediol.
5. A method for preparing 1,2-propanediol comprising culturing a microorganism having a modification to produce 1,2-propanediol according to claim 1 in culture medium that includes a source of carbon and recovering the 1,2-propanediol.