Recombinant bacteria comprising novel sucrose transporters
Recombinant bacteria capable of metabolizing sucrose are described. The recombinant bacteria comprise in their genome or on at least one recombinant construct, a novel nucleotide sequence encoding a polypeptide having sucrose transporter activity and a nucleotide sequence encoding a polypeptide having sucrose hydrolase activity. These nucleotide sequences are each operably linked to the same or a different promoter. Recombinant bacteria capable of metabolizing sucrose to produce glycerol and/or glycerol-derived products such as 1,3-propanediol and 3-hydroxypropionic acid are also described.
1. A recombinant bacterium comprising in its genome or in at least one recombinant construct:
(a) a nucleotide sequence encoding a polypeptide having sucrose transporter activity, said polypeptide having at least 95% sequence identity, based on a Clustal W method of alignment, when compared to an amino acid sequence selected from the group consisting of SEQ ID NO:78 and SEQ ID NO:82; and
(b) a nucleotide sequence encoding a polypeptide having sucrose hydrolase catalytic activity, wherein the polypeptide having sucrose hydrolase catalytic activity is classified as EC 3.2.1.26 or EC 2.4.1.7;
wherein (a) and (b) are each operably linked to the same or a different promoter, wherein each of (a) and (b): (i) is in a recombinant construct; (ii) encodes a polypeptide comprising a non-native polypeptide; (iii) is a foreign gene; or (iv) is any combination of (i), (ii), or (iii), and further wherein said recombinant bacterium is capable of metabolizing sucrose.
2. The recombinant bacterium of claim 1 further comprising in its genome or in at least one recombinant construct, a nucleotide sequence encoding a polypeptide having fructokinase catalytic activity, wherein the polypeptide having fructokinase catalytic activity is classified as EC 2.7.1.4, EC 2.7.1.3, or EC 2.7.1.1.
3. The recombinant bacterium of claim 1 wherein said bacterium is selected from the group consisting of the genera: Escherichia, Klebsiella, Citrobacter , and Aerobacter.
4. The recombinant bacterium of claim 3 wherein said bacterium is Escherichia coli.
5. The recombinant bacterium of claim 1 wherein the recombinant bacterium produces 1,3-propanediol, glycerol, and/or 3-hydroxypropionic acid.
6. A process for making glycerol, 1,3-propanediol and/or 3-hydroxypropionic acid from sucrose comprising:
(a) culturing the recombinant bacterium of claim 5 in the presence of sucrose; and
(b) optionally, recovering the glycerol, 1,3-propanediol and/or 3-hydroxypropionic acid produced.