Recombinant bacteria having the ability to metabolize sucrose
Recombinant bacteria capable of metabolizing sucrose are described. The recombinant bacteria comprise in their genome or on at least one recombinant construct: a nucleotide sequence from Bacillus licheniformis ATCC® 14580 encoding a polypeptide having sucrose transporter activity and a nucleotide sequence from Bacillus licheniformis ATCC® 14580 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 on at least one recombinant construct:
(a) a nucleotide sequence encoding a polypeptide having sucrose transporter activity, said polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:24, based on a Clustal V method of alignment; and
(b) a nucleotide sequence encoding a polypeptide having sucrose hydrolase activity, said polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:26, based on a Clustal V method of alignment;
wherein (a) and (b) are each operably linked to the same or a different promoter, wherein said recombinant bacterium has the ability to metabolize sucrose and wherein the recombinant bacterium produces 1,3-propanediol, glycerol, and/or 3-hydroxypropionic acid.
2. The recombinant bacterium of claim 1 , wherein the nucleotide sequence encoding the polypeptide having sucrose transporter activity has at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:23, based on BLASTN method of alignment.
3. The recombinant bacterium of claim 1 , wherein the nucleotide sequence encoding the polypeptide having sucrose hydrolase activity has at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:25, based on BLASTN method of alignment.
4. The recombinant bacterium of claim 1 , further comprising in its genome or on at least one recombinant construct, a nucleotide sequence encoding a polypeptide having fructokinase activity.
5. The recombinant bacterium of claim 4 , wherein the polypeptide having fructokinase activity has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, or SEQ ID NO:46, based on the Clustal V method of alignment.
6. The recombinant bacterium of claim 4 , wherein the polypeptide having fructokinase activity has the amino acid sequence of SEQ ID NO:40.
7. The recombinant bacterium of claim 1 , wherein said bacterium is selected from the group consisting of the genera: Escherichia, Klebsiella, Citrobacter , and Aerobacter.
8. The recombinant bacterium of claim 7 , wherein said bacterium is Escherichia coli.
9. A process for making glycerol, 1,3-propanediol and/or 3-hydroxypropionic acid from sucrose comprising:
a) culturing the recombinant bacterium of claim 1 in the presence of sucrose;
and b) recovering the glycerol, 1,3-propanediol and/or 3-hydroxypropionic acid produced.