Nucleic acids, compositions and uses thereof
View Patent ↗The subject invention provides at least one nucleic acid sequence encoding an aldouronate-utilization regulon isolated from Paenibacillus sp. strain JDR-2, a bacterium which efficiently utilizes xylan and metabolizes aldouronates (methylglucuronoxylosaccharides). The subject invention also provides a means for providing a coordinately regulated process in which xylan depolymerization and product assimilation are coupled in Paenibacillus sp. strain JDR-2 to provide a favorable system for the conversion of lignocellulosic biomass to biobased products. Additionally, the nucleic acid sequences encoding the aldouronate-utilization regulon can be used to transform other bacteria to form organisms capable of producing a desired product (e.g., ethanol, 1-butanol, acetoin, 2,3-butanediol, 1,3-propanediol, succinate, lactate, acetate, malate or alanine) from lignocellulosic biomass.
1. An isolated nucleic acid comprising a polynucleotide that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 12, wherein said polypeptide has α-glucuronidase activity.
2. The isolated nucleic acid of claim 1 , wherein said polynucleotide comprises the nucleotide sequence of SEQ ID NO: 11.
3. A genetic construct comprising the polynucleotide of claim 1 .
4. The genetic construct of claim 3 , wherein said polynucleotide comprises the nucleotide sequence of SEQ ID NO: 11.
5. A vector comprising the polynucleotide of claim 1 .
6. The vector of claim 5 , wherein said polynucleotide comprises the nucleotide sequence of SEQ ID NO: 11.
7. A host cell transformed with the nucleic acid of claim 1 .
8. The host cell of claim 7 , wherein said nucleic acid comprises a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 11.
9. The host cell of claim 7 , wherein said host cell is a Bacillus spp.
10. The host cell of claim 9 , wherein said Bacillus spp is Bacillus coagulans.
11. A probe that hybridizes with the nucleic acid of claim 1 .
12. The probe of claim 11 , wherein said probe hybridizes with the nucleotide sequence of SEQ ID NO: 11.
13. An isolated polynucleotide that hybridizes under intermediate stringency or high stringency conditions to the nucleic acid of claim 1 , wherein said intermediate or high stringency conditions comprise a hybridization step at 60° C. in the presence of a 5×SSC buffer and washes performed in a solution containing 2×SSC at 50° C. or a hybridization step at 65° C. in the presence of SSC buffer, 1×SSC corresponding to 0.15M NaC1 and 0.05 M Na citrate, washes at 37° C. for 1 hour in a solution containing 2×SSC, 0.01% PVP, 0.01% Ficoll, and 0.01% B SA, followed by a wash in 0.1×SSC at 50° C. for 45 minutes, respectively.
14. The isolated polynucleotide of claim 13 , wherein said nucleic acid comprises the nucleotide sequence of SEQ ID NO: 11.
15. A composition comprising a host cell transformed with a polynucleotide that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 12, wherein said polypeptide has α-glucuronidase activity.
16. The composition of claim 15 , wherein said host cell is contained in a solid medium.
17. The composition of claim 16 , wherein said medium is a liquid medium.
18. An isolated nucleic acid that comprises a polynucleotide, wherein said polynucleotide is the full complement of the polynucleotide of claim 1 .