Isoprene synthase and method of preparing isoprene using thereof
Provided are a novel isoprene synthase derived from sweet potato and a method of preparing isoprene using the same, and more specifically, a novel isoprene synthase derived from sweet potato, a gene encoding the isoprene synthase, a host cell transformed with the gene, and a method of preparing isoprene using the same. The isoprene synthase of the present invention may have higher isoprene productivity as compared to isoprene synthases known in the related art to thereby be effectively used in isoprene biosynthesis and preparation of an isoprene polymer using the same.
1. A recombinant vector comprising a codon-optimized polynucleotide encoding an isoprene synthase operably introduced therein, wherein the codon-optimized polynucleotide has a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 3 or SEQ ID NO: 33.
2. The recombinant vector of claim 1 , wherein the recombinant vector comprises a promoter for expressing said polynucleotide.
3. The recombinant vector of claim 2 , wherein the promoter is selected from the group consisting of psbA2, trc, rbcL, petJ, psaA, psaB, tac, cpcB, petC and lac for expressing in cyanobacteria, wherein the promoter is selected from the group consisting of PGK1, TPI1, TDH, PDC1, FBA1, ENO1, ENO2, PYK1, ADH1, and TEF1 for expressing in yeast, wherein the promoter is selected from the group consisting of cbhl, gpdA, glaA, pdc and exlA for expressing in filamentous fungi, or wherein the promoter is CMV35S for expressing in plant cells.
4. The recombinant vector of claim 1 , wherein the codon-optimized polynucleotide has the nucleotide sequence of SEQ ID NO: 3.
5. The recombinant vector of claim 1 , wherein the codon-optimized polynucleotide has the nucleotide sequence of SEQ ID NO: 33.
6. A recombinant host cell comprising a recombinant vector comprising a codon-optimized polynucleotide encoding an isoprene synthase operably introduced therein, wherein the codon-optimized polynucleotide has a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 3 or SEQ ID NO: 33.
7. The recombinant host cell of claim 6 , wherein the host cell is selected from the group consisting of cyanobacteria, yeast, filamentous fungi and plant cell.
8. The recombinant host cell of claim 7 , wherein the cyanobacterium is unicellular cyanobacterium or multicellular cyanobacterium.
9. The recombinant host cell of claim 8 , wherein the unicellular cyanobacterium is Synechocystis sp. or Synechococcus sp.
10. The recombinant host cell of claim 8 , wherein the multicellular cyanobacterium is Gloeocapsa sp. or filamentous cyanobacterium sp.
11. The recombinant host cell of claim 10 , wherein the filamentous cyanobacterium is selected from the group consisting of Nostoc sp., Anabaena sp., and Arthrospira sp.
12. The recombinant host cell of claim 7 , wherein the yeast is selected from the group consisting of Saccharomyces sp., Pichia sp., Candida sp., Kazachstania sp., Kluyveromyces sp., Hansenula sp., Rhodosporidium sp., Cryptococcus sp., and Yarrowia sp.
13. The recombinant host cell of claim 7 , wherein the filamentous fungus is selected from the group consisting of Trichoderma sp., Mucor sp., Mortierella sp., Neurospora sp., and Aspergillus sp.
14. The recombinant host cell of claim 7 , wherein the plant cell is selected from the group consisting of Nicotiana sp., Catharantus sp., and Hyoscyamus sp.
15. The recombinant host cell of claim 6 , further comprising one or more genes, which are introduced thereto, selected from the group of:
(i) genes coding for the mevalonate pathway components, selected from the group of gene coding for mevalonate kinase (EC2.7.1.36), phosphomevalonate kinase (EC2.7.4.2), pyrophoshomevalonate decarboxylase (EC 4.1.1.33), acetoacetyl-CoA thiolase (EC2.3.1.9), HMG-CoA synthase (EC2.3.3.10), and HMG-CoA reductase (EC1.1.1.34); and
(ii) genes coding for isopentenyl-diphosphate delta-isomerase (EC 5.3.3.2).
16. The recombinant host cell of claim 6 , further comprising a gene which is introduced thereto, wherein the gene is an idi gene coding for isopentenyl-diphosphate delta-isomerase (EC 5.3.3.2) and/or a gene coding for HMG-CoA reductase (EC1.1.1.34).
17. A method for preparing isoprene comprising:
(a) culturing the recombinant host cell of claim 6 , thereby producing isoprene; and
(b) recovering the isoprene.
18. The method of claim 17 , wherein the culturing of step (a) is performed on a medium comprising a carbon source selected from the group consisting of CO 2 , bicarbonate, glucose, glycerol, glycerin, dihydroxyacetone, yeast extract, biomass, molasses, sucrose, galactose, sorbose, sorbitol, xylose, arabinose, cellulose, xylan, lactose and oil.