Back to results
US 2021/0324362 A1Published application
Method for producing isoprene
AssigneeIndustry-Academic Cooperation Foundation Gyeongsang National University
InventorsSeon Won KIM, Jung Hun KIM, Myeong Seok CHA, Hui Jeong JANG
Life of this patent
Estimating the term…
Abstract
A method for producing isoprene includes culturing E. coli, which has isoprene productivity and in which a gene encoding a recA protein is attenuated or deleted, in a medium containing a carbon source. Therefore, a great amount of isoprene may be produced within a short period of time, and thereby considerably decreasing isoprene production unit costs.
Claims29
1:
- A method for producing isoprene, comprising:
- culturing E. coli, which has isoprene productivity and in which a gene encoding a recA protein is attenuated or deleted, in a medium containing a carbon source.
2:
- The method according to claim 1, wherein the E. coli is DH5α, MG1655, BL21(DE), S17-1, XL1-Blue, BW25113 or a combination thereof.
3:
- The method according to claim 1, wherein the E. coli is MG1655.
4:
- The method according to claim 1, wherein the gene encoding a recA protein has a nucleotide sequence of SEQ ID NO:
- 76.
5:
- The method according to claim 1, wherein the E. coli expresses isoprene synthase and an enzyme of Enterococcus genus or Streptococcus genus mevalonate pathway.
6:
- The method according to claim 1, wherein the E. coli has a gene encoding isoprene synthase derived from Populus trichocarpa of SEQ ID NO:
- 1 intrinsically or by introduction therein.
7:
- The method according to claim 6, wherein the gene is introduced into a plasmid having a translation initiation rate value of 3,000 au or more in a ribosomal binding site sequence corresponding thereto.
8:
- The method according to claim 1, wherein the E. coli includes:
- a gene of SEQ ID NO:
- 2 encoding an enzyme with functions of acetyacetyl-CoA synthase derived from Enterococcus faecalis and HMG-CoA reductase, simultaneously;
- a gene of SEQ ID NO:
- 3 encoding HMG-CoA synthase derived from Enterococcus faecalis;
- a gene of SEQ ID NO:
- 4 encoding mevalonate kinase derived from Streptococcus pneumoniae;
- a gene SEQ ID NO:
- 5 encoding mevalonate diphosphate carboxylase derived from Streptococcus pneumoniae;
- a gene of SEQ ID NO:
- 6 encoding phosphomevalonate kinase derived from Streptococcus pneumoniae;
- and
- a gene SEQ ID NO:
- 7 encoding isoprenyl pyrophosphate isomerase derived from Escherichia coli MG 1655,
- which are contained intrinsically or by introduction therein.
12 more elements
9:
- The method according to claim 8, wherein the E. coli further includes a gene selected from a gene of SEQ ID NO:
- 8 encoding isoprenyl pyrophosphate isomerase derived from Synechocystis sp. PCC6803;
- a gene of SEQ ID NO:
- 9 encoding isoprenyl pyrophosphate isomerase derived from Streptococcus pneumoniae;
- and
- a gene of SEQ ID NO:
- 10 encoding isoprenyl pyrophosphate isomerase derived from Haematococcus plavialis,
- which is included intrinsically or by introduction therein.
5 more elements
10:
- The method according to claim 1, wherein the E. coli includes:
- a gene of SEQ ID NO:
- 11 encoding a fusion protein of isoprene synthase derived from Populus trichocarpa and isoprenyl pyrophosphate isomerase derived from E. coli MG1655;
- a gene of SEQ ID NO:
- 12 encoding a fusion protein of isoprenyl pyrophosphate isomerase derived from E. coli MG1655 and isoprene synthase derived from Populus trichocarpa;
- a gene of SEQ ID NO:
- 13 encoding a fusion protein of isoprene synthase derived from Populus trichocarpa and Synechocystis isoprenyl pyrophosphate isomerase;
- or
- a gene of SEQ ID NO:
- 14 encoding a fusion protein of Synechocystis isoprenyl pyrophosphate isomerase and isoprene synthase derived from Populus trichocarpa.
7 more elements
11:
- The method according to claim 1, wherein the E. coli is characterized in that at least one gene selected from the group consisting of dld, atoD, atoA and pps is attenuated or deleted.
12:
- The method according to claim 1, wherein the E. coli is characterized in that at least one gene selected from the group consisting of ackA-pta, poxB, adhE and ldhA is attenuated or deleted.
13:
- The method according to claim 1, wherein the E. coli is characterized in that a gene encoding NudB protein is attenuated or deleted.
14:
- The method according to claim 13, wherein the gene has a nucleotide sequence of SEQ ID NO:
- 77.
15:
- The method according to claim 1, wherein the E. coli is characterized in that flagella are inactivated or removed.
16:
- The method according to claim 15, wherein the E. coli is characterized in that at least one gene selected from the group consisting of fliF, fliG, fliH, fliI, fliJ and fliK is deleted or inactivated.
17:
- The method according to claim 1, wherein the medium contains lactose.
18:
- The method according to claim 1, wherein the medium contains Mg2+.
19:
- E. coli for producing isoprene, characterized by having isoprene productivity, wherein a gene encoding a recA protein is attenuated or deleted.
20:
- The E. coli according to claim 19, wherein the E. coli is DH5α, MG1655, BL21(DE), S17-1, XL1-Blue, BW25113 or a combination thereof.
21:
- The E. coli according to claim 19, wherein the E. coli is MG1655.
22:
- The E. coli according to claim 19, wherein the gene encoding a recA protein has a nucleotide sequence of SEQ ID NO:
- 76.
23:
- The E. coli according to claim 19, wherein the E. coli includes:
- a gene of SEQ ID NO:
- 1 encoding isoprene synthase derived from Populus trichocarpa;
- a gene of SEQ ID NO:
- 2 encoding an enzyme with functions of acetyacetyl-CoA synthase derived from Enterococcus faecalis and HMG-CoA reductase, simultaneously;
- a gene of SEQ ID NO:
- 3 encoding HMG-CoA synthase derived from Enterococcus faecalis;
- a gene of SEQ ID NO:
- 4 encoding mevalonate kinase derived from Streptococcus pneumoniae;
- a gene of SEQ ID NO:
- 5 encoding mevalonate diphosphate carboxylase derived from Streptococcus pneumoniae;
- a gene of SEQ ID NO:
- 6 encoding phosphomevalonate kinase derived from Streptococcus pneumoniae;
- and
- a gene of SEQ ID NO:
- 7 encoding isoprenyl pyrophosphate isomerase derived from Escherichia coli MG 1655,
- which are contained intrinsically or by introduction therein.
14 more elements
24:
- The E. coli according to claim 19, wherein the E. coli includes:
- a gene of SEQ ID NO:
- 11 encoding a fusion protein of isoprene synthase derived from Populus trichocarpa and isoprenyl pyrophosphate isomerase derived from E. coli MG1655;
- a gene of SEQ ID NO:
- 12 encoding a fusion protein of isoprenyl pyrophosphate isomerase derived from E. coli MG1655 and isoprene synthase derived from Populus trichocarpa;
- a gene of SEQ ID NO:
- 13 encoding a fusion protein of isoprene synthase derived from Populus trichocarpa and Synechocystis isoprenyl pyrophosphate isomerase;
- or
- a gene of SEQ ID NO:
- 14 encoding a fusion protein of Synechocystis isoprenyl pyrophosphate isomerase and isoprene synthase derived from Populus trichocarpa.
7 more elements
25:
- The E. coli according to claim 19, wherein the E. coli is characterized in that at least one gene selected from the group consisting of dld, atoD, atoA and pps is attenuated or deleted.
26:
- The E. coli according to claim 19, wherein the E. coli is characterized in that a gene encoding NudB protein is attenuated or deleted.
27:
- The E. coli according to claim 19, wherein the gene has a nucleotide sequence of SEQ ID NO:
- 77.
28:
- The E. coli according to claim 19, wherein the E. coli is characterized in that flagella are inactivated or removed.
29:
- The E. coli according to claim 19, wherein the E. coli is characterized in that at least one gene selected from the group consisting of fliF, fliG, fliH, fliI, fliJ and fliK is deleted or inactivated