Application US20200291434

METABOLIC ENGINEERING OF E. COLI FOR THE BIOSYNTHESIS OF CANNABINOID PRODUCTS
[+] Show Details

US20200291434

1. An expression cassette comprising a heterologous promoter operably linked to a nucleic acid encoding a bifunctional ispDF enzyme. (9) (0)

2. The expression cassette of claim 1, wherein the bifunctional ispDF enzyme differs by at least one amino acid from the following ispDF enzymes H. pylori HP1020, H. pylori J99 jhp0404, H. pylori HPAG1 HPAG1_0427, H. hepaticus HH1582, H. acinonychis st. Sheeba Hac_1124, W. succinogenes DSM 1740 WS1940, S. denitrificans DSM 1251 Suden_1487, C. jejuni subsp. jejuni NCTC 11168 Cj1607, C. jejuni RM1221 CJE1779, C. jejuni subsp. jejuni 81-176 CJJ81176_1594, and C. fetus subsp. fetus 82-40 CFF8240_0409. (0)

3. The expression cassette of claim 1, wherein the promoter operably linked to the nucleic acid encoding the bifunctional ispDF enzyme is a constitutive promoter. (0)

4. The expression cassette of claim 1, wherein the promoter operably linked to the nucleic acid encoding the bifunctional ispDF enzyme is an inducible promoter. (0)

5. The expression cassette of claim 1, wherein the bifunctional ispDF enzyme comprises an amino acid sequence at least 50% or 55% identical to 300 contiguous amino acids of SEQ ID No. 1, SEQ ID No. 2, or SEQ ID No. 3. (0)

6. The expression cassette of claim 1, wherein the nucleic acid encoding the bifunctional ispDF enzyme is codon optimized (0)

7. The expression cassette of claim 1, wherein the expression cassette further comprises a nucleic acid encoding one or more, two or more, or all of the enzymes selected from the group consisting of dxs, idi, and ispE. (0)

8. The expression cassette of claim 1, wherein the expression cassette further comprises a nucleic acid encoding dxs and idi. (0)

9. The expression cassette of claim 1, wherein the expression cassette further comprises a nucleic acid encoding dxs, idi, and ispE. (0)

10. The expression cassette of claim 1, wherein the bifunctional ispDF is no more than 31% similar to CJ-ispDF. (0)

22. A host cell comprising: (7) (2)

a. an expression cassette comprising a heterologous promoter operably linked to a nucleic acid encoding a bifunctional ispDF enzyme; and

b. an expression cassette comprising a heterologous promoter operably linked to a nucleic acid encoding a terpenoid synthase.

23. The host cell of claim 22, wherein the terpenoid synthase is an isoprene synthase. (0)

24. The host cell of claim 22, wherein the terpenoid synthase is a component of a lycopene synthesis pathway. (0)

25. The host cell of claim 22, wherein the terpenoid synthase is a cannabinoid synthase. (1) (0)

28. The host cell of claim 22, comprising an expression cassette comprising a heterologous promoter operably linked to a nucleic acid encoding a GPP synthase. (0)

29. The host cell of claim 22, wherein the expression cassette of a) or b) further comprises a nucleic acid encoding a GPP synthase. (0)

30. The host cell of claim 22, wherein the host cell exhibits higher flux through the MEP pathway as compared to a control cell that does not comprise at least one of the one or more expression cassette(s). (0)

31. The host cell of claim 22, wherein the host cell does not comprise: a heterologous nucleic acid encoding ispC, ispE, ispG, or ispH; a combination thereof; or all thereof. (0)

View Abstract and Specification Size

PDF with Images and Document Face >

Full Text Publication >

Application US20200291434

METABOLIC ENGINEERING OF E. COLI FOR THE BIOSYNTHESIS OF CANNABINOID PRODUCTS

Patent Matrix® Search