Methods and compositions for production of blue pigment indigoidine

Patent metadata

Field	Value
Publication number	US-9243269-B2
Application number	US-201314080024-A
Country	US
Kind code	B2
Filing date	Nov 14, 2013
Priority date	Nov 20, 2012
Publication date	Jan 26, 2016
Grant date	Jan 26, 2016

Abstract

The present disclosure provides for an expression system for the production of blue pigment indigoidine. The system comprises a heterologous host cell, a DNA sequence encoding a Sc-IndB protein, and a DNA sequence encoding a Sc-IndC protein. The system may be configured for the co-expression of the Sc-IndB and Sc-IndC. DNA sequences encoding the Sc-IndB and Sc-IndC may be provided on at least one vector. Alternatively, the DNA sequences encoding the Sc-IndB and Sc-IndC may optionally be integrated into the genome of the heterologous host genome. The expression system may further comprise a sfp gene or a PPTase.

First claim

What is claimed is:

1. An expression system for the production of blue pigment indigoidine, comprising:
(i) a heterologous host cell,
(ii) a DNA sequence encoding a Sc-IndB protein and having at least 85% identity to the amino acid sequence set forth in SEQ ID NO:2,
(iii) a DNA sequence encoding a Sc-IndC protein and having at least 85% identity to the amino acid sequence set forth in SEQ ID NO:4,
wherein the system is configured for the co-expression of the Sc-IndB and Sc-IndC.

2. The expression system of claim 1 , wherein the DNA sequences encoding the Sc-IndB and Sc-IndC are provided on at least one vector.

3. The expression system of claim 2 , wherein the DNA sequences are provided on a single vector.

4. The expression system of claim 1 , wherein at least one of the DNA sequences encoding the Sc-IndB and Sc-IndC are integrated into the genome of the heterologous host genome.

5. The expression system of claim 1 , further comprising a Bacillus subtilis -derived sfp gene.

6. The expression system of claim 1 , further comprising a 4′-phosphopantetheinyl transferase (PPTase).

7. The expression system of claim 6 , wherein the PPTase is an endogenous host PPTase.

8. The expression system of claim 6 , wherein the PPTase is an exogenous PPTase.

9. The expression system of claim 1 , wherein the DNA sequences encoding the Sc-IndB and Sc-IndC are operatively linked to at least one promoter.

10. The expression system of claim 9 , wherein the DNA sequences encoding the Sc-IndB and Sc-IndC are operatively linked to a single promoter.

11. The expression system of claim 1 , wherein the DNA sequence encoding the Sc-IndB protein is a sequence at least 85% identical to the sequence set forth in SEQ ID NO:1.

12. The expression system of claim 1 , wherein the DNA sequence encoding the Sc-IndC protein is a sequence at least 85% identical to the sequence set forth in SEQ ID NO:3.

13. A method for synthesizing a blue pigment indigoidine, the method comprising:
co-expressing a Sc-IndB protein and a Sc-IndC protein in a heterologous host cell,
wherein the Sc-IndB protein has at least 85% identity to the amino acid sequence set forth in SEQ ID NO:2, and the Sc-IndC protein has at least 85% identity to the amino acid sequence set forth in SEQ ID NO:4.

14. The method of claim 13 , wherein the heterologous host is a bacterium.

15. The method of claim 13 , wherein the heterologous host further comprises a nucleotide sequence that encodes a Bacillus subtilis -derived sfp gene.

16. The method of claim 13 , wherein the heterologous host expresses a phosphopantetheinyl transferase (PPTase).

17. The expression system of claim 1 , wherein:
the Sc-IndB protein is encoded by the amino acid sequence set forth in SEQ ID NO:2, and
the Sc-IndC protein is encoded by the amino acid sequence set forth in SEQ ID NO:4.

18. The expression system of claim 1 , wherein:
the DNA sequence encoding the Sc-IndB protein is the sequence set forth in SEQ ID NO:1, and
the DNA sequence encoding the Sc-IndC protein is the sequence set forth in SEQ ID NO:3.

Assignees

• Zhan Jixun
• Univ Utah State

Inventors

• Zhan Jixun

Classifications

• C07D213/90

  having more than three double bonds between ring members or between ring members and non-ring members · CPC title

• C12N1/00

  Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12P17/165 Primary

  Heterorings having nitrogen atoms as the only ring heteroatoms · CPC title