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An indigoidine biosynthetic gene cluster from Streptomyces chromofuscus ATCC 49982 contains an unusual IndB homologue

  • Dayu Yu
  • Fuchao Xu
  • Jonathan Valiente
  • Siyuan Wang
  • Jixun ZhanEmail author
Natural Products

Abstract

A putative indigoidine biosynthetic gene cluster was located in the genome of Streptomyces chromofuscus ATCC 49982. The silent 9.4-kb gene cluster consists of five open reading frames, named orf1, Sc-indC, Sc-indA, Sc-indB, and orf2, respectively. Sc-IndC was functionally characterized as an indigoidine synthase through heterologous expression of the enzyme in both Streptomyces coelicolor CH999 and Escherichia coli BAP1. The yield of indigoidine in E. coli BAP1 reached 2.78 g/l under the optimized conditions. The predicted protein product of Sc-indB is unusual and much larger than any other reported IndB-like protein. The N-terminal portion of this enzyme resembles IdgB and the C-terminal portion is a hypothetical protein. Sc-IndA and/or Sc-IndB were co-expressed with Sc-IndC in E. coli BAP1, which demonstrated the involvement of Sc-IndB, but not Sc-IndA, in the biosynthetic pathway of indigoidine. The yield of indigoidine was dramatically increased by 41.4 % (3.93 g/l) when Sc-IndB was co-expressed with Sc-IndC in E. coli BAP1. Indigoidine is more stable at low temperatures.

Keywords

Indigoidine Streptomyces chromofuscus Sc-IndC Sc-IndB Heterologous expression Blue pigment 

Abbreviations

PKS

Polyketide synthase

NRPS

Non-ribosomal peptide synthetase

PPTase

4′-Phosphopantetheinyl transferase

LDLR

Low-density lipoprotein receptor

ORF

Open reading frame

ATCC

American Type Culture Collection

PCR

Polymerase chain reaction

A

Adenylation

T

Thiolation

TE

Thioesterase

Ox

Oxidation

IPTG

Isopropyl-1-thio-β-d-galactopyranoside

DMSO

Dimethyl sulfoxide

Notes

Acknowledgments

This work was supported by a National Scientist Development Grant (09SDG2060080) from the American Heart Association and a grant from the National Natural Science Foundation of China (31170763). We thank Dr. Chaitan Khosla, Stanford University for kindly providing S. coelicolor CH999 and E. coli BAP1.

Supplementary material

10295_2012_1207_MOESM1_ESM.docx (997 kb)
Supplementary material 1 (DOCX 997 kb)

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2012

Authors and Affiliations

  • Dayu Yu
    • 1
    • 2
  • Fuchao Xu
    • 1
    • 2
  • Jonathan Valiente
    • 2
  • Siyuan Wang
    • 2
  • Jixun Zhan
    • 2
    Email author
  1. 1.Department of Applied Chemistry and Biological Engineering, College of Chemical EngineeringNortheast Dianli UniversityJilinChina
  2. 2.Department of Biological EngineeringUtah State UniversityLoganUSA

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