Enhanced heterologous production of desosaminyl macrolides and their hydroxylated derivatives by overexpression of the pikD regulatory gene in Streptomyces Venezuelae

Seok Jung Won, Jeong Jeong Soon, Ryeol Park Sung, Yong Choi Cha, Chul Park Byoung, Je Won Park, Joon Yoon Yeo

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

To elevate the production level of heterologous polyketide in Streptomyces venezuelae, an additional copy of the positive regulatory gene pikD was introduced into the pikromycin (Pik) polyketide synthase (PKS) deletion mutant of S. venezuelae ATCC 15439 expressing tylosin PKS genes. The resulting mutant strain showed enhanced production of both tylactone (TL) and desosaminyl tylactone (DesTL) of 2.7- and 17.1-fold, respectively. The notable increase in DesTL production strongly suggested that PikD upregulates the expression of the desosamine (des) biosynthetic gene cluster. In addition, two hydroxylated forms of DesTL were newly detected from the extract of this mutant. These hydroxylated forms presumably resulted from a PikD-dependent increase in expression of the pikC gene that encodes P450 hydroxylase. Gene expression analysis by reverse transcriptase PCR and bioconversion experiments of 10-deoxymethynolide, narbonolide, and TL into the corresponding desosaminyl macrolides indicated that PikD is a positive regulator of the des and pikC genes, as well as the Pik PKS genes. These results demonstrate the role of PikD as a pathway-specific positive regulator of the entire Pik biosynthetic pathway and its usefulness in the development of a host-vector system for efficient heterologous production of desosaminyl macrolides and novel hydroxylated compounds.

Original languageEnglish
Pages (from-to)1972-1979
Number of pages8
JournalApplied and Environmental Microbiology
Volume74
Issue number7
DOIs
Publication statusPublished - 2008 Apr 1
Externally publishedYes

Fingerprint

Streptomyces venezuelae
polyketide synthases
macrolides
Macrolides
Streptomyces
Regulator Genes
regulator genes
Polyketide Synthases
chemical derivatives
mutants
gene
polyketides
gene expression
tylosin
genes
biotransformation
multigene family
biochemical pathways
Tylosin
Genes

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Cite this

Enhanced heterologous production of desosaminyl macrolides and their hydroxylated derivatives by overexpression of the pikD regulatory gene in Streptomyces Venezuelae. / Won, Seok Jung; Soon, Jeong Jeong; Sung, Ryeol Park; Cha, Yong Choi; Byoung, Chul Park; Park, Je Won; Yeo, Joon Yoon.

In: Applied and Environmental Microbiology, Vol. 74, No. 7, 01.04.2008, p. 1972-1979.

Research output: Contribution to journalArticle

Won, Seok Jung ; Soon, Jeong Jeong ; Sung, Ryeol Park ; Cha, Yong Choi ; Byoung, Chul Park ; Park, Je Won ; Yeo, Joon Yoon. / Enhanced heterologous production of desosaminyl macrolides and their hydroxylated derivatives by overexpression of the pikD regulatory gene in Streptomyces Venezuelae. In: Applied and Environmental Microbiology. 2008 ; Vol. 74, No. 7. pp. 1972-1979.
@article{117e8e953b694c5294ba43c7c0ec70af,
title = "Enhanced heterologous production of desosaminyl macrolides and their hydroxylated derivatives by overexpression of the pikD regulatory gene in Streptomyces Venezuelae",
abstract = "To elevate the production level of heterologous polyketide in Streptomyces venezuelae, an additional copy of the positive regulatory gene pikD was introduced into the pikromycin (Pik) polyketide synthase (PKS) deletion mutant of S. venezuelae ATCC 15439 expressing tylosin PKS genes. The resulting mutant strain showed enhanced production of both tylactone (TL) and desosaminyl tylactone (DesTL) of 2.7- and 17.1-fold, respectively. The notable increase in DesTL production strongly suggested that PikD upregulates the expression of the desosamine (des) biosynthetic gene cluster. In addition, two hydroxylated forms of DesTL were newly detected from the extract of this mutant. These hydroxylated forms presumably resulted from a PikD-dependent increase in expression of the pikC gene that encodes P450 hydroxylase. Gene expression analysis by reverse transcriptase PCR and bioconversion experiments of 10-deoxymethynolide, narbonolide, and TL into the corresponding desosaminyl macrolides indicated that PikD is a positive regulator of the des and pikC genes, as well as the Pik PKS genes. These results demonstrate the role of PikD as a pathway-specific positive regulator of the entire Pik biosynthetic pathway and its usefulness in the development of a host-vector system for efficient heterologous production of desosaminyl macrolides and novel hydroxylated compounds.",
author = "Won, {Seok Jung} and Soon, {Jeong Jeong} and Sung, {Ryeol Park} and Cha, {Yong Choi} and Byoung, {Chul Park} and Park, {Je Won} and Yeo, {Joon Yoon}",
year = "2008",
month = "4",
day = "1",
doi = "10.1128/AEM.02296-07",
language = "English",
volume = "74",
pages = "1972--1979",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "7",

}

TY - JOUR

T1 - Enhanced heterologous production of desosaminyl macrolides and their hydroxylated derivatives by overexpression of the pikD regulatory gene in Streptomyces Venezuelae

AU - Won, Seok Jung

AU - Soon, Jeong Jeong

AU - Sung, Ryeol Park

AU - Cha, Yong Choi

AU - Byoung, Chul Park

AU - Park, Je Won

AU - Yeo, Joon Yoon

PY - 2008/4/1

Y1 - 2008/4/1

N2 - To elevate the production level of heterologous polyketide in Streptomyces venezuelae, an additional copy of the positive regulatory gene pikD was introduced into the pikromycin (Pik) polyketide synthase (PKS) deletion mutant of S. venezuelae ATCC 15439 expressing tylosin PKS genes. The resulting mutant strain showed enhanced production of both tylactone (TL) and desosaminyl tylactone (DesTL) of 2.7- and 17.1-fold, respectively. The notable increase in DesTL production strongly suggested that PikD upregulates the expression of the desosamine (des) biosynthetic gene cluster. In addition, two hydroxylated forms of DesTL were newly detected from the extract of this mutant. These hydroxylated forms presumably resulted from a PikD-dependent increase in expression of the pikC gene that encodes P450 hydroxylase. Gene expression analysis by reverse transcriptase PCR and bioconversion experiments of 10-deoxymethynolide, narbonolide, and TL into the corresponding desosaminyl macrolides indicated that PikD is a positive regulator of the des and pikC genes, as well as the Pik PKS genes. These results demonstrate the role of PikD as a pathway-specific positive regulator of the entire Pik biosynthetic pathway and its usefulness in the development of a host-vector system for efficient heterologous production of desosaminyl macrolides and novel hydroxylated compounds.

AB - To elevate the production level of heterologous polyketide in Streptomyces venezuelae, an additional copy of the positive regulatory gene pikD was introduced into the pikromycin (Pik) polyketide synthase (PKS) deletion mutant of S. venezuelae ATCC 15439 expressing tylosin PKS genes. The resulting mutant strain showed enhanced production of both tylactone (TL) and desosaminyl tylactone (DesTL) of 2.7- and 17.1-fold, respectively. The notable increase in DesTL production strongly suggested that PikD upregulates the expression of the desosamine (des) biosynthetic gene cluster. In addition, two hydroxylated forms of DesTL were newly detected from the extract of this mutant. These hydroxylated forms presumably resulted from a PikD-dependent increase in expression of the pikC gene that encodes P450 hydroxylase. Gene expression analysis by reverse transcriptase PCR and bioconversion experiments of 10-deoxymethynolide, narbonolide, and TL into the corresponding desosaminyl macrolides indicated that PikD is a positive regulator of the des and pikC genes, as well as the Pik PKS genes. These results demonstrate the role of PikD as a pathway-specific positive regulator of the entire Pik biosynthetic pathway and its usefulness in the development of a host-vector system for efficient heterologous production of desosaminyl macrolides and novel hydroxylated compounds.

UR - http://www.scopus.com/inward/record.url?scp=42049102997&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=42049102997&partnerID=8YFLogxK

U2 - 10.1128/AEM.02296-07

DO - 10.1128/AEM.02296-07

M3 - Article

VL - 74

SP - 1972

EP - 1979

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 7

ER -