Proteomic approach to enhance doxorubicin production in panK-integrated Streptomyces peucetius ATCC 27952

Eunjung Song, Sailesh Malla, Yung Hun Yang, Kwangwon Lee, Eun Jung Kim, Hei Chan Lee, Jae Kyung Sohng, Min-Kyu Oh, Byung Gee Kim

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Biosynthesis of polyketide compounds depends upon the starter and extender units of coenzyme A derivatives of carboxylic acids present in the host organism. To increase the coenzyme A (CoA) pool, pantothenate kinase (panK) gene from Escherichia coli was integrated into S. peucetius ATCC 27952 (panK-integrated strain, BG200), which resulted in increase in aglycone polyketide ε-rhodomycinone (RHO), but decrease in the desired product, i.e., doxorubicin (DXR). To reduce RHO accumulation by synthesizing daunorubicin (DNR) from RHO more efficiently, glycosyltransferase (dnrQS) was overexpressed (pIBR25::dnrQS in panK-integrated strain, BG201). However, DnrQS overexpression still resulted in less production of DXR compared with the parental strain. To understand the results in detail by investigating the proteome changes in the panK-integrated strain, two-dimensional (2D) gel electrophoresis was performed. Among the several proteins that are up- or downregulated in BG200, efflux protein DrrA was our main target of interest, because it is directly related to DXR/DNR production in S. peucetius. DXR transporter DrrAB was additionally introduced in BG200 to enhance secretion of toxic DXR. Compared with S. peucetius ATCC 27952, BG204 (pIBR25::drrAB in panK-integrated strain), produced two times higher amount of DXR, which is 9.4-fold higher than that of panK-integrated strain BG200. The results show that the proteomic approach is quite useful in host development of Streptomyces and understanding cell physiology for antibiotic production.

Original languageEnglish
Pages (from-to)1245-1253
Number of pages9
JournalJournal of Industrial Microbiology and Biotechnology
Volume38
Issue number9
DOIs
Publication statusPublished - 2011 Sep 1

Fingerprint

Streptomyces
Proteomics
Doxorubicin
Polyketides
Daunorubicin
Electrophoresis, Gel, Two-Dimensional
Coenzyme A
Cell Physiological Phenomena
Glycosyltransferases
Poisons
Proteome
Carboxylic Acids
pantothenate kinase
Proteins
Down-Regulation
Escherichia coli
Anti-Bacterial Agents
Genes
rhodomycinone

Keywords

  • 2-DE
  • nLC-MS/MS
  • Pantothenate kinase
  • Streptomyces peucetius

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Proteomic approach to enhance doxorubicin production in panK-integrated Streptomyces peucetius ATCC 27952. / Song, Eunjung; Malla, Sailesh; Yang, Yung Hun; Lee, Kwangwon; Kim, Eun Jung; Lee, Hei Chan; Sohng, Jae Kyung; Oh, Min-Kyu; Kim, Byung Gee.

In: Journal of Industrial Microbiology and Biotechnology, Vol. 38, No. 9, 01.09.2011, p. 1245-1253.

Research output: Contribution to journalArticle

Song, Eunjung ; Malla, Sailesh ; Yang, Yung Hun ; Lee, Kwangwon ; Kim, Eun Jung ; Lee, Hei Chan ; Sohng, Jae Kyung ; Oh, Min-Kyu ; Kim, Byung Gee. / Proteomic approach to enhance doxorubicin production in panK-integrated Streptomyces peucetius ATCC 27952. In: Journal of Industrial Microbiology and Biotechnology. 2011 ; Vol. 38, No. 9. pp. 1245-1253.
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AU - Kim, Eun Jung

AU - Lee, Hei Chan

AU - Sohng, Jae Kyung

AU - Oh, Min-Kyu

AU - Kim, Byung Gee

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AB - Biosynthesis of polyketide compounds depends upon the starter and extender units of coenzyme A derivatives of carboxylic acids present in the host organism. To increase the coenzyme A (CoA) pool, pantothenate kinase (panK) gene from Escherichia coli was integrated into S. peucetius ATCC 27952 (panK-integrated strain, BG200), which resulted in increase in aglycone polyketide ε-rhodomycinone (RHO), but decrease in the desired product, i.e., doxorubicin (DXR). To reduce RHO accumulation by synthesizing daunorubicin (DNR) from RHO more efficiently, glycosyltransferase (dnrQS) was overexpressed (pIBR25::dnrQS in panK-integrated strain, BG201). However, DnrQS overexpression still resulted in less production of DXR compared with the parental strain. To understand the results in detail by investigating the proteome changes in the panK-integrated strain, two-dimensional (2D) gel electrophoresis was performed. Among the several proteins that are up- or downregulated in BG200, efflux protein DrrA was our main target of interest, because it is directly related to DXR/DNR production in S. peucetius. DXR transporter DrrAB was additionally introduced in BG200 to enhance secretion of toxic DXR. Compared with S. peucetius ATCC 27952, BG204 (pIBR25::drrAB in panK-integrated strain), produced two times higher amount of DXR, which is 9.4-fold higher than that of panK-integrated strain BG200. The results show that the proteomic approach is quite useful in host development of Streptomyces and understanding cell physiology for antibiotic production.

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