TY - JOUR
T1 - Proteomic approach to enhance doxorubicin production in panK-integrated Streptomyces peucetius ATCC 27952
AU - Song, Eunjung
AU - Malla, Sailesh
AU - Yang, Yung Hun
AU - Lee, Kwangwon
AU - Kim, Eun Jung
AU - Lee, Hei Chan
AU - Sohng, Jae Kyung
AU - Oh, Min Kyu
AU - Kim, Byung Gee
N1 - Funding Information:
This work was supported by WCU program (R322009000102130), Basic Science Research Program (2010-0009942), Priority Research Centers Program (2009-0094021), and NRL Program (20090083035) through the National Research Foundation (NRF) grant funded by the Korea Government (MEST).
PY - 2011/9
Y1 - 2011/9
N2 - 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.
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.
KW - 2-DE
KW - Pantothenate kinase
KW - Streptomyces peucetius
KW - nLC-MS/MS
UR - http://www.scopus.com/inward/record.url?scp=80052482741&partnerID=8YFLogxK
U2 - 10.1007/s10295-010-0903-6
DO - 10.1007/s10295-010-0903-6
M3 - Article
C2 - 21308395
AN - SCOPUS:80052482741
SN - 1367-5435
VL - 38
SP - 1245
EP - 1253
JO - Journal of Industrial Microbiology and Biotechnology
JF - Journal of Industrial Microbiology and Biotechnology
IS - 9
ER -