caa a22 4500 606211861 CHVBK 20210128101040.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s10295-014-1546-9 doi (NATIONALLICENCE)springer-10.1007/s10295-014-1546-9 Characterization of negative regulatory genes for the biosynthesis of rapamycin in Streptomyces rapamycinicus and its application for improved production [Elektronische Daten] [Young Yoo, Jae-yeon Hwang, Hea-luyung Shin, Heqing Cui, Jinwon Lee, Yeo Yoon] Sequence analysis of the rapamycin biosynthetic gene cluster in Streptomyces rapamycinicus ATCC 29253 identified several putative regulatory genes. The deduced product of rapY, rapR, and rapS showed high sequence similarity to the TetR family transcription regulators, response regulators and histidine kinases of two-component systems, respectively. Overexpression of each of the three genes resulted in a significant reduction in rapamycin production, while in-frame deletion of rapS and rapY from the S. rapamycinicus chromosome improved the levels of rapamycin production by approximately 4.6-fold (33.9mg l−1) and 3.7-fold (26.7mg l−1), respectively, compared to that of the wild-type strain. Gene expression analysis by semi-quantitative reverse transcription-PCR (RT-PCR) in the wild-type and mutant strains indicated that most of the rapamycin biosynthetic genes are regulated negatively by rapS (probably through its partner response regulator RapR) and rapY. Interestingly, RapS negatively regulates the expression of the rapY gene, and in turn, rapX encoding an ABC-transporter is negatively controlled by RapY. Finally, overexpression of rapX in the rapS deletion mutant resulted in a 6.7-fold (49mg l−1) increase in rapamycin production compared to that of wild-type strain. These results demonstrate the role of RapS/R and RapY as negative regulators of rapamycin biosynthesis and provide valuable information to both understand the complex regulatory mechanism in S. rapamycinicus and exploit the regulatory genes to increase the level of rapamycin production in industrial strains. Society for Industrial Microbiology and Biotechnology, 2014 Rapamycin nationallicence Negative regulator nationallicence Streptomyces rapamycinicus nationallicence ABC-transporter nationallicence Yoo Young Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut Hwang Jae-yeon Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut Shin Hea-luyung Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut Cui Heqing Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut Lee Jinwon Department of Chemical and Biomolecular Engineering, Sogang University, 121-742, Seoul, Republic of Korea aut Yoon Yeo Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/1(2015-01-01), 125-135 1367-5435 42:1<125 2015 42 10295 https://doi.org/10.1007/s10295-014-1546-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-014-1546-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yoo Young Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Hwang Jae-yeon Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Shin Hea-luyung Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Cui Heqing Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Lee Jinwon Department of Chemical and Biomolecular Engineering, Sogang University, 121-742, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Yoon Yeo Department of Chemistry and Nano Science, Ewha Global Top5 Research Program, Ewha Womans University, 120-750, Seoul, Republic of Korea aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/1(2015-01-01), 125-135 1367-5435 42:1<125 2015 42 10295