Early Steps in the Biosynthetic Pathway of Rishirilide B
The biological active compound rishirilide B is produced by <i>Streptomyces bottropensis</i>. The cosmid cos4 contains the complete rishirilide B biosynthesis gene cluster. Its heterologous expression in the host <i>Streptomyces albus</i> J1074 led to the production of rishir...
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MDPI AG
2020-04-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/8/1955 |
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| author | Philipp Schwarzer Olga Tsypik Chijian Zuo Ahmad Alali Julia Wunsch-Palasis Tanja Heitzler Jana Derochefort Mirjam Bernhardt Xiaohui Yan Thomas Paululat Andreas Bechthold |
| author_facet | Philipp Schwarzer Olga Tsypik Chijian Zuo Ahmad Alali Julia Wunsch-Palasis Tanja Heitzler Jana Derochefort Mirjam Bernhardt Xiaohui Yan Thomas Paululat Andreas Bechthold |
| author_sort | Philipp Schwarzer |
| collection | DOAJ |
| description | The biological active compound rishirilide B is produced by <i>Streptomyces bottropensis</i>. The cosmid cos4 contains the complete rishirilide B biosynthesis gene cluster. Its heterologous expression in the host <i>Streptomyces albus</i> J1074 led to the production of rishirilide B as a major compound and to small amounts of rishirilide A, rishirilide D and lupinacidin A. In order to gain more insights into the biosynthesis, gene inactivation experiments and gene expression experiments were carried out. This study lays the focus on the functional elucidation of the genes involved in the early biosynthetic pathway. A total of eight genes were deleted and six gene cassettes were generated. Rishirilide production was not strongly affected by mutations in <i>rslO2, rslO6</i> and <i>rslH</i>. The deletion of <i>rslK4</i> and <i>rslO3</i> led to the formation of polyketides with novel structures. These results indicated that RslK4 and RslO3 are involved in the generation or selection of the starter unit for rishirilide biosynthesis. In the <i>rslO10</i> mutant strain, two novel compounds were detected, which were also produced by a strain containing solely the genes <i>rslK1</i>, <i>rslK2</i>, <i>rslK3</i>, <i>rslK4</i>, and <i>rslA</i>. <i>rslO1</i> and <i>rslO4</i> mutants predominately produce galvaquinones. Therefore, the ketoreductase RslO10 is involved in an early step of rishirilide biosynthesis and the oxygenases RslO1 and RslO4 are most probably acting on an anthracene moiety. This study led to the functional elucidation of several genes of the rishirilide pathway, including <i>rslK4</i>, which is involved in selecting the unusual starter unit for polyketide synthesis. |
| first_indexed | 2024-03-10T20:17:41Z |
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| institution | Directory Open Access Journal |
| issn | 1420-3049 |
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| publishDate | 2020-04-01 |
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| spelling | doaj.art-c05ab7b0c9e943e59b63209d672bf58c2023-11-19T22:27:25ZengMDPI AGMolecules1420-30492020-04-01258195510.3390/molecules25081955Early Steps in the Biosynthetic Pathway of Rishirilide BPhilipp Schwarzer0Olga Tsypik1Chijian Zuo2Ahmad Alali3Julia Wunsch-Palasis4Tanja Heitzler5Jana Derochefort6Mirjam Bernhardt7Xiaohui Yan8Thomas Paululat9Andreas Bechthold10Institute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyOrganic Chemsitry II, Universität Siegen, Adolf-Reichwein-Strasse 2, 57068 Siegen, GermanyInstitute for Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität, Stefan-Meier-Straße 19, 79104 Freiburg, GermanyThe biological active compound rishirilide B is produced by <i>Streptomyces bottropensis</i>. The cosmid cos4 contains the complete rishirilide B biosynthesis gene cluster. Its heterologous expression in the host <i>Streptomyces albus</i> J1074 led to the production of rishirilide B as a major compound and to small amounts of rishirilide A, rishirilide D and lupinacidin A. In order to gain more insights into the biosynthesis, gene inactivation experiments and gene expression experiments were carried out. This study lays the focus on the functional elucidation of the genes involved in the early biosynthetic pathway. A total of eight genes were deleted and six gene cassettes were generated. Rishirilide production was not strongly affected by mutations in <i>rslO2, rslO6</i> and <i>rslH</i>. The deletion of <i>rslK4</i> and <i>rslO3</i> led to the formation of polyketides with novel structures. These results indicated that RslK4 and RslO3 are involved in the generation or selection of the starter unit for rishirilide biosynthesis. In the <i>rslO10</i> mutant strain, two novel compounds were detected, which were also produced by a strain containing solely the genes <i>rslK1</i>, <i>rslK2</i>, <i>rslK3</i>, <i>rslK4</i>, and <i>rslA</i>. <i>rslO1</i> and <i>rslO4</i> mutants predominately produce galvaquinones. Therefore, the ketoreductase RslO10 is involved in an early step of rishirilide biosynthesis and the oxygenases RslO1 and RslO4 are most probably acting on an anthracene moiety. This study led to the functional elucidation of several genes of the rishirilide pathway, including <i>rslK4</i>, which is involved in selecting the unusual starter unit for polyketide synthesis.https://www.mdpi.com/1420-3049/25/8/1955rishirilide biosynthesisgene deletionstreptomycesoxygenaseketoreductaseλ-Red/ET mediated recombination |
| spellingShingle | Philipp Schwarzer Olga Tsypik Chijian Zuo Ahmad Alali Julia Wunsch-Palasis Tanja Heitzler Jana Derochefort Mirjam Bernhardt Xiaohui Yan Thomas Paululat Andreas Bechthold Early Steps in the Biosynthetic Pathway of Rishirilide B Molecules rishirilide biosynthesis gene deletion streptomyces oxygenase ketoreductase λ-Red/ET mediated recombination |
| title | Early Steps in the Biosynthetic Pathway of Rishirilide B |
| title_full | Early Steps in the Biosynthetic Pathway of Rishirilide B |
| title_fullStr | Early Steps in the Biosynthetic Pathway of Rishirilide B |
| title_full_unstemmed | Early Steps in the Biosynthetic Pathway of Rishirilide B |
| title_short | Early Steps in the Biosynthetic Pathway of Rishirilide B |
| title_sort | early steps in the biosynthetic pathway of rishirilide b |
| topic | rishirilide biosynthesis gene deletion streptomyces oxygenase ketoreductase λ-Red/ET mediated recombination |
| url | https://www.mdpi.com/1420-3049/25/8/1955 |
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