Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous Host
Atratumycin is a cyclodepsipeptide with activity against <i>Mycobacteria tuberculosis</i> isolated from deep-sea derived <i>Streptomyces atratus</i> SCSIO ZH16NS-80S. Analysis of the atratumycin biosynthetic gene cluster (<i>atr</i>) revealed that its biosynthesis...
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| Format: | Article |
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MDPI AG
2019-09-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/17/10/560 |
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| author | Zhijie Yang Xin Wei Jianqiao He Changli Sun Jianhua Ju Junying Ma |
| author_facet | Zhijie Yang Xin Wei Jianqiao He Changli Sun Jianhua Ju Junying Ma |
| author_sort | Zhijie Yang |
| collection | DOAJ |
| description | Atratumycin is a cyclodepsipeptide with activity against <i>Mycobacteria tuberculosis</i> isolated from deep-sea derived <i>Streptomyces atratus</i> SCSIO ZH16NS-80S. Analysis of the atratumycin biosynthetic gene cluster (<i>atr</i>) revealed that its biosynthesis is regulated by multiple factors, including two LuxR regulatory genes (<i>atr1</i> and <i>atr2</i>), two ABC transporter genes (<i>atr29</i> and <i>atr30</i>) and one <i>Streptomyces</i> antibiotic regulatory gene (<i>atr32</i>). In this work, three regulatory and two transporter genes were unambiguously determined to provide positive, negative and self-protective roles during biosynthesis of atratumycin through bioinformatic analyses, gene inactivations and <i>trans</i>-complementation studies. Notably, an unusual <i>Streptomyces</i> antibiotic regulatory protein Atr32 was characterized as a negative regulator; the function of Atr32 is distinct from previous studies. Five over-expression mutant strains were constructed by rational application of the regulatory and transporter genes; the resulting strains produced significantly improved titers of atratumycin that were ca. 1.7−2.3 fold greater than wild-type (WT) producer. Furthermore, the atratumycin gene cluster was successfully expressed in <i>Streptomyces coelicolor</i> M1154, thus paving the way for the transfer and recombination of large DNA fragments. Overall, this finding sets the stage for understanding the unique biosynthesis of pharmaceutically important atratumycin and lays the foundation for generating anti-tuberculosis lead compounds possessing novel structures. |
| first_indexed | 2024-04-12T19:32:37Z |
| format | Article |
| id | doaj.art-0a3257a21f164c52afacfc4cd92a783f |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-04-12T19:32:37Z |
| publishDate | 2019-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-0a3257a21f164c52afacfc4cd92a783f2022-12-22T03:19:18ZengMDPI AGMarine Drugs1660-33972019-09-01171056010.3390/md17100560md17100560Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous HostZhijie Yang0Xin Wei1Jianqiao He2Changli Sun3Jianhua Ju4Junying Ma5CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, ChinaCAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, ChinaAtratumycin is a cyclodepsipeptide with activity against <i>Mycobacteria tuberculosis</i> isolated from deep-sea derived <i>Streptomyces atratus</i> SCSIO ZH16NS-80S. Analysis of the atratumycin biosynthetic gene cluster (<i>atr</i>) revealed that its biosynthesis is regulated by multiple factors, including two LuxR regulatory genes (<i>atr1</i> and <i>atr2</i>), two ABC transporter genes (<i>atr29</i> and <i>atr30</i>) and one <i>Streptomyces</i> antibiotic regulatory gene (<i>atr32</i>). In this work, three regulatory and two transporter genes were unambiguously determined to provide positive, negative and self-protective roles during biosynthesis of atratumycin through bioinformatic analyses, gene inactivations and <i>trans</i>-complementation studies. Notably, an unusual <i>Streptomyces</i> antibiotic regulatory protein Atr32 was characterized as a negative regulator; the function of Atr32 is distinct from previous studies. Five over-expression mutant strains were constructed by rational application of the regulatory and transporter genes; the resulting strains produced significantly improved titers of atratumycin that were ca. 1.7−2.3 fold greater than wild-type (WT) producer. Furthermore, the atratumycin gene cluster was successfully expressed in <i>Streptomyces coelicolor</i> M1154, thus paving the way for the transfer and recombination of large DNA fragments. Overall, this finding sets the stage for understanding the unique biosynthesis of pharmaceutically important atratumycin and lays the foundation for generating anti-tuberculosis lead compounds possessing novel structures.https://www.mdpi.com/1660-3397/17/10/560atratumycinluxrsarpabc transporterheterologous expression |
| spellingShingle | Zhijie Yang Xin Wei Jianqiao He Changli Sun Jianhua Ju Junying Ma Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous Host Marine Drugs atratumycin luxr sarp abc transporter heterologous expression |
| title | Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous Host |
| title_full | Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous Host |
| title_fullStr | Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous Host |
| title_full_unstemmed | Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous Host |
| title_short | Characterization of the Noncanonical Regulatory and Transporter Genes in Atratumycin Biosynthesis and Production in a Heterologous Host |
| title_sort | characterization of the noncanonical regulatory and transporter genes in atratumycin biosynthesis and production in a heterologous host |
| topic | atratumycin luxr sarp abc transporter heterologous expression |
| url | https://www.mdpi.com/1660-3397/17/10/560 |
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