Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis
A single cycle of strain improvement was performed in Saccharopolyspora erythraea mutB and 15 genotypes influencing erythromycin production were found. Genotypes generated by transposon mutagenesis appeared in the screen at a frequency of ∼3%. Mutations affecting central metabolism and regulatory ge...
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| Veröffentlicht in: | FEMS microbiology letters 2015-11, Vol.362 (22), p.fnv180 |
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| creator | Fedashchin, Andrij Cernota, William H. Gonzalez, Melissa C. Leach, Benjamin I. Kwan, Noelle Wesley, Roy K. Weber, J. Mark |
| description | A single cycle of strain improvement was performed in Saccharopolyspora erythraea mutB and 15 genotypes influencing erythromycin production were found. Genotypes generated by transposon mutagenesis appeared in the screen at a frequency of ∼3%. Mutations affecting central metabolism and regulatory genes were found, as well as hydrolases, peptidases, glycosyl transferases and unknown genes. Only one mutant retained high erythromycin production when scaled-up from micro-agar plug fermentations to shake flasks. This mutant had a knockout of the cwh1 gene (SACE_1598), encoding a cell-wall-associated hydrolase. The cwh1 knockout produced visible growth and morphological defects on solid medium. This study demonstrated that random transposon mutagenesis uncovers strain improvement-related genes potentially useful for strain engineering.
A gene influencing the production level of the antibiotic erythromycin was uncovered using a random transposon mutagenesis procedure. |
| doi_str_mv | 10.1093/femsle/fnv180 |
| format | Article |
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A gene influencing the production level of the antibiotic erythromycin was uncovered using a random transposon mutagenesis procedure.</description><identifier>ISSN: 1574-6968</identifier><identifier>ISSN: 0378-1097</identifier><identifier>EISSN: 1574-6968</identifier><identifier>DOI: 10.1093/femsle/fnv180</identifier><identifier>PMID: 26468041</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Anti-Bacterial Agents - biosynthesis ; Antibiotics ; Biosynthesis ; Biosynthetic Pathways - genetics ; Biotechnology & Synthetic Biology ; Cell culture ; DNA Transposable Elements ; Erythromycin ; Erythromycin - biosynthesis ; Flasks ; Genes ; Genes, Bacterial ; Genome, Bacterial ; Genomes ; Genotypes ; Hydrolase ; Metabolic Engineering - methods ; Microbiology ; Mutagenesis ; Mutagenesis, Insertional ; Mutation ; Peptidases ; Research Letter ; Saccharopolyspora - genetics ; Saccharopolyspora - metabolism ; Saccharopolyspora erythraea ; Transposon mutagenesis ; Transposons</subject><ispartof>FEMS microbiology letters, 2015-11, Vol.362 (22), p.fnv180</ispartof><rights>FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com 2015</rights><rights>FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.</rights><rights>FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com</rights><rights>FEMS 2015. All rights reserved. For permissions, please e-mail: 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c409t-17d02b9b959b324a9977c476316587fbf07e69813ffcf522bb0a97fb002185483</citedby><cites>FETCH-LOGICAL-c409t-17d02b9b959b324a9977c476316587fbf07e69813ffcf522bb0a97fb002185483</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,1584,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26468041$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Silver, Simon</contributor><creatorcontrib>Fedashchin, Andrij</creatorcontrib><creatorcontrib>Cernota, William H.</creatorcontrib><creatorcontrib>Gonzalez, Melissa C.</creatorcontrib><creatorcontrib>Leach, Benjamin I.</creatorcontrib><creatorcontrib>Kwan, Noelle</creatorcontrib><creatorcontrib>Wesley, Roy K.</creatorcontrib><creatorcontrib>Weber, J. Mark</creatorcontrib><title>Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis</title><title>FEMS microbiology letters</title><addtitle>FEMS Microbiol Lett</addtitle><description>A single cycle of strain improvement was performed in Saccharopolyspora erythraea mutB and 15 genotypes influencing erythromycin production were found. Genotypes generated by transposon mutagenesis appeared in the screen at a frequency of ∼3%. Mutations affecting central metabolism and regulatory genes were found, as well as hydrolases, peptidases, glycosyl transferases and unknown genes. Only one mutant retained high erythromycin production when scaled-up from micro-agar plug fermentations to shake flasks. This mutant had a knockout of the cwh1 gene (SACE_1598), encoding a cell-wall-associated hydrolase. The cwh1 knockout produced visible growth and morphological defects on solid medium. This study demonstrated that random transposon mutagenesis uncovers strain improvement-related genes potentially useful for strain engineering.
A gene influencing the production level of the antibiotic erythromycin was uncovered using a random transposon mutagenesis procedure.</description><subject>Anti-Bacterial Agents - biosynthesis</subject><subject>Antibiotics</subject><subject>Biosynthesis</subject><subject>Biosynthetic Pathways - genetics</subject><subject>Biotechnology & Synthetic Biology</subject><subject>Cell culture</subject><subject>DNA Transposable Elements</subject><subject>Erythromycin</subject><subject>Erythromycin - biosynthesis</subject><subject>Flasks</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Genome, Bacterial</subject><subject>Genomes</subject><subject>Genotypes</subject><subject>Hydrolase</subject><subject>Metabolic Engineering - methods</subject><subject>Microbiology</subject><subject>Mutagenesis</subject><subject>Mutagenesis, Insertional</subject><subject>Mutation</subject><subject>Peptidases</subject><subject>Research Letter</subject><subject>Saccharopolyspora - genetics</subject><subject>Saccharopolyspora - metabolism</subject><subject>Saccharopolyspora erythraea</subject><subject>Transposon mutagenesis</subject><subject>Transposons</subject><issn>1574-6968</issn><issn>0378-1097</issn><issn>1574-6968</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkUtv1TAQRi1ERUthyRZZYsMmdOw4TrxBQlV5SJUqtbC2nGR8r6vEDnZypfwBfjcuuVRlxcqvM2c8-gh5w-ADA1VeWBzTgBfWH1gDz8gZq2pRSCWb50_2p-RlSvcAIDjIF-SUSyEbEOyM_Lo1vg8jnaPxaQopeDous9mhx-QSDZbOe6R3puv2JoYpDGumoqEY13kfDRqa0TAijXhAMyRq-t7NLngz0D8S6rwdFvSd87tjVRjXfKKtC2n1WZ8bvSInNlfj6-N6Tn58vvp--bW4vvny7fLTddEJUHPB6h54q1pVqbbkwihV152oZclk1dS2tVCjVA0rre1sxXnbglH5HoCzphJNeU4-bt5paUfsO_R58EFP0Y0mrjoYp_998W6vd-GgRQNKKZYF746CGH4umGZ9H5aYp02al1BVFS_lA1VsVBdDShHtYwcG-iE2vcWmt9gy__bptx7pvzll4P0GhGX6j-s3HoGokw</recordid><startdate>20151101</startdate><enddate>20151101</enddate><creator>Fedashchin, Andrij</creator><creator>Cernota, William H.</creator><creator>Gonzalez, Melissa C.</creator><creator>Leach, Benjamin I.</creator><creator>Kwan, Noelle</creator><creator>Wesley, Roy K.</creator><creator>Weber, J. 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Mark</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c409t-17d02b9b959b324a9977c476316587fbf07e69813ffcf522bb0a97fb002185483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Anti-Bacterial Agents - biosynthesis</topic><topic>Antibiotics</topic><topic>Biosynthesis</topic><topic>Biosynthetic Pathways - genetics</topic><topic>Biotechnology & Synthetic Biology</topic><topic>Cell culture</topic><topic>DNA Transposable Elements</topic><topic>Erythromycin</topic><topic>Erythromycin - biosynthesis</topic><topic>Flasks</topic><topic>Genes</topic><topic>Genes, Bacterial</topic><topic>Genome, Bacterial</topic><topic>Genomes</topic><topic>Genotypes</topic><topic>Hydrolase</topic><topic>Metabolic Engineering - methods</topic><topic>Microbiology</topic><topic>Mutagenesis</topic><topic>Mutagenesis, Insertional</topic><topic>Mutation</topic><topic>Peptidases</topic><topic>Research Letter</topic><topic>Saccharopolyspora - genetics</topic><topic>Saccharopolyspora - metabolism</topic><topic>Saccharopolyspora erythraea</topic><topic>Transposon mutagenesis</topic><topic>Transposons</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fedashchin, Andrij</creatorcontrib><creatorcontrib>Cernota, William H.</creatorcontrib><creatorcontrib>Gonzalez, Melissa C.</creatorcontrib><creatorcontrib>Leach, Benjamin I.</creatorcontrib><creatorcontrib>Kwan, Noelle</creatorcontrib><creatorcontrib>Wesley, Roy K.</creatorcontrib><creatorcontrib>Weber, J. 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Mark</au><au>Silver, Simon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis</atitle><jtitle>FEMS microbiology letters</jtitle><addtitle>FEMS Microbiol Lett</addtitle><date>2015-11-01</date><risdate>2015</risdate><volume>362</volume><issue>22</issue><spage>fnv180</spage><pages>fnv180-</pages><issn>1574-6968</issn><issn>0378-1097</issn><eissn>1574-6968</eissn><abstract>A single cycle of strain improvement was performed in Saccharopolyspora erythraea mutB and 15 genotypes influencing erythromycin production were found. Genotypes generated by transposon mutagenesis appeared in the screen at a frequency of ∼3%. Mutations affecting central metabolism and regulatory genes were found, as well as hydrolases, peptidases, glycosyl transferases and unknown genes. Only one mutant retained high erythromycin production when scaled-up from micro-agar plug fermentations to shake flasks. This mutant had a knockout of the cwh1 gene (SACE_1598), encoding a cell-wall-associated hydrolase. The cwh1 knockout produced visible growth and morphological defects on solid medium. This study demonstrated that random transposon mutagenesis uncovers strain improvement-related genes potentially useful for strain engineering.
A gene influencing the production level of the antibiotic erythromycin was uncovered using a random transposon mutagenesis procedure.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>26468041</pmid><doi>10.1093/femsle/fnv180</doi><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; OUP_牛津大学出版社现刊; Alma/SFX Local Collection |
| subjects | Anti-Bacterial Agents - biosynthesis Antibiotics Biosynthesis Biosynthetic Pathways - genetics Biotechnology & Synthetic Biology Cell culture DNA Transposable Elements Erythromycin Erythromycin - biosynthesis Flasks Genes Genes, Bacterial Genome, Bacterial Genomes Genotypes Hydrolase Metabolic Engineering - methods Microbiology Mutagenesis Mutagenesis, Insertional Mutation Peptidases Research Letter Saccharopolyspora - genetics Saccharopolyspora - metabolism Saccharopolyspora erythraea Transposon mutagenesis Transposons |
| title | Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis |
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