Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907
Micafungin (FK463) is a widely used treatment for life-threatening, deep-seated fungal infections. It is an echinocandin-like lipopeptide derived from the chemical modification of deacylated FR901379, a type of lipopeptide antibiotic produced by Coleophoma empetri F-11899. The palmitoyl moiety of FR...
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creator | Ueda, Satoshi Kinoshita, Masayoshi Tanaka, Fumihiro Tsuboi, Masaru Shimizu, Shiho Oohata, Nobutaka Hino, Motohiro Yamada, Masato Isogai, Yasuhiro Hashimoto, Seiji |
description | Micafungin (FK463) is a widely used treatment for life-threatening, deep-seated fungal infections. It is an echinocandin-like lipopeptide derived from the chemical modification of deacylated FR901379, a type of lipopeptide antibiotic produced by
Coleophoma empetri F-11899. The palmitoyl moiety of FR901379 is deacylated by FR901379 acylase produced by
Streptomyces sp. no. 6907. In this study, our goal was to generate an improved strain of
Streptomyces sp. no. 6907 capable of hyperproducing the FR901379-acylase enzyme. To accomplish this goal, modified strains of
Streptomyces sp. no. 6907 were generated using UV-irradiation mutagenesis, and strain selection was performed using an agar-plate screening method to efficiently select an acylase-hyperproducing strain. Three marker indices were shown to correlate with elevated acylase production: decreased candidacidal activity of FR901379, decreased proteolytic activity on skim milk, and phenotypic characteristics. Cloning and subsequent sequencing of the acylase gene from the hyperproducing mutant revealed no mutations in either the acylase structural gene or the 5′-flanking region required for gene expression. The growth medium was also modified to maximize acylase production. We successfully increased acylase activity approximately 65-fold, compared with the original growth conditions (wild strain cultured in the original unmodified medium). To minimize formation of excess foam during the fermentation process, we optimized the parameters of agitation speed, as calculated from the discharge flow rate. Using our improved strain and the optimized medium and growth conditions, we have developed an improved and highly reproducible method for stable large-scale production of FR901379-acylase. |
doi_str_mv | 10.1016/j.jbiosc.2011.06.002 |
format | Article |
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Coleophoma empetri F-11899. The palmitoyl moiety of FR901379 is deacylated by FR901379 acylase produced by
Streptomyces sp. no. 6907. In this study, our goal was to generate an improved strain of
Streptomyces sp. no. 6907 capable of hyperproducing the FR901379-acylase enzyme. To accomplish this goal, modified strains of
Streptomyces sp. no. 6907 were generated using UV-irradiation mutagenesis, and strain selection was performed using an agar-plate screening method to efficiently select an acylase-hyperproducing strain. Three marker indices were shown to correlate with elevated acylase production: decreased candidacidal activity of FR901379, decreased proteolytic activity on skim milk, and phenotypic characteristics. Cloning and subsequent sequencing of the acylase gene from the hyperproducing mutant revealed no mutations in either the acylase structural gene or the 5′-flanking region required for gene expression. The growth medium was also modified to maximize acylase production. We successfully increased acylase activity approximately 65-fold, compared with the original growth conditions (wild strain cultured in the original unmodified medium). To minimize formation of excess foam during the fermentation process, we optimized the parameters of agitation speed, as calculated from the discharge flow rate. Using our improved strain and the optimized medium and growth conditions, we have developed an improved and highly reproducible method for stable large-scale production of FR901379-acylase.</description><identifier>ISSN: 1389-1723</identifier><identifier>EISSN: 1347-4421</identifier><identifier>DOI: 10.1016/j.jbiosc.2011.06.002</identifier><identifier>PMID: 21752710</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Agitation ; Amidohydrolases - biosynthesis ; Amidohydrolases - metabolism ; antibiotics ; Antifungal Agents - chemistry ; Antifungal Agents - metabolism ; Bioconversion ; Bioconversions. Hemisynthesis ; Biological and medical sciences ; Biotechnology ; culture media ; Culture Media - chemistry ; Echinocandins - chemistry ; Echinocandins - metabolism ; Fermentation ; foams ; FR901379 acylase ; Fundamental and applied biological sciences. Psychology ; fungi ; gene expression ; Lipopeptides - chemistry ; Lipopeptides - metabolism ; Medium improvement ; Methods. Procedures. Technologies ; micafungin ; Microbial engineering. Fermentation and microbial culture technology ; mutagenesis ; mutants ; Peptides, Cyclic - chemistry ; Peptides, Cyclic - metabolism ; proteolysis ; Scale-up ; screening ; skim milk ; Strain breeding ; Streptomyces ; Streptomyces - classification ; Streptomyces - enzymology ; structural genes</subject><ispartof>Journal of bioscience and bioengineering, 2011-10, Vol.112 (4), p.409-414</ispartof><rights>2011 The Society for Biotechnology, Japan</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c513t-6a2593b9d62878067e8a4c8cf997f0be7214148e470a2f810428379e2e7604a83</citedby><cites>FETCH-LOGICAL-c513t-6a2593b9d62878067e8a4c8cf997f0be7214148e470a2f810428379e2e7604a83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jbiosc.2011.06.002$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24720025$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21752710$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ueda, Satoshi</creatorcontrib><creatorcontrib>Kinoshita, Masayoshi</creatorcontrib><creatorcontrib>Tanaka, Fumihiro</creatorcontrib><creatorcontrib>Tsuboi, Masaru</creatorcontrib><creatorcontrib>Shimizu, Shiho</creatorcontrib><creatorcontrib>Oohata, Nobutaka</creatorcontrib><creatorcontrib>Hino, Motohiro</creatorcontrib><creatorcontrib>Yamada, Masato</creatorcontrib><creatorcontrib>Isogai, Yasuhiro</creatorcontrib><creatorcontrib>Hashimoto, Seiji</creatorcontrib><title>Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907</title><title>Journal of bioscience and bioengineering</title><addtitle>J Biosci Bioeng</addtitle><description>Micafungin (FK463) is a widely used treatment for life-threatening, deep-seated fungal infections. It is an echinocandin-like lipopeptide derived from the chemical modification of deacylated FR901379, a type of lipopeptide antibiotic produced by
Coleophoma empetri F-11899. The palmitoyl moiety of FR901379 is deacylated by FR901379 acylase produced by
Streptomyces sp. no. 6907. In this study, our goal was to generate an improved strain of
Streptomyces sp. no. 6907 capable of hyperproducing the FR901379-acylase enzyme. To accomplish this goal, modified strains of
Streptomyces sp. no. 6907 were generated using UV-irradiation mutagenesis, and strain selection was performed using an agar-plate screening method to efficiently select an acylase-hyperproducing strain. Three marker indices were shown to correlate with elevated acylase production: decreased candidacidal activity of FR901379, decreased proteolytic activity on skim milk, and phenotypic characteristics. Cloning and subsequent sequencing of the acylase gene from the hyperproducing mutant revealed no mutations in either the acylase structural gene or the 5′-flanking region required for gene expression. The growth medium was also modified to maximize acylase production. We successfully increased acylase activity approximately 65-fold, compared with the original growth conditions (wild strain cultured in the original unmodified medium). To minimize formation of excess foam during the fermentation process, we optimized the parameters of agitation speed, as calculated from the discharge flow rate. Using our improved strain and the optimized medium and growth conditions, we have developed an improved and highly reproducible method for stable large-scale production of FR901379-acylase.</description><subject>Agitation</subject><subject>Amidohydrolases - biosynthesis</subject><subject>Amidohydrolases - metabolism</subject><subject>antibiotics</subject><subject>Antifungal Agents - chemistry</subject><subject>Antifungal Agents - metabolism</subject><subject>Bioconversion</subject><subject>Bioconversions. Hemisynthesis</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>culture media</subject><subject>Culture Media - chemistry</subject><subject>Echinocandins - chemistry</subject><subject>Echinocandins - metabolism</subject><subject>Fermentation</subject><subject>foams</subject><subject>FR901379 acylase</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>fungi</subject><subject>gene expression</subject><subject>Lipopeptides - chemistry</subject><subject>Lipopeptides - metabolism</subject><subject>Medium improvement</subject><subject>Methods. Procedures. Technologies</subject><subject>micafungin</subject><subject>Microbial engineering. Fermentation and microbial culture technology</subject><subject>mutagenesis</subject><subject>mutants</subject><subject>Peptides, Cyclic - chemistry</subject><subject>Peptides, Cyclic - metabolism</subject><subject>proteolysis</subject><subject>Scale-up</subject><subject>screening</subject><subject>skim milk</subject><subject>Strain breeding</subject><subject>Streptomyces</subject><subject>Streptomyces - classification</subject><subject>Streptomyces - enzymology</subject><subject>structural genes</subject><issn>1389-1723</issn><issn>1347-4421</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtv1TAQhSMEoqXwDxB4g7pKmHF8_dggVRUFpEpIlK4tx5lUvsrjYieV7r_HIRfYwcoj-ZtzjuYUxWuECgHl-321b8KUfMUBsQJZAfAnxTnWQpVCcHy6ztqUqHh9VrxIaQ-AChQ-L844qh1XCOfFw90cXRhZop78HKaRubFlybueyuXAOooDjbP79dNNkd18M4C1Msz5Y-8SsUOc2mXbbI4sq9Fhnoajp8TSoWLjVDFpQL0snnWuT_Tq9F4U9zcfv19_Lm-_fvpyfXVb-h3Wcykd35m6Ma3kWmmQirQTXvvOGNVBQ4qjQKFJKHC80wiC65yGOCkJwun6orjcdHOuHwul2Q4heep7N9K0JGuAZx6Q_5fURmdpqUQmxUb6OKUUqbOHGAYXjxbBrl3Yvd26sGsXFqTNXeS1NyeDpRmo_bP0-_gZeHcC3HrwLrrRh_SXE4pnnV3m3m5c5ybrHmJm7u-yk4TVRiJm4sNGUD7tY6Bokw80empDzLXadgr_zvoTxpOvPg</recordid><startdate>20111001</startdate><enddate>20111001</enddate><creator>Ueda, Satoshi</creator><creator>Kinoshita, Masayoshi</creator><creator>Tanaka, Fumihiro</creator><creator>Tsuboi, Masaru</creator><creator>Shimizu, Shiho</creator><creator>Oohata, Nobutaka</creator><creator>Hino, Motohiro</creator><creator>Yamada, Masato</creator><creator>Isogai, Yasuhiro</creator><creator>Hashimoto, Seiji</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7QL</scope><scope>7QO</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20111001</creationdate><title>Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907</title><author>Ueda, Satoshi ; Kinoshita, Masayoshi ; Tanaka, Fumihiro ; Tsuboi, Masaru ; Shimizu, Shiho ; Oohata, Nobutaka ; Hino, Motohiro ; Yamada, Masato ; Isogai, Yasuhiro ; Hashimoto, Seiji</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c513t-6a2593b9d62878067e8a4c8cf997f0be7214148e470a2f810428379e2e7604a83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Agitation</topic><topic>Amidohydrolases - biosynthesis</topic><topic>Amidohydrolases - metabolism</topic><topic>antibiotics</topic><topic>Antifungal Agents - chemistry</topic><topic>Antifungal Agents - metabolism</topic><topic>Bioconversion</topic><topic>Bioconversions. Hemisynthesis</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>culture media</topic><topic>Culture Media - chemistry</topic><topic>Echinocandins - chemistry</topic><topic>Echinocandins - metabolism</topic><topic>Fermentation</topic><topic>foams</topic><topic>FR901379 acylase</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>fungi</topic><topic>gene expression</topic><topic>Lipopeptides - chemistry</topic><topic>Lipopeptides - metabolism</topic><topic>Medium improvement</topic><topic>Methods. Procedures. Technologies</topic><topic>micafungin</topic><topic>Microbial engineering. Fermentation and microbial culture technology</topic><topic>mutagenesis</topic><topic>mutants</topic><topic>Peptides, Cyclic - chemistry</topic><topic>Peptides, Cyclic - metabolism</topic><topic>proteolysis</topic><topic>Scale-up</topic><topic>screening</topic><topic>skim milk</topic><topic>Strain breeding</topic><topic>Streptomyces</topic><topic>Streptomyces - classification</topic><topic>Streptomyces - enzymology</topic><topic>structural genes</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ueda, Satoshi</creatorcontrib><creatorcontrib>Kinoshita, Masayoshi</creatorcontrib><creatorcontrib>Tanaka, Fumihiro</creatorcontrib><creatorcontrib>Tsuboi, Masaru</creatorcontrib><creatorcontrib>Shimizu, Shiho</creatorcontrib><creatorcontrib>Oohata, Nobutaka</creatorcontrib><creatorcontrib>Hino, Motohiro</creatorcontrib><creatorcontrib>Yamada, Masato</creatorcontrib><creatorcontrib>Isogai, Yasuhiro</creatorcontrib><creatorcontrib>Hashimoto, Seiji</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of bioscience and bioengineering</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ueda, Satoshi</au><au>Kinoshita, Masayoshi</au><au>Tanaka, Fumihiro</au><au>Tsuboi, Masaru</au><au>Shimizu, Shiho</au><au>Oohata, Nobutaka</au><au>Hino, Motohiro</au><au>Yamada, Masato</au><au>Isogai, Yasuhiro</au><au>Hashimoto, Seiji</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907</atitle><jtitle>Journal of bioscience and bioengineering</jtitle><addtitle>J Biosci Bioeng</addtitle><date>2011-10-01</date><risdate>2011</risdate><volume>112</volume><issue>4</issue><spage>409</spage><epage>414</epage><pages>409-414</pages><issn>1389-1723</issn><eissn>1347-4421</eissn><abstract>Micafungin (FK463) is a widely used treatment for life-threatening, deep-seated fungal infections. It is an echinocandin-like lipopeptide derived from the chemical modification of deacylated FR901379, a type of lipopeptide antibiotic produced by
Coleophoma empetri F-11899. The palmitoyl moiety of FR901379 is deacylated by FR901379 acylase produced by
Streptomyces sp. no. 6907. In this study, our goal was to generate an improved strain of
Streptomyces sp. no. 6907 capable of hyperproducing the FR901379-acylase enzyme. To accomplish this goal, modified strains of
Streptomyces sp. no. 6907 were generated using UV-irradiation mutagenesis, and strain selection was performed using an agar-plate screening method to efficiently select an acylase-hyperproducing strain. Three marker indices were shown to correlate with elevated acylase production: decreased candidacidal activity of FR901379, decreased proteolytic activity on skim milk, and phenotypic characteristics. Cloning and subsequent sequencing of the acylase gene from the hyperproducing mutant revealed no mutations in either the acylase structural gene or the 5′-flanking region required for gene expression. The growth medium was also modified to maximize acylase production. We successfully increased acylase activity approximately 65-fold, compared with the original growth conditions (wild strain cultured in the original unmodified medium). To minimize formation of excess foam during the fermentation process, we optimized the parameters of agitation speed, as calculated from the discharge flow rate. Using our improved strain and the optimized medium and growth conditions, we have developed an improved and highly reproducible method for stable large-scale production of FR901379-acylase.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>21752710</pmid><doi>10.1016/j.jbiosc.2011.06.002</doi><tpages>6</tpages></addata></record> |
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source | MEDLINE; Elsevier ScienceDirect Journals Complete |
subjects | Agitation Amidohydrolases - biosynthesis Amidohydrolases - metabolism antibiotics Antifungal Agents - chemistry Antifungal Agents - metabolism Bioconversion Bioconversions. Hemisynthesis Biological and medical sciences Biotechnology culture media Culture Media - chemistry Echinocandins - chemistry Echinocandins - metabolism Fermentation foams FR901379 acylase Fundamental and applied biological sciences. Psychology fungi gene expression Lipopeptides - chemistry Lipopeptides - metabolism Medium improvement Methods. Procedures. Technologies micafungin Microbial engineering. Fermentation and microbial culture technology mutagenesis mutants Peptides, Cyclic - chemistry Peptides, Cyclic - metabolism proteolysis Scale-up screening skim milk Strain breeding Streptomyces Streptomyces - classification Streptomyces - enzymology structural genes |
title | Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907 |
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