{"_buckets": {"deposit": "4f1539e4-ec5f-4790-8778-f802ca92ebba"}, "_deposit": {"id": "19989", "owners": [], "pid": {"revision_id": 0, "type": "depid", "value": "19989"}, "status": "published"}, "_oai": {"id": "oai:soar-ir.repo.nii.ac.jp:00019989", "sets": ["1016:1017"]}, "author_link": ["105946", "105947", "105948", "105949", "105950", "105951", "105952"], "item_1628147817048": {"attribute_name": "\u51fa\u7248\u30bf\u30a4\u30d7", "attribute_value_mlt": [{"subitem_version_resource": "http://purl.org/coar/version/c_ab4af688f83e57aa", "subitem_version_type": "AM"}]}, "item_6_biblio_info_6": {"attribute_name": "\u66f8\u8a8c\u60c5\u5831", "attribute_value_mlt": [{"bibliographicIssueDates": {"bibliographicIssueDate": "2017-10", "bibliographicIssueDateType": "Issued"}, "bibliographicIssueNumber": "19", "bibliographicPageStart": "UNSP e01322-17", "bibliographicVolumeNumber": "83", "bibliographic_titles": [{"bibliographic_title": "APPLIED AND ENVIRONMENTAL MICROBIOLOGY"}]}]}, "item_6_description_20": {"attribute_name": "\u6284\u9332", "attribute_value_mlt": [{"subitem_description": "For fatty acid biosynthesis, Corynebacterium glutamicum uses two type I fatty acid synthases (FAS-I), FasA and FasB, in addition to acetyl-coenzyme A (CoA) carboxylase (ACC) consisting of AccBC, AccD1, and AccE. The in vivo roles of the enzymes in supplying precursors for biotin and alpha-lipoic acid remain unclear. Here, we report genetic evidence demonstrating that the biosynthesis of these cofactors is linked to fatty acid biosynthesis through the FAS-I pathway. For this study, we used wild-type C. glutamicum and its derived biotin vitamer producer BFI-5, which was engineered to express Escherichia coli bioBF and Bacillus subtilis bioI. Disruption of either fasA or fasB in strain BFI-5 led to decreased production of biotin vitamers, whereas its amplification contributed to increased production, with a larger impact of fasA in both cases. Double disruptions of fasA and fasB resulted in no biotin vitamer production. The acc genes showed a positive effect on production when amplified simultaneously. Augmented fatty acid biosynthesis was also reflected in pimelic acid production when carbon flow was blocked at the BioF reaction. These results indicate that carbon flow down the FAS-I pathway is destined for channeling into the biotin biosynthesis pathway, and that FasA in particular has a significant impact on precursor supply. In contrast, fasB disruption resulted in auxotrophy for lipoic acid or its precursor octanoic acid in both wild-type and BFI-5 strains. The phenotypes were fully complemented by plasmid-mediated expression of fasB but not fasA. These results reveal that FasB plays a specific physiological role in lipoic acid biosynthesis in C. glutamicum. IMPORTANCE For the de novo biosynthesis of fatty acids, C. glutamicum exceptionally uses a eukaryotic multifunctional type I fatty acid synthase (FAS-I) system comprising FasA and FasB, in contrast to most bacteria, such as E. coli and B. subtilis, which use an individual nonaggregating type II fatty acid synthase (FAS-II) system. In this study, we reported genetic evidence demonstrating that the FAS-I system is the source of the biotin precursor in vivo in the engineered biotin-prototrophic C. glutamicum strain. This study also uncovered the important physiological role of FasB in lipoic acid biosynthesis. Here, we present an FAS-I enzyme that functions in supplying the lipoic acid precursor, although its biosynthesis has been believed to exclusively depend on FAS-II in organisms. 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The in vivo roles of the enzymes in supplying precursors for biotin and alpha-lipoic acid remain unclear. Here, we report genetic evidence demonstrating that the biosynthesis of these cofactors is linked to fatty acid biosynthesis through the FAS-I pathway. For this study, we used wild-type C. glutamicum and its derived biotin vitamer producer BFI-5, which was engineered to express Escherichia coli bioBF and Bacillus subtilis bioI. Disruption of either fasA or fasB in strain BFI-5 led to decreased production of biotin vitamers, whereas its amplification contributed to increased production, with a larger impact of fasA in both cases. Double disruptions of fasA and fasB resulted in no biotin vitamer production. The acc genes showed a positive effect on production when amplified simultaneously. Augmented fatty acid biosynthesis was also reflected in pimelic acid production when carbon flow was blocked at the BioF reaction. These results indicate that carbon flow down the FAS-I pathway is destined for channeling into the biotin biosynthesis pathway, and that FasA in particular has a significant impact on precursor supply. In contrast, fasB disruption resulted in auxotrophy for lipoic acid or its precursor octanoic acid in both wild-type and BFI-5 strains. The phenotypes were fully complemented by plasmid-mediated expression of fasB but not fasA. These results reveal that FasB plays a specific physiological role in lipoic acid biosynthesis in C. glutamicum. IMPORTANCE For the de novo biosynthesis of fatty acids, C. glutamicum exceptionally uses a eukaryotic multifunctional type I fatty acid synthase (FAS-I) system comprising FasA and FasB, in contrast to most bacteria, such as E. coli and B. subtilis, which use an individual nonaggregating type II fatty acid synthase (FAS-II) system. In this study, we reported genetic evidence demonstrating that the FAS-I system is the source of the biotin precursor in vivo in the engineered biotin-prototrophic C. glutamicum strain. This study also uncovered the important physiological role of FasB in lipoic acid biosynthesis. Here, we present an FAS-I enzyme that functions in supplying the lipoic acid precursor, although its biosynthesis has been believed to exclusively depend on FAS-II in organisms. The findings obtained here provide new insights into the metabolic engineering of this industrially important microorganism to produce these compounds effectively."}]}, "item_creator": {"attribute_name": "\u8457\u8005", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "Ikeda, Masato"}], "nameIdentifiers": [{"nameIdentifier": "105946", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Nagashima, Takashi"}], "nameIdentifiers": [{"nameIdentifier": "105947", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Nakamura, Eri"}], "nameIdentifiers": [{"nameIdentifier": "105948", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Kato, Ryosuke"}], "nameIdentifiers": [{"nameIdentifier": "105949", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Ohshita, Masakazu"}], "nameIdentifiers": [{"nameIdentifier": "105950", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Hayashi, Mikiro"}], "nameIdentifiers": [{"nameIdentifier": "105951", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Takeno, Seiki"}], "nameIdentifiers": [{"nameIdentifier": "105952", "nameIdentifierScheme": "WEKO"}]}]}, "item_files": {"attribute_name": "\u30d5\u30a1\u30a4\u30eb\u60c5\u5831", "attribute_type": "file", "attribute_value_mlt": [{"accessrole": "open_date", "date": [{"dateType": "Available", "dateValue": "2018-07-24"}], "displaytype": "detail", "download_preview_message": "", "file_order": 0, "filename": "In_vivo_roles_of_fatty_acid-biosynthetic_enzymes_in_biosynthesis_3_of_biotin.pdf", "filesize": [{"value": "783.2 kB"}], "format": "application/pdf", "future_date_message": "", "is_thumbnail": false, "licensefree": "Copyright \u00a9 2017 American Society for Microbiology.", "licensetype": "license_note", "mimetype": "application/pdf", "size": 783200.0, "url": {"label": "In_vivo_roles_of_fatty_acid-biosynthetic_enzymes_in_biosynthesis_3_of_biotin.pdf", "url": "https://soar-ir.repo.nii.ac.jp/record/19989/files/In_vivo_roles_of_fatty_acid-biosynthetic_enzymes_in_biosynthesis_3_of_biotin.pdf"}, "version_id": "71e15239-d6a6-4d7a-b6fc-0f8665ec5d3b"}]}, "item_keyword": {"attribute_name": "\u30ad\u30fc\u30ef\u30fc\u30c9", "attribute_value_mlt": [{"subitem_subject": "Corynebacterium glutamicum", "subitem_subject_scheme": "Other"}, {"subitem_subject": "biotin", "subitem_subject_scheme": "Other"}, {"subitem_subject": "fatty acid biosynthesis", "subitem_subject_scheme": "Other"}, {"subitem_subject": "lipoic acid", "subitem_subject_scheme": "Other"}, {"subitem_subject": "metabolic engineering", "subitem_subject_scheme": "Other"}]}, "item_language": {"attribute_name": "\u8a00\u8a9e", "attribute_value_mlt": [{"subitem_language": "eng"}]}, "item_resource_type": {"attribute_name": "\u8cc7\u6e90\u30bf\u30a4\u30d7", "attribute_value_mlt": [{"resourcetype": "journal article", "resourceuri": "http://purl.org/coar/resource_type/c_6501"}]}, "item_title": "In Vivo Roles of Fatty Acid Biosynthesis Enzymes in Biosynthesis of Biotin and alpha-Lipoic Acid in Corynebacterium glutamicum", "item_titles": {"attribute_name": "\u30bf\u30a4\u30c8\u30eb", "attribute_value_mlt": [{"subitem_title": "In Vivo Roles of Fatty Acid Biosynthesis Enzymes in Biosynthesis of Biotin and alpha-Lipoic Acid in Corynebacterium glutamicum", "subitem_title_language": "en"}]}, "item_type_id": "6", "owner": "1", "path": ["1016/1017"], "permalink_uri": "http://hdl.handle.net/10091/00020750", "pubdate": {"attribute_name": "PubDate", "attribute_value": "2018-07-24"}, "publish_date": "2018-07-24", "publish_status": "0", "recid": "19989", "relation": {}, "relation_version_is_last": true, "title": ["In Vivo Roles of Fatty Acid Biosynthesis Enzymes in Biosynthesis of Biotin and alpha-Lipoic Acid in Corynebacterium glutamicum"], "weko_shared_id": -1}