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Development of high-throughput quantitative analytical method for l-cysteine-containing dipeptides by LC–MS/MS toward its fermentative production
http://hdl.handle.net/2241/00159701
http://hdl.handle.net/2241/001597018344f01c-7531-4874-9013-3378d2d8551d
名前 / ファイル | ライセンス | アクション |
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AMB_9 (1.4 MB)
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Item type | Journal Article(1) | |||||
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公開日 | 2020-03-05 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | Development of high-throughput quantitative analytical method for l-cysteine-containing dipeptides by LC–MS/MS toward its fermentative production | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源 | http://purl.org/coar/resource_type/c_6501 | |||||
タイプ | journal article | |||||
著者 |
河野, 祐介
× 河野, 祐介× 大津, 厳生× Shiroyama, Maeka× Kanazawa, Koji× Suzuki, Yasushi A. |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | l-Cysteine (Cys) is metabolically fundamental sulfur compound and important components in various cellular fac-tors. Interestingly, free-form Cys itself as a simple monomeric amino acid was recently shown to function in a novel antioxidative system (cysteine/cystine shuttle system) in Escherichia coli. However, as for Cys-containing dipeptides, the biological functions, effects, and even contents have still remained largely elusive. The potential functions should be a part of cellular redox system and important in basic and applied biology. For its progress, establishment of reli-able quantitation method is the first. However, such accurate analysis is unexpectedly difficult even in Cys, because thiol compounds convert through disulfide-exchange and air oxidation during sample preparation. Addressing this problem, in this study, thiol molecules like Cys-containing dipeptides were derivatized by using monobromobimane (thiol-specific alkylating reagent) and detected as S-bimanyl derivatives by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS). Sample separation was processed with a C18 column (2.1 mm × 150 mm, 1.7 μm) and with water-acetonitrile gradient mobile phase containing 0.1% (v/v) formic acid at flow rate of 0.25 ml/min. The mass spectrometer was operated in the multiple reaction monitoring in positive/negative mode with electrospray ionization. The derivatization could indeed avoid the unfavorable reactions, namely, developed the method reflecting their correct contents on sampling. Furthermore, the method was successfully applied to monitoring Cys-containing dipeptides in E. coli Cys producer overexpressing bacD gene. This is the first report of the quantitative analysis of Cys-containing dipeptides, which should be useful for further study of fermentative production of Cys-containing dipeptides. | |||||
書誌情報 |
en : AMB Express 巻 9, p. 91, 発行日 2019-06 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 2191-0855 | |||||
PubMed番号 | ||||||
識別子タイプ | PMID | |||||
関連識別子 | 31227937 | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1186/s13568-019-0817-2 | |||||
権利 | ||||||
権利情報 | © The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. | |||||
著者版フラグ | ||||||
値 | publisher | |||||
出版者 | ||||||
出版者 | BioMed Central Ltd |