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Functional Characterization of F3H Gene and Optimization of Dihydrokaempferol Biosynthesis in ISaccharomyces cerevisiae/I
The 1092 bp F3H gene from Trapa bispinosa Roxb., which was named TbF3H, was cloned and it encodes 363 amino acids. Bioinformatic and phylogenetic tree analyses revealed the high homology of TbF3H with flavanone 3-hydroxylase from other plants. A functional analysis showed that TbF3H of Trapa bispino...
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Published in: | Molecules (Basel, Switzerland) Switzerland), 2024-05, Vol.29 (10) |
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container_title | Molecules (Basel, Switzerland) |
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creator | Chen, Qinyi Song, Dandan Sun, Xiaoyan Tian, Yulong Yan, Zecheng Min, Ting Wang, Hongxun Wang, Limei |
description | The 1092 bp F3H gene from Trapa bispinosa Roxb., which was named TbF3H, was cloned and it encodes 363 amino acids. Bioinformatic and phylogenetic tree analyses revealed the high homology of TbF3H with flavanone 3-hydroxylase from other plants. A functional analysis showed that TbF3H of Trapa bispinosa Roxb. encoded a functional flavanone 3-hydroxylase; it catalyzed the formation of dihydrokaempferol (DHK) from naringenin in S. cerevisiae. The promoter strengths were compared by fluorescence microscopy and flow cytometry detection of the fluorescence intensity of the reporter genes initiated by each constitutive promoter (FITC), and DHK production reached 216.7 mg/L by the promoter adjustment strategy and the optimization of fermentation conditions. The results presented in this study will contribute to elucidating DHK biosynthesis in Trapa bispinosa Roxb. |
doi_str_mv | 10.3390/molecules29102196 |
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Bioinformatic and phylogenetic tree analyses revealed the high homology of TbF3H with flavanone 3-hydroxylase from other plants. A functional analysis showed that TbF3H of Trapa bispinosa Roxb. encoded a functional flavanone 3-hydroxylase; it catalyzed the formation of dihydrokaempferol (DHK) from naringenin in S. cerevisiae. The promoter strengths were compared by fluorescence microscopy and flow cytometry detection of the fluorescence intensity of the reporter genes initiated by each constitutive promoter (FITC), and DHK production reached 216.7 mg/L by the promoter adjustment strategy and the optimization of fermentation conditions. 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The results presented in this study will contribute to elucidating DHK biosynthesis in Trapa bispinosa Roxb.</description><subject>Amino acids</subject><subject>Biosynthesis</subject><subject>Genetic aspects</subject><issn>1420-3049</issn><issn>1420-3049</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid/><recordid>eNptj09LAzEQxYMoWKsfwFvAc2uySXeTY632DxR6sPcyzk7a6G5SNlth_fSu6KEHmcMb3vzeg2HsXoqxUlY81rEiPFWUMitFJm1-wQZSZ2KkhLaXZ_s1u0npXfSMlpMB6-angK2PASo-O0AD2FLjv-DH4tHxuVryBQXiEEq-Oba-Pjs--0NXNvEDqD46amLFn3xMXWgPlHziPvDVKyD2tbHukBJHaujTJw_0uLplVw6qRHd_OmTb-ct2thytN4vVbLoe7fPCjIxw1mSo0UiHDlWphZLOCQ2lNBkYZXIU4Gw5KSXY3CA4YXMBKLV9A8jUkD381u6hop0PLrb9k7VPuJsWdqKLQhjTU-N_qH5Kqj3GQM73_lngG72wclE</recordid><startdate>20240501</startdate><enddate>20240501</enddate><creator>Chen, Qinyi</creator><creator>Song, Dandan</creator><creator>Sun, Xiaoyan</creator><creator>Tian, Yulong</creator><creator>Yan, Zecheng</creator><creator>Min, Ting</creator><creator>Wang, Hongxun</creator><creator>Wang, Limei</creator><general>MDPI AG</general><scope/></search><sort><creationdate>20240501</creationdate><title>Functional Characterization of F3H Gene and Optimization of Dihydrokaempferol Biosynthesis in ISaccharomyces cerevisiae/I</title><author>Chen, Qinyi ; Song, Dandan ; Sun, Xiaoyan ; Tian, Yulong ; Yan, Zecheng ; Min, Ting ; Wang, Hongxun ; Wang, Limei</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-g678-80f982c4c81fcfc3d4031ff04ad182a8386c0af9d5d1a968caf0960ac149baa23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Amino acids</topic><topic>Biosynthesis</topic><topic>Genetic aspects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Qinyi</creatorcontrib><creatorcontrib>Song, Dandan</creatorcontrib><creatorcontrib>Sun, Xiaoyan</creatorcontrib><creatorcontrib>Tian, Yulong</creatorcontrib><creatorcontrib>Yan, Zecheng</creatorcontrib><creatorcontrib>Min, Ting</creatorcontrib><creatorcontrib>Wang, Hongxun</creatorcontrib><creatorcontrib>Wang, Limei</creatorcontrib><jtitle>Molecules (Basel, Switzerland)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Qinyi</au><au>Song, Dandan</au><au>Sun, Xiaoyan</au><au>Tian, Yulong</au><au>Yan, Zecheng</au><au>Min, Ting</au><au>Wang, Hongxun</au><au>Wang, Limei</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Functional Characterization of F3H Gene and Optimization of Dihydrokaempferol Biosynthesis in ISaccharomyces cerevisiae/I</atitle><jtitle>Molecules (Basel, Switzerland)</jtitle><date>2024-05-01</date><risdate>2024</risdate><volume>29</volume><issue>10</issue><issn>1420-3049</issn><eissn>1420-3049</eissn><abstract>The 1092 bp F3H gene from Trapa bispinosa Roxb., which was named TbF3H, was cloned and it encodes 363 amino acids. Bioinformatic and phylogenetic tree analyses revealed the high homology of TbF3H with flavanone 3-hydroxylase from other plants. A functional analysis showed that TbF3H of Trapa bispinosa Roxb. encoded a functional flavanone 3-hydroxylase; it catalyzed the formation of dihydrokaempferol (DHK) from naringenin in S. cerevisiae. The promoter strengths were compared by fluorescence microscopy and flow cytometry detection of the fluorescence intensity of the reporter genes initiated by each constitutive promoter (FITC), and DHK production reached 216.7 mg/L by the promoter adjustment strategy and the optimization of fermentation conditions. The results presented in this study will contribute to elucidating DHK biosynthesis in Trapa bispinosa Roxb.</abstract><pub>MDPI AG</pub><doi>10.3390/molecules29102196</doi></addata></record> |
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title | Functional Characterization of F3H Gene and Optimization of Dihydrokaempferol Biosynthesis in ISaccharomyces cerevisiae/I |
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