Enzymatic Characterization of <i>SpPAL</i> Genes in <i>S. polyrhiza</i> and Overexpression of the <i>SpPAL3</i>
Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) catalyzes the deamination of phenylalanine, which is the initial step in the biosynthesis of phenylpropanoids. It serves as a crucial enzyme that facilitates the transfer of carbon from primary to secondary metabolism in plants. Duckweed is regarded as a...
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2024-09-01
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| author | Xiaoxue Li Yinxing Zhang Chunfeng Zhu Pufan Zheng Cunkun Chen Na Zhang Haipeng Ji Chenghu Dong Jinze Yu Jie Ren Yerong Zhu Yong Wang |
| author_facet | Xiaoxue Li Yinxing Zhang Chunfeng Zhu Pufan Zheng Cunkun Chen Na Zhang Haipeng Ji Chenghu Dong Jinze Yu Jie Ren Yerong Zhu Yong Wang |
| author_sort | Xiaoxue Li |
| collection | DOAJ |
| description | Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) catalyzes the deamination of phenylalanine, which is the initial step in the biosynthesis of phenylpropanoids. It serves as a crucial enzyme that facilitates the transfer of carbon from primary to secondary metabolism in plants. Duckweed is regarded as a promising chassis plant in synthetic biology research and application, due to its being rich in secondary metabolites and other advantages. The genes encoding PAL in <i>Spirodela polyrhiza</i> (L.) Schleid, the giant duckweed, were investigated in this study. Three <i>SpPAL</i> genes (<i>SpPAL1–SpPAL3</i>) were identified and cloned. All of them were successfully expressed in <i>E. coli</i>, and their recombinant proteins all showed PAL activity. In addition, SpPAL1 and SpPAL2 proteins could also utilize tyrosine as substrate, although the activity was low. A qRT-PCR analysis demonstrated that the expression of <i>SpPAL3</i> was most pronounced in young fronds. It was found that the expression of <i>SpPAL1</i> and <i>SpPAL3</i> was significantly induced by MeJA treatment. Overexpression of <i>SpPAL3</i> in <i>Lemna turionifera</i> inhibited the growth of fronds and adventitious roots in the transgenic plants, indicating the importance of <i>SpPAL3</i> in duckweed besides its involvement in the secondary metabolism. |
| format | Article |
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| institution | OA Journals |
| issn | 2223-7747 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | MDPI AG |
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| series | Plants |
| spelling | doaj-art-e4b75d360f014be1b664df4cb70e5da92025-08-20T01:55:48ZengMDPI AGPlants2223-77472024-09-011318260710.3390/plants13182607Enzymatic Characterization of <i>SpPAL</i> Genes in <i>S. polyrhiza</i> and Overexpression of the <i>SpPAL3</i>Xiaoxue Li0Yinxing Zhang1Chunfeng Zhu2Pufan Zheng3Cunkun Chen4Na Zhang5Haipeng Ji6Chenghu Dong7Jinze Yu8Jie Ren9Yerong Zhu10Yong Wang11Institute of Agricultural Products Preservation and Processing Technology, National Engineering Technology Research Center for Preservation of Agriculture Product, Tianjin Academy of Agricultural Sciences, Tianjin 300384, ChinaSchool of Life Science, Tianjin University, Tianjin 300072, ChinaSchool of Life Science, Tianjin University, Tianjin 300072, ChinaInstitute of Agricultural Products Preservation and Processing Technology, National Engineering Technology Research Center for Preservation of Agriculture Product, Tianjin Academy of Agricultural Sciences, Tianjin 300384, ChinaInstitute of Agricultural Products Preservation and Processing Technology, National Engineering Technology Research Center for Preservation of Agriculture Product, Tianjin Academy of Agricultural Sciences, Tianjin 300384, ChinaInstitute of Agricultural Products Preservation and Processing Technology, National Engineering Technology Research Center for Preservation of Agriculture Product, Tianjin Academy of Agricultural Sciences, Tianjin 300384, ChinaInstitute of Agricultural Products Preservation and Processing Technology, National Engineering Technology Research Center for Preservation of Agriculture Product, Tianjin Academy of Agricultural Sciences, Tianjin 300384, ChinaInstitute of Agricultural Products Preservation and Processing Technology, National Engineering Technology Research Center for Preservation of Agriculture Product, Tianjin Academy of Agricultural Sciences, Tianjin 300384, ChinaInstitute of Agricultural Products Preservation and Processing Technology, National Engineering Technology Research Center for Preservation of Agriculture Product, Tianjin Academy of Agricultural Sciences, Tianjin 300384, ChinaCollege of Food Science and Biological Engineering, Tianjin Agricultural University, Tianjin 300392, ChinaCollege of Life Science, Nankai University, Tianjin 300071, ChinaCollege of Life Science, Nankai University, Tianjin 300071, ChinaPhenylalanine ammonia-lyase (PAL, EC 4.3.1.5) catalyzes the deamination of phenylalanine, which is the initial step in the biosynthesis of phenylpropanoids. It serves as a crucial enzyme that facilitates the transfer of carbon from primary to secondary metabolism in plants. Duckweed is regarded as a promising chassis plant in synthetic biology research and application, due to its being rich in secondary metabolites and other advantages. The genes encoding PAL in <i>Spirodela polyrhiza</i> (L.) Schleid, the giant duckweed, were investigated in this study. Three <i>SpPAL</i> genes (<i>SpPAL1–SpPAL3</i>) were identified and cloned. All of them were successfully expressed in <i>E. coli</i>, and their recombinant proteins all showed PAL activity. In addition, SpPAL1 and SpPAL2 proteins could also utilize tyrosine as substrate, although the activity was low. A qRT-PCR analysis demonstrated that the expression of <i>SpPAL3</i> was most pronounced in young fronds. It was found that the expression of <i>SpPAL1</i> and <i>SpPAL3</i> was significantly induced by MeJA treatment. Overexpression of <i>SpPAL3</i> in <i>Lemna turionifera</i> inhibited the growth of fronds and adventitious roots in the transgenic plants, indicating the importance of <i>SpPAL3</i> in duckweed besides its involvement in the secondary metabolism.https://www.mdpi.com/2223-7747/13/18/2607<i>Spirodela polyrhiza</i>phenylalanine ammonia-lyasecloningcharacterizationoverexpression |
| spellingShingle | Xiaoxue Li Yinxing Zhang Chunfeng Zhu Pufan Zheng Cunkun Chen Na Zhang Haipeng Ji Chenghu Dong Jinze Yu Jie Ren Yerong Zhu Yong Wang Enzymatic Characterization of <i>SpPAL</i> Genes in <i>S. polyrhiza</i> and Overexpression of the <i>SpPAL3</i> Plants <i>Spirodela polyrhiza</i> phenylalanine ammonia-lyase cloning characterization overexpression |
| title | Enzymatic Characterization of <i>SpPAL</i> Genes in <i>S. polyrhiza</i> and Overexpression of the <i>SpPAL3</i> |
| title_full | Enzymatic Characterization of <i>SpPAL</i> Genes in <i>S. polyrhiza</i> and Overexpression of the <i>SpPAL3</i> |
| title_fullStr | Enzymatic Characterization of <i>SpPAL</i> Genes in <i>S. polyrhiza</i> and Overexpression of the <i>SpPAL3</i> |
| title_full_unstemmed | Enzymatic Characterization of <i>SpPAL</i> Genes in <i>S. polyrhiza</i> and Overexpression of the <i>SpPAL3</i> |
| title_short | Enzymatic Characterization of <i>SpPAL</i> Genes in <i>S. polyrhiza</i> and Overexpression of the <i>SpPAL3</i> |
| title_sort | enzymatic characterization of i sppal i genes in i s polyrhiza i and overexpression of the i sppal3 i |
| topic | <i>Spirodela polyrhiza</i> phenylalanine ammonia-lyase cloning characterization overexpression |
| url | https://www.mdpi.com/2223-7747/13/18/2607 |
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