Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6
Bacteria-derived enzymes that can modify specific lignin substructures are potential targets to engineer plants for better biomass processability. The Gram-negative bacterium Sphingobium sp. SYK-6 possesses a Ca-dehydrogenase (LigD) enzyme that has been shown to oxidize the a-hydroxy functionalities...
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my.utm.560152017-09-12T08:51:25Z http://eprints.utm.my/id/eprint/56015/ Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6 Tsuji, Yukiko Vanholme, Ruben Tobimatsu, Yuki Ishikawa, Yasuyuki Foster, Clifton E. Kamimura, Naofumi Hishiyama, Shojiro Hashimoto, Saki Shino, Amiu Hara, Hirofumi Kanna, Sato-Izawa Oyarce, Paula Goeminne, Geert Morreel, Kris Kikuchi Morreel, Jun Takano, Toshiyuki Fukuda, Masao Katayama, Yoshihiro Boerjan, Wout John, Ralph Masai, Eiji Kajita, Shinya T Technology (General) Bacteria-derived enzymes that can modify specific lignin substructures are potential targets to engineer plants for better biomass processability. The Gram-negative bacterium Sphingobium sp. SYK-6 possesses a Ca-dehydrogenase (LigD) enzyme that has been shown to oxidize the a-hydroxy functionalities in ß-O-4-linked dimers into a-keto analogues that are more chemically labile. Here, we show that recombinant LigD can oxidize an even wider range of ß-O-4-linked dimers and oligomers, including the genuine dilignols, guaiacylglycerol-ß-coniferyl alcohol ether and syringylglycerol-ß-sinapyl alcohol ether. We explored the possibility of using LigD for biosynthetically engineering lignin by expressing the codon-optimized ligD gene in Arabidopsis thaliana. The ligD cDNA, with or without a signal peptide for apoplast targeting, has been successfully expressed, and LigD activity could be detected in the extracts of the transgenic plants. UPLC-MS/MS-based metabolite profiling indicated that levels of oxidized guaiacyl (G) ß-O-4-coupled dilignols and analogues were significantly elevated in the LigD transgenic plants regardless of the signal peptide attachment to LigD. In parallel, 2D NMR analysis revealed a 2.1- to 2.8-fold increased level of G-type a-keto-ß-O-4 linkages in cellulolytic enzyme lignins isolated from the stem cell walls of the LigD transgenic plants, indicating that the transformation was capable of altering lignin structure in the desired manner Blackwell Publishing 2015-08 Article PeerReviewed Tsuji, Yukiko and Vanholme, Ruben and Tobimatsu, Yuki and Ishikawa, Yasuyuki and Foster, Clifton E. and Kamimura, Naofumi and Hishiyama, Shojiro and Hashimoto, Saki and Shino, Amiu and Hara, Hirofumi and Kanna, Sato-Izawa and Oyarce, Paula and Goeminne, Geert and Morreel, Kris and Kikuchi Morreel, Jun and Takano, Toshiyuki and Fukuda, Masao and Katayama, Yoshihiro and Boerjan, Wout and John, Ralph and Masai, Eiji and Kajita, Shinya (2015) Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6. Plant Biotechnology Journal, 13 (6). pp. 821-832. ISSN 1467-7644 http://dx.doi.org/0.1111/pbi.12316 DOI:0.1111/pbi.12316 |
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T Technology (General) Tsuji, Yukiko Vanholme, Ruben Tobimatsu, Yuki Ishikawa, Yasuyuki Foster, Clifton E. Kamimura, Naofumi Hishiyama, Shojiro Hashimoto, Saki Shino, Amiu Hara, Hirofumi Kanna, Sato-Izawa Oyarce, Paula Goeminne, Geert Morreel, Kris Kikuchi Morreel, Jun Takano, Toshiyuki Fukuda, Masao Katayama, Yoshihiro Boerjan, Wout John, Ralph Masai, Eiji Kajita, Shinya Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6 |
| description |
Bacteria-derived enzymes that can modify specific lignin substructures are potential targets to engineer plants for better biomass processability. The Gram-negative bacterium Sphingobium sp. SYK-6 possesses a Ca-dehydrogenase (LigD) enzyme that has been shown to oxidize the a-hydroxy functionalities in ß-O-4-linked dimers into a-keto analogues that are more chemically labile. Here, we show that recombinant LigD can oxidize an even wider range of ß-O-4-linked dimers and oligomers, including the genuine dilignols, guaiacylglycerol-ß-coniferyl alcohol ether and syringylglycerol-ß-sinapyl alcohol ether. We explored the possibility of using LigD for biosynthetically engineering lignin by expressing the codon-optimized ligD gene in Arabidopsis thaliana. The ligD cDNA, with or without a signal peptide for apoplast targeting, has been successfully expressed, and LigD activity could be detected in the extracts of the transgenic plants. UPLC-MS/MS-based metabolite profiling indicated that levels of oxidized guaiacyl (G) ß-O-4-coupled dilignols and analogues were significantly elevated in the LigD transgenic plants regardless of the signal peptide attachment to LigD. In parallel, 2D NMR analysis revealed a 2.1- to 2.8-fold increased level of G-type a-keto-ß-O-4 linkages in cellulolytic enzyme lignins isolated from the stem cell walls of the LigD transgenic plants, indicating that the transformation was capable of altering lignin structure in the desired manner |
| format |
Article |
| author |
Tsuji, Yukiko Vanholme, Ruben Tobimatsu, Yuki Ishikawa, Yasuyuki Foster, Clifton E. Kamimura, Naofumi Hishiyama, Shojiro Hashimoto, Saki Shino, Amiu Hara, Hirofumi Kanna, Sato-Izawa Oyarce, Paula Goeminne, Geert Morreel, Kris Kikuchi Morreel, Jun Takano, Toshiyuki Fukuda, Masao Katayama, Yoshihiro Boerjan, Wout John, Ralph Masai, Eiji Kajita, Shinya |
| author_facet |
Tsuji, Yukiko Vanholme, Ruben Tobimatsu, Yuki Ishikawa, Yasuyuki Foster, Clifton E. Kamimura, Naofumi Hishiyama, Shojiro Hashimoto, Saki Shino, Amiu Hara, Hirofumi Kanna, Sato-Izawa Oyarce, Paula Goeminne, Geert Morreel, Kris Kikuchi Morreel, Jun Takano, Toshiyuki Fukuda, Masao Katayama, Yoshihiro Boerjan, Wout John, Ralph Masai, Eiji Kajita, Shinya |
| author_sort |
Tsuji, Yukiko |
| title |
Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6 |
| title_short |
Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6 |
| title_full |
Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6 |
| title_fullStr |
Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6 |
| title_full_unstemmed |
Introduction of chemically labile substructures into Arabidopsis lignin through the use of LigD, the C-dehydrogenase from Sphingobium sp strain SYK-6 |
| title_sort |
introduction of chemically labile substructures into arabidopsis lignin through the use of ligd, the c-dehydrogenase from sphingobium sp strain syk-6 |
| publisher |
Blackwell Publishing |
| publishDate |
2015 |
| url |
http://eprints.utm.my/id/eprint/56015/ http://dx.doi.org/0.1111/pbi.12316 |
| _version_ |
1643653969236459520 |
| score |
13.211869 |