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. 2022 Apr 28;13:840397. doi: 10.3389/fpls.2022.840397

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Copyright © 2022 Li, Zhang, Zhao, Zhao, Xie, Guo, Wang, Li, Guo, Zhu, Zhang, Jia and Miao.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of the cinnamyl alcohol dehydrogenase (CAD) catalytic activities of GhCAD35, GhCAD45, and GhCAD43. (A–C) HPLC analyses of in vitro incubation of GhCAD35, GhCAD45, and GhCAD43 with coniferaldehyde, sinapaldehyde and p-coumaraldehyde, to yield corresponding monolignols, coniferyl alcohol, sinapyl alcohol, and p-coumaryl alcohol, respectively. C1-C6 indicate coniferyl alcohol, coniferaldehyde, sinapyl alcohol, sinapaldehyde, p-coumaryl alcohol, and p-coumaraldehyde, respectively. Controls in A-C were incubated with GST protein. (D) Western blot showing the levels of recombinant GhCAD35, GhCAD45, or GhCAD43 protein fused with a GST tag. GST antibody was used. (E) The catalytic parameters of GhCAD35, GhCAD45, and GhCAD43 toward the substrates p-coumaraldehyde, coniferaldehyde, and sinapaldehyde, respectively.