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4-Coumarate:coenzyme A ligase in black locust (Robinia pseudoacacia) catalyses the conversion of sinapate to sinapoyl-CoA

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Abstract

4-Coumarate:coenzyme A (CoA) ligase (4CL, EC 6.2.1.12) in crude enzyme preparation from the developing xylem of black locust (Robinia pseudoacacia) converted sinapate to sinapoyl CoA. The sinapate-converting activity was not inhibited by other cinnamate derivatives, such as p-coumarate, caffeate or ferulate, in the mixed-substrate assay. The crude extract prepared from the developing xylem was separated by anion-exchange chromatography into three different 4CL isoforms. The isoform 4CL1 had a strong substrate preference for p-coumarate, but lacked the activity for ferulate and sinapate. On the other hand, 4CL2 and 4CL3 displayed activity toward sinapate and also possessed high activity toward caffeate as well as p-coumarate. The crude extract from the shoots exhibited a very similar substrate preference to that of the developing xylem; therefore, 4CL2 may be a major isoform in both crude enzyme preparations. These results support the hypothesis that sinapate-converting 4CL isoform is constitutively expressed in lignin-forming cells.

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Acknowledgments

The authors are very grateful to Professor C.J. Douglas of the University of British Columbia, Canada, for the kind gift of 4CL antiserum.

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Authors and Affiliations

  1. Department of Forest Resources Science, Faculty of Agriculture, Shizuoka University, Shizuoka, Japan

    Katsuyoshi Hamada & Tomoaki Nishida

  2. Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan

    Kazuchika Yamauchi & Kazuhiko Fukushima

  3. Department of Forest and Forest Product Sciences, Faculty of Agriculture, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka 812-8581, Japan

    Ryuichiro Kondo & Yuji Tsutsumi

Authors
  1. Katsuyoshi Hamada
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  2. Tomoaki Nishida
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  3. Kazuchika Yamauchi
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  4. Kazuhiko Fukushima
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  5. Ryuichiro Kondo
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  6. Yuji Tsutsumi
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Correspondence to Yuji Tsutsumi.

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Hamada, K., Nishida, T., Yamauchi, K. et al. 4-Coumarate:coenzyme A ligase in black locust (Robinia pseudoacacia) catalyses the conversion of sinapate to sinapoyl-CoA. J Plant Res 117, 303–310 (2004). https://doi.org/10.1007/s10265-004-0159-1

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  • DOI: https://doi.org/10.1007/s10265-004-0159-1

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