Isolation and functional characterization of a Lonicera japonica hydroxycinnamoyl transferase involved in chlorogenic acid synthesis

Gaobin Pu; Bingqian Zhou; Fengning Xiang

doi:10.1515/biolog-2017-0069

Article

Isolation and functional characterization of a Lonicera japonica hydroxycinnamoyl transferase involved in chlorogenic acid synthesis

Gaobin Pu , Bingqian Zhou and Fengning Xiang

Published/Copyright: June 30, 2017

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From the journal Biologia Volume 72 Issue 6

Abstract

The flower buds of Lonicera japonica are a rich source of antioxidants, comprising mostly flavonoids and phenolic acids, and in particular chlorogenic acid (5-caffeoylquinic acid, CGA), dicaffeoylquinic acid and caffeic acid. Hydroxycinnamoyl-CoA transferases are important for the synthesis of CGA. Here, the isolation and characterization of a Lonicera japonica gene encoding a hydroxycinnamoyl-CoA quinate transferase (HQT) is described. The recombinant LjHQT, produced by heterologously expressing the gene in Escherichia coli, favored p-coumaroyl-CoA over caffeoyl-CoA as its acyl donor, and expressed a higher affinity for quinate over shikimate as its its acyl acceptor. Southern blot analysis showed that the L. japonica genome harbors at least two copies of LjHQT. LjHQT transcript abundance was promoted both by spraying the leaves with salicylic acid or abscisic acid, and exposing them to a period of low temperature. The constitutive expression of LjHQT in Arabidopsis thaliana resulted in the production of CGA in the leaf and an higher level of resistance to the bacterial pathogen Pseudomonas syringae. CGA synthesis was positively correlated with LjHQT transcript abundance. When a fusion between the LjHQT promoter and GUS was expressed in A. thaliana, GUS expression was observed mainly in the sepal, vascular bundle, stigma and axillary bud.

Key words: Lonicera japonica; hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase; chlorogenic acid; enzyme assay; gene expression

Acknowledgements

This research was supported by the Shandong Province Key Research Program (grant no. 2016GSF202008), Shandong Province Science and Technology Program of Higher Education (grant no. J15LM55), the National Natural Science Foundation of China (grant nos. 30970243, 31200226), International technology cooperation project of Shandong Province of China (grant no. 2011176) and the Chinese Natural Education Ministry Doctor Station Foundation Fellowship (grant no. 913111006).

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Received: 2016-12-9

Accepted: 2017-2-27

Published Online: 2017-6-30

Published in Print: 2017-6-27

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Articles in the same Issue

https://doi.org/10.1515/biolog-2017-0069

Keywords for this article

Lonicera japonica; hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase; chlorogenic acid; enzyme assay; gene expression