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

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

Gaobin Pu , Bingqian Zhou und Fengning Xiang

Veröffentlicht/Copyright: 30. Juni 2017

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Aus der Zeitschrift Biologia Band 72 Heft 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).

References

Choi C.W., Jung H.A., Kang S.S. & Choi J.S. 2007. Antioxidant constituents and a new triterpenoid glycoside from flos Lonicerae. Arch. Pharm. Res. 30: 1–7.10.1007/BF02977770Suche in Google Scholar

Clifford M.N. 2000. Chlorogenic acids and other cinnamates-nature, occurrence, dietary burden, absorption and metabolism. J. Sci. Food Agr. 80: 1033–1043.10.1002/(SICI)1097-0010(20000515)80:7<1033::AID-JSFA595>3.0.CO;2-TSuche in Google Scholar

Franke R., Hemm M.R., Denault J.W., Ruegger M.O., Humphreys J.M. & Chapple C. 2002. Changes in secondary metabolism and deposition of an unusual lignin in the ref8 mutant of Arabidopsis. Plant J. 30: 47–59.10.1046/j.1365-313X.2002.01267.xSuche in Google Scholar

He L., Xu X., Li Y., Li C., Zhu Y., Yan H., Sun Z., Sun C., Song J., Bi Y., Shen J., Cheng R., Wang Z., Xiao W. & Chen S. 2013. Transcriptome analysis of buds and leaves using 454 pyrosequencing to discover genes associated with the biosynthesis of active ingredients in Lonicera japonica Thunb. PLoS One. 8: e62922.10.1371/journal.pone.0062922Suche in Google Scholar

Hoffmann L., Maury S., Martz F., Geoffroy P. & Legrand M. 2003. Purification, cloning, and properties of an acyltransferase controlling shikimate and quinate ester intermediates in phenylpropanoid metabolism. J. Biol. Chem. 278: 95–103.10.1074/jbc.M209362200Suche in Google Scholar

Jefferson R.A., Kavanagh T.A. & Bevan M.W. 1987. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907.10.1002/j.1460-2075.1987.tb02730.xSuche in Google Scholar

Ku S.K., Seo B.I., Park J.H., Park G.Y., Seo Y.B., Kim J.S., Lee H.S. & Roh S.S. 2009. Effect of Lonicerae Flos extracts on reflux esophagitis with antioxidant activity. World J. Gastroenterol. 15: 4799–4805.10.3748/wjg.15.4799Suche in Google Scholar

Kumar N., Singh B., Bhandari P., Gupta A.P., Uniyal S.K. & Kaul V.K. 2005. Biflavonoids from Lonicera japonica. Phytochem. 66: 2740–2744.10.1016/j.phytochem.2005.10.002Suche in Google Scholar

Kwak W.J., Han C.K., Chang H.W., Kim H.P., Kang S.S. & Son K.H. 2003. Loniceroside C, an antiinflammatory saponin from Lonicera japonica. Chem. Pharm. Bull. 51: 333–335.10.1248/cpb.51.333Suche in Google Scholar

Lepelley M., Cheminade G., Tremillon N., Simkin A., Cailet V., et al. 2007. Chlorogenic acid synthesis in coffee: an analysis of CGA content and real-time RT-PCR expression of HCT, HQT, C3H1, and CCoAOMTl genes during grain development in C. canephora. Plant Sci. 172: 978–996.10.1016/j.plantsci.2007.02.004Suche in Google Scholar

Li H., Zhang Z. & Li P. 2002. Comparative study on volatile oils in flower and stem of Lonicera japonica. Zhong Yao Cai 25: 476–477.Suche in Google Scholar

Lu H.T., Jiang Y. & Chen F. 2004. Application of preparative high-speed counter-current chromatography for separation of chlorogenic acid from Flos Lonicerae. J Chromatogr. A 1026: 185–190.10.1016/j.chroma.2003.11.004Suche in Google Scholar PubMed

National Pharmacopoeia Committee 2015. Chinese Pharmacopoeia. China Medical Science Press, Beijing.Suche in Google Scholar

Navarre D.A., Payyavula R.S., Shakya R. & Knowles N.R. 2013. Changes in potato phenylpropanoid metabolism during tuber development. Plant Physiol. Biochem. 65: 89–101.10.1016/j.plaphy.2013.01.007Suche in Google Scholar PubMed

Niggeweg R., Michael A.J. & Martin C. 2004. Engineering plants with increased levels of the antioxidant chlorogenic acid. Nat. Biotechnol. 22: 746–754.10.1038/nbt966Suche in Google Scholar PubMed

Payyavula R.S., Shakya R., Sengoda V.G., Munyaneza J.E., Swamy P. & Navarre D.A. 2014. Synthesis and regulation of chlorogenic acid in potato: rerouting phenylpropanoid flux in HQT-silenced lines. Plant Biotechnol. J. 13: 551–564.10.1111/pbi.12280Suche in Google Scholar PubMed

Peng X., Li W., Wang W. & Bai G. 2010. Cloning and characterization of a cDNA coding a hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase involved in chlorogenic acid biosynthesis in Lonicera japonica. Planta Med. 76: 1921–1926.10.1055/s-0030-1250020Suche in Google Scholar PubMed

Pu G., Wang P., Zhou B., Liu Z. & Xiang F. 2013. Cloning and characterization of Lonicera japonica p-coumaroyl ester 3’- hydroxylase which is involved in the biosynthesis of chlorogenic acid. Biosci. Biotechnol. Biochem. 77: 1403–1409.10.1271/bbb.130011Suche in Google Scholar PubMed

Qian Z.M., Li H.J., Li P., Chen J. & Tang D. 2007. Simultaneous quantification of seven bioactive components in Caulis Lonicerae japonicae by high performance liquid chromatography. Biomed. Chromatogr. 21: 649–654.10.1002/bmc.803Suche in Google Scholar PubMed

Sambrook J. & Russell D.W. 2006. Southern blotting: simultaneous transfer of DNA from a single agarose gel to two membranes. CSH Protoc. 10.1101/pdb.prot4043.Suche in Google Scholar PubMed

Schlotzhauer W.S., Pair S.D. & Horvat R.J. 1996. Volatile constituents from the flowers of Japanese Honeysuckle (Lonicera japonica). J. Agric. Food Chem. 44: 206–209.10.1021/jf950275bSuche in Google Scholar

Schoch G., Goepfert S., Morant M., Hehn A., Meyer D., Ullmann P. & Werck-Reichhart D. 2001. CYP98A3 from Arabidopsis thaliana is a 3’-hydroxylase of phenolic esters, a missing link in the phenylpropanoid pathway. J. Biol. Chem. 276: 36566–36574.10.1074/jbc.M104047200Suche in Google Scholar PubMed

Sonnante G., D’Amore R., Blanco E., Pierri C.L., De Palma M., Luo J., Tucci M. & Cathie Martin C. 2010. Novel hydroxycinnamoyl-coenzyme A quinate transferase genes from artichoke are involved in the synthesis of chlorogenic acid. Plant Physiol. 153: 1224–1238.10.1104/pp.109.150144Suche in Google Scholar PubMed PubMed Central

Terauchi R. & Kahl G. 2000. Rapid isolation of promoter sequences by TAIL-PCR: the 5’-flanking regions of Pal and Pgi genes from yams (Dioscorea). Mol. Gen. Genet. 263: 554–560.10.1007/s004380051201Suche in Google Scholar PubMed

Received: 2016-12-9

Accepted: 2017-2-27

Published Online: 2017-6-30

Published in Print: 2017-6-27

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

Schlagwörter für diesen Artikel

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