Skip to main content
Log in

Characterization of a multifunctional methyltransferase from the orchid Vanilla planifolia

  • Physiology and Biochemistry
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

The final enzymatic step in the synthesis of the flavor compound vanillin (4-hydroxy-3-methoxybenzaldehyde) is believed to be methylation of 3,4-dihydroxybenzaldehyde. We have isolated and functionally characterized a cDNA that encodes a multifunctional methyltransferase from Vanilla planifolia tissue cultures that can catalyze the conversion of 3,4-dihydroxybenzaldehyde to vanillin, although 3,4-dihydroxybenzaldehyde is not the preferred substrate. The higher catalytic efficiency of the purified recombinant enzyme with the substrates caffeoyl aldehyde and 5-OH-coniferaldehyde, and its tissue distribution, suggest this methyltransferase may primarily function in lignin biosynthesis. However, since the enzyme characterized here does have 3,4-dihydroxybenzaldehyde-O-methyltransferase activity, it may be useful in engineering strategies for the synthesis of natural vanillin from alternate sources.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

COMT :

Caffeic acid O-methyltransferase

DOMT :

3,4-Dihydroxybenzaldehyde-O-methyltransferase

OMTs :

O-Methyltransferases

SAM :

S-adenosyl-l-methionine

References

  • Angiosperm Phylogeny Group (1998) An ordinal classification for the families of flowering plants. Ann Mo Bot Gard 85:531–553

    Google Scholar 

  • Attieh JM, Hanson AD, Saini HS (1995) Characterization of a novel methyltransferase responsible for biosynthesis of halomethanes and methanethiol in Brassica oleracea. J Biol Chem 270:9250–9257

    Article  CAS  PubMed  Google Scholar 

  • Bout S, Vermerris W (2003) A candidate-gene approach to clone the sorghum brown midrib gene encoding caffeic acid O-methltransferase. Mol Genet Genomics 269:205–214

    CAS  PubMed  Google Scholar 

  • Bugos RC, Chiang VL, Campbell WH (1991) cDNA cloning, sequence analysis and seasonal expression of lignin-bispecific caffeic acid/5-hydroxyferulic acid O-methyltransferase of aspen. Plant Mol Biol 17:1203–1215

    CAS  PubMed  Google Scholar 

  • Collazo P, Montoliu L, Puigdomenech P, Rigau J (1992) Structure and expression of the lignin O-methyltransferase gene from Zea mays L. Plant Mol Biol 20:857–867

    CAS  PubMed  Google Scholar 

  • Dignum MJW, Kerler J, Verpoorte R (2001) Vanilla production: technological, chemical, and biosynthetic aspects. Food Rev Int 17:199–219

    Article  CAS  Google Scholar 

  • Dixon RA, Chen F, Guo D, Parvathi K (2001) The biosynthesis of monolignols: a “metabolic grid”, or independent pathways to guaiacyl and syringyl units? Phytochemistry 57:1069–1084

    Article  CAS  PubMed  Google Scholar 

  • Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS (1991) “Touchdown” PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res 19:4008

    CAS  PubMed  Google Scholar 

  • Frick S, Kutchan TM (1999) Molecular cloning and functional expression of O-methyltransferases common to isoquinoline alkaloid and phenylpropanoid biosynthesis. Plant J 17:329–339

    Article  CAS  PubMed  Google Scholar 

  • Gang DR, Lavid N, Zubieta C, Chen F, Beuerle T, Lewinsohn E, Noel JP, Pichersky E (2002) Characterization of phenylpropene O-methyltransferases from sweet basil: facile change of substrate specificity and convergent evolution within a plant O-methyltransferase family. Plant Cell 14:505–519

    Article  CAS  PubMed  Google Scholar 

  • Garcia-Mas J, Messeguer R, Arus P, Puigdomenech P (1995) The caffeic acid O-methyltransferase from Prunus amygdalus (GenBank X83217). Plant Physiol 108:1341

    PubMed  Google Scholar 

  • Gowri G, Bugos RC, Campbell WH, Maxwell CA, Dixon RA (1991) Stress responses in alfalfa (Medicago sativa L.) X. Molecular cloning and expression of S-adenosyl-l-methionine:caffeic acid 3-O-methyltransferase, a key enzyme of lignin biosynthesis. Plant Physiol 97:7–14

    CAS  Google Scholar 

  • Guo D, Chen F, Inoue K, Blount JW, Dixon RA (2001) Downregulation of caffeic acid 3-O-methyltransferase and caffeoyl CoA3-O-methyltransferase in transgenic alfalfa: impacts on lignin structure and implications for the biosynthesis of G and S lignin. Plant Cell 13:73–88

    CAS  PubMed  Google Scholar 

  • Havkin-Frenkel D, Podstolski A, Knorr D (1996) Effect of light on vanillin precursors formation by in vitro cultures of Vanilla planifolia. Plant Cell Tissue Organ Cult 46:169–170

    Google Scholar 

  • Havkin-Frenkel D, Podstolski A, Witkowska E, Molecki P, Mlkolajczyk M (1999) Vanillin biosynthetic pathways: an overview. In: Fu TJ, Singh G, Curtis WR (eds) Plant cell and tissue culture for the production of food ingredients. Kluwer/Plenum, Dordrecht/New York, pp 35–43

  • Havkin-Frenkel D, French JC, Graft NM, Joel DM, Pak FE, Frenkel C (2003) Interrelation of curing and botany in vanilla (Vanilla planifolia) bean. Acta Hortic 629:93–102

    Google Scholar 

  • Herz LE (2000) Dynamics and regulatory control of the vanillin biosynthetic pathway in V. planifolia embryo cultures. PhD thesis, Rutgers University, N.J.

  • Ibrahim RK, Bruneau A, Bantignies B (1998) Plant O-methyltransferases: molecular analysis, common signature and classification. Plant Mol Biol 36:1–10

    Article  CAS  PubMed  Google Scholar 

  • Jaeck E, Martz F, Stiefel V, Fritig B, Legrand M (1996) Expression of class I O-methyltransferase in healthy and TMV-infected tobacco. Mol Plant Microbe Interact 9:681–688

    CAS  PubMed  Google Scholar 

  • Jang CS, Kim JY, Haam JW, Lee MS, Kim DS, Li YW, Seo YW (2003) Expressed sequence tags from a wheat-rye translocation line (2BS/2RL) infested by larvae of Hessian fly [Mayetiola destructor (Say)]. Plant Cell Rep 22:150–158

    Article  CAS  PubMed  Google Scholar 

  • Joel DM, French JC, Graft N, Kourteva G, Dixon RA, Havkin-Frenkel D (2003) A hairy tissue produces vanillin. Isr J Plant Sci 51:157–159

    CAS  Google Scholar 

  • Knorr D, Caster C, Dorneburg H, Dorn R, Graf S, Havkin-Frenkel D, Podstolski A, Werrman T (1993) Biosynthesis and yield improvement of food ingredients from plant cell and tissue cultures. Food Technol 47:57–63

    CAS  Google Scholar 

  • Kuhl JC, Cheung F, Yuan Q, Martin W, Zewdie Y, McCallum J, Catanach A, Rutherford P, Sink KC, Jenderek M, Prince J, Town CD, Havey MJ (2004) A unique set of 11,008 onion expressed sequence tags reveals expressed sequence and genomic differences between the monocot orders Asparagales and Poales. Plant Cell 16:114–125

    Article  PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    PubMed  Google Scholar 

  • Li L, Popko JL, Umezawa T, Chiang VL (2000) 5-Hydroxyconiferyl aldehyde modulates enzymatic methylation for syringyl monolignol formation, a new view of monolignol biosynthesis in angiosperms. J Biol Chem 275:6537–6545

    CAS  PubMed  Google Scholar 

  • Maury S, Geoffroy P, Legrand M (1999) Tobacco O-methyltransferases involved in phenylpropanoid metabolism. The different caffeoyl-coenzyme A/5-hydroxyferuloyl-coenzyme A 3/5-O-methltransferse and caffeic acid/5-hydroxyferulic acid 3/5-O-methyltransferase classes have distinct substrate specificities and expression patterns. Plant Physiol 121:215–223

    PubMed  Google Scholar 

  • McAlister FM, Jenkins CLD, Watson JM (1998) Sequence and expression of a stem-abundant caffeic acid O-methyltransferase cDNA from perennial ryegrass (Lolium perenne). Aust J Plant Physiol 25:225–235

    CAS  Google Scholar 

  • Osakabe K, Tsao CC, Li L, Popko JL, Umezawa T, Carraway DT (1999) Coniferyl aldehyde 5-hydroxylation and methylation direct syringyl lignin biosynthesis in angiosperms. Proc Natl Acad Sci USA 96:8955–8960

    CAS  PubMed  Google Scholar 

  • Parvathi K, Chen F, Guo D, Blount JW, Dixon RA (2001) Substrate preferences of O-methyltransferases in alfalfa suggest new pathways for 3-O-methylation of monolignols. Plant J 25:193–202

    CAS  PubMed  Google Scholar 

  • Podstolski A, Havkin-Frenkel D, Malinowski J, Blount JW, Kourteva G, Dixon RA (2002) Unusual 4-hydroxybenzaldehyde synthase activity from tissue cultures of the vanilla orchid Vanilla planifolia. Phytochemistry 61:611–620

    Article  CAS  PubMed  Google Scholar 

  • Poeydomenge O, Boudet AM, Grima-Pettenati J (1994) A cDNA encoding S-adenosyl-l-methionine:caffeic acid 3-O-methyltransferase from Eucalyptus. Plant Physiol 105:749–750

    Article  CAS  PubMed  Google Scholar 

  • Rao SR, Ravishankar GA (2000) Vanilla flavour: production by conventional and biotechnological routes. J Sci Food Agric 80:289–304

    Article  Google Scholar 

  • Schroder G, Wehinger E, Schroder J (2002) Predicting the substrates of cloned plant O-methyltransferases. Phytochemistry 59:1–8

    Article  CAS  PubMed  Google Scholar 

  • Schwab W (2003) Metabolome diversity: too few genes, too many metabolites? Phytochemistry 62:837–849

    Article  CAS  PubMed  Google Scholar 

  • Selman-Housein G, Lopez M, Hernandez D, Civardi L, Miranda F, Rigau J, Puigdomenech P (1999) Molecular cloning of cDNAs coding for three sugarcane enzymes involved in lignification. Plant Sci 143:163–171

    Article  CAS  Google Scholar 

  • Short U, Fernandez JM, Sorge JA, Huse WD (1988) λZAP: a bacteriophage λ expression vector with in vivo excision properties. Nucleic Acids Res 16:7583–7599

    CAS  PubMed  Google Scholar 

  • Walton NJ, Mayer MJ, Narbad A (2003) Vanillin. Phytochemistry 63:505–515

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Pichersky E (1997) Nucleotide sequence of S-adenosyl-l-methionine:caffeic acid 3-O-methyltransferase from Clarkia breweri (accession no. AF006009). Plant Physiol 114:1567

    Article  PubMed  Google Scholar 

  • Wang J, Pichersky E (1998) Characterization of S-adenosyl-l-methionine:(iso)eugenol O-methyltransferase involved in floral scent production in Clarkia breweri. Arch Biochem Biophys 349:153–160

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Dudareva N, Bhakta S, Raguso RA, Pichersky E (1997) Floral scent production in Clarkia breweri (Onagraceae) II. Localization and developmental modulation of the enzyme S-adenosyl-l-methionine:(iso)eugenol O-methyltransferase and phenylpropanoid emission. Plant Physiol 114:213–221

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Dudareva N, Kish CM, Simon JE, Lewinsohn E, Pichersky E (1999) Nucleotide sequences of two cDNAs encoding caffeic acid O-methyltransferases (accession nos. AF154917 and AF154918) from sweet basil (Ocimum basilicum). Plant Physiol 120:1205

    Article  PubMed  Google Scholar 

  • Wein M, Lavid N, Lunkenbein S, Lewinsohn E, Schwab W, Kaldenhoff R (2002) Isolation, cloning and expression of a multifunctional O-methyltransferase capable of forming 2,5-dimethyl-4-methoxy-3(2H)-furanone, one of the key aroma compounds in strawberry fruits. Plant J 32:755–765

    Article  Google Scholar 

  • Xue Z-T, Brodelius PE (1998) Kinetin-induced caffeic acid O-methyltransferases in cell suspension cultures of Vanilla planifolia Andr. and isolation of caffeic acid O-methyltransferase cDNAs. Plant Physiol Biochem 36:779–788

    Article  CAS  Google Scholar 

  • Zubieta C, Kota P, Ferrer J-L, Dixon RA, Noel JP (2002) Structural basis for the modulation of lignin monomer methylation by caffeic acid/5-hydroxyferulic acid 3/5-O-methyltransferase. Plant Cell 14:1265–1277

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr. Richard Dixon, Noble Foundation, for the sequencing and Dr. Eran Pichersky, University of Michigan, for advice on the affinity chromatography. This work was supported by a grant from David Michael & Company.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to F. C. Belanger.

Additional information

Communicated by H. Barz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pak, F.E., Gropper, S., Dai, W.D. et al. Characterization of a multifunctional methyltransferase from the orchid Vanilla planifolia . Plant Cell Rep 22, 959–966 (2004). https://doi.org/10.1007/s00299-004-0795-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-004-0795-x

Keywords

Navigation