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Characterization of basic p-coumaryl and coniferyl alcohol oxidizing peroxidases from a lignin-forming Picea abies suspension culture

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Abstract

A Norway spruce (Picea abies) tissue culture line that produces extracellular lignin into the culture medium has been used as a model system to study the enzymes involved in lignin polymerization. We report here the purification of two highly basic culture medium peroxidases, PAPX4 and PAPX5, and isolation of the corresponding cDNAs. Both isoforms had high affinity to monolignols with apparent K m values in µM range. PAPX4 favoured coniferyl alcohol with a six-fold higher catalytic efficiency (V max/K m) and PAPX5 p-coumaryl alcohol with a two-fold higher catalytic efficiency as compared to the other monolignol. Thus coniferyl and p-coumaryl alcohol could be preferentially oxidized by different peroxidase isoforms in this suspension culture, which may reflect a control mechanism for the incorporation of different monolignols into the cell wall. Dehydrogenation polymers produced by the isoforms were structurally similar. All differed from the released suspension culture lignin and milled wood lignin, in accordance with previous observations on the major effects that e.g. cell wall context, rate of monolignol feeding and other proteins have on polymerisation. Amino acid residues shown to be involved in monolignol binding in the lignification-related Arabidopsis ATPA2 peroxidase were nearly identical in PAPX4 and PAPX5. This similarity extended to other peroxidases involved in lignification, suggesting that a preferential structural organization of the substrate access channel for monolignol oxidation might exist in both angiosperms and gymnosperms.

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Abbreviations

CA:

coniferyl alcohol

DHP:

dehydrogenation polymer

FA:

ferulic acid

HRP:

horseradish peroxidase

IEF:

isoelectric focusing

MWL:

milled wood lignin

pCA:

p-coumaryl alcohol

RSCL:

released suspension culture lignin

SA:

sinapyl alcohol

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Authors and Affiliations

  1. Department of Applied Biology, University of Helsinki, P.O. Box 27, 00014UH, Finland

    Sanna Koutaniemi & Teemu H. Teeri

  2. Institute of Biotechnology, University of Helsinki, P.O. Box 65, 00014UH, Finland

    Merja M. Toikka & Ilkka A. Kilpeläinen

  3. Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 56, 00014UH, Finland

    Anna Kärkönen & Maaret Mustonen

  4. Department of Applied Chemistry and Microbiology, University of Helsinki, P.O. Box 56, 00014UH, Finland

    Taina Lundell & Liisa K. Simola

  5. Department of Chemistry, University of Helsinki, P.O. Box 55, 00014UH, Finland

    Ilkka A. Kilpeläinen

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  1. Sanna Koutaniemi
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Koutaniemi, S., Toikka, M.M., Kärkönen, A. et al. Characterization of basic p-coumaryl and coniferyl alcohol oxidizing peroxidases from a lignin-forming Picea abies suspension culture. Plant Mol Biol 58, 141–157 (2005). https://doi.org/10.1007/s11103-005-5345-6

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