Identification and partial characterisation of a coniferyl alcohol oxidase from lignifying xylem of Sitka spruce (Picea sitchensis)

A. Richardson, D. Stewart, G.J. McDougall

    Research output: Contribution to journalArticle

    Abstract

    Oxidase activity in the developing xylem of branches of Sitka spruce [Picea sitchensis] (Bong) Carr. was expressed in synchrony with the deposition of lignin. The activity was closely associated with the cell wall but it could be extracted by elution with salt solutions such as 1?M?NaCl or CaCl2. A number of different oxidase isoforms with isoelectric points in the range 8–5 were present in these cell wall extracts. These enzymes displayed a marked preference for the oxidation of coniferyl alcohol and efficiently initiated polymerization of coniferyl alcohol into insoluble, lignin-like polymers. They also had a substrate preference and profile of sensitivity to inhibitors that was dissimilar to those reported for classical catechol oxidase or laccase-type polyphenol oxidases. A novel procedure that combines extraction and affinity chromatography on Concanavalin-A to select high-mannose-type glycoproteins provided oxidase activity at higher purity and yield than previously used methods. A single band of oxidase activity (apparent Mr approx. 84 kDa) which was capable of oxidizing a-naphthol/N,N,N'N'-tetramethyl p-phenylene diamine in the absence of added hydrogen peroxide was detected in these cell wall extracts using non-denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The addition of hydrogen peroxide did not intensify the staining of this band but it confirmed the presence of a true peroxidase band of apparent Mr approx. 40 kDa. The properties of this coniferyl alcohol oxidase are different from those of laccase-type polyphenol oxidases (EC 1.10.3.2) previously implicated in lignin deposition in tree species, and their possible roles in this process are discussed.
    © Springer-Verlag
    Original languageEnglish
    Pages (from-to)35-43
    Number of pages9
    JournalPlanta
    Volume203
    Issue number1
    DOIs
    Publication statusPublished - 1997

    Fingerprint

    alcohol oxidase
    coniferyl alcohol
    laccase
    Picea sitchensis
    catechol oxidase
    xylem
    lignin
    cell walls
    hydrogen peroxide
    naphthols
    diamines
    extracts
    affinity chromatography
    concanavalin A
    isoelectric point
    mannose
    polymerization
    purity
    polyacrylamide gel electrophoresis
    glycoproteins

    Keywords

    • cell wall
    • coniferyl alcohol oxidase
    • lignification
    • peroxidase
    • picea (lignification)
    • xylem

    Cite this

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    abstract = "Oxidase activity in the developing xylem of branches of Sitka spruce [Picea sitchensis] (Bong) Carr. was expressed in synchrony with the deposition of lignin. The activity was closely associated with the cell wall but it could be extracted by elution with salt solutions such as 1?M?NaCl or CaCl2. A number of different oxidase isoforms with isoelectric points in the range 8–5 were present in these cell wall extracts. These enzymes displayed a marked preference for the oxidation of coniferyl alcohol and efficiently initiated polymerization of coniferyl alcohol into insoluble, lignin-like polymers. They also had a substrate preference and profile of sensitivity to inhibitors that was dissimilar to those reported for classical catechol oxidase or laccase-type polyphenol oxidases. A novel procedure that combines extraction and affinity chromatography on Concanavalin-A to select high-mannose-type glycoproteins provided oxidase activity at higher purity and yield than previously used methods. A single band of oxidase activity (apparent Mr approx. 84 kDa) which was capable of oxidizing a-naphthol/N,N,N'N'-tetramethyl p-phenylene diamine in the absence of added hydrogen peroxide was detected in these cell wall extracts using non-denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The addition of hydrogen peroxide did not intensify the staining of this band but it confirmed the presence of a true peroxidase band of apparent Mr approx. 40 kDa. The properties of this coniferyl alcohol oxidase are different from those of laccase-type polyphenol oxidases (EC 1.10.3.2) previously implicated in lignin deposition in tree species, and their possible roles in this process are discussed.{\circledC} Springer-Verlag",
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    Identification and partial characterisation of a coniferyl alcohol oxidase from lignifying xylem of Sitka spruce (Picea sitchensis). / Richardson, A.; Stewart, D.; McDougall, G.J.

    In: Planta, Vol. 203, No. 1, 1997, p. 35-43.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Identification and partial characterisation of a coniferyl alcohol oxidase from lignifying xylem of Sitka spruce (Picea sitchensis)

    AU - Richardson, A.

    AU - Stewart, D.

    AU - McDougall, G.J.

    PY - 1997

    Y1 - 1997

    N2 - Oxidase activity in the developing xylem of branches of Sitka spruce [Picea sitchensis] (Bong) Carr. was expressed in synchrony with the deposition of lignin. The activity was closely associated with the cell wall but it could be extracted by elution with salt solutions such as 1?M?NaCl or CaCl2. A number of different oxidase isoforms with isoelectric points in the range 8–5 were present in these cell wall extracts. These enzymes displayed a marked preference for the oxidation of coniferyl alcohol and efficiently initiated polymerization of coniferyl alcohol into insoluble, lignin-like polymers. They also had a substrate preference and profile of sensitivity to inhibitors that was dissimilar to those reported for classical catechol oxidase or laccase-type polyphenol oxidases. A novel procedure that combines extraction and affinity chromatography on Concanavalin-A to select high-mannose-type glycoproteins provided oxidase activity at higher purity and yield than previously used methods. A single band of oxidase activity (apparent Mr approx. 84 kDa) which was capable of oxidizing a-naphthol/N,N,N'N'-tetramethyl p-phenylene diamine in the absence of added hydrogen peroxide was detected in these cell wall extracts using non-denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The addition of hydrogen peroxide did not intensify the staining of this band but it confirmed the presence of a true peroxidase band of apparent Mr approx. 40 kDa. The properties of this coniferyl alcohol oxidase are different from those of laccase-type polyphenol oxidases (EC 1.10.3.2) previously implicated in lignin deposition in tree species, and their possible roles in this process are discussed.© Springer-Verlag

    AB - Oxidase activity in the developing xylem of branches of Sitka spruce [Picea sitchensis] (Bong) Carr. was expressed in synchrony with the deposition of lignin. The activity was closely associated with the cell wall but it could be extracted by elution with salt solutions such as 1?M?NaCl or CaCl2. A number of different oxidase isoforms with isoelectric points in the range 8–5 were present in these cell wall extracts. These enzymes displayed a marked preference for the oxidation of coniferyl alcohol and efficiently initiated polymerization of coniferyl alcohol into insoluble, lignin-like polymers. They also had a substrate preference and profile of sensitivity to inhibitors that was dissimilar to those reported for classical catechol oxidase or laccase-type polyphenol oxidases. A novel procedure that combines extraction and affinity chromatography on Concanavalin-A to select high-mannose-type glycoproteins provided oxidase activity at higher purity and yield than previously used methods. A single band of oxidase activity (apparent Mr approx. 84 kDa) which was capable of oxidizing a-naphthol/N,N,N'N'-tetramethyl p-phenylene diamine in the absence of added hydrogen peroxide was detected in these cell wall extracts using non-denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The addition of hydrogen peroxide did not intensify the staining of this band but it confirmed the presence of a true peroxidase band of apparent Mr approx. 40 kDa. The properties of this coniferyl alcohol oxidase are different from those of laccase-type polyphenol oxidases (EC 1.10.3.2) previously implicated in lignin deposition in tree species, and their possible roles in this process are discussed.© Springer-Verlag

    KW - cell wall

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    KW - lignification

    KW - peroxidase

    KW - picea (lignification)

    KW - xylem

    U2 - 10.1007/s00050162

    DO - 10.1007/s00050162

    M3 - Article

    VL - 203

    SP - 35

    EP - 43

    JO - Planta

    JF - Planta

    SN - 0032-0935

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    ER -