Transgenic rice as a novel production system for Melanocarpus and Pycnoporus laccases

Chris de Wilde, Eva Uzan, Zhongyi Zhou, Kristiina Kruus, Martina Andberg, Johanna Buchert, Eric Record, Marcel Asther, Anne Lomascolo (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

21 Citations (Scopus)

Abstract

Laccases have numerous biotechnological applications, among them food processing. The widespread use of laccases has increased the demand for an inexpensive and safe source of recombinant enzyme. We explored the use of a rice-based system for the production of two fungal laccases derived from the ascomycete Melanocarpus albomyces and the basidiomycete Pycnoporus cinnabarinus. High-expression levels of active recombinant laccases were achieved by targeting expression to the endosperm of rice seeds. The laccase cDNAs were fused to a plant-derived signal sequence for targeting to the secretory pathway, and placed under the control of a constitutive seed-specific promoter fused to an intron for enhanced expression. This construct enabled the recovery of on average 0.1–1% of soluble laccase in total soluble proteins (TSP). The highest yields of recombinant laccases obtained in rice seeds were 13 and 39 ppm for riceMaL and ricePycL, respectively. The rice-produced laccases were purified and characterized. The wild-type and the recombinant proteins showed similar biochemical features in terms of molecular mass, pI, temperature and optimal pH and the N-terminus was correctly processed. Although presenting lower kinetic parameters, the rice-produced laccases were also suitable for the oxidative cross-linking of a food model substrate [maize-bran feruloylated arabinoxylans (AX)].
Original languageEnglish
Pages (from-to)515 - 527
Number of pages13
JournalTransgenic Research
Volume17
Issue number4
DOIs
Publication statusPublished - 2008
MoE publication typeA1 Journal article-refereed

Fingerprint

Pycnoporus
Laccase
laccase
production technology
genetically modified organisms
rice
Seeds
seeds
Oryza
Basidiomycota
Ascomycota
Endosperm
Food Handling
arabinoxylan
Secretory Pathway
bran
Protein Sorting Signals
signal peptide
crosslinking
food processing

Keywords

  • Laccase
  • Melanocarpus albomyces
  • Pycnoporus cinnabarinus
  • Oryza sativa
  • Heterologous protein production
  • Recombinant fungal enzyme
  • Rice

Cite this

de Wilde, C., Uzan, E., Zhou, Z., Kruus, K., Andberg, M., Buchert, J., ... Lomascolo, A. (2008). Transgenic rice as a novel production system for Melanocarpus and Pycnoporus laccases. Transgenic Research, 17(4), 515 - 527. https://doi.org/10.1007/s11248-007-9124-9
de Wilde, Chris ; Uzan, Eva ; Zhou, Zhongyi ; Kruus, Kristiina ; Andberg, Martina ; Buchert, Johanna ; Record, Eric ; Asther, Marcel ; Lomascolo, Anne. / Transgenic rice as a novel production system for Melanocarpus and Pycnoporus laccases. In: Transgenic Research. 2008 ; Vol. 17, No. 4. pp. 515 - 527.
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abstract = "Laccases have numerous biotechnological applications, among them food processing. The widespread use of laccases has increased the demand for an inexpensive and safe source of recombinant enzyme. We explored the use of a rice-based system for the production of two fungal laccases derived from the ascomycete Melanocarpus albomyces and the basidiomycete Pycnoporus cinnabarinus. High-expression levels of active recombinant laccases were achieved by targeting expression to the endosperm of rice seeds. The laccase cDNAs were fused to a plant-derived signal sequence for targeting to the secretory pathway, and placed under the control of a constitutive seed-specific promoter fused to an intron for enhanced expression. This construct enabled the recovery of on average 0.1–1{\%} of soluble laccase in total soluble proteins (TSP). The highest yields of recombinant laccases obtained in rice seeds were 13 and 39 ppm for riceMaL and ricePycL, respectively. The rice-produced laccases were purified and characterized. The wild-type and the recombinant proteins showed similar biochemical features in terms of molecular mass, pI, temperature and optimal pH and the N-terminus was correctly processed. Although presenting lower kinetic parameters, the rice-produced laccases were also suitable for the oxidative cross-linking of a food model substrate [maize-bran feruloylated arabinoxylans (AX)].",
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de Wilde, C, Uzan, E, Zhou, Z, Kruus, K, Andberg, M, Buchert, J, Record, E, Asther, M & Lomascolo, A 2008, 'Transgenic rice as a novel production system for Melanocarpus and Pycnoporus laccases', Transgenic Research, vol. 17, no. 4, pp. 515 - 527. https://doi.org/10.1007/s11248-007-9124-9

Transgenic rice as a novel production system for Melanocarpus and Pycnoporus laccases. / de Wilde, Chris; Uzan, Eva; Zhou, Zhongyi; Kruus, Kristiina; Andberg, Martina; Buchert, Johanna; Record, Eric; Asther, Marcel; Lomascolo, Anne (Corresponding Author).

In: Transgenic Research, Vol. 17, No. 4, 2008, p. 515 - 527.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Transgenic rice as a novel production system for Melanocarpus and Pycnoporus laccases

AU - de Wilde, Chris

AU - Uzan, Eva

AU - Zhou, Zhongyi

AU - Kruus, Kristiina

AU - Andberg, Martina

AU - Buchert, Johanna

AU - Record, Eric

AU - Asther, Marcel

AU - Lomascolo, Anne

PY - 2008

Y1 - 2008

N2 - Laccases have numerous biotechnological applications, among them food processing. The widespread use of laccases has increased the demand for an inexpensive and safe source of recombinant enzyme. We explored the use of a rice-based system for the production of two fungal laccases derived from the ascomycete Melanocarpus albomyces and the basidiomycete Pycnoporus cinnabarinus. High-expression levels of active recombinant laccases were achieved by targeting expression to the endosperm of rice seeds. The laccase cDNAs were fused to a plant-derived signal sequence for targeting to the secretory pathway, and placed under the control of a constitutive seed-specific promoter fused to an intron for enhanced expression. This construct enabled the recovery of on average 0.1–1% of soluble laccase in total soluble proteins (TSP). The highest yields of recombinant laccases obtained in rice seeds were 13 and 39 ppm for riceMaL and ricePycL, respectively. The rice-produced laccases were purified and characterized. The wild-type and the recombinant proteins showed similar biochemical features in terms of molecular mass, pI, temperature and optimal pH and the N-terminus was correctly processed. Although presenting lower kinetic parameters, the rice-produced laccases were also suitable for the oxidative cross-linking of a food model substrate [maize-bran feruloylated arabinoxylans (AX)].

AB - Laccases have numerous biotechnological applications, among them food processing. The widespread use of laccases has increased the demand for an inexpensive and safe source of recombinant enzyme. We explored the use of a rice-based system for the production of two fungal laccases derived from the ascomycete Melanocarpus albomyces and the basidiomycete Pycnoporus cinnabarinus. High-expression levels of active recombinant laccases were achieved by targeting expression to the endosperm of rice seeds. The laccase cDNAs were fused to a plant-derived signal sequence for targeting to the secretory pathway, and placed under the control of a constitutive seed-specific promoter fused to an intron for enhanced expression. This construct enabled the recovery of on average 0.1–1% of soluble laccase in total soluble proteins (TSP). The highest yields of recombinant laccases obtained in rice seeds were 13 and 39 ppm for riceMaL and ricePycL, respectively. The rice-produced laccases were purified and characterized. The wild-type and the recombinant proteins showed similar biochemical features in terms of molecular mass, pI, temperature and optimal pH and the N-terminus was correctly processed. Although presenting lower kinetic parameters, the rice-produced laccases were also suitable for the oxidative cross-linking of a food model substrate [maize-bran feruloylated arabinoxylans (AX)].

KW - Laccase

KW - Melanocarpus albomyces

KW - Pycnoporus cinnabarinus

KW - Oryza sativa

KW - Heterologous protein production

KW - Recombinant fungal enzyme

KW - Rice

U2 - 10.1007/s11248-007-9124-9

DO - 10.1007/s11248-007-9124-9

M3 - Article

VL - 17

SP - 515

EP - 527

JO - Transgenic Research

JF - Transgenic Research

SN - 0962-8819

IS - 4

ER -