Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-α-D-mannosidase

Marleen Maras, Nico Callewaert, Kathleen Piens, Marc Claeyssens, Wim Martinet, Sylviane Dewaele, Hans Contreras, Isabelle Dewerte, Merja Penttilä, Roland Contreras

Research output: Contribution to journalArticleScientificpeer-review

51 Citations (Scopus)

Abstract

A cDNA encoding 1,2-α-D-mannosidase mds1 from Trichoderma reesei was cloned. The largest open reading frame occupied 1571 bp. The predicted sequence contains 523 amino acid residues for a calculated molecular mass of 56 266 Da and shows high similarity to the amino acid sequences of 1,2-α-D-mannosidases from Aspergillus saitoi and Penicillium citrinum (51.6 and 51.0% identity, respectively). T. reesei mannosidase was produced as a recombinant enzyme in the yeast Pichia pastoris. Replacement of the N-terminal part with the prepro-signal peptide of the Saccharomyces cerevisiae α-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analysed. The enzyme was characterized as a class-I mannosidase. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)255-263
Number of pages9
JournalJournal of Biotechnology
Volume77
Issue number2-3
DOIs
Publication statusPublished - 17 Feb 2000
MoE publication typeA1 Journal article-refereed

Fingerprint

Mannosidases
Trichoderma
Cloning
Molecular Cloning
Enzymes
mannosyl-oligosaccharide 1,2-alpha-mannosidase
Mating Factor
Yeast
Amino acids
Amino Acids
Pichia
Aspergillus
Penicillium
Molecular mass
Protein Sorting Signals
Open Reading Frames
Purification
Saccharomyces cerevisiae
Amino Acid Sequence
Complementary DNA

Keywords

  • Fungus
  • Glycosidase
  • Mannosidase
  • Oligosaccharide
  • Trichoderma reesei
  • Yeast

Cite this

Maras, M., Callewaert, N., Piens, K., Claeyssens, M., Martinet, W., Dewaele, S., ... Contreras, R. (2000). Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-α-D-mannosidase. Journal of Biotechnology, 77(2-3), 255-263. https://doi.org/10.1016/S0168-1656(99)00222-9
Maras, Marleen ; Callewaert, Nico ; Piens, Kathleen ; Claeyssens, Marc ; Martinet, Wim ; Dewaele, Sylviane ; Contreras, Hans ; Dewerte, Isabelle ; Penttilä, Merja ; Contreras, Roland. / Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-α-D-mannosidase. In: Journal of Biotechnology. 2000 ; Vol. 77, No. 2-3. pp. 255-263.
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abstract = "A cDNA encoding 1,2-α-D-mannosidase mds1 from Trichoderma reesei was cloned. The largest open reading frame occupied 1571 bp. The predicted sequence contains 523 amino acid residues for a calculated molecular mass of 56 266 Da and shows high similarity to the amino acid sequences of 1,2-α-D-mannosidases from Aspergillus saitoi and Penicillium citrinum (51.6 and 51.0{\%} identity, respectively). T. reesei mannosidase was produced as a recombinant enzyme in the yeast Pichia pastoris. Replacement of the N-terminal part with the prepro-signal peptide of the Saccharomyces cerevisiae α-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analysed. The enzyme was characterized as a class-I mannosidase. Copyright (C) 2000 Elsevier Science B.V.",
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Maras, M, Callewaert, N, Piens, K, Claeyssens, M, Martinet, W, Dewaele, S, Contreras, H, Dewerte, I, Penttilä, M & Contreras, R 2000, 'Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-α-D-mannosidase', Journal of Biotechnology, vol. 77, no. 2-3, pp. 255-263. https://doi.org/10.1016/S0168-1656(99)00222-9

Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-α-D-mannosidase. / Maras, Marleen; Callewaert, Nico; Piens, Kathleen; Claeyssens, Marc; Martinet, Wim; Dewaele, Sylviane; Contreras, Hans; Dewerte, Isabelle; Penttilä, Merja; Contreras, Roland.

In: Journal of Biotechnology, Vol. 77, No. 2-3, 17.02.2000, p. 255-263.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Maras, Marleen

AU - Callewaert, Nico

AU - Piens, Kathleen

AU - Claeyssens, Marc

AU - Martinet, Wim

AU - Dewaele, Sylviane

AU - Contreras, Hans

AU - Dewerte, Isabelle

AU - Penttilä, Merja

AU - Contreras, Roland

PY - 2000/2/17

Y1 - 2000/2/17

N2 - A cDNA encoding 1,2-α-D-mannosidase mds1 from Trichoderma reesei was cloned. The largest open reading frame occupied 1571 bp. The predicted sequence contains 523 amino acid residues for a calculated molecular mass of 56 266 Da and shows high similarity to the amino acid sequences of 1,2-α-D-mannosidases from Aspergillus saitoi and Penicillium citrinum (51.6 and 51.0% identity, respectively). T. reesei mannosidase was produced as a recombinant enzyme in the yeast Pichia pastoris. Replacement of the N-terminal part with the prepro-signal peptide of the Saccharomyces cerevisiae α-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analysed. The enzyme was characterized as a class-I mannosidase. Copyright (C) 2000 Elsevier Science B.V.

AB - A cDNA encoding 1,2-α-D-mannosidase mds1 from Trichoderma reesei was cloned. The largest open reading frame occupied 1571 bp. The predicted sequence contains 523 amino acid residues for a calculated molecular mass of 56 266 Da and shows high similarity to the amino acid sequences of 1,2-α-D-mannosidases from Aspergillus saitoi and Penicillium citrinum (51.6 and 51.0% identity, respectively). T. reesei mannosidase was produced as a recombinant enzyme in the yeast Pichia pastoris. Replacement of the N-terminal part with the prepro-signal peptide of the Saccharomyces cerevisiae α-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analysed. The enzyme was characterized as a class-I mannosidase. Copyright (C) 2000 Elsevier Science B.V.

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JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

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Maras M, Callewaert N, Piens K, Claeyssens M, Martinet W, Dewaele S et al. Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-α-D-mannosidase. Journal of Biotechnology. 2000 Feb 17;77(2-3):255-263. https://doi.org/10.1016/S0168-1656(99)00222-9