Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-alfa-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

Abstract

A cDNA encoding 1,2-alpha-D-mannosidase mds 1 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-alpha-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 alpha-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analyzed. The enzyme was characterized as a class-I mannosidase.
Original languageEnglish
Pages (from-to)255-263
JournalJournal of Biotechnology
Volume77
Issue number2-3
Publication statusPublished - 2000
MoE publication typeA1 Journal article-refereed

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

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-alfa-D-mannosidase. Journal of Biotechnology, 77(2-3), 255-263.
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-alfa-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-alpha-D-mannosidase mds 1 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-alpha-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 alpha-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analyzed. The enzyme was characterized as a class-I mannosidase.",
author = "Marleen Maras and Nico Callewaert and Kathleen Piens and Marc Claeyssens and Wim Martinet and Sylviane Dewaele and Hans Contreras and Isabelle Dewerte and Merja Penttil{\"a} and Roland Contreras",
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journal = "Journal of Biotechnology",
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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-alfa-D-mannosidase', Journal of Biotechnology, vol. 77, no. 2-3, pp. 255-263.

Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-alfa-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, 2000, p. 255-263.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Molecular cloning and enzymatic characterization of a Trichoderma reesei 1,2-alfa-D-mannosidase

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

Y1 - 2000

N2 - A cDNA encoding 1,2-alpha-D-mannosidase mds 1 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-alpha-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 alpha-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analyzed. The enzyme was characterized as a class-I mannosidase.

AB - A cDNA encoding 1,2-alpha-D-mannosidase mds 1 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-alpha-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 alpha-mating factor resulted in high amounts of secreted enzyme. A three-step purification protocol was designed and the enzymatic properties were analyzed. The enzyme was characterized as a class-I mannosidase.

M3 - Article

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JO - Journal of Biotechnology

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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-alfa-D-mannosidase. Journal of Biotechnology. 2000;77(2-3):255-263.