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

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

Vlaams Instituut voor Biotechnologie (VIB)

Research output: Contribution to journal › Article › Scientific › peer-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 language	English
Pages (from-to)	255-263
Journal	Journal of Biotechnology
Volume	77
Issue number	2-3
Publication status	Published - 2000
MoE publication type	A1 Journal article-refereed

Cite this

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

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",

year = "2000",

language = "English",

volume = "77",

pages = "255--263",

journal = "Journal of Biotechnology",

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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

SN - 0168-1656

VL - 77

SP - 255

EP - 263

JO - Journal of Biotechnology

JF - Journal of Biotechnology

IS - 2-3

ER -

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

Abstract

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