The Experts below are selected from a list of 186 Experts worldwide ranked by ideXlab platform
Yasushi Morikawa - One of the best experts on this subject based on the ideXlab platform.
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Cloning and heterologous expression of the exo-β-d-Glucosaminidase-encoding gene (gls93) from a filamentous fungus, Trichoderma reesei PC-3-7
Applied Microbiology and Biotechnology, 2006Co-Authors: Masakazu Ike, Masahiro Nogawa, Hirofumi Okada, Koji Isami, Yoshio Tanabe, Wataru Ogasawara, Yasushi MorikawaAbstract:We have previously reported on purification and characterization of an exo-β- d -Glucosaminidase (Gls93) from culture filtrate of Trichoderma reesei PC-3-7 grown on N -acetyl- d -glucosamine (GlcNAc). The corresponding gene of Gls93 was cloned and characterized in this work. To our knowledge, this is the first report on cloning of the gene encoding fungal exo-β- d -Glucosaminidase. This gene has no introns and encodes a polypeptide of 892 amino acids (aa) containing a secretion signal of 28 amino acids. Comparison of the amino acid sequence to known proteins and phylogenetic analysis indicated that gls93 belongs to the glycoside hydrolase family (GHF) 2 and should be further classified into a new subgroup, exo-β- d -Glucosaminidase subgroup. The gls93 transcription was biphasic when T. reesei was grown on GlcNAc, suggesting that the expression of this gene may be regulated by a complex mechanism, in which multiple regulatory proteins are involved. Furthermore, gls93 could be expressed in Pichia pastoris (ca. 0.49-mg/ml culture). The recombinant Gls93 had the two molecular forms, ca. 105 and 100 kDa, whose difference is caused by N -glycosylation. Both of them had the same properties such as specific activity and substrate specificity and showed only the activity of exo-β- d -Glucosaminidase but not those of β-galactosidase, β-glucuronidase, and β-mannosidase belonging to GHF2.
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Purification and Characterization of Exo-β-d-Glucosaminidase from a Cellulolytic Fungus,Trichoderma reesei PC-3-7
Applied and environmental microbiology, 1998Co-Authors: Masahiro Nogawa, Hiroya Takahashi, Aya Kashiwagi, Kenji Ohshima, Hirofumi Okada, Yasushi MorikawaAbstract:Chitosan-degrading activities induced by glucosamine (GlcN) or N-acetylglucosamine (GlcNAc) were found in a culture filtrate of Trichoderma reesei PC-3-7. One of the chitosan-degrading enzymes was purified to homogeneity by precipitation with ammonium sulfate followed by anion-exchange and hydrophobic-interaction chromatographies. The enzyme was monomeric, and its molecular mass was 93 kDa. The optimum pH and temperature of the enzyme were 4.0 and 50°C, respectively. The activity was stable in the pH range 6.0 to 9.0 and at a temperature below 50°C. Reaction product analysis from the viscosimetric assay and thin-layer chromatography and 1H nuclear magnetic resonance spectroscopy clearly indicated that the enzyme was an exo-type chitosanase, exo-β-d-Glucosaminidase, that releases GlcN from the nonreducing end of the chitosan chain. 1H nuclear magnetic resonance spectroscopy also showed that the exo-β-d-Glucosaminidase produced a β-form of GlcN, demonstrating that the enzyme is a retaining glycanase. Time-dependent liberation of the reducing sugar from partially acetylated chitosan with exo-β-d-Glucosaminidase and the partially purified exo-β-d-N-acetylGlucosaminidase from T. reesei PC-3-7 suggested that the exo-β-d-Glucosaminidase cleaves the glycosidic link of either GlcN-β(1→4)-GlcN or GlcN-β(1→4)-GlcNAc.
Masahiro Nogawa - One of the best experts on this subject based on the ideXlab platform.
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Cloning and heterologous expression of the exo-β-d-Glucosaminidase-encoding gene (gls93) from a filamentous fungus, Trichoderma reesei PC-3-7
Applied Microbiology and Biotechnology, 2006Co-Authors: Masakazu Ike, Masahiro Nogawa, Hirofumi Okada, Koji Isami, Yoshio Tanabe, Wataru Ogasawara, Yasushi MorikawaAbstract:We have previously reported on purification and characterization of an exo-β- d -Glucosaminidase (Gls93) from culture filtrate of Trichoderma reesei PC-3-7 grown on N -acetyl- d -glucosamine (GlcNAc). The corresponding gene of Gls93 was cloned and characterized in this work. To our knowledge, this is the first report on cloning of the gene encoding fungal exo-β- d -Glucosaminidase. This gene has no introns and encodes a polypeptide of 892 amino acids (aa) containing a secretion signal of 28 amino acids. Comparison of the amino acid sequence to known proteins and phylogenetic analysis indicated that gls93 belongs to the glycoside hydrolase family (GHF) 2 and should be further classified into a new subgroup, exo-β- d -Glucosaminidase subgroup. The gls93 transcription was biphasic when T. reesei was grown on GlcNAc, suggesting that the expression of this gene may be regulated by a complex mechanism, in which multiple regulatory proteins are involved. Furthermore, gls93 could be expressed in Pichia pastoris (ca. 0.49-mg/ml culture). The recombinant Gls93 had the two molecular forms, ca. 105 and 100 kDa, whose difference is caused by N -glycosylation. Both of them had the same properties such as specific activity and substrate specificity and showed only the activity of exo-β- d -Glucosaminidase but not those of β-galactosidase, β-glucuronidase, and β-mannosidase belonging to GHF2.
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Purification and Characterization of Exo-β-d-Glucosaminidase from a Cellulolytic Fungus,Trichoderma reesei PC-3-7
Applied and environmental microbiology, 1998Co-Authors: Masahiro Nogawa, Hiroya Takahashi, Aya Kashiwagi, Kenji Ohshima, Hirofumi Okada, Yasushi MorikawaAbstract:Chitosan-degrading activities induced by glucosamine (GlcN) or N-acetylglucosamine (GlcNAc) were found in a culture filtrate of Trichoderma reesei PC-3-7. One of the chitosan-degrading enzymes was purified to homogeneity by precipitation with ammonium sulfate followed by anion-exchange and hydrophobic-interaction chromatographies. The enzyme was monomeric, and its molecular mass was 93 kDa. The optimum pH and temperature of the enzyme were 4.0 and 50°C, respectively. The activity was stable in the pH range 6.0 to 9.0 and at a temperature below 50°C. Reaction product analysis from the viscosimetric assay and thin-layer chromatography and 1H nuclear magnetic resonance spectroscopy clearly indicated that the enzyme was an exo-type chitosanase, exo-β-d-Glucosaminidase, that releases GlcN from the nonreducing end of the chitosan chain. 1H nuclear magnetic resonance spectroscopy also showed that the exo-β-d-Glucosaminidase produced a β-form of GlcN, demonstrating that the enzyme is a retaining glycanase. Time-dependent liberation of the reducing sugar from partially acetylated chitosan with exo-β-d-Glucosaminidase and the partially purified exo-β-d-N-acetylGlucosaminidase from T. reesei PC-3-7 suggested that the exo-β-d-Glucosaminidase cleaves the glycosidic link of either GlcN-β(1→4)-GlcN or GlcN-β(1→4)-GlcNAc.
Hirofumi Okada - One of the best experts on this subject based on the ideXlab platform.
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Cloning and heterologous expression of the exo-β-d-Glucosaminidase-encoding gene (gls93) from a filamentous fungus, Trichoderma reesei PC-3-7
Applied Microbiology and Biotechnology, 2006Co-Authors: Masakazu Ike, Masahiro Nogawa, Hirofumi Okada, Koji Isami, Yoshio Tanabe, Wataru Ogasawara, Yasushi MorikawaAbstract:We have previously reported on purification and characterization of an exo-β- d -Glucosaminidase (Gls93) from culture filtrate of Trichoderma reesei PC-3-7 grown on N -acetyl- d -glucosamine (GlcNAc). The corresponding gene of Gls93 was cloned and characterized in this work. To our knowledge, this is the first report on cloning of the gene encoding fungal exo-β- d -Glucosaminidase. This gene has no introns and encodes a polypeptide of 892 amino acids (aa) containing a secretion signal of 28 amino acids. Comparison of the amino acid sequence to known proteins and phylogenetic analysis indicated that gls93 belongs to the glycoside hydrolase family (GHF) 2 and should be further classified into a new subgroup, exo-β- d -Glucosaminidase subgroup. The gls93 transcription was biphasic when T. reesei was grown on GlcNAc, suggesting that the expression of this gene may be regulated by a complex mechanism, in which multiple regulatory proteins are involved. Furthermore, gls93 could be expressed in Pichia pastoris (ca. 0.49-mg/ml culture). The recombinant Gls93 had the two molecular forms, ca. 105 and 100 kDa, whose difference is caused by N -glycosylation. Both of them had the same properties such as specific activity and substrate specificity and showed only the activity of exo-β- d -Glucosaminidase but not those of β-galactosidase, β-glucuronidase, and β-mannosidase belonging to GHF2.
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Purification and Characterization of Exo-β-d-Glucosaminidase from a Cellulolytic Fungus,Trichoderma reesei PC-3-7
Applied and environmental microbiology, 1998Co-Authors: Masahiro Nogawa, Hiroya Takahashi, Aya Kashiwagi, Kenji Ohshima, Hirofumi Okada, Yasushi MorikawaAbstract:Chitosan-degrading activities induced by glucosamine (GlcN) or N-acetylglucosamine (GlcNAc) were found in a culture filtrate of Trichoderma reesei PC-3-7. One of the chitosan-degrading enzymes was purified to homogeneity by precipitation with ammonium sulfate followed by anion-exchange and hydrophobic-interaction chromatographies. The enzyme was monomeric, and its molecular mass was 93 kDa. The optimum pH and temperature of the enzyme were 4.0 and 50°C, respectively. The activity was stable in the pH range 6.0 to 9.0 and at a temperature below 50°C. Reaction product analysis from the viscosimetric assay and thin-layer chromatography and 1H nuclear magnetic resonance spectroscopy clearly indicated that the enzyme was an exo-type chitosanase, exo-β-d-Glucosaminidase, that releases GlcN from the nonreducing end of the chitosan chain. 1H nuclear magnetic resonance spectroscopy also showed that the exo-β-d-Glucosaminidase produced a β-form of GlcN, demonstrating that the enzyme is a retaining glycanase. Time-dependent liberation of the reducing sugar from partially acetylated chitosan with exo-β-d-Glucosaminidase and the partially purified exo-β-d-N-acetylGlucosaminidase from T. reesei PC-3-7 suggested that the exo-β-d-Glucosaminidase cleaves the glycosidic link of either GlcN-β(1→4)-GlcN or GlcN-β(1→4)-GlcNAc.
Masakazu Ike - One of the best experts on this subject based on the ideXlab platform.
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Cloning and heterologous expression of the exo-β-d-Glucosaminidase-encoding gene (gls93) from a filamentous fungus, Trichoderma reesei PC-3-7
Applied Microbiology and Biotechnology, 2006Co-Authors: Masakazu Ike, Masahiro Nogawa, Hirofumi Okada, Koji Isami, Yoshio Tanabe, Wataru Ogasawara, Yasushi MorikawaAbstract:We have previously reported on purification and characterization of an exo-β- d -Glucosaminidase (Gls93) from culture filtrate of Trichoderma reesei PC-3-7 grown on N -acetyl- d -glucosamine (GlcNAc). The corresponding gene of Gls93 was cloned and characterized in this work. To our knowledge, this is the first report on cloning of the gene encoding fungal exo-β- d -Glucosaminidase. This gene has no introns and encodes a polypeptide of 892 amino acids (aa) containing a secretion signal of 28 amino acids. Comparison of the amino acid sequence to known proteins and phylogenetic analysis indicated that gls93 belongs to the glycoside hydrolase family (GHF) 2 and should be further classified into a new subgroup, exo-β- d -Glucosaminidase subgroup. The gls93 transcription was biphasic when T. reesei was grown on GlcNAc, suggesting that the expression of this gene may be regulated by a complex mechanism, in which multiple regulatory proteins are involved. Furthermore, gls93 could be expressed in Pichia pastoris (ca. 0.49-mg/ml culture). The recombinant Gls93 had the two molecular forms, ca. 105 and 100 kDa, whose difference is caused by N -glycosylation. Both of them had the same properties such as specific activity and substrate specificity and showed only the activity of exo-β- d -Glucosaminidase but not those of β-galactosidase, β-glucuronidase, and β-mannosidase belonging to GHF2.
Hiroya Takahashi - One of the best experts on this subject based on the ideXlab platform.
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Purification and Characterization of Exo-β-d-Glucosaminidase from a Cellulolytic Fungus,Trichoderma reesei PC-3-7
Applied and environmental microbiology, 1998Co-Authors: Masahiro Nogawa, Hiroya Takahashi, Aya Kashiwagi, Kenji Ohshima, Hirofumi Okada, Yasushi MorikawaAbstract:Chitosan-degrading activities induced by glucosamine (GlcN) or N-acetylglucosamine (GlcNAc) were found in a culture filtrate of Trichoderma reesei PC-3-7. One of the chitosan-degrading enzymes was purified to homogeneity by precipitation with ammonium sulfate followed by anion-exchange and hydrophobic-interaction chromatographies. The enzyme was monomeric, and its molecular mass was 93 kDa. The optimum pH and temperature of the enzyme were 4.0 and 50°C, respectively. The activity was stable in the pH range 6.0 to 9.0 and at a temperature below 50°C. Reaction product analysis from the viscosimetric assay and thin-layer chromatography and 1H nuclear magnetic resonance spectroscopy clearly indicated that the enzyme was an exo-type chitosanase, exo-β-d-Glucosaminidase, that releases GlcN from the nonreducing end of the chitosan chain. 1H nuclear magnetic resonance spectroscopy also showed that the exo-β-d-Glucosaminidase produced a β-form of GlcN, demonstrating that the enzyme is a retaining glycanase. Time-dependent liberation of the reducing sugar from partially acetylated chitosan with exo-β-d-Glucosaminidase and the partially purified exo-β-d-N-acetylGlucosaminidase from T. reesei PC-3-7 suggested that the exo-β-d-Glucosaminidase cleaves the glycosidic link of either GlcN-β(1→4)-GlcN or GlcN-β(1→4)-GlcNAc.