Expression of Two Novel β-Glucosidases from Chaetomium atrobrunneum in Trichoderma reesei and Characterization of the Heterologous Protein Products

Ana C. Colabardini, Mari Valkonen, Anne Huuskonen, Matti Siika-aho, Anu Koivula, Gustavo H. Goldman, Markku Saloheimo

Research output: Contribution to journalArticleScientificpeer-review

7 Citations (Scopus)

Abstract

Two novel GH3 family thermostable β-glucosidases from the filamentous fungus Chaetomium atrobrunneum (CEL3a and CEL3b) were expressed in Trichoderma reesei, purified by two-step ion exchange chromatography, and characterized. Both enzymes were active over a wide range of pH as compared to Neurospora crassa β-glucosidase GH3-3, which was also expressed in T. reesei and purified. The optimum temperature of both C. atrobrunneum enzymes was around 60 °C at pH 5, and both enzymes had better thermal and pH stability and higher resistance to metallic compounds and to glucose inhibition than GH3-3. They also showed higher activity against oligosaccharides composed of glucose units and linked with β-1,4-glycosidic bonds and moreover, had higher affinity for cellotriose over cellobiose. In hydrolysis tests against Avicel cellulose and steam-exploded sugarcane bagasse, performed at 45 °C, particularly the CEL3a enzyme performed similarly to N. crassa GH3-3 β-glucosidase. Taking into account the thermal stability of the C. atrobrunneum β-glucosidases, they both represent promising alternatives as enzyme mixture components for improved cellulose saccharification at elevated temperatures.

Original languageEnglish
Pages (from-to)821-831
Number of pages11
JournalMolecular Biotechnology
Volume58
Issue number12
DOIs
Publication statusPublished - 1 Dec 2016
MoE publication typeA1 Journal article-refereed

Fingerprint

Chaetomium
Glucosidases
Trichoderma
Enzymes
Proteins
Cellulose
Neurospora crassa
Glucose
Hot Temperature
Metallic compounds
Cellobiose
Saccharification
Oligosaccharides
Saccharum
Bagasse
Temperature
Ion Exchange Chromatography
Steam
Chromatography
Fungi

Keywords

  • Bioethanol
  • Chaetomium atrobrunneum
  • Hydrolysis
  • Thermostability
  • β-glucosidase

Cite this

@article{9550f54afdbf47a69e7ec457f7774d6a,
title = "Expression of Two Novel β-Glucosidases from Chaetomium atrobrunneum in Trichoderma reesei and Characterization of the Heterologous Protein Products",
abstract = "Two novel GH3 family thermostable β-glucosidases from the filamentous fungus Chaetomium atrobrunneum (CEL3a and CEL3b) were expressed in Trichoderma reesei, purified by two-step ion exchange chromatography, and characterized. Both enzymes were active over a wide range of pH as compared to Neurospora crassa β-glucosidase GH3-3, which was also expressed in T. reesei and purified. The optimum temperature of both C. atrobrunneum enzymes was around 60 °C at pH 5, and both enzymes had better thermal and pH stability and higher resistance to metallic compounds and to glucose inhibition than GH3-3. They also showed higher activity against oligosaccharides composed of glucose units and linked with β-1,4-glycosidic bonds and moreover, had higher affinity for cellotriose over cellobiose. In hydrolysis tests against Avicel cellulose and steam-exploded sugarcane bagasse, performed at 45 °C, particularly the CEL3a enzyme performed similarly to N. crassa GH3-3 β-glucosidase. Taking into account the thermal stability of the C. atrobrunneum β-glucosidases, they both represent promising alternatives as enzyme mixture components for improved cellulose saccharification at elevated temperatures.",
keywords = "Bioethanol, Chaetomium atrobrunneum, Hydrolysis, Thermostability, β-glucosidase",
author = "Colabardini, {Ana C.} and Mari Valkonen and Anne Huuskonen and Matti Siika-aho and Anu Koivula and Goldman, {Gustavo H.} and Markku Saloheimo",
year = "2016",
month = "12",
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language = "English",
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pages = "821--831",
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Expression of Two Novel β-Glucosidases from Chaetomium atrobrunneum in Trichoderma reesei and Characterization of the Heterologous Protein Products. / Colabardini, Ana C.; Valkonen, Mari; Huuskonen, Anne; Siika-aho, Matti; Koivula, Anu; Goldman, Gustavo H.; Saloheimo, Markku.

In: Molecular Biotechnology, Vol. 58, No. 12, 01.12.2016, p. 821-831.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Expression of Two Novel β-Glucosidases from Chaetomium atrobrunneum in Trichoderma reesei and Characterization of the Heterologous Protein Products

AU - Colabardini, Ana C.

AU - Valkonen, Mari

AU - Huuskonen, Anne

AU - Siika-aho, Matti

AU - Koivula, Anu

AU - Goldman, Gustavo H.

AU - Saloheimo, Markku

PY - 2016/12/1

Y1 - 2016/12/1

N2 - Two novel GH3 family thermostable β-glucosidases from the filamentous fungus Chaetomium atrobrunneum (CEL3a and CEL3b) were expressed in Trichoderma reesei, purified by two-step ion exchange chromatography, and characterized. Both enzymes were active over a wide range of pH as compared to Neurospora crassa β-glucosidase GH3-3, which was also expressed in T. reesei and purified. The optimum temperature of both C. atrobrunneum enzymes was around 60 °C at pH 5, and both enzymes had better thermal and pH stability and higher resistance to metallic compounds and to glucose inhibition than GH3-3. They also showed higher activity against oligosaccharides composed of glucose units and linked with β-1,4-glycosidic bonds and moreover, had higher affinity for cellotriose over cellobiose. In hydrolysis tests against Avicel cellulose and steam-exploded sugarcane bagasse, performed at 45 °C, particularly the CEL3a enzyme performed similarly to N. crassa GH3-3 β-glucosidase. Taking into account the thermal stability of the C. atrobrunneum β-glucosidases, they both represent promising alternatives as enzyme mixture components for improved cellulose saccharification at elevated temperatures.

AB - Two novel GH3 family thermostable β-glucosidases from the filamentous fungus Chaetomium atrobrunneum (CEL3a and CEL3b) were expressed in Trichoderma reesei, purified by two-step ion exchange chromatography, and characterized. Both enzymes were active over a wide range of pH as compared to Neurospora crassa β-glucosidase GH3-3, which was also expressed in T. reesei and purified. The optimum temperature of both C. atrobrunneum enzymes was around 60 °C at pH 5, and both enzymes had better thermal and pH stability and higher resistance to metallic compounds and to glucose inhibition than GH3-3. They also showed higher activity against oligosaccharides composed of glucose units and linked with β-1,4-glycosidic bonds and moreover, had higher affinity for cellotriose over cellobiose. In hydrolysis tests against Avicel cellulose and steam-exploded sugarcane bagasse, performed at 45 °C, particularly the CEL3a enzyme performed similarly to N. crassa GH3-3 β-glucosidase. Taking into account the thermal stability of the C. atrobrunneum β-glucosidases, they both represent promising alternatives as enzyme mixture components for improved cellulose saccharification at elevated temperatures.

KW - Bioethanol

KW - Chaetomium atrobrunneum

KW - Hydrolysis

KW - Thermostability

KW - β-glucosidase

U2 - 10.1007/s12033-016-9981-7

DO - 10.1007/s12033-016-9981-7

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AN - SCOPUS:84990858995

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JO - Molecular Biotechnology

JF - Molecular Biotechnology

SN - 1073-6085

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ER -