Cloning of cellulase genes from Melanocarpus albomyces and their efficient expression in Trichoderma reesei

Heli Haakana, Arja Miettinen-Oinonen (Corresponding Author), Vesa Joutsjoki, Arja Mäntylä, Pirkko Suominen, Jari Vehmaanperä

Research output: Contribution to journalArticleScientificpeer-review

57 Citations (Scopus)

Abstract

In our previous study, three purified cellulases of Melanocarpus albomyces proved to be effective in biostoning application at neutral pH [Enzyme Microb. Technol., accepted for publication]. We cloned and sequenced three genes of M. albomyces, which encode a 20 kDa and two 50-kDa polypeptides. The 20-kDa protein (Cel45A) and one of the 50-kDa proteins (Cel7A) are endoglucanases of the glycosyl hydrolase families 45 and 7, respectively. The other 50-kDa protein (Cel7B) is a family 7 cellobiohydrolase. None of the cellulases harbors a cellulose binding domain (CBD). These genes were expressed in Trichoderma reesei under the control of the T. reesei cbh1 promoter and the proteins detected in the culture medium. The endoglucanase production levels of the cel45A- and cel7A-transformants were several times higher than those of the parental M. albomyces strain. The sizes of Cel45A, Cel7A and Cel7B proteins produced by the transformants were the same as the sizes of the corresponding proteins purified from M. albomyces. Cellulase preparations produced by the cel45A transformants performed well at neutral pH in stone-washing of denim fabric and caused considerably less backstaining as compared to the acid cellulase product of T. reesei.
Original languageEnglish
Pages (from-to)159-167
JournalEnzyme and Microbial Technology
Volume34
Issue number2
DOIs
Publication statusPublished - 2004
MoE publication typeA1 Journal article-refereed

Fingerprint

Trichoderma
Cellulase
Cloning
Organism Cloning
Genes
Proteins
Cellulases
Cellulose 1,4-beta-Cellobiosidase
Hydrolases
Polypeptides
Ports and harbors
Washing
Cellulose
Culture Media
Publications
Enzymes
Peptides
Acids

Keywords

  • Cloning
  • Endoglucanase
  • Heterologous expression
  • Biostoning

Cite this

Haakana, H., Miettinen-Oinonen, A., Joutsjoki, V., Mäntylä, A., Suominen, P., & Vehmaanperä, J. (2004). Cloning of cellulase genes from Melanocarpus albomyces and their efficient expression in Trichoderma reesei. Enzyme and Microbial Technology, 34(2), 159-167. https://doi.org/10.1016/j.enzmictec.2003.10.009
Haakana, Heli ; Miettinen-Oinonen, Arja ; Joutsjoki, Vesa ; Mäntylä, Arja ; Suominen, Pirkko ; Vehmaanperä, Jari. / Cloning of cellulase genes from Melanocarpus albomyces and their efficient expression in Trichoderma reesei. In: Enzyme and Microbial Technology. 2004 ; Vol. 34, No. 2. pp. 159-167.
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Haakana, H, Miettinen-Oinonen, A, Joutsjoki, V, Mäntylä, A, Suominen, P & Vehmaanperä, J 2004, 'Cloning of cellulase genes from Melanocarpus albomyces and their efficient expression in Trichoderma reesei', Enzyme and Microbial Technology, vol. 34, no. 2, pp. 159-167. https://doi.org/10.1016/j.enzmictec.2003.10.009

Cloning of cellulase genes from Melanocarpus albomyces and their efficient expression in Trichoderma reesei. / Haakana, Heli; Miettinen-Oinonen, Arja (Corresponding Author); Joutsjoki, Vesa; Mäntylä, Arja; Suominen, Pirkko; Vehmaanperä, Jari.

In: Enzyme and Microbial Technology, Vol. 34, No. 2, 2004, p. 159-167.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Cloning of cellulase genes from Melanocarpus albomyces and their efficient expression in Trichoderma reesei

AU - Haakana, Heli

AU - Miettinen-Oinonen, Arja

AU - Joutsjoki, Vesa

AU - Mäntylä, Arja

AU - Suominen, Pirkko

AU - Vehmaanperä, Jari

PY - 2004

Y1 - 2004

N2 - In our previous study, three purified cellulases of Melanocarpus albomyces proved to be effective in biostoning application at neutral pH [Enzyme Microb. Technol., accepted for publication]. We cloned and sequenced three genes of M. albomyces, which encode a 20 kDa and two 50-kDa polypeptides. The 20-kDa protein (Cel45A) and one of the 50-kDa proteins (Cel7A) are endoglucanases of the glycosyl hydrolase families 45 and 7, respectively. The other 50-kDa protein (Cel7B) is a family 7 cellobiohydrolase. None of the cellulases harbors a cellulose binding domain (CBD). These genes were expressed in Trichoderma reesei under the control of the T. reesei cbh1 promoter and the proteins detected in the culture medium. The endoglucanase production levels of the cel45A- and cel7A-transformants were several times higher than those of the parental M. albomyces strain. The sizes of Cel45A, Cel7A and Cel7B proteins produced by the transformants were the same as the sizes of the corresponding proteins purified from M. albomyces. Cellulase preparations produced by the cel45A transformants performed well at neutral pH in stone-washing of denim fabric and caused considerably less backstaining as compared to the acid cellulase product of T. reesei.

AB - In our previous study, three purified cellulases of Melanocarpus albomyces proved to be effective in biostoning application at neutral pH [Enzyme Microb. Technol., accepted for publication]. We cloned and sequenced three genes of M. albomyces, which encode a 20 kDa and two 50-kDa polypeptides. The 20-kDa protein (Cel45A) and one of the 50-kDa proteins (Cel7A) are endoglucanases of the glycosyl hydrolase families 45 and 7, respectively. The other 50-kDa protein (Cel7B) is a family 7 cellobiohydrolase. None of the cellulases harbors a cellulose binding domain (CBD). These genes were expressed in Trichoderma reesei under the control of the T. reesei cbh1 promoter and the proteins detected in the culture medium. The endoglucanase production levels of the cel45A- and cel7A-transformants were several times higher than those of the parental M. albomyces strain. The sizes of Cel45A, Cel7A and Cel7B proteins produced by the transformants were the same as the sizes of the corresponding proteins purified from M. albomyces. Cellulase preparations produced by the cel45A transformants performed well at neutral pH in stone-washing of denim fabric and caused considerably less backstaining as compared to the acid cellulase product of T. reesei.

KW - Cloning

KW - Endoglucanase

KW - Heterologous expression

KW - Biostoning

U2 - 10.1016/j.enzmictec.2003.10.009

DO - 10.1016/j.enzmictec.2003.10.009

M3 - Article

VL - 34

SP - 159

EP - 167

JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

SN - 0141-0229

IS - 2

ER -