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

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58 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

Keywords

  • Cloning
  • Endoglucanase
  • Heterologous expression
  • Biostoning

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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, 34(2), 159-167. https://doi.org/10.1016/j.enzmictec.2003.10.009