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Stachybotrys atra BP-A produces alkali-resistant and thermostable cellulases

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Abstract

A cellulose-degrading fungal strain has been isolated from a rotten rag. Morphological characterization and ITS1, 5.8S and ITS2 rDNA sequencing showed that the strain is a new isolate of Stachybotrys atra. The strain secreted high cellulase activity in media supplemented with rice straw. However, cellulases were not produced in glucose-supplemented media. The crude cellulase showed the highest activity on amorphous celluloses such as carboxymethyl cellulose, while activity on crystalline celluloses such as Avicel was lower. The optimal temperature and pH for CMCase activity were 70°C and pH 5 respectively, although a second peak of activity was found at pH 8. Activity was strongly inhibited by Cu2+, Mn2+ and Hg2+. Analysis by SDS-PAGE, isoelectric focusing and zymography showed that the strain secretes a complex cellulase system comprising several enzymes. Most of these enzymes are alkali-resistant CMCases that remained stable at pH 9 and 65°C for at least 1 h. Cellulose binding assays showed notable differences among the CMCases. While some CMCase bands did not bind Avicel, other bands bound to this polymer and were eluted either with NaCl or by boiling with SDS. Analysis by two-dimensional electrophoresis showed that the band eluted by SDS boiling contained at least 4 different polypeptides. The complex set of cellulases produced by the strain, and their activity and stability at alkaline pH and a high temperature indicate that both the isolated strain and the cellulases identified are good candidates for biotechnological applications involving cellulose modification.

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Acknowledgements

This work was partially supported by the Spanish Ministry of Education and Science (CICYT), projects ref. CTQ2004-07560-C02-02 and CTQ2007-68003-CO2-02/PPQ; and the Centre de Referència en Biotecnologia (CeRBa). Pere Picart held an FPI grant ref. FP-2001-1668 from the Spanish Ministry of Science and Technology.

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  1. Department of Microbiology, Faculty of Biology, University of Barcelona, Avinguda Diagonal 645, 08028, Barcelona, Spain

    Pere Picart, Pilar Diaz & F. I. Javier Pastor

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  1. Pere Picart
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  2. Pilar Diaz
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  3. F. I. Javier Pastor
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Correspondence to F. I. Javier Pastor.

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Picart, P., Diaz, P. & Pastor, F.I.J. Stachybotrys atra BP-A produces alkali-resistant and thermostable cellulases. Antonie van Leeuwenhoek 94, 307–316 (2008). https://doi.org/10.1007/s10482-008-9248-9

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