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Electrophoresis
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Electrophoresis
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Electrophoresis
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Reliable simultaneous zymographic method of characterization of cellulolytic enzymes from fungal cellulase complex

Authors: Dojnov, Biljana; Grujić, Marica; Vujčić, Zoran;

Reliable simultaneous zymographic method of characterization of cellulolytic enzymes from fungal cellulase complex

Abstract

A method for zymographic detection of specific cellulases in a complex (endocellulase, exocellulase, and cellobiase) from crude fermentation extracts, after a single electrophoretic separation, is described in this paper. Cellulases were printed onto a membrane and, subsequently, substrate gel. Cellobiase isoforms were detected on the membrane using esculine as substrate, endocellulase isoforms on substrate gel with copolymerized carboxymethyl cellulose (CMC), while exocellulase isoforms were detected in electrophoresis gel with 4-methylumbelliferyl--d-cellobioside (MUC). This can be a useful additional tool for monitoring and control of fungal cellulase production in industrial processes and fundamental research, screening for particular cellulase producers, or testing of new lignocellulose substrates.

This is the peer-reviewed version of the following article: Dojnov, B.; Grujić, M.; Vujčić, Z. Reliable Simultaneous Zymographic Method of Characterization of Cellulolytic Enzymes from Fungal Cellulase Complex. Electrophoresis 2015, 36 (15), 1724–1727. [https://doi.org/10.1002/elps.201400541]

[http://cer.ihtm.bg.ac.rs/handle/123456789/1713]

Country
Serbia
Subjects by Vocabulary

Microsoft Academic Graph classification: Cellulase medicine Zymography chemistry.chemical_classification Gel electrophoresis biology Substrate (chemistry) Carboxymethyl cellulose Electrophoresis Enzyme chemistry Biochemistry biology.protein Fermentation medicine.drug

Keywords

beta -Glucosidase, Clinical Biochemistry, Biochemistry, Analytical Chemistry, Fungal Proteins, Cellulase, Zymogram, SSF, Trichoderma, Endocellulase, Carboxymethylcellulose Sodium, Fermentation, Exocellulase, Isoelectric Focusing

11 references, page 1 of 2

[1] Singhania, R. R., Sukumaran, R. K., Patel, A. K., Larroche, C., Pandey, A., Enzyme Microb Tech 2010, 46, 541-549.

[2] Zhou, J., Wang, Y. H., Chu, J., Zhuang, Y. P., Zhang, S. L., Yin, P., Bioresour Technol 2008, 99, 6826-6833.

[3] Kubicek, C. P., Mikus, M., Schuster, A., Schmoll, M., Seiboth, B., Biotechnol Biofuels 2009, 2, 19.

[4] Kubicek, C. P., Fungi and lignocellulosic biomass, Wiley-BlackWell, Ames, Iowa, USA 2013.

[5] Martinez, D., Berka, R. M., Henrissat, B., Saloheimo, M., Arvas, M., Baker, S. E., Chapman, J., Chertkov, O., Coutinho, P. M., Cullen, D., Danchin, E. G., Grigoriev, I. V., Harris, P., Jackson, M., Kubicek, C. P., Han, C. S., Ho, I., Larrondo, L. F., de Leon, A. L., Magnuson, J. K., Merino, S., Misra, M., Nelson, B., Putnam, N., Robbertse, B., Salamov, A. A., Schmoll, M., Terry, A., Thayer, N., Westerholm-Parvinen, A., Schoch, C. L., Yao, J., Barabote, R., Nelson, M. A., Detter, C., Bruce, D., Kuske, C. R., Xie, G., Richardson, P., Rokhsar, D. S., Lucas, S. M., Rubin, E. M., Dunn-Coleman, N., Ward, M., Brettin, T. S., Nature biotechnology 2008, 26, 553-560.

[6] Seiboth, B., Ivanova, C., Seidl-Seiboth, V., in: Bernardes, M. A. D. S. (Ed.), Biofuel Production-Recent Developments and Prospects, InTech, Rijeka, Croatia 2011.

[7] Eriksson, K.-E., Pettersson, B., Anal Biochem 1973, 56, 2.

[8] Biely, P., Markovič, O., Mislovičová, D., Anal Biochem 1985, 144, 147-151.

[9] Purwardaria, T., Annales Bogorienses 1990, 1, 4.

[10] Holt, S., Hartman, P., Journal of Industrial Microbiology 1994, 13, 3.

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This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
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