The ascending application of enzymes in organic synthesis creates a growing demand for novel biocatalysts. The applied methods for their identification range from microbial enrichment cultures over metagenome screenings to solely computational methods. In this communication, we demonstrate a straightforward screening approach for the detection of novel biocatalysts in fungi belonging to the phylum Basidiomycota. It basically relies on mincing of the whole fruit bodies of freshly collected mushrooms with subsequent direct screening. Suitability was demonstrated with eight different mushrooms which were investigated for carbonyl reductase activity on sterically demanding carbonyl compounds. The results indicate the presence of potentially useful carbonyl reductases (KREDs) in all tested fungi. Closer characterization of the preparation from pigskin poison puffball (Scleroderma citrinum) showed the presence of KRED exhibiting a broad substrate range. Thus, applicability of this low-tech screening approach could be verified in this study.
Bornscheuer, U.T., Huisman, G.W., Kazlauskas, R.J., Lutz, S., Moore, J.C. and Robins, K. (2012) Engineering the Third Wave of Biocatalysis. Nature, 485, 185-194.
http://dx.doi.org/10.1038/nature11117
Sanchez, S. and Demain, A.L. (2011) Enzymes and Bioconversions of Industrial, Pharmaceutical, and Biotechnological Significance. Organic Process Research & Development, 15, 224-230. http://dx.doi.org/10.1021/op100302x
Ni, Y. and Xu, J.H. (2012) Biocatalytic Ketone Reduction: A Green and Efficient Access to Enantiopure Alcohols. Biotechnology Advances, 30, 1279-1288.
http://dx.doi.org/10.1016/j.biotechadv.2011.10.007
Gabor, E.M., de Vries, E.J. and Janssen, D.B. (2004) Construction, Characterization, and Use of Small-Insert Gene Banks of DNA Isolated from Soil and Enrichment Cultures for the Recovery of Novel Amidases. Environmental Microbiology, 6, 948-958.
http://dx.doi.org/10.1111/j.1462-2920.2004.00643.x
Voget, S., Leggewie, C., Uesbeck, A., Raasch, C., Jaeger, K.E. and Streit, W.R. (2003) Prospecting for Novel Biocatalysts in a Soil Metagenome. Applied and Environmental Microbiology, 69, 6235-6242. http://dx.doi.org/10.1128/AEM.69.10.6235-6242.2003
Wasser, P. and Weis, L. (1999) Medicinal Properties of Substances Occurring in Higher Basidiomycetes Mushrooms: Current Perspectives. International Journal of Medicinal Mushrooms, 1, 31-62. http://dx.doi.org/10.1615/IntJMedMushrooms.v1.i1.30
Buttner, E., Ullrich, R., Strittmatter, E., Piontek, K., Plattner, D.A., Hofrichter, M. and Liers, C. (2015) Oxidation and Nitration of Mononitrophenols by a DyP-Type Peroxidase. Archives of Biochemistry and Biophysics, 574, 86-92.
http://dx.doi.org/10.1016/j.abb.2015.03.003
Dashtban, M., Schraft, H., Syed, T.A. and Qin, W. (2010) Fungal Biodegradation and Enzymatic Modification of Lignin. International Journal of Biochemistry and Molecular Biology, 1, 36-50.
Bertrand, T., Jolivalt, C., Briozzo, P., Caminade, E., Joly, N., Madzak, C. and Mougin, C. (2002) Crystal Structure of a Four-Copper Laccase Complexed with an Arylamine: Insights into Substrate Recognition and Correlation with Kinetics. Biochemistry, 41, 7325-7333.
http://dx.doi.org/10.1021/bi0201318
Evans, C.S. (1985) Laccase Activity in Lignin Degradation by Coriolus versicolor in Vivo and in Vitro Studies. FEMS Microbiology Letters, 27, 339-343.
http://dx.doi.org/10.1111/j.1574-6968.1985.tb00693.x
Ullah, M.A., Bedford, C.T. and Evans, C.S. (2000) Reactions of Pentachlorophenol with Laccase from Coriolus versicolor. Applied Microbiology and Biotechnology, 53, 230-234.
http://dx.doi.org/10.1007/s002530050013
Couto, R.S. and Herrera, T.J.L. (2006) Industrial and Biotechnological Applications of Laccases: A Review. Biotechnology Advances, 24, 500-513.
http://dx.doi.org/10.1016/j.biotechadv.2006.04.003
Peláez, F., Martínez, M.J. and Martínez, A.T. (1995) Screening of 68 Species of Basidiomycetes for Enzymes Involved in Lignin Degradation. Mycological Research, 99, 37-42.
http://dx.doi.org/10.1016/S0953-7562(09)80313-4
Breuer, M., Ditrich, K., Habicher, T., Hauer, B., Keβeler, M., Stürmer, R. and Zelinski, T. (2004) Industrielle Verfahren zur Herstellung von optisch aktiven Zwischenprodukten. Angewandte Chemie, 116, 806-843. http://dx.doi.org/10.1002/ange.200300599
Hoyos, P., Sinisterra, J.V., Molinari, F., Alcántara, A.R. and Domínguez de María, P. (2009) Biocatalytic Strategies for the Asymmetric Synthesis of Alpha-Hydroxy Ketones. Accounts of Chemical Research, 43, 288-299. http://dx.doi.org/10.1021/ar900196n
Loderer, C., Dhoke, G.V., Davari, M.D., Kroutil, W., Schwaneberg, U., Bocola, M. and Ansorge-Schumacher, M.B. (2015) Investigation of Structural Determinants for the Substrate Specificity in the Zinc-Dependent Alcohol Dehydrogenase CPCR2 from Candida parapsilosis. Chembiochem, 16, 1512-1519. http://dx.doi.org/10.1002/cbic.201500100
Park, Y.J., Baek, J.H., Lee, S., Kim, C., Rhee, H., Kim, H., Seo, J.S., Park, H.R., Yoon, D.E., Nam, J.Y., Kim, H.I., Kim, J.G., Yoon, H., Kang, H.W., Cho, J.Y., Song, E.S., Sung, G.H., Yoo, Y.B., Lee, C.S., Lee, B.M. and Kong, W.S. (2014) Whole Genome and Global Gene Expression Analyses of the Model Mushroom Flammulina velutipes Reveal a High Capacity for Lignocellulose Degradation. PLoS ONE, 9, e93560.
http://dx.doi.org/10.1371/journal.pone.0093560
Peters, J., Minuth, T. and Kula, M.R. (1993) A Novel NADH-Dependent Carbonyl Reductase with an Extremely Broad Substrate Range from Candida parapsilosis: Purification and Characterization. Enzyme and Microbial Technology, 15, 950-958.
http://dx.doi.org/10.1016/0141-0229(93)90171-W
Jakoblinnert, A., Bocola, M., Bhattacharjee, M., Steinsiek, S., Bonitz-Dulat, M., Schwaneberg, U. and Ansorge-Schumacher, M.B. (2012) Who’s Who? Allocation of Carbonyl Reductase Isoenzymes from Candida parapsilosis by Combining Bio- and Computational Chemistry. Chembiochem, 13, 803-809. http://dx.doi.org/10.1002/cbic.201200023