Fungal β-N-acetylhexosaminidases are enzymes that are used in the chemoenzymatic synthesis of biologically interesting oligosaccharides. The enzyme from Aspergillus oryzae was produced and purified from its natural source and crystallized using the hanging-drop vapour-diffusion method. Diffraction data from two crystal forms (primitive monoclinic and primitive tetragonal) were collected to resolutions of 3.2 and 2.4 Å, respectively. Electrophoretic and quantitative N-terminal protein-sequencing analyses confirmed that the crystals are formed by a complete biologically active enzyme consisting of a glycosylated catalytic unit and a noncovalently attached propeptide.

Institute of Microbiology Academy of Sciences of the Czech Republic Vídeňská 1083 14220 Prague Czech Republic

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Berman, H. M., Westbrook, J., Feng, Z., Gilliland, G., Bhat, T. N., Weissig, H., Shindyalov, I. N. & Bourne, P. E. (2000). Nucleic Acids Res. 28, 235–242. PubMed PMC

Brune, D. C., Hampton, B., Kobayashi, R., Leone, J. W., Linse, K. D., Pohl, J., Thoma, R. S. & Denslow, N. D. (2007). J. Biomol. Tech. 18, 306–320. PubMed PMC

Bulawa, C. E. (1993). Annu. Rev. Microbiol. 47, 505–534. PubMed

Cheng, Q., Li, H., Merdek, K. & Park, J. T. (2000). J. Bacteriol. 182, 4836–4840. PubMed PMC

Cohen, E. (2001). Pest Manag. Sci. 57, 946–950. PubMed

Collaborative Computational Project, Number 4 (1994). Acta Cryst. D50, 760–763. PubMed

Ettrich, R., Kopecký, V., Hofbauerová, K., Baumruk, V., Novák, P., Pompach, P., Man, P., Plíhal, O., Kutý, M., Kulik, N., Sklenář, J., Ryšlavá, H., Křen, V. & Bezouška, K. (2007). BMC Struct. Biol. 7, 32. PubMed PMC

Evans, P. (2006). Acta Cryst. D62, 72–82. PubMed

Gooday, G. W., Zhu, W.-Y. & O’Donnell, R. W. (1992). FEMS Microbiol. Lett. 100, 387–391.

Huňková, Z., Křen, V., Ščigelová, M., Weignerova, L., Scheel, O. & Thiem, J. (1996). Biotechnol. Lett. 18, 725–730.

Křen, V., Ščigelová, M., Přikrylova, V., Havliček, V. & Sedmera, P. (1994). Biocatalysis, 10, 181–193.

Krist, P., Herkommerová-Rajnochová, E., Rauvolfová, J., Semeňuk, T., Vavrusková, P., Pavlíček, J., Bezouška, K., Petrus, L. & Křen, V. (2001). Biochem. Biophys. Res. Commun. 287, 11–20. PubMed

Leslie, A. G. W. (1999). Acta Cryst. D55, 1696–1702. PubMed

Mahuran, D. J. (1999). Biochim. Biophys. Acta, 1455, 105–138. PubMed

Mark, B. L., Mahuran, D. J., Cherney, M. M., Zhao, D., Knapp, S. & James, M. N. (2003). J. Mol. Biol. 327, 1093–1109. PubMed PMC

Mark, B. L., Vocadlo, D. J., Zhao, D., Knapp, S., Withers, S. G. & James, M. N. (2001). J. Biol. Chem. 276, 42131–42137. PubMed

Matthews, B. W. (1968). J. Mol. Biol. 33, 491–497. PubMed

Minor, W., Cymborowski, M., Otwinowski, Z. & Chruszcz, M. (2006). Acta Cryst. D62, 859–866. PubMed

Plíhal, O., Sklenář, J., Hofbauerová, K., Novák, P., Man, P., Pompach, P., Kavan, D., Ryšlavá, H., Weignerová, L., Charvátová-Pišvejcová, A., Křen, V. & Bezouška, K. (2007). Biochemistry, 46, 2719–2734. PubMed

Pompach, P., Man, P., Kavan, D., Hofbauerová, K., Kumar, V., Bezouška, K., Havlíček, V. & Novák, P. (2009). J. Mass Spectrom. 44, 1571–1578. PubMed

Prag, G., Papanikolau, Y., Tavlas, G., Vorgias, C. E., Petratos, K. & Oppenheim, A. B. (2000). J. Mol. Biol. 300, 611–617. PubMed

Rajnochová, E., Dvořakova, J., Huňková, Z. & Křen, V. (1997). Biotechnol. Lett. 19, 869–872.

Slámová, K., Bojarová, P., Petrásková, L. & Křen, V. (2010). Biotechnol. Adv. 28, 682–693. PubMed

Weignerová, L., Vavrušková, P., Pišvejcová, A., Thiem, J. & Křen, V. (2003). Carbohydr. Res. 338, 1003–1008. PubMed

Weiss, M. S. (2001). J. Appl. Cryst. 34, 130–135.

Computational study of β-N-acetylhexosaminidase from Talaromyces flavus, a glycosidase with high substrate flexibility