Crystallization of carbohydrate oxidase from Microdochium nivale.
Microdochium nivale carbohydrate oxidase was produced by heterologous recombinant expression in Aspergillus oryzae, purified and crystallized. The enzyme crystallizes with varying crystal morphologies depending on the crystallization conditions. Several different crystal forms were obtained using the hanging-drop vapour-diffusion method, two of which were used for diffraction measurements. Hexagon-shaped crystals (form I) diffracted to 2.66 A resolution, with unit-cell parameters a = b = 55.7, c = 610.4 A and apparent space group P6(2)22. Analysis of the data quality showed almost perfect twinning of the crystals. Attempts to solve the structure by molecular replacement did not give satisfactory results. Recently, clusters of rod-shaped crystals (form II) were grown in a solution containing PEG MME 550. These crystals belonged to the monoclinic system C2, with unit-cell parameters a = 132.9, b = 56.6, c = 86.5 A, beta = 95.7 degrees . Data sets were collected to a resolution of 2.4 A. The structure was solved by the molecular-replacement method. Model refinement is currently in progress.
Dusková
Jarmila
J
Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám. 2, Praha 6, Czech Republic. duskova@imc.cas.cz
Dohnálek
Jan
J
Skálová
Tereza
T
Østergaard
Lars Henrik
LH
Fuglsang
Claus Crone
CC
Kolenko
Petr
P
Stepánková
Andrea
A
Hasek
Jindrich
J
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000235:genetics; D002237:Carbohydrate Dehydrogenases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D003625:Data Collection; D005690:Galactose / Q000378:metabolism; D005822:Genetic Vectors; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017386:Sequence Homology, Amino Acid; D013223:Statistics as Topic; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors; D014961:X-Ray Diffraction; D013098:Xylariales / Q000201:enzymology* / Q000235:genetics; D014994:Xylose / Q000378:metabolism
D014994:Xylose; D002237:Carbohydrate Dehydrogenases; D005690:Galactose