Crystallization and preliminary X-ray crystallographic studies of the thermoactive pullulanase type I, hydrolyzing alpha-1,6 glycosidic linkages, from Fervidobacterium pennivorans Ven5.
Crystals of the thermoactive recombinant F. pennivorans type I pullulanase, purified from the supernatant of a Bacillus subtilis culture, have been obtained by the vapour-diffusion method in the presence of the inhibitor beta-cyclodextrin (2 mM) by mixing protein (15 mg ml(-1)) with an equal volume of crystallization solution containing 0.1 M bis-tris propane pH 6.5, 50 mM MgCl(2) and 15% polyethylene glycol 3350. Crystals diffracted to 3.0 A using conventional Cu Kalpha radiation and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 76.8, b = 96.2, c = 98. 5 A. The asymmetric unit contains one monomer. A preliminary 26% complete data set has been collected at 2.2 A resolution using synchrotron radiation.
Lebbink
J H
JH
Center for Structural Biochemistry, NOVUM, Karolinska Institutet, 14157 Huddinge, Sweden. joyce.lebbink@csb.ki.se
Bertoldo
C
C
Tibbelin
G
G
Andersen
J T
JT
Duffner
F
F
Antranikian
G
G
Ladenstein
R
R
D001419:Bacteria / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D006026:Glycoside Hydrolases / Q000737:chemistry*; D006027:Glycosides / Q000737:chemistry*; D006868:Hydrolysis; D011487:Protein Conformation
D006027:Glycosides; D006026:Glycoside Hydrolases; C100181:pullulanase