Preparation and crystallization of selenomethionyl dextranase from Penicillium minioluteum expressed in Pichia pastoris.
Dextranase from the fungus Penicillium minioluteum hydrolyses alpha-1,6-glycosidic bonds in dextran polymers. The enzyme has been expressed in Pichia pastoris in the presence of selenomethionine (SeMet). The level of SeMet incorporation was estimated by amino-acid analysis to be 50%. The protein has been crystallized in space group P2(1)2(1)2, with unit-cell parameters a = 103.6, b = 115.3, c = 49.8 A and one molecule per asymmetric unit. The crystals diffract to 2.0 A and the presence of SeMet in the crystals has been confirmed by an X-ray absorption spectrum.
Larsson
Anna M
AM
Department of Cell and Molecular Biology, Uppsala University, Biomedical Centre, Box 596, SE-751 24 Uppsala, Sweden.
Ståhlberg
Jerry
J
Jones
T Alwyn
TA
D003460:Crystallization; D018360:Crystallography, X-Ray; D003910:Dextranase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D010407:Penicillium / Q000201:enzymology*; D010843:Pichia / Q000235:genetics; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012645:Selenomethionine / Q000737:chemistry*
D011994:Recombinant Proteins; D012645:Selenomethionine; D003910:Dextranase