Expression, purification, crystallization and preliminary X-ray diffraction analysis of Thermotoga neapolitana beta-glucosidase B.
Beta-glucosidases belong to families 1, 3 and 9 of the glycoside hydrolases and act on cello-oligosaccharides. Family 1 and 3 enzymes are retaining and are reported to have transglycosylation activity, which can be used to produce oligosaccharides and glycoconjugates. Family 3 enzymes are less well characterized than their family 1 homologues and to date only two crystal structures have been solved. Here, the expression, purification, crystallization and X-ray diffraction data of a family 3 beta-glucosidase from the hyperthermophilic bacterium Thermotoga neapolitana are reported. Crystals of selenomethionine-substituted protein have also been grown. The crystals belong to space group C222(1), with unit-cell parameters a = 74.9, b = 127.0, c = 175.2 A. Native data have been collected to 2.4 A resolution and the structure has been solved to 2.7 A using the selenomethionine MAD method. Model building and refinement of the structure are under way.
Turner
Pernilla
P
Department of Biotechnology, Centre for Chemistry and Chemical Engineering, Lund University, Box 124, S-221 00 Lund, Sweden.
Pramhed
Anna
A
Kanders
Erik
E
Hedström
Martin
M
Karlsson
Eva Nordberg
EN
Logan
Derek T
DT
D000595:Amino Acid Sequence; D044602:Cellulases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003460:Crystallization; D004926:Escherichia coli; D016965:Gram-Negative Anaerobic Straight, Curved, and Helical Rods / Q000201:enzymology*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012645:Selenomethionine / Q000378:metabolism; D016415:Sequence Alignment; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; D012645:Selenomethionine; D044602:Cellulases