Crystallization and preliminary X-ray diffraction analysis of a thermostable endo-1,5-alpha-L-arabinanase from Bacillus thermodenitrificans TS-3.
A thermostable endo-1,5-alpha-L-arabinanase ABN-TS from Bacillus thermodenitrificans TS-3 with a molecular weight of 35 kDa was crystallized by the hanging-drop vapour-diffusion method using sodium citrate as a precipitant. The crystals were loop-mounted in a cryoprotectant solution containing 28%(w/v) sucrose and 1 M sodium citrate pH 6.0 and flash-cooled. Sucrose was selected as the most suitable cryoprotectant. The crystal belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 40.3, b = 77.8, c = 89.7 angstroms. The calculated VM based on one molecule per asymmetric unit was 2.0 angstroms3 Da(-1). A complete data set from a frozen crystal was collected to 1.9 angstroms resolution using synchrotron radiation at SPring-8. A molecular-replacement solution was obtained using the structure of alpha-arabinanase 43A from Cellvibrio japonicus.
Yamaguchi
Asako
A
Research Institute for Advanced Science and Technology, Osaka Prefecture University, Sakai, Osaka 599-8570, Japan. asako-y@biochem.osakafu-u.ac.jp
Tada
Toshiji
T
Nakaniwa
Tetsuko
T
Kitatani
Tomoya
T
Takao
Makoto
M
Sakai
Takuo
T
Nishimura
Keiichiro
K
D001407:Bacillus / Q000201:enzymology*; D020633:Cellvibrio / Q000378:metabolism; D002951:Citrates / Q000737:chemistry / Q000494:pharmacology; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D005990:Glycerol / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000737:chemistry*; D007700:Kinetics; D000077559:Sodium Citrate; D013395:Sucrose / Q000737:chemistry / Q000494:pharmacology; D014961:X-Ray Diffraction / Q000379:methods*
D002951:Citrates; D000077559:Sodium Citrate; D013395:Sucrose; D006026:Glycoside Hydrolases; C082526:arabinan endo-1,5-alpha-L-arabinosidase; D005990:Glycerol