Crystallization and preliminary X-ray diffraction analysis of levansucrase (LsdA) from Gluconacetobacter diazotrophicus SRT4.
The endophytic bacterium Gluconacetobacter diazotrophicus SRT4 secretes a constitutively expressed levansucrase (LsdA; EC 2.4.1.10), which converts sucrose to fructo-oligosaccharides and levan. Fully active LsdA was purified to high homogeneity by non-denaturing reversed-phase HPLC and was crystallized at room temperature by the hanging-drop vapour-diffusion method using ammonium sulfate and ethanol as precipitants. The crystals are extremely sensitive, but native data have been collected to 2.5 A under cryogenic conditions using synchrotron radiation. LsdA crystals belong to the orthorhombic space group P22(1)2(1) or P2(1)2(1)2, with unit-cell parameters a = 53.80, b = 119.39, c = 215.10 A.
Martínez-Fleites
Carlos
C
Physical Chemistry Division, Center for Genetic Engineering and Biotechnology, PO Box 6162, Havana 10600, Cuba.
Tarbouriech
Nicolas
N
Ortiz-Lombardia
Miguel
M
Taylor
Edward
E
Rodríguez
Armando
A
Ramírez
Ricardo
R
Hernández
Lázaro
L
Davies
Gideon J
GJ
D003460:Crystallization; D018360:Crystallography, X-Ray; D041821:Gluconacetobacter / Q000201:enzymology*; D006602:Hexosyltransferases / Q000737:chemistry*
D006602:Hexosyltransferases; C021513:levansucrase