Crystallization and preliminary X-ray analysis of 4-pyridoxolactonase from Mesorhizobium loti.
4-pyridoxolactonase from Mesorhizobium loti MAFF303099 has been overexpressed in Escherichia coli. The recombinant enzyme was purified and was crystallized by the sitting-drop vapour-diffusion method using PEG 4000 and ammonium sulfate as precipitants. Crystals of the free enzyme (form I) and of the 5-pyridoxolactone-bound enzyme (form II) grew under these conditions. Crystals of form I diffracted to 2.0 A resolution and belonged to the monoclinic space group C2, with unit-cell parameters a = 77.93, b = 38.88, c = 81.60 A, beta = 117.33 degrees. Crystals of form II diffracted to 1.9 A resolution and belonged to the monoclinic space group C2, with unit-cell parameters a = 86.24, b = 39.35, c = 82.68 A, beta = 118.02 degrees. The calculated V(M) values suggested that the asymmetric unit contains one molecule in both crystal forms.
Matsuda
Sayoko
S
Faculty of Agriculture, Graduate School of Integral Arts and Science, Kochi University, Monobe, Nankoku, Kochi, Japan.
Yokochi
Nana
N
Yoshikane
Yu
Y
Kobayashi
Jun
J
Huy
Chu Nhat
CN
Baba
Seiki
S
Kuramitsu
Seiki
S
Mikami
Bunzo
B
Yagi
Toshiharu
T
D020561:Alphaproteobacteria / Q000201:enzymology*; D055162:Biocatalysis; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel
C050712:4-pyridoxolactonase; D002265:Carboxylic Ester Hydrolases