Cloning, expression, purification, crystallization and preliminary X-ray diffraction studies of the catalytic domain of a hyperthermostable endo-1,4-beta-D-mannanase from Thermotoga petrophila RKU-1.
Endo-1,4-beta-D-mannanases play key roles in seed germination and fruit ripening and have recently received much attention owing to their potential applications in the food, detergent and kraft pulp industries. In order to delineate their structural determinants for specificity and stability, X-ray crystallographic investigations combined with detailed functional studies are being performed. In this work, crystals of the catalytic domain of a hyperthermostable endo-1,4-beta-D-mannanase from Thermotoga petrophila RKU-1 were obtained from three different conditions, resulting in two crystalline forms. Crystals from conditions with phosphate or citrate salts as precipitant (CryP) belonged to space group P2(1)2(1)2(1), with unit-cell parameters a=58.76, b=87.99, c=97.34 A, while a crystal from a condition with ethanol as precipitant (CryE) belonged to space group I2(1)2(1)2(1), with unit-cell parameters a=91.03, b=89.97, c=97.89 A. CryP and CryE diffracted to resolutions of 1.40 and 1.45 A, respectively.
Santos
Camila Ramos
CR
Laboratório Nacional de Biociências (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, 13083-970 Campinas-SP, Brazil.
Squina
Fabio Marcio
FM
Navarro
Andreia Meza
AM
Ruller
Roberto
R
Prade
Rolf
R
Murakami
Mario Tyago
MT
D020134:Catalytic Domain; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004795:Enzyme Stability; D015870:Gene Expression; D016965:Gram-Negative Anaerobic Straight, Curved, and Helical Rods / Q000201:enzymology*; D008361:Mannosidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
D008361:Mannosidases; C041867:endo-1,4-beta-D-mannanase
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