Expression, purification, crystallization and preliminary X-ray crystallographic studies of the trehalulose synthase MutB from Pseudomonas mesoacidophila MX-45

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Stéphanie RavaudNushin Aghajari

Abstract

The trehalulose synthase (MutB) from Pseudomonas mesoacidophila MX-45, belonging to glycoside hydrolase family 13, catalyses the isomerization of sucrose to trehalulose (alpha-D-glucosylpyranosyl-1,1-D-fructofuranose) and isomaltulose (alpha-D-glucosylpyranosyl-1,6-D-fructofuranose) as main products and glucose and fructose in residual amounts from the hydrolytic reaction. To date, a three-dimensional structure of a sucrose isomerase that produces mainly trehalulose, as is the case for MutB, has been lacking. Crystallographic studies of this 64 kDa enzyme have therefore been initiated in order to contribute to the understanding of the molecular basis of sucrose decomposition, isomerization and of the selectivity of this enzyme that leads to the formation of different products. The MutB protein has been overexpressed, purified and crystallized using the hanging-drop vapour-diffusion method. Two different crystal forms have been obtained: one diffracts X-rays to 1.6 A resolution using synchrotron radiation and belongs to space group P1, with unit-cell parameters a = 63.8, b = 72.0, c = 82.2 A, alpha = 67.5, beta = 73.1, gamma = 70.8 degrees, while the other form diffracts to 1.8 A resolution using synchrotron radiation and belong...Continue Reading

References

Apr 28, 1968·Journal of Molecular Biology·B W Matthews
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Sep 22, 2001·Acta Crystallographica. Section D, Biological Crystallography·J Navaza
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Jun 24, 2003·The Journal of Biological Chemistry·Daohai ZhangKunchithapadam Swaminathan

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Citations

Mar 3, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Stéphanie RavaudNushin Aghajari
May 29, 2014·Applied Microbiology and Biotechnology·Wanmeng MuBo Jiang
Feb 7, 2013·Acta Crystallographica. Section D, Biological Crystallography·Alexandra LipskiNushin Aghajari
Jun 29, 2007·The Journal of Biological Chemistry·Stéphanie RavaudNushin Aghajari

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