Overexpression, crystallization and preliminary X-ray crystallographic analysis of D-ribose-5-phosphate isomerase from Clostridium thermocellum.

Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Junho JungLin Woo Kang

Abstract

Rare sugars are used for many industrial and medical purposes and are produced by the interconversion between aldoses and ketoses catalyzed by sugar and sugar-phosphate isomerases. Recently, Clostridium thermocellum d-ribose-5-phosphate isomerase (CTRPI), an aldose-ketose isomerase, was cloned in order to synthesize d-allose and its substrate specificity was further characterized for industrial usage. CTRPI has a novel substrate specificity that differs from those of other isomerases, which have broad substrate specificities. CTRPI prefers aldose substrates such as l-talose, d-ribose and d-allose. CTRPI was purified and crystallized in order to determine its three-dimensional structure and thus to elucidate its enzymatic reaction mechanism and understand its substrate specificity. The crystal belonged to the trigonal space group P3(2)21, with unit-cell parameters a = b = 69.5, c = 154.4 angstrom, and diffracted to 1.9 angstrom resolution. According to Matthews coefficient calculations, the crystallographic structure consists of a dimer in the asymmetric unit, with a V(M) of 3.2 angstrom(3) Da(-1) and a solvent content of 61.7%.

References

Apr 28, 1968·Journal of Molecular Biology·B W Matthews
Dec 1, 1996·Glycobiology·S MuniruzzamanA D Elbein
May 25, 2002·Journal of Nutritional Science and Vitaminology·Tatsuhiro MatsuoKen Izumori
Jan 4, 2003·Journal of Medicinal Food·Gilbert V Levin
Jul 6, 2004·Journal of Hepato-biliary-pancreatic Surgery·Mohammad Akram HossainHajime Maeta
Nov 6, 2008·Journal of Biotechnology·Ran-Young YoonDeok-Kun Oh

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Citations

Dec 7, 2010·Applied Microbiology and Biotechnology·Soo-Jin YeomDeok-Kun Oh
Apr 10, 2012·Biochemical and Biophysical Research Communications·Preet Kamal KaurSushma Singh

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