02105naa a2200241 a 450000100080000000500110000800800410001910000170006024501050007726000090018252013850019165000250157670000230160170000210162470000220164570000220166770000140168970000190170370000210172270000190174370000250176277300760178719132532023-02-24       2011    bl uuuu     u00u1 u    #d1 aMULINARI, F.  aCharacterization of JBURE-IIb isoform of Canavalia ensiformis (L.) DC urease.h[electronic resource]  c2011  aUreases, nickel-dependent enzymes that catalyze the hydrolysis of urea into ammonia and bicarbonate, are widespread in plants, bacteria, and fungi. Previously, we cloned a cDNA encoding a Canavalia ensiformis urease isoform named JBURE-II, corresponding to a putative smaller urease protein (78 kDa) when compared to other plant ureases. Aiming to produce the recombinant protein, we obtained jbure-IIb, with different 3? and 5? ends, encoding a 90 kDa urease. Three peptides unique to the JBURE-II/-IIb protein were detected by mass spectrometry in seed extracts, indicating that jbure-II/-IIb is a functional gene. Comparative modeling indicates that JBURE-IIb urease has an overall shape almost identical to C. ensiformis major urease JBURE-I with all residues critical for urease activity. The cDNA was cloned into the pET101 vector and the recombinant protein was produced in Escherichia coli. The JBURE-IIb protein, although enzymatically inactive presumably due to the absence of Ni atoms in its active site, impaired the growth of a phytopathogenic fungus and showed entomotoxic properties, inhibiting diuresis of Rhodnius prolixus isolated Malpighian tubules, in concentrations similar to those reported for JBURE-I and canatoxin. The antifungal and entomotoxic properties of the recombinant JBURE-IIb apourease are consistent with a protective role of ureases in plants.  aCanavalia Ensiformis1 aBECKER-RITT, A. B.1 aDEMARTINI, D. R.1 aLIGABUE-BRAUN, R.1 aSTANISÇUASKI, F.1 aVERLI, H.1 aFRAGOSO, R. R.1 aSCHROEDER, E. K.1 aCARLINI, C. R.1 aGROSSI-de-SÁ, M. F.  tBiochimica et Biophysica Actagv. 1814, n. 12, p. 1758-1768, Dec. 2011.