Soybean ubiquitous urease with purification facilitator: An addition to the moonlighting studies toolbox

BROLL, V.; LIGABUE-BRAUN, R.; MOYETTA, N.R.; SILVA, M.M.; CARLINI, C.R.; LOPES, F.C.; KAPPAUN, K. ; FRUTTERO, L.L.; POSTAL, M.; PASQUALI, G.; MARTINELLI, A.H.S.; DEFFERRARI, M.S.; TICHOTA, D.M.; DEMARTINI, D.R.; BECKER-RITT, A.B.

PROCESS BIOCHEMISTRY - (Print)

ELSEVIER SCI LTD

Lugar: Amsterdam; Año: 2017 vol. 53 p. 245 - 258

1359-5113

Ureases are nickel-dependent enzymes that catalyze the hydrolysis of urea to ammoniaand carbon dioxide. In soybean (Glycine max), the embryo-specific urease (eSBU), theubiquitous urease (uSBU), and a third isoform (SBU-III) are synthesized. Our group haspreviously demonstrated that eSBU, purified from seeds, has antifungal propertiesagainst phytopathogenic fungi, entomotoxicity against Dysdercus peruvianus, theability to induce blood platelet aggregation, and these properties are independent of itsenzymatic activity. Here we describe the biological properties of apo-uSBU fused toglutathione S-transferase (GST) produced in Escherichia coli. Removal of GST affectedapo-uSBU stability. We performed a Response Surface Methodology to optimize GSTuSBUproduction to 5 mg per liter and then bioassays were carried out. Therecombinant protein exhibited inhibitory effects on filamentous fungi and affectedfungal secondary metabolism. Candida albicans and C. tropicalis were also susceptibleto GST-uSBU and formed pseudo-hyphae. The fusion protein was toxic againstRhodnius prolixus, with the toxicity being accompanied by in vivo and in vitrohemocyte aggregation. Rabbit platelet also aggregated in the presence of GST-uSBU.Thus, uSBU displayed similar biological properties as previously described for eSBUeven when fused to GST, reinforcing the proposed role of ureases in plant defense.