1887

Abstract

The urease from the picoplanktonic oceanic sp. strain PCC 9511 was purified 900-fold to a specific activity of 94.6 μmol urea min (mg protein) by heat treatment and liquid chromatography methods. The enzyme, with a molecular mass of 168 kDa as determined by gel filtration, is the smallest urease known to date. Three different subunits with apparent molecular masses of 11 kDa (γ or UreA; predicted molecular mass 11 kDa), 13 kDa (β or UreB; predicted molecular mass 12 kDa) and 63 kDa (α or UreC; predicted molecular mass 62 kDa) were detected in the native enzyme, suggesting a quaternary structure of (αβγ). The of the purified enzyme was determined as being 023 mM urea. The urease activity was inhibited by HgCl, acetohydroxamic acid and EDTA but neither by boric acid nor by L-methionine-DL-sulfoximine. Degenerate primers were designed to amplify a conserved region of the gene. The amplification product was then used as a probe to clone a 57 kbp fragment of the sp. strain PCC 9511 genome. The nucleotide sequence of this DNA fragment revealed two divergently orientated gene clusters, and , encoding the urease subunits, UreA, UreB and UreC, and the urease accessory molecules UreD, UreE, UreF and UreG. A putative NtcA-binding site was found upstream from , indicating that this gene cluster might be under nitrogen control.

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2000-12-01
2019-12-08
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