"5"^^ . "0"^^ . "5"^^ . "Structure, function and regulation of FerB, a broad-specificity bacterial oxidoreductase of a potential ecotechnological relevance"@en . . . "The FerB enzyme of Paracoccus denitrificans was originally discovered by our group as an NAD(P)H-dependent oxidoreductase able to reduce ferric complexes, quinones and chromate. Given the potential of using FerB for bioremediation of chromium(VI) and possibly also other heavy metals, we now propose to undertake a systematic investigation of this enzyme by conventional biochemistry, structural and molecular biology. The gene encoding FerB will be amplified and cloned based on 27 known N-terminal aminoacid residues of the protein and the already published P. denitrificans whole genome sequence. Heterologous expression of the ferB gene is expected to yield large quantities of pure recombinant protein for studies on substrate specificity, stoichiometry of electron transfer and redox kinetics of the bound flavin coenzyme. Trials will also set out to crystallise FerB and collect diffraction data for structural analysis. Suicide vectors and triparental mating will serve for construction of mutant strains"@en . "2009-12-31+01:00"^^ . "1"^^ . . . "NAD(P)H:acceptor oxidoreductase; flavin; chromate; bioremediation; Paracoccus denitrificans"@en . "1070"^^ . . "1070"^^ . . "Enzym FerB z Paracoccus denitrificans byl\u00A0 p\u016Fvodn\u011B objeven na\u0161\u00ED skupinou jako NAD(P)H z\u00E1visl\u00E1 oxidoreduktasa schopn\u00E1 redukovat \u017Eelezit\u00E9 komplexy, chinony a chroman. Vzhledem k mo\u017En\u00E9mu vyu\u017Eit\u00ED FerB pro bioremediaci \u0161estimocn\u00E9ho chromu a snad i jin\u00FDch t\u011B\u017Ek\u00FDch kov\u016F navrhujeme nyn\u00ED systematick\u00FD v\u00FDzkum tohoto enzymu metodami obecn\u00E9 biochemie a strukturn\u00ED a molekul\u00E1rn\u00ED biologie. Gen k\u00F3duj\u00EDc\u00ED FerB bude amplifikov\u00E1n a klonov\u00E1n na z\u00E1klad\u011B zn\u00E1m\u00FDch 27 aminokyselinov\u00FDch zbytk\u016F na N-konci proteinu a ji\u017E zve\u0159ejn\u011Bn\u00E9 sekvence cel\u00E9ho genomu P. denitrificans. O\u010Dek\u00E1v\u00E1me, \u017Ee heterologn\u00ED exprese genu ferB poskytne velk\u00E1 mno\u017Estv\u00ED \u010Dist\u00E9ho rekombinantn\u00EDho proteinu pro studie zam\u011B\u0159en\u00E9 na substr\u00E1tovou specifitu, stechiometrii p\u0159enosu elektron\u016F a redoxn\u00ED kinetiku v\u00E1zan\u00E9ho flavinov\u00E9ho koenzymu. Budou tak\u00E9 zah\u00E1jeny pokusy o krystalizaci FerB a sb\u011Br difrak\u010Dn\u00EDch dat pro strukturn\u00ED anal\u00FDzu. Sebevra\u017Eedn\u00E9 vektory a triparent\u00E1ln\u00ED k\u0159\u00ED\u017Een\u00ED poslou\u017E\u00ED k sestrojen\u00ED mutantn\u00EDch kmen\u016F bez FerB a report\u00E9rov\u00FDch kmen\u016F nesouc\u00EDch\u00A0"@cs . . "Struktura, funkce a regulace FerB, \u0161irokospecifick\u00E9 bakteri\u00E1ln\u00ED oxidoreduktasy s mo\u017En\u00FDm v\u00FDznamem pro ekotechnologii"@cs . . . . "2007-01-01+01:00"^^ . . . "0"^^ .