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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Structure and peroxidase activity of ferric Streptomyces clavuligerus orf10-encoded protein P450CLA: UV-visible, CD, MCD and EPR spectroscopic characterization

Texto completo
Autor(es):
Goto, Leandro S. [1] ; Hokka, Carlos O. [1] ; Lima, Jose F. [2] ; Prieto, Tatiana [2] ; Araujo, Ana Paula U. [2] ; Nantes, Iseli L. [3] ; Nascimento, Otaciro R. [2, 3]
Número total de Autores: 7
Afiliação do(s) autor(es):
[1] Univ Fed Sao Carlos, Dept Engn Quim, Grp Engn Bioquim, BR-13565905 Sao Carlos, SP - Brazil
[2] Univ Sao Paulo, Inst Fis Sao Carlos, Grp Biofis Mol Sergio Mascarenhas, BR-13560970 Sao Carlos, SP - Brazil
[3] Univ Fed ABC, Ctr Ciencias Nat & Humanas, Grp Nanoestruturas Biol & Mat Avancados, BR-09210170 Santo Andre, SP - Brazil
Número total de Afiliações: 3
Tipo de documento: Artigo Científico
Fonte: Journal of the Brazilian Chemical Society; v. 23, n. 5, p. 913-920, MAY 2012.
Citações Web of Science: 2
Resumo

The present study reports the spectroscopic characterization by UV-visible absorption spectroscopy, magnetic circular dichroism (MCD) and electron paramagnetic resonance (EPR) of the recombinant orf10-encoded P450-camphor like protein (P450CLA)of Streptomyces clavuligerus expressed in Escherichia coli Rosetta in the native form and associated to external ligands containing the β-lactam, oxazole and alkylamine-derived (alcohol) moieties of the clavulamic acid. Considering the diversity of potential applications for the enzyme, the reactivity with tert-butylhydroperoxide (tert-BuOOH) was also characterized. P450CLA presents a covalently bound heme group and exhibited the UV-visible, CD and MCD spectral features of P450CAM including the fingerprint Soret band at 450 nm generated by the ferrous CO-complex. P450CLA was converted to high valence species by tert-BuOOH and promoted homolytic scission of the O-O bond. The radical profile of the reaction was tert-butyloxyl as primary and methyl and butylperoxyl as secondary radicals. The secondary methyl and butylperoxyl radicals resulted respectively from the β-scission of the alkoxyl radical and from the reaction of methyl radical with molecular oxygen. (AU)

Processo FAPESP: 06/59693-1 - Caracterização de produtos gênicos pertinentes à via de síntese do ácido clavulânico de Streptomyces clavuligerus ATCC 27064
Beneficiário:Leandro Seiji Goto
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
Processo FAPESP: 08/04948-0 - Estrutura e reatividade de hemoproteínas, hemopeptídeos e porfirinas em meios homogêneos e heterogêneos: aspectos básicos e aplicados à nanotecnologia
Beneficiário:Iseli Lourenço Nantes Cardoso
Modalidade de apoio: Auxílio à Pesquisa - Temático