Identification of oxidoreductases from the petroleum Bacillus safensis strain

Francine S A da Fonseca; Célio F F Angolini; Marco A Zezzi Arruda; Cícero A L Junior; Clelton A Santos; Antonio M Saraiva; Eduardo Pilau; Anete P Souza; Prianda R Laborda; Patrícia F L de Oliveira; Valéria M de Oliveira; Francisco de Assis M Reis; Anita J Marsaioli

doi:10.1016/j.btre.2015.09.001

Identification of oxidoreductases from the petroleum Bacillus safensis strain

Biotechnol Rep (Amst). 2015 Sep 5:8:152-159. doi: 10.1016/j.btre.2015.09.001. eCollection 2015 Dec.

Affiliations

¹ Chemistry Institute, University of Campinas, P.B 6154, 13083-970 Campinas, São Paulo, Brazil; Institute of Agricultural Sciences, Federal University of Minas Gerais, 39404-547 Minas Gerais, Brazil.
² Chemistry Institute, University of Campinas, P.B 6154, 13083-970 Campinas, São Paulo, Brazil.
³ Center for Molecular Biology and Genetic Engineering, University of Campinas, 13083-875 São Paulo, Brazil.
⁴ Chemistry Institute, University of Campinas, P.B 6154, 13083-970 Campinas, São Paulo, Brazil; National Institute of Metrology, Quality and Technology - INMETRO, Xerém, Rio de Janeiro, Brazil.
⁵ Microbial Resources Division, Research Center for Chemistry, Biology and Agriculture (CPQBA), University of Campinas, CP 6171, CEP 13081-970, Campinas, SP, Brazil.

Abstract

A gram-positive bacterium, denominated CFA-06, was isolated from Brazilian petroleum in the Campos Basin and is responsible for the degradation of aromatic compounds and petroleum aromatic fractions. The CFA-06 strain was identified as Bacillus safensis using the 16S rRNA and gyrase B sequence. Enzymatic assays revealed the presence of two oxidoreductases: a catalase and a new oxidoreductase. The oxidoreductases were enzymatically digested and analyzed via ESI-LTQ-Orbitrap mass spectrometry. The mass data revealed a novel oxidoreductase (named BsPMO) containing 224 amino acids and 89% homology with a hypothetic protein from B. safensis (CFA-06) and a catalase (named BsCat) with 491 amino acids and 60% similarity with the catalase from Bacillus pumilus (SAFR-032). The new protein BsPMO contains iron atom(s) and shows catalytic activity toward a monooxygenase fluorogenic probe in the presence of cofactors (NADH, NADPH and NAD). This study enhances our knowledge of the biodegradation process of petroleum by B. safensis.

Keywords: Bacillus safensis; Catalase; Monooxygenase.