Baeyer-Villiger monooxygenases (BVMOs) are flavin-containing enzymes that catalyse a wide variety of oxidative reactions specifically on the carbonyl moiety of substrates, many of which are difficult to achieve using chemical approaches. Moreover, these proteins have great value in the areas of bioremediation and green chemistry. A novel BVMO gene in Acinetobacter radioresistens S13 was identified and using Clustal V its amino acid sequence was aligned with known BVMOs. Phylogenetic analysis demonstrated that it clustered together with BVMO from Mycobacterium, distant from the classical cycloalkanone MOs. After cloning and successful expression in E.coli, the BVMO protein was purified in high yields (20mg/L culture) using Nickel affinity chromatography. Preliminary characterisation showed a 57 kDa soluble protein with a tightly but non-covalently bound FAD. Due to the large substrate specificity of BVMOs, a 3D homology model of this enzyme was generated in order to select its putative substrates using in silico docking. Well-defined substrate profiles are available for several BVMOs and together with our model this may allow careful predictions for substrates of this novel enzyme.

A novel bacterial flavoenzyme (BVMO) for biotechnological applications

ZGRABLIC, IVAN;SADEGHI, JILA;MINERDI, Daniela;GILARDI, Gianfranco
2010

Abstract

Baeyer-Villiger monooxygenases (BVMOs) are flavin-containing enzymes that catalyse a wide variety of oxidative reactions specifically on the carbonyl moiety of substrates, many of which are difficult to achieve using chemical approaches. Moreover, these proteins have great value in the areas of bioremediation and green chemistry. A novel BVMO gene in Acinetobacter radioresistens S13 was identified and using Clustal V its amino acid sequence was aligned with known BVMOs. Phylogenetic analysis demonstrated that it clustered together with BVMO from Mycobacterium, distant from the classical cycloalkanone MOs. After cloning and successful expression in E.coli, the BVMO protein was purified in high yields (20mg/L culture) using Nickel affinity chromatography. Preliminary characterisation showed a 57 kDa soluble protein with a tightly but non-covalently bound FAD. Due to the large substrate specificity of BVMOs, a 3D homology model of this enzyme was generated in order to select its putative substrates using in silico docking. Well-defined substrate profiles are available for several BVMOs and together with our model this may allow careful predictions for substrates of this novel enzyme.
55th National Congress of the Italian Society of Biochemistry and Molecular Biology.
Milan
14-17 Sept
55th National Congress of the Italian Society of Biochemistry and Molecular Biology.
University of Milan Press
2010
83
83
9788884538208
BVMO; protein engineering; computer modelling; enzyme catalysis
I. Zgrablic; S.J. Sadeghi; D. Minerdi; G. Gilardi
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2318/78344
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