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Cloning, expression and characterization of a versatile Baeyer-Villiger monooxygenase from Dietzia sp. D5.

Bisagni, Serena LU ; Hatti-Kaul, Rajni LU and Mamo, Gashaw LU (2014) In AMB Express 4.
Abstract
A novel BVMO encoding gene was identified from a draft genome sequence of a newly isolated strain of Dietzia. Analysis of the protein sequence revealed that it belongs to a group of BVMOs whose most characterized member is cyclopentadecanone monooxygenase (CPDMO). The gene was PCR amplified, cloned and successfully expressed in E. coli. The expressed recombinant enzyme was purified using metal affinity chromatography. Characterization of the purified enzyme revealed that it has a broad substrate scope and oxidized different compounds including substituted and unsubstituted alicyclic, bicyclic-, aliphatic-ketones, ketones with an aromatic moiety, and sulfides. The highest activities were measured for 2- and 3-methylcyclohexanone,... (More)
A novel BVMO encoding gene was identified from a draft genome sequence of a newly isolated strain of Dietzia. Analysis of the protein sequence revealed that it belongs to a group of BVMOs whose most characterized member is cyclopentadecanone monooxygenase (CPDMO). The gene was PCR amplified, cloned and successfully expressed in E. coli. The expressed recombinant enzyme was purified using metal affinity chromatography. Characterization of the purified enzyme revealed that it has a broad substrate scope and oxidized different compounds including substituted and unsubstituted alicyclic, bicyclic-, aliphatic-ketones, ketones with an aromatic moiety, and sulfides. The highest activities were measured for 2- and 3-methylcyclohexanone, phenylacetone, bicyclo-[3.2.0]-hept-2-en-6-one and menthone. The enzyme was optimally active at pH 7.5 and 35°C, a temperature at which its half-life was about 20 hours. The stability studies have shown that this enzyme is more stable than all other reported BVMOs except the phenylacetone monooxygenase from the thermophilic organism Thermobifida fusca. (Less)
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publication status
published
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in
AMB Express
volume
4
publisher
Springer Open
external identifiers
  • pmid:24949258
  • scopus:84899645132
ISSN
2191-0855
DOI
10.1186/s13568-014-0023-1
language
English
LU publication?
yes
id
ff12e32f-d0e8-442e-a278-40f8dabe1c73 (old id 4527858)
date added to LUP
2014-08-08 14:56:41
date last changed
2017-05-21 04:00:31
@article{ff12e32f-d0e8-442e-a278-40f8dabe1c73,
  abstract     = {A novel BVMO encoding gene was identified from a draft genome sequence of a newly isolated strain of Dietzia. Analysis of the protein sequence revealed that it belongs to a group of BVMOs whose most characterized member is cyclopentadecanone monooxygenase (CPDMO). The gene was PCR amplified, cloned and successfully expressed in E. coli. The expressed recombinant enzyme was purified using metal affinity chromatography. Characterization of the purified enzyme revealed that it has a broad substrate scope and oxidized different compounds including substituted and unsubstituted alicyclic, bicyclic-, aliphatic-ketones, ketones with an aromatic moiety, and sulfides. The highest activities were measured for 2- and 3-methylcyclohexanone, phenylacetone, bicyclo-[3.2.0]-hept-2-en-6-one and menthone. The enzyme was optimally active at pH 7.5 and 35°C, a temperature at which its half-life was about 20 hours. The stability studies have shown that this enzyme is more stable than all other reported BVMOs except the phenylacetone monooxygenase from the thermophilic organism Thermobifida fusca.},
  articleno    = {23},
  author       = {Bisagni, Serena and Hatti-Kaul, Rajni and Mamo, Gashaw},
  issn         = {2191-0855},
  language     = {eng},
  publisher    = {Springer Open},
  series       = {AMB Express},
  title        = {Cloning, expression and characterization of a versatile Baeyer-Villiger monooxygenase from Dietzia sp. D5.},
  url          = {http://dx.doi.org/10.1186/s13568-014-0023-1},
  volume       = {4},
  year         = {2014},
}