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Cloning and expression of a Baeyer-Villiger monooxygenase oxidizing linear aliphatic ketones from Dietzia sp. D5

Bisagni, Serena LU ; Smus, Justyna; Chávez, Georgina LU ; Hatti-Kaul, Rajni LU and Mamo, Gashaw LU (2014) In Journal of Molecular Catalysis B: Enzymatic 109. p.161-169
Abstract
A Baeyer-Villiger monooxygenase has been identified in the genome sequence of Dietzia sp. D5. Sequence similarity search revealed that the enzyme belongs to a group of BVMOs that are closely related to ethionamide monooxygenase from Mycobacterium tuberculosis (EthA). The BVMO was expressed in E. coli BL21-CodonPlus(DE3)-RP and the best expression was achieved when the E. coli cells were cultivated in terrific broth (TB) at 15 degrees C and induced with 0.1 mM of IPTG. Since the purified enzyme did not show any measurable activity, the substrate scope of the BVMO has been determined using whole-cell and crude cell extract systems. The enzyme was most active towards linear aliphatic substrates. However, it has shown a moderate degree of... (More)
A Baeyer-Villiger monooxygenase has been identified in the genome sequence of Dietzia sp. D5. Sequence similarity search revealed that the enzyme belongs to a group of BVMOs that are closely related to ethionamide monooxygenase from Mycobacterium tuberculosis (EthA). The BVMO was expressed in E. coli BL21-CodonPlus(DE3)-RP and the best expression was achieved when the E. coli cells were cultivated in terrific broth (TB) at 15 degrees C and induced with 0.1 mM of IPTG. Since the purified enzyme did not show any measurable activity, the substrate scope of the BVMO has been determined using whole-cell and crude cell extract systems. The enzyme was most active towards linear aliphatic substrates. However, it has shown a moderate degree of conversion for cyclobutanone, 2-methylcyclohexanone, bicyclo[3.2.0]hept-2-en-6-one, phenylacetone and thioanisole. There was no detectable conversion of ethionamide, cyclohexanone and acetophenone. (C) 2014 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
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Contribution to journal
publication status
published
subject
keywords
Dietzia, Baeyer-Villiger monooxygenase, Aliphatic ketones, Ethionamide
in
Journal of Molecular Catalysis B: Enzymatic
volume
109
pages
161 - 169
publisher
Elsevier
external identifiers
  • wos:000343854100022
  • scopus:84907202816
ISSN
1873-3158
DOI
10.1016/j.molcatb.2014.08.020
language
English
LU publication?
yes
id
7b60da80-0870-48b9-bfce-882231f8027b (old id 4780758)
date added to LUP
2014-11-20 09:20:53
date last changed
2017-09-03 03:26:59
@article{7b60da80-0870-48b9-bfce-882231f8027b,
  abstract     = {A Baeyer-Villiger monooxygenase has been identified in the genome sequence of Dietzia sp. D5. Sequence similarity search revealed that the enzyme belongs to a group of BVMOs that are closely related to ethionamide monooxygenase from Mycobacterium tuberculosis (EthA). The BVMO was expressed in E. coli BL21-CodonPlus(DE3)-RP and the best expression was achieved when the E. coli cells were cultivated in terrific broth (TB) at 15 degrees C and induced with 0.1 mM of IPTG. Since the purified enzyme did not show any measurable activity, the substrate scope of the BVMO has been determined using whole-cell and crude cell extract systems. The enzyme was most active towards linear aliphatic substrates. However, it has shown a moderate degree of conversion for cyclobutanone, 2-methylcyclohexanone, bicyclo[3.2.0]hept-2-en-6-one, phenylacetone and thioanisole. There was no detectable conversion of ethionamide, cyclohexanone and acetophenone. (C) 2014 Elsevier B.V. All rights reserved.},
  author       = {Bisagni, Serena and Smus, Justyna and Chávez, Georgina and Hatti-Kaul, Rajni and Mamo, Gashaw},
  issn         = {1873-3158},
  keyword      = {Dietzia,Baeyer-Villiger monooxygenase,Aliphatic ketones,Ethionamide},
  language     = {eng},
  pages        = {161--169},
  publisher    = {Elsevier},
  series       = {Journal of Molecular Catalysis B: Enzymatic},
  title        = {Cloning and expression of a Baeyer-Villiger monooxygenase oxidizing linear aliphatic ketones from Dietzia sp. D5},
  url          = {http://dx.doi.org/10.1016/j.molcatb.2014.08.020},
  volume       = {109},
  year         = {2014},
}