PMID- 22286514
OWN - NLM
STAT- MEDLINE
DCOM- 20130122
LR  - 20121108
IS  - 1432-0614 (Electronic)
IS  - 0175-7598 (Linking)
VI  - 96
IP  - 2
DP  - 2012 Oct
TI  - Completing the series of BVMOs involved in camphor metabolism of Pseudomonas 
      putida NCIMB 10007 by identification of the two missing genes, their functional 
      expression in E. coli, and biochemical characterization.
PG  - 419-29
AB  - The camphor-degrading Baeyer-Villiger monooxygenases (BVMOs) from Pseudomonas 
      putida NCIMB 10007 have been of interest for over 40 years. So far the FMN- and 
      NADH-dependent type II BVMO 3,6-diketocamphane 1,6-monooxygenase (3,6-DKCMO) and 
      the FAD- and NADPH-dependent type I BVMO 
      2-oxo-∆3-4,5,5-trimethylcyclopentenylacetyl-CoA monooxygenase (OTEMO) have not 
      been entirely studied, since it was not possible to produce those enzymes in 
      satisfactory amounts and purity. In this study, we were able to clone and 
      recombinantly express both enzymes and subsequently use them as biocatalysts for 
      various mono- and bicyclic ketones. Full conversion could be reached with both 
      enzymes towards (+/-)-cis-bicyclo[3.2.0]hept-2-en-6-one and with 3,6-DKCMO towards 
      (-)-camphor. Further OTEMO gave full conversion with norcamphor. OTEMO was found 
      to have a pH optimum of 9 and a temperature optimum of 20  degrees C and converted 
      (+/-)-cis-bicyclo[3.2.0]hept-2-en-6-one with a k cat/K M value of 49.3 mM-1 s-1.
FAU - Kadow, Maria
AU  - Kadow M
AD  - Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, 
      Greifswald University, Felix-Hausdorff-Str. 4, 17487 Greifswald, Germany.
FAU - Loschinski, Kathleen
AU  - Loschinski K
FAU - Sass, Stefan
AU  - Sass S
FAU - Schmidt, Marlen
AU  - Schmidt M
FAU - Bornscheuer, Uwe T
AU  - Bornscheuer UT
LA  - eng
SI  - GENBANK/JQ034404
SI  - GENBANK/JQ034405
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Appl Microbiol Biotechnol
JT  - Applied microbiology and biotechnology
JID - 8406612
RN  - 0 (Bacterial Proteins)
RN  - 76-22-2 (Camphor)
RN  - EC 1.- (Mixed Function Oxygenases)
SB  - IM
MH  - Bacterial Proteins/*chemistry/*genetics/metabolism
MH  - Camphor/chemistry/*metabolism
MH  - Enzyme Stability
MH  - Escherichia coli/*genetics/metabolism
MH  - Gene Expression
MH  - Kinetics
MH  - Mixed Function Oxygenases/*chemistry/*genetics/metabolism
MH  - Molecular Sequence Data
MH  - Phylogeny
MH  - Pseudomonas putida/chemistry/classification/*enzymology/genetics
MH  - Substrate Specificity
EDAT- 2012/01/31 06:00
MHDA- 2013/01/23 06:00
CRDT- 2012/01/31 06:00
PHST- 2011/11/18 00:00 [received]
PHST- 2011/12/22 00:00 [accepted]
PHST- 2011/12/20 00:00 [revised]
PHST- 2012/01/31 06:00 [entrez]
PHST- 2012/01/31 06:00 [pubmed]
PHST- 2013/01/23 06:00 [medline]
AID - 10.1007/s00253-011-3859-1 [doi]
PST - ppublish
SO  - Appl Microbiol Biotechnol. 2012 Oct;96(2):419-29. doi: 10.1007/s00253-011-3859-1.