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Reaction and strain engineering for improved stereo-selective whole-cell reduction of a bicyclic diketone

Johanson, Ted LU ; Carlquist, Magnus LU ; Olsson, Cecilia LU ; Rudolf, Andreas LU ; Frejd, Torbjörn LU and Gorwa-Grauslund, Marie-Francoise LU (2008) In Applied Microbiology and Biotechnology 77(5). p.1111-1118
Abstract
Reduction of bicyclo[2.2.2]octane-2,6-dione to (1R, 4S, 6S)-6-hydroxy-bicyclo[2.2.2]octane-2-one by whole cells of Saccharomyces cerevisiae was improved using an engineered recombinant strain and process design. The substrate inhibition followed a Han-Levenspiel model showing an effective concentration window between 12 and 22 g/l, in which the activity was kept above 95%. Yeast growth stage, substrate concentration and a stable pH were shown to be important parameters for effective conversion. The over-expression of the reductase gene YDR368w significantly improved diastereoselectivity compared to previously reported results. Using strain TMB4110 expressing YDR368w in batch reduction with pH control, complete conversion of 40 g/l (290 mM)... (More)
Reduction of bicyclo[2.2.2]octane-2,6-dione to (1R, 4S, 6S)-6-hydroxy-bicyclo[2.2.2]octane-2-one by whole cells of Saccharomyces cerevisiae was improved using an engineered recombinant strain and process design. The substrate inhibition followed a Han-Levenspiel model showing an effective concentration window between 12 and 22 g/l, in which the activity was kept above 95%. Yeast growth stage, substrate concentration and a stable pH were shown to be important parameters for effective conversion. The over-expression of the reductase gene YDR368w significantly improved diastereoselectivity compared to previously reported results. Using strain TMB4110 expressing YDR368w in batch reduction with pH control, complete conversion of 40 g/l (290 mM) substrate was achieved with 97% diastereomeric excess (de) and >99 enantiomeric excess (ee), allowing isolation of the optically pure ketoalcohol in 84% yield. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Whole-cell - Bioreduction - Reductase - Yeast - Dicarbonyl - Process optimisation - Toxicity - Substrate inhibition - Diastereoselectivity
in
Applied Microbiology and Biotechnology
volume
77
issue
5
pages
1111 - 1118
publisher
Springer
external identifiers
  • wos:000251606900015
  • scopus:37249019632
ISSN
1432-0614
DOI
10.1007/s00253-007-1240-1
language
English
LU publication?
yes
id
607bbba2-6029-48a3-8f6c-aa48c60e75e1 (old id 756835)
date added to LUP
2007-12-18 09:50:26
date last changed
2017-05-21 03:38:12
@article{607bbba2-6029-48a3-8f6c-aa48c60e75e1,
  abstract     = {Reduction of bicyclo[2.2.2]octane-2,6-dione to (1R, 4S, 6S)-6-hydroxy-bicyclo[2.2.2]octane-2-one by whole cells of Saccharomyces cerevisiae was improved using an engineered recombinant strain and process design. The substrate inhibition followed a Han-Levenspiel model showing an effective concentration window between 12 and 22 g/l, in which the activity was kept above 95%. Yeast growth stage, substrate concentration and a stable pH were shown to be important parameters for effective conversion. The over-expression of the reductase gene YDR368w significantly improved diastereoselectivity compared to previously reported results. Using strain TMB4110 expressing YDR368w in batch reduction with pH control, complete conversion of 40 g/l (290 mM) substrate was achieved with 97% diastereomeric excess (de) and >99 enantiomeric excess (ee), allowing isolation of the optically pure ketoalcohol in 84% yield.},
  author       = {Johanson, Ted and Carlquist, Magnus and Olsson, Cecilia and Rudolf, Andreas and Frejd, Torbjörn and Gorwa-Grauslund, Marie-Francoise},
  issn         = {1432-0614},
  keyword      = {Whole-cell - Bioreduction - Reductase - Yeast - Dicarbonyl - Process optimisation - Toxicity - Substrate inhibition - Diastereoselectivity},
  language     = {eng},
  number       = {5},
  pages        = {1111--1118},
  publisher    = {Springer},
  series       = {Applied Microbiology and Biotechnology},
  title        = {Reaction and strain engineering for improved stereo-selective whole-cell reduction of a bicyclic diketone},
  url          = {http://dx.doi.org/10.1007/s00253-007-1240-1},
  volume       = {77},
  year         = {2008},
}