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

Ted Johanson; Magnus Carlquist; Cecilia Olsson; Andreas Rudolf; Torbjörn Frejd; Marie-Francoise Gorwa-Grauslund

doi:10.1007/s00253-007-1240-1

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

Ted Johanson, Magnus Carlquist, Cecilia Olsson, Andreas Rudolf, Torbjörn Frejd, Marie-Francoise Gorwa-Grauslund

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	1111-1118
Journal	Applied Microbiology and Biotechnology
Volume	77
Issue number	5
DOIs	https://doi.org/10.1007/s00253-007-1240-1
Publication status	Published - 2008

Bibliographical note

The information about affiliations in this record was updated in December 2015.
The record was previously connected to the following departments: Applied Microbiology (LTH) (011001021), Organic chemistry (S/LTH) (011001240), Center for Chemistry and Chemical Engineering (011001000), Chemical Engineering (011001014)

Subject classification (UKÄ)

Chemical Sciences
Industrial Biotechnology
Organic Chemistry

Free keywords

Whole-cell - Bioreduction - Reductase - Yeast - Dicarbonyl - Process optimisation - Toxicity - Substrate inhibition - Diastereoselectivity

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10.1007/s00253-007-1240-1

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

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.",

keywords = "Whole-cell - Bioreduction - Reductase - Yeast - Dicarbonyl - Process optimisation - Toxicity - Substrate inhibition - Diastereoselectivity",

author = "Ted Johanson and Magnus Carlquist and Cecilia Olsson and Andreas Rudolf and Torbj{\"o}rn Frejd and Marie-Francoise Gorwa-Grauslund",

note = "The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Applied Microbiology (LTH) (011001021), Organic chemistry (S/LTH) (011001240), Center for Chemistry and Chemical Engineering (011001000), Chemical Engineering (011001014)",

year = "2008",

doi = "10.1007/s00253-007-1240-1",

language = "English",

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journal = "Applied Microbiology and Biotechnology",

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AU - Johanson, Ted

AU - Carlquist, Magnus

AU - Olsson, Cecilia

AU - Rudolf, Andreas

AU - Frejd, Torbjörn

AU - Gorwa-Grauslund, Marie-Francoise

N1 - The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Applied Microbiology (LTH) (011001021), Organic chemistry (S/LTH) (011001240), Center for Chemistry and Chemical Engineering (011001000), Chemical Engineering (011001014)

PY - 2008

Y1 - 2008

N2 - 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.

AB - 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.

KW - Whole-cell - Bioreduction - Reductase - Yeast - Dicarbonyl - Process optimisation - Toxicity - Substrate inhibition - Diastereoselectivity

UR - https://www.scopus.com/pages/publications/37249019632

U2 - 10.1007/s00253-007-1240-1

DO - 10.1007/s00253-007-1240-1

M3 - Article

C2 - 17962934

SN - 1432-0614

VL - 77

SP - 1111

EP - 1118

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

IS - 5

ER -

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

Abstract

Bibliographical note

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Free keywords

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