Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum

Journal Article (2020)
Author(s)

Meera Haridas (TU Delft - ChemE/Product and Process Engineering)

Carolin Bisterfeld (TU Delft - BT/Biocatalysis)

Le Min Chen (Student TU Delft)

S.R. Marsden (TU Delft - BT/Biocatalysis)

F. Tonin (TU Delft - BT/Biocatalysis)

R Médici (TU Delft - BT/Biocatalysis)

Adolfo Iribarren (Universidad Nacional de Quilmes, Bernal and Concet)

Elizabeth Lewkowicz (Universidad Nacional de Quilmes, Bernal and Concet)

PL Hagedoorn (TU Delft - BT/Biocatalysis)

Ulf Hanefeld (TU Delft - BT/Biocatalysis)

Eman Abdelraheem (TU Delft - BT/Biocatalysis)

Research Group
ChemE/Product and Process Engineering
Copyright
© 2020 M. Haridas, C. Bisterfeld, Le Min Chen, S.R. Marsden, F. Tonin, R. Medici, Adolfo Iribarren, Elizabeth Lewkowicz, P.L. Hagedoorn, U. Hanefeld, E.M.M. Abdelraheem
DOI related publication
https://doi.org/10.3390/catal10080883
More Info
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Publication Year
2020
Language
English
Copyright
© 2020 M. Haridas, C. Bisterfeld, Le Min Chen, S.R. Marsden, F. Tonin, R. Medici, Adolfo Iribarren, Elizabeth Lewkowicz, P.L. Hagedoorn, U. Hanefeld, E.M.M. Abdelraheem
Research Group
ChemE/Product and Process Engineering
Issue number
8
Volume number
10
Pages (from-to)
1-10
Reuse Rights

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Abstract

DERA (2-Deoxy-D-ribose 5-phosphate aldolase) is the only known aldolase that accepts two aldehyde substrates, which makes it an attractive catalyst for the synthesis of a chiral polyol motif that is present in several pharmaceuticals, such as atorvastatin and pravastatin. However, inactivation of the enzyme in the presence of aldehydes hinders its practical application. Whole cells of Pectobacterium atrosepticum were reported to exhibit good tolerance toward acetaldehyde and to afford 2-deoxyribose 5-phosphate with good yields. The DERA gene (PaDERA) was identified, and both the wild-type and a C49M mutant were heterologously expressed in Escherichia coli. The purification protocol was optimized and an initial biochemical characterization was conducted. Unlike other DERAs, which show a maximal activity between pH 4.0 and 7.5, PaDERA presented an optimum pH in the alkaline range between 8.0 and 9.0. This could warrant its use for specific syntheses in the future. PaDERA also displayed fourfold higher specific activity than DERA from E. coli (EcDERA) and displayed a promising acetaldehyde resistance outside the whole-cell environment. The C49M mutation, which was previously identified to increase acetaldehyde tolerance in EcDERA, also led to significant improvements in the acetaldehyde tolerance of PaDERA.