A highly productive, whole-cell DERA chemoenzymatic process for production of key lactonized side-chain intermediates in statin synthesis.
Reads0
Chats0
TLDR
To the authors' knowledge, this is the first asymmetric aldol condensation process achieved with whole-cell DERA catalysis and it simplifies and extends previously developed DERA-catalyzed approaches based on the isolated enzyme.Abstract:
Employing DERA (2-deoxyribose-5-phosphate aldolase), we developed the first whole-cell biotransformation process for production of chiral lactol intermediates useful for synthesis of optically pure super-statins such as rosuvastatin and pitavastatin. Herein, we report the development of a fed-batch, high-density fermentation with Escherichia coli BL21 (DE3) overexpressing the native E. coli deoC gene. High activity of this biomass allows direct utilization of the fermentation broth as a whole-cell DERA biocatalyst. We further show a highly productive bioconversion processes with this biocatalyst for conversion of 2-substituted acetaldehydes to the corresponding lactols. The process is evaluated in detail for conversion of acetyloxy-acetaldehyde with the first insight into the dynamics of reaction intermediates, side products and enzyme activity, allowing optimization of the feeding strategy of the aldehyde substrates for improved productivities, yields and purities. The resulting process for production of ((2S,4R)-4,6-dihydroxytetrahydro-2H-pyran-2-yl)methyl acetate (acetyloxymethylene-lactol) has a volumetric productivity exceeding 40 g L−1 h−1 (up to 50 g L−1 h−1) with >80% yield and >80% chromatographic purity with titers reaching 100 g L−1. Stereochemical selectivity of DERA allows excellent enantiomeric purities (ee >99.9%), which were demonstrated on downstream advanced intermediates. The presented process is highly cost effective and environmentally friendly. To our knowledge, this is the first asymmetric aldol condensation process achieved with whole-cell DERA catalysis and it simplifies and extends previously developed DERA-catalyzed approaches based on the isolated enzyme. Finally, applicability of the presented process is demonstrated by efficient preparation of a key lactol precursor, which fits directly into the lactone pathway to optically pure super-statins.read more
Citations
More filters
Journal ArticleDOI
Exploiting Enzymatic Dynamic Reductive Kinetic Resolution (DYRKR) in Stereocontrolled Synthesis.
TL;DR: The scope and limitations of such dehydrogenase-mediated processes are overviewed, as are future possibilities for the evolution of enzymatic DYRKR.
Journal ArticleDOI
Shortening Synthetic Routes to Small Molecule Active Pharmaceutical Ingredients Employing Biocatalytic Methods
TL;DR: In this paper, a review of large-scale synthetic routes containing biocatalytic key steps toward >130 APIs of approved drugs and drug candidates are compared with the corresponding chemical protocols.
Journal ArticleDOI
Engineering a short, aldolase-based pathway for (R)-1,3-butanediol production in Escherichia coli.
Kayla Nemr,Jonas E.N. Müller,Jeong Chan Joo,Pratish Gawand,Ruhi Choudhary,Burton Mendonca,Shuyi Lu,Xiuyan Yu,Alexander F. Yakunin,Radhakrishnan Mahadevan +9 more
TL;DR: This study implements a short, aldolase-based pathway in Escherichia coli to produce (R)-1,3-BDO from glucose, an essential component of pharmaceutical products and cosmetics and highlights the potential of the a Aldolase chemistry to synthesize diverse products directly from renewable resources in microbes.
Journal ArticleDOI
2-Deoxy-D-ribose-5-phosphate aldolase (DERA): applications and modifications.
TL;DR: The present review is aimed to provide a brief overview of DERA, its history, and progress made in understanding the functioning of the enzyme.
Journal ArticleDOI
Biocatalyzed Synthesis of Statins: A Sustainable Strategy for the Preparation of Valuable Drugs
TL;DR: There is a growing interest in developing more sustainable protocols for preparation of statins, and the introduction of biocatalyzed steps into the synthetic pathways is highly advantageous—synthetic routes are conducted under mild reaction conditions, at ambient temperature, and can use water as a reaction medium in many cases.
References
More filters
Journal ArticleDOI
Industrial biocatalysis today and tomorrow
TL;DR: Biocatalytic processes can now be carried out in organic solvents as well as aqueous environments, so that apolar organic compounds aswell as water-soluble compounds can be modified selectively and efficiently with enzymes and bioc atalytically active cells.
Journal ArticleDOI
Structural mechanism for statin inhibition of HMG-CoA reductase.
Eva S. Istvan,Johann Deisenhofer +1 more
TL;DR: The structures of the catalytic portion of human HMGR complexed with six different statins are determined, which show several catalytically relevant residues are disordered in the enzyme-statin complexes.
Book
Chemical kinetics and dynamics
TL;DR: The transition from the macroscopic to the microscopic level is discussed in this article, where the transition state theory is applied to the transition from macroscopy to the microscopic level.
Journal ArticleDOI