Improvement of aldehyde tolerance and sequential aldol condensation activity of deoxyriboaldolase via immobilization on interparticle pore type mesoporous silica

Nara, Takayuki; Togashi, Hideaki; Ono, Seigo; Egami, Miki; Sekikawa, Chisato; Suzuki, Yohei; Masuda, Isao; Ogawa, Jun; Horinouchi, Nobuyuki; Shimizu, Sakayu; Mizukami, Fujio; Tsunoda, Tetsuto

doi:10.1016/j.molcatb.2010.10.008

Cited by 28 publications

(41 citation statements)

References 35 publications

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“…The present work studied the consecutive aldol reaction system catalyzed by DERA Tm , in which the acceptor substrate 2 reacts in a first step with the donor substrate 1 to give the adduct 3 , which subsequently reacts with a second equivalent of the donor 1 to result in the key product 4 (Fig. ) .…”

Section: Resultsmentioning

confidence: 99%

“…DERA accepts a broad range of acceptor aldehydes, has a relatively large donor tolerance for compounds having up to four carbon atoms, and generates ( R )‐configured chiral centers . Therefore, it is not surprising that much attention recently has been given to its application for the synthesis of valuable chiral molecules …”

Section: Introductionmentioning

confidence: 99%

“…By using achiral substrates, such as chloroacetaldehyde and acetaldehyde, DERA catalyzes a sequential aldol addition that results in an enantiomerically pure lactol, (3 R ,5 R )‐6‐chloro‐2,4,6‐trideoxyhexose . This 2,4,6‐trideoxyhexose is a valuable chiral synthon for HMG‐CoA reductase inhibitors, collectively known as statins, and used for the production of cholesterol‐lowering drugs, such as atorvastatin and rosuvastatin . Those statins have c .…”

Section: Introductionmentioning

confidence: 99%

“…DERA shows poor resistance to high aldehyde concentrations and low catalytic efficiency towards the sequential aldol addition of unnatural substrates . Thus, large quantities of the enzyme are necessary to obtain industrially useful product yields . This issue can be circumvented by improving its resistance and catalytic activity using a molecular evolution approach, or by applying a cell‐free lysate or a whole‐cell biocatalyst instead of an isolated enzyme or by immobilization on appropriate supports …”

Section: Introductionmentioning

confidence: 99%

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2‐Deoxyribose‐5‐phosphate aldolase from Thermotoga maritima in the synthesis of a statin side‐chain precursor: characterization, modeling and optimization

Švarc

Blažević

Vasić-Rački

et al. 2019

J of Chemical Tech & Biotech

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BACKGROUND Producing statin side‐chains by sequential aldol condensation catalyzed by 2‐deoxyribose‐5‐phosphate aldolase (DERA) (EC 4.1.2.4) is the best‐known and most promising synthetic route. The advantage of this process is the formation of two stereocenters in a single step starting from inexpensive achiral components, acetaldehyde and chloroacetaldehyde. The limitation for the industrial‐scale application is enzyme inactivation caused by substrates as well by the intermediate produced by single aldol addition. RESULTS The DERA enzyme from Thermotoga maritima (DERATm) was investigated extensively in this work. The influence of aldehydes on DERATm stability was studied in detail. Mathematical models in different reactor configurations were developed based on experimentally determined kinetic parameters of all reactions included in the synthesis of statin side‐chain. Models also included enzyme inactivation by all compounds with negative impact on its stability. Mathematical model‐based optimization enabled optimization of process conditions and reactor configuration. The fed‐batch reactor proved to be the most suitable choice achieving product concentration of 78 g L−1, productivity of 56 g L day−1 and yield of 95% under an optimal condition. CONCLUSION The validated mathematical model and applied methodology can be exploited for further process improvement and for process design of similar systems. © 2019 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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2‐Deoxyribose‐5‐phosphate aldolase from Thermotoga maritima in the synthesis of a statin side‐chain precursor: characterization, modeling and optimization

Švarc

Blažević

Vasić-Rački

et al. 2019

J of Chemical Tech & Biotech

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“…This was also the case in pure physical adsorption on porous silica nanoparticles. Again, part of the enzyme surface is protected by the carrier (Nara et al 2011). A recent structural study demonstrated that immobilization via the C-terminal his tag is impossible as the C-terminus is folded into the active site.…”

Section: Acetaldehyde Resistance Of Deramentioning

confidence: 99%

2-Deoxy-d-ribose-5-phosphate aldolase (DERA): applications and modifications

Haridas

Abdelraheem

Hanefeld

2018

Appl Microbiol Biotechnol

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2-Deoxy-d-ribose-5-phosphate aldolase (DERA) is a class I aldolase that offers access to several building blocks for organic synthesis. It catalyzes the stereoselective C–C bond formation between acetaldehyde and numerous other aldehydes. However, the practical application of DERA as a biocatalyst is limited by its poor tolerance towards industrially relevant concentrations of aldehydes, in particular acetaldehyde. Therefore, the development of proper experimental conditions, including protein engineering and/or immobilization on appropriate supports, is required. The present review is aimed to provide a brief overview of DERA, its history, and progress made in understanding the functioning of the enzyme. Furthermore, the current understanding regarding aldehyde resistance of DERA and the various optimizations carried out to modify this property are discussed.

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Covalently Immobilized 2‐Deoxyribose‐5‐phosphate Aldolase (DERA) for Biocatalysis in Flow: Utilization of the 3‐Hydroxyaldehyde Intermediate in Reaction Cascades

et al. 2022

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C−C bond forming enzymes, such as 2‐deoxyribose‐5‐phosphate aldolase (DERA), are an important class of enzymes in synthesis. The use of DERA, with its reaction routes to the double aldol product, is an established method for producing statin side chains in industry. Herein, the selective synthesis of 3‐hydroxyaldehydes as single aldol products of DERA is presented. Successful covalent immobilization of a suitable Escherichia coli (Ec) DERA variants with C47M and C47V exchanges, respectively, enabled their use in continuous synthesis. Precise reaction control using flow chemistry made the selective formation of 3‐hydroxyoctanal from hexanal possible. For optimization, a systematic approach of design of experiment (DoE) was used, resulting in 90 % conversion. Furthermore, five two‐step reaction systems have been established to scavenge the reactive single aldol product combining biocatalysis in flow mode and a consecutive chemical reactions step either in flow (continuous flow mode) or in batch (semi‐continuous flow mode).

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Improvement of aldehyde tolerance and sequential aldol condensation activity of deoxyriboaldolase via immobilization on interparticle pore type mesoporous silica

Cited by 28 publications

References 35 publications

2‐Deoxyribose‐5‐phosphate aldolase from Thermotoga maritima in the synthesis of a statin side‐chain precursor: characterization, modeling and optimization

2‐Deoxyribose‐5‐phosphate aldolase from Thermotoga maritima in the synthesis of a statin side‐chain precursor: characterization, modeling and optimization

2-Deoxy-d-ribose-5-phosphate aldolase (DERA): applications and modifications

Covalently Immobilized 2‐Deoxyribose‐5‐phosphate Aldolase (DERA) for Biocatalysis in Flow: Utilization of the 3‐Hydroxyaldehyde Intermediate in Reaction Cascades

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