A Multi-Enzymatic Cascade Reaction for the Stereoselective Production of γ-Oxyfunctionalyzed Amino Acids

Enoki, Junichi; Meisborn, Jaqueline; Müller, A.-K.; Kourist, Robert

doi:10.3389/fmicb.2016.00425

Cited by 22 publications

(19 citation statements)

References 21 publications

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“…Due to the choice of the vectors, all of the C. crescentus enzymes were N-terminally histidine tagged. The recombinant L-isoleucine dioxygenase from B. thuringiensis (BtDO) contained a C-terminal 6× histidine tag 47 .…”

Section: Methodsmentioning

confidence: 99%

“…The L-isoleucine dioxygenase from Bacillus thuringiensis (BtDO) was previously reported to catalyse the conversion of different amino acids including L-isoleucine to the corresponding hydroxyl amino acids 45,46 , using α-ketoglutarate as a reductant. By combination of the C. crescentus Weimberg enzyme cascade with the dioxygenase from B. thuringiensis 47 Supplementary Fig. 28).…”

Section: Model Validation In One-pot Cascade Perturbation Experimentsmentioning

confidence: 99%

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A combined experimental and modelling approach for the Weimberg pathway optimisation

et al. 2020

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The oxidative Weimberg pathway for the five-step pentose degradation to α-ketoglutarate is a key route for sustainable bioconversion of lignocellulosic biomass to added-value products and biofuels. The oxidative pathway from Caulobacter crescentus has been employed in in-vivo metabolic engineering with intact cells and in in-vitro enzyme cascades. The performance of such engineering approaches is often hampered by systems complexity, caused by non-linear kinetics and allosteric regulatory mechanisms. Here we report an iterative approach to construct and validate a quantitative model for the Weimberg pathway. Two sensitive points in pathway performance have been identified as follows: (1) product inhibition of the dehydrogenases (particularly in the absence of an efficient NAD + recycling mechanism) and (2) balancing the activities of the dehydratases. The resulting model is utilized to design enzyme cascades for optimized conversion and to analyse pathway performance in C. cresensus cellfree extracts.

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Section: Methodsmentioning

confidence: 99%

Section: Model Validation In One-pot Cascade Perturbation Experimentsmentioning

confidence: 99%

A combined experimental and modelling approach for the Weimberg pathway optimisation

et al. 2020

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“…Moreover, the incompatibility of the second step with a component of the first step prohibits sequential cascades. A typical example is the inactivation or inhibition of enzymes by metal ions . The removal of metal ions from an aqueous solution is fairly practical for any envisioned cascade reaction.…”

Section: Chemoenzymatic Cascade Reactionsmentioning

confidence: 99%

“…Atypical example is the inactivation or inhibition of enzymes by metal ions. [8] The removal of metal ions from an aqueous solution is fairly practical for any envisioned cascade reaction. Grçgerd emonstrated that spatial separation of two reactions can overcome this limitation (Scheme 3a).…”

Section: Concurrent Cascades In Organic Mediamentioning

confidence: 99%

Overcoming the Incompatibility Challenge in Chemoenzymatic and Multi‐Catalytic Cascade Reactions

Schmidt

Castiglione

Kourist

2017

Chemistry A European J

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Multi-catalytic cascade reactions bear a great potential to minimize downstream and purification steps, leading to a drastic reduction of the produced waste. In many examples, the compatibility of chemo- and biocatalytic steps could be easily achieved. Problems associated with the incompatibility of the catalysts and their reactions, however, are very frequent. Cascade-like reactions can hardly occur in this way. One possible solution to combine, in principle, incompatible chemo- and biocatalytic reactions is the defined control of the microenvironment by compartmentalization or scaffolding. Current methods for the control of the microenvironment of biocatalysts go far beyond classical enzyme immobilization and are thus believed to be very promising tools to overcome incompatibility issues and to facilitate the synthetic application of cascade reactions. In this Minireview, we will summarize recent synthetic examples of (chemo)enzymatic cascade reactions and outline promising methods for their spatial control either by using bio-derived or synthetic systems.

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“…The hydroxylation of Lisoleucine by this enzyme produces (2S,3R,4S)-4-hydroxyisoleucine which was previously identified from fenugreek seeds (Trigonella foenum-graecum) and is a natural nonproteinogenic amino acid holding insulinotropic biological activity and also anti-obesity effects (Broca et al, 1999;Broca et al, 2000;Handa et al, 2005). IDO is also able to catalyze highly enantioselective sulfoxidation of Lmethionine and L-ethionine into (S)-sulfoxides (Enoki et al, 2016;Hibi et al, 2013b). SadA from Burkholderia ambifaria AMMD converts N-succinyl-L-leucine…”

Section: Introductionmentioning

confidence: 99%

Cloning and characterization of a new delta-specific l-leucine dioxygenase from Anabaena variabilis

Cordeiro

Enoki

Büsch

et al. 2018

Journal of Biotechnology

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Optically pure hydroxy amino acids show several bioactivities and are valuable building blocks for the pharmaceutical industry. Fe(II)/α-ketoglutarate dependent dioxygenases catalyze the hydroxylation or sulfoxidation of l-amino acids with high regio- and stereoselectivity. While several β- and γ-specific enzymes have been described, only one δ-specific hydroxylase has been reported so far. Based on its similarity to the known l-leucine 5-hydroxylase from Nostoc punctiforme, an open reading frame from the cyanobacterium Anabaena variabilis was identified as putative l-leucine dioxygenase (AvLDO). Here we report the cloning and characterization of this dioxygenase. The enzyme showed a high preference for acidic conditions and moderate reaction temperatures. AvLDO catalyzed the regio- and stereoselective hydroxylation of several aliphatic amino acids in δ-position. In case of the sulfoxidation of l-methionine, AvLDO produced the opposite diastereomer than isoleucine dioxygenase. AvLDO is thus an interesting addition to the toolbox of Fe(II)/α-ketoglutarate dependent dioxygenases. On the genomic DNA of Anabaena variabilis ATCC 29413, the avldo gene is located on a gene cluster involved (2S,4S)-4-methylproline biosynthesis, which is contained in bioactive peptides often found from cyanobacteria. This fact suggests the metabolic functional role of this amino acid dioxygenase in cyanobacteria.

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A Multi-Enzymatic Cascade Reaction for the Stereoselective Production of γ-Oxyfunctionalyzed Amino Acids

Cited by 22 publications

References 21 publications

A combined experimental and modelling approach for the Weimberg pathway optimisation

A combined experimental and modelling approach for the Weimberg pathway optimisation

Overcoming the Incompatibility Challenge in Chemoenzymatic and Multi‐Catalytic Cascade Reactions

Cloning and characterization of a new delta-specific l-leucine dioxygenase from Anabaena variabilis

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