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Use of ADH as a racemization catalyst in a dynamic kinetic resolution by lipase (Gruber et al. 2007; Musa et al. 2015; Karume et al. 2016; Popłoński et al. 2018)
Source publication
Alcohol dehydrogenases (ADHs) catalyze the reversible reduction of a carbonyl group to its corresponding alcohol. ADHs are widely employed for organic synthesis due to their lack of harm to the environment, broad substrate acceptance, and high enantioselectivity. This review focuses on the impact and relevance of ADH enantioselectivities on their b...
Citations
... Biocatalysts play an important role in the development of a sustainable society due to their environmentally friendly nature and several advantageous properties, including safety, chemo-, regio-, and enantioselectivity, and high efficiency. Consequently, they have found successful applications in various industries such as chemical [1,2], pharmaceutical [3], and food [1]. To enhance the versatility of biocatalysts, immobilization has emerged as a crucial strategy [4][5][6]. ...
The development of green catalysts, specifically biocatalysts, is crucial for building a sustainable society. To enhance the versatility of biocatalysts, the immobilization of enzymes plays a vital role as it improves their recyclability and robustness. As target enzymes to immobilize, glucose dehydrogenases and carboxylases are particularly important among various kinds of enzymes due to their involvement in two significant reactions: regeneration of the reduced form of coenzyme required for various reactions, and carboxylation reactions utilizing CO2 as a substrate, respectively. In this study, we immobilized Thermoplasma acidophilum glucose dehydrogenase (TaGDH) and T. acidophilum isocitrate dehydrogenase (TaIDH) using a previously reported method involving the formation of enzyme-inorganic hybrid nanocrystals, in the course of our continuing study focusing on carboxylation catalyzed by the free form of TaGDH and TaIDH. Subsequently, we investigated the properties of the resulting immobilized enzymes. Our results indicate the successful immobilization of TaGDH and TaIDH through the formation of hybrid nanocrystals utilizing Mn²⁺. The immobilization process enhanced TaIDH activity, up to 211%, while TaGDH retained 71% of its original activity. Notably, the immobilized TaGDH exhibited higher activity at temperatures exceeding 87 °C than the free TaGDH. Moreover, these immobilized enzymes could be recycled. Finally, we successfully utilized the immobilized enzymes for the carboxylation of 2-ketoglutaric acid under 1 MPa CO2. In conclusion, this study represents the first immobilization of TaGDH and TaIDH using the hybrid nanocrystal forming method. Furthermore, we achieved significant activity enhancement of TaIDH through immobilization and demonstrated the recyclability of the immobilized enzymes.
... Compared with natural enzymes such as alcohol dehydrogenase (ADHs), ADHs exhibit high reactivity and enantioselectivity, efficiently catalyzing both dehydrogenation of alcohols and hydrogenation of ketones. [13] However, their substrate scope is limited. On the other hand, our artificial [Mn]-hydrogenase displays a broader substrate scope, efficiently catalyzing the transfer hydrogenation of various ketones. ...
Artificial (transfer) hydrogenases have been developed for organic synthesis, but they rely on precious metals. Native hydrogenases use Earth‐abundant metals, but these cannot be applied for organic synthesis due, in part, to their substrate specificity. Herein, we report the design and development of manganese transfer hydrogenases based on the biotin‐streptavidin technology. By incorporating bio‐mimetic Mn(I) complexes into the binding cavity of streptavidin, and through chemo‐genetic optimization, we have obtained artificial enzymes that hydrogenate ketones with nearly quantitative yield and up to 98 % enantiomeric excess (ee). These enzymes exhibit broad substrate scope and high functional‐group tolerance. According to QM/MM calculations and X‐ray crystallography, the S112Y mutation, combined with the appropriate chemical structure of the Mn cofactor plays a critical role in the reactivity and enantioselectivity of the artificial metalloenzyme (ArMs). Our work highlights the potential of ArMs incorporating base‐meal cofactors for enantioselective organic synthesis.
... [1][2][3][4][5][6][7] Due to their unique properties, enzymes can help in many synthetic aspects: from reducing the number of steps by avoiding protection and deprotection, [8] to the easy and efficient synthesis of enantiopure products. [9][10][11] Certainly, enzymes have found their niche in the synthesis of chemically complex molecules. This traction in the use of these biocatalysts, due to their excellent chemo, regio-and enantioselectivity is also a driving force for the preparation of better, more resistant, and easier to use enzymatic preparations. ...
Protein bioinformatics has been applied to a myriad of opportunities in biocatalysis from enzyme engineering to enzyme discovery, but its application in enzyme immobilization is still very limited. Enzyme immobilization brings clear advantages in terms of sustainability and cost‐efficiency but is still limited in its implementation. This, because it is a technique that remains tied to a quasi‐blind protocol of trial and error, and therefore, is regarded as a time‐intensive and costly approach. Here, we present the use of a set of bioinformatic tools to rationalize the results of protein immobilization that have been previously described. The study of proteins with these new tools allows the discovery of key driving forces in the process of immobilization that explain the obtained results, moving us a step closer to the final goal: predictive enzyme immobilization protocols.
... The 3-keto-DON was transformed into 3-epi-DON and DON simultaneously, which were catalyzed by aldehyde-ketone reductases from yeast, as their reduction reactions can form two alcohol configurations (S type, R type). Usually, these two types of en-zymes are expressed simultaneously in one strain [68]. For example, Candida parapsilosis CCTCC M20301 [46,47] contained both (R)-and (S)-aldo-keto reductases (RCR and SCR), which produced both (R)-phenyl glycol and (S)-phenyl glycol when acted on substrates 2hydroxyacetophenone. ...
Deoxynivalenol (DON), one of the main mycotoxins with enteric toxicity, genetic toxicity, and immunotoxicity, and is widely found in corn, barley, wheat, and rye. In order to achieve effective detoxification of DON, the least toxic 3-epi-DON (1/357th of the toxicity of DON) was chosen as the target for degradation. Quinone-dependent dehydrogenase (QDDH) reported from Devosia train D6-9 detoxifies DON by converting C3-OH to a ketone group with toxicity of less than 1/10 that of DON. In this study, the recombinant plasmid pPIC9K-QDDH was constructed and successfully expressed in Pichia pastoris GS115. Within 12 h, recombinant QDDH converted 78.46% of the 20 μg/mL DON to 3-keto-DON. Candida parapsilosis ACCC 20221 was screened for its activity in reducing 86.59% of 3-keto-DON within 48 h; its main products were identified as 3-epi-DON and DON. In addition, a two-step method was performed for epimerizing DON: 12 h catalysis by recombinant QDDH and 6 h transformation of the C. parapsilosis ACCC 20221 cell catalyst. The production rates of 3-keto-DON and 3-epi-DON were 51.59% and 32.57%, respectively, after manipulation. Through this study, effective detoxification of 84.16% of DON was achieved, with the products being mainly 3-keto-DON and 3-epi-DON.
... Several enzymes associated with oxidation reactions have been applied by pharmaceutical and chemical industries for the oxidation of alcohols, sulfides, aldehydes, Baeyer-Villiger oxidation, and hydroxylation [122][123][124][125][126][127]. G. candidum was discovered to produce dehydrogenases with broad applications in organic synthesis [125,[128][129][130][131][132][133][134]. For instance, G. candidum NBRC 4597 (GcAPRD) was discovered to produce a novel alcohol dehydrogenase (ALDH), acetophenone reductase, an enzyme with broad spectrum activity regarding the oxidation of aldehydes to carboxylic acids and selective activity for the oxidation of dialdehydes to aldehydic acids [129,135,136]. ...
Fungi make a fundamental contribution to several biotechnological processes, including brewing, winemaking, and the production of enzymes, organic acids, alcohols, antibiotics, and pharmaceuticals. The present review explores the biotechnological importance of the filamentous yeast-like fungus Geotrichum candidum, a ubiquitous species known for its use as a starter in the dairy industry. To uncover G. candidum's biotechnological role, we performed a search for related work through the scientific indexing internet services, Web of Science and Google Scholar. The following query was used: Geotrichum candidum, producing about 6500 scientific papers from 2017 to 2022. From these, approximately 150 that were associated with industrial applications of G. candidum were selected. Our analysis revealed that apart from its role as a starter in the dairy and brewing industries, this species has been administered as a probiotic nutritional supplement in fish, indicating improvements in developmental and immunological parameters. Strains of this species produce a plethora of biotechnologically important enzymes, including cellulases, β-glucanases, xylanases, lipases, proteases, and α-amylases. Moreover, strains that produce antimicrobial compounds and that are capable of bioremediation were identified. The findings of the present review demonstrate the importance of G. candidum for agrifood-and bio-industries and provide further insights into its potential future biotechnological roles.
... Various biological functions are observed within this family (Sirota et al. 2021), including polyol dehydrogenases catalyzing the conversion between sugar and sugar alcohol (Lu et al. 2019), cinnamyl alcohol dehydrogenases (Larroy et al. 2002;Pick et al. 2013), and glutathione-dependent formaldehyde dehydrogenases (Gutheil et al. 1992;Sanghani et al. 2000;Achkor et al. 2003), which play an important part in the detoxification of formaldehyde (Vorholt 2002). Additionally, ADHs provide numerous advantageous properties for organic synthesis, including high enantioselectivity and applicability under mild reaction conditions (Koesoema et al. 2020). Consequently, they are now employed in numerous biotechnological applications such as the preparation of chiral alcohols (Zhang et al. 2015), rare sugars (Lu et al. 2019), fine chemicals, as well as the synthesis of building blocks for various essential pharmaceuticals (Hall and Bommarius 2011;Zheng et al. 2017). ...
Marine algae produce complex polysaccharides, which can be degraded by marine heterotrophic bacteria utilizing carbohydrate-active enzymes. The red algal polysaccharide porphyran contains the methoxy sugar 6-O-methyl-d-galactose (G6Me). In the degradation of porphyran, oxidative demethylation of this monosaccharide towards d-galactose and formaldehyde occurs, which is catalyzed by a cytochrome P450 monooxygenase and its redox partners. In direct proximity to the genes encoding for the key enzymes of this oxidative demethylation, genes encoding for zinc-dependent alcohol dehydrogenases (ADHs) were identified, which seem to be conserved in porphyran utilizing marine Flavobacteriia. Considering the fact that dehydrogenases could play an auxiliary role in carbohydrate degradation, we aimed to elucidate the physiological role of these marine ADHs. Although our results reveal that the ADHs are not involved in formaldehyde detoxification, a knockout of the ADH gene causes a dramatic growth defect of Zobellia galactanivorans with G6Me as a substrate. This indicates that the ADH is required for G6Me utilization. Complete biochemical characterizations of the ADHs from Formosa agariphila KMM 3901T (FoADH) and Z. galactanivorans DsijT (ZoADH) were performed, and the substrate screening revealed that these enzymes preferentially convert aromatic aldehydes. Additionally, we elucidated the crystal structures of FoADH and ZoADH in complex with NAD⁺ and showed that the strict substrate specificity of these new auxiliary enzymes is based on a narrow active site.
Key points
• Knockout of the ADH-encoding gene revealed its role in 6-O-methyl-D-galactose utilization, suggesting a new auxiliary activity in marine carbohydrate degradation.
• Complete enzyme characterization indicated no function in a subsequent reaction of the oxidative demethylation, such as formaldehyde detoxification.
• These marine ADHs preferentially convert aromatic compounds, and their strict substrate specificity is based on a narrow active site.
... Good activities and excellent enantioselectivities with 99% ee towards structurally related aromatic ketones were achieved by some of the examined marine fungi [27]. Although substantial progress has been achieved, most of the known biocatalysts, whole cells or isolated enzymes used in the asymmetric reduction of prochiral ketones to chiral alcohols generally follow Prelog's rule [28,29]. Ketone reductases or cells with anti-Prelog stereospecificity are still very limited in number, and anti-Prelog chiral alcohols are in great demand in organic synthesis and the pharmaceutical industry [30]. ...
... Molecules 2023, 28 ...
The biocatalytic asymmetric reduction of prochiral ketones for the production of enantiopure alcohols is highly desirable due to its inherent advantages over chemical methods. In this study, a new bacterial strain capable of transforming ketones to corresponding alcohols with high activity and excellent enantioselectivity was discovered in a soil sample. The strain was subsequently identified as Bacillus cereus TQ-2 based on its physiological characteristics and 16S rDNA sequence analysis. Under optimized reaction conditions, the resting cells of B. cereus TQ-2 converted acetophenone to enantioenriched (R)-1-phenylethanol with 99% enantiometric excess following anti-Prelog’s rule, which is scarce in biocatalytic ketone reduction. The optimum temperature for the cells was 30 °C, and considerable catalytic activity was observed over a broad pH range from 5.0 to 9.0. The cells showed enhanced catalytic activity in the presence of 15% (v/v) glycerol as a co-substrate. The catalytic activity can also be substantially improved by adding Ca2+ or K+ ions. Moreover, the B. cereus TQ-2 cell was highly active in reducing several structurally diverse ketones and aldehydes to form corresponding alcohols with good to excellent conversion. Our study provides a versatile whole-cell biocatalyst that can be used in the asymmetric reduction of ketones for the production of chiral alcohol, thereby expanding the biocatalytic toolbox for potential practical applications.
... They are ubiquitous enzymes that can be found in virtually all organisms. Still, most of the commercial ADHs originate from microorganisms, or, when coming from a mammalian source, they are recombinant in Escherichia coli or other host strains to avoid ethical issues, such as in the production of the widely used ADH from horse liver (HLADH) [11,12]. With the advent and consolidation of protein engineering techniques, wild-type ADHs have been tailored to address reaction and process constraints in terms of substrate scope, substrate and/or product inhibition, solvent and thermostability, activity, and selectivity. ...
Alcohol dehydrogenases (ADHs) catalyze the reversible oxidation of primary and secondary alcohols to the corresponding aldehydes or carboxylic acids and ketones, respectively, under mild conditions, which may minimize the environmental impact compared to the traditional chemical counterpart. In this short review, we briefly report recent advances in the development of ADH-catalyzed oxidation employed both as single biocatalysts but also in multienzyme cascades of reactions. Current bottlenecks, prospects, and future outlooks in the field are addressed.
... This approach is exemplified in biocatalyzed stereoselective reduction reactions. Alcohol dehydrogenases and enereductases, catalyze the asymmetric reduction of carbonyl groups and activated carbon-carbon double bonds, respectively (Toogood and Scrutton, 2014) (Koesoema et al., 2020). These enzyme families have been studied extensively and are used in industrial biocatalysis (de Gonzalo and Paul, 2021) (Toogood and Scrutton, 2018). ...
The discovery of new enzymes, alongside the push to make chemical processes more sustainable, has resulted in increased industrial interest in the use of biocatalytic processes to produce high-value and chiral precursor chemicals. Huge strides in protein engineering methodology and in silico tools have facilitated significant progress in the discovery and production of enzymes for biocatalytic processes. However, there are significant gaps in our knowledge of the relationship between enzyme structure and function. This has demonstrated the need for improved computational methods to model mechanisms and understand structure dynamics. Here, we explore efforts to rationally modify enzymes toward changing aspects of their catalyzed chemistry. We highlight examples of enzymes where links between enzyme function and structure have been made, thus enabling rational changes to the enzyme structure to give predictable chemical outcomes. We look at future directions the field could take and the technologies that will enable it.
... Therefore, an effort to use environmentally benign oxidants such as H 2 O 2 and O 2 has been made (Bryliakov 2017;Liu et al. 2020). On the other hand, biocatalysts have been considered to be sustainable catalysts (Matsuda 2017;Dong et al. 2018;Sheldon and Woodley 2018;Birolli et al. 2019;Wu et al. 2020;Koesoema et al. 2020). For biocatalytic BVO, Baeyer-Villiger monooxygenase (BVMO) can use oxygen in the air as an oxidant, producing water as a byproduct ( Fig. 1), so that the reactions are not explosive (Morii et al. 1999;Kyte et al. 2004;Rehdorf et al. 2007;Franceschini et al. 2012;Leipold et al. 2012Leipold et al. , 2013Fürst et al. 2017Fürst et al. , 2019Nguyen et al. 2017;Fordwour et al. 2018;Woo et al. 2018). ...
Objective
A novel biocatalyst for Baeyer–Villiger oxidations is necessary for pharmaceutical and chemical industries, so this study aims to find a Baeyer–Villiger monooxygenase (BVMO) and to improve its stability by immobilization.
Results
Acetone, the simplest ketone, was selected as the only carbon source for the screening of microorganisms with a BVMO. A eukaryote, Fusarium sp. NBRC 109816, with a BVMO (FBVMO), was isolated from a soil sample. FBVMO was overexpressed in E. coli and successfully immobilized by the organic–inorganic nanocrystal formation method. The immobilization improved the thermostability of FBVMO. Substrate specificity investigation revealed that both free and immobilized FBVMO were found to show catalytic activities not only for Baeyer–Villiger oxidation of ketones to esters but also for oxidation of sulfides to sulfoxides. Furthermore, a preparative scale reaction using immobilized FBVMO was successfully conducted.
Conclusions
FBVMO was discovered from an environmental sample, overexpressed in E. coli, and immobilized by the organic–inorganic nanocrystal formation method. The immobilization successfully improved its thermostability.
