Biocatalytic Synthesis of Natural Dihydrocoumarin by Microbial Reduction of Coumarin

Serra, Stefano; Castagna, Antonio; Valentino, Mario

doi:10.3390/catal9080665

Cited by 13 publications

(10 citation statements)

References 37 publications

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“…The second reason concerns its biocatalytic potential. As described in our previous research [16], Kluyveromyces marxianus possesses high resistance to coumarin toxicity.…”

Section: Resultsmentioning

confidence: 53%

“…Therefore, we recently decided to select new microorganisms to be employed for dihydrocoumarin production. Accordingly, we first screened a variety of yeasts and filamentous fungi, isolated from different sources, in order to evaluate their ability to reduce selectively the conjugated double bond of coumarin [16]. Out of eighteen selected strains, the yeasts Torulaspora delbrueckii, Kluyveromyces marxianus and the fungus Penicillium camemberti displayed the higher activity and selectivity in the substrate transformation.…”

Section: Introductionmentioning

confidence: 99%

“…Overall, our previous results indicated that the problem of substrate concentration could be partially solved employing specific strains, specially selected for their resistance to coumarin toxicity. In addition, our previous studies on the enzymatic mechanism of coumarin reduction [13,16] suggest that an enzymatic process could be a valid alternative to the microbial biotransformation.…”

Section: Introductionmentioning

confidence: 99%

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Two Biotechnological Approaches to the Preparative Synthesis of Natural Dihydrocoumarin

Serra¹,

Marzorati²,

Valentino³

2021

Catalysts

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In this work, we describe two different biotechnological processes that provide the natural flavour dihydrocoumarin in preparative scale. Both the presented approaches are based on the enzyme-mediated reduction of natural coumarin. The first one is a whole-cell process exploiting the reductive activity of the yeast Kluyveromyces marxianus, a Generally Recognized As Safe (GRAS) microorganism that possesses high resistance to the substrate toxicity. Differently, the second is based on the reduction of natural coumarin by nicotinamide adenine dinucleotide phosphate (NADPH) and using the Old Yellow Enzyme reductase OYE2 as catalyst. NADPH is used in catalytic amount since the co-factor regeneration is warranted employing an enzymatic system based on glucose oxidation, in turn catalysed by a further enzyme, namely glucose dehydrogenase (GDH). Both processes compare favourably over the previously reported industrial method as they work with higher coumarin concentration (up to 3 g/L for the enzymatic process) yet allowing the complete conversion of the substrate. Furthermore, the two approaches have significant differences. The microbial reduction is experimentally simple but the isolated dihydrocoumarin yield does not exceed 60%. On the contrary, the enzymatic approach requires the use of two specially prepared recombinant enzymes, however, it is more efficient, affording the product in 90% of isolated yield.

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“…The second reason concerns its biocatalytic potential. As described in our previous research [16], Kluyveromyces marxianus possesses high resistance to coumarin toxicity.…”

Section: Resultsmentioning

confidence: 53%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Two Biotechnological Approaches to the Preparative Synthesis of Natural Dihydrocoumarin

Serra¹,

Marzorati²,

Valentino³

2021

Catalysts

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“…Coumarin is banned because of its safety risk, but dihydrocoumarin has been used as a food flavouring agent for more than twenty years (Serra et al, 2019). Dihydrocoumarin is safe for humans at a dose of 100 ppm (0.1 g/L) and can be found in puddings and frozen dairy products (Häser et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

The potential of Myosoton aquaticum extracts and compounds to control barnyardgrass (Echinochloa crus‐galli)

et al. 2021

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Although cover crops are widely used for weed suppression in the agroecosystem, little research is available regarding the possibility that the decomposed weed solution may control weeds. In this study, we tested the allelopathic effect of decomposed weed solution on barnyardgrass (Echinochloa crus-galli). In two separate tests, we found that the solution of decomposed Myosoton aquaticum completely suppressed the germination of barnyardgrass seeds at a concentration of 20 g l-1. We then isolated and identified the predominant inhibitory substance in the decomposed M. aquaticum solution. The solution was subjected to organic solvent extraction and then separated on a silica gel column before the final identification of its components by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/ MS). Dihydrocoumarin was isolated from the M2 and M3 fractions using a bioassayguided method and was found to be the most active of eleven candidate allelochemical compounds. Dihydrocoumarin is typically used as a food additive. In this study, barnyardgrass germination was completely inhibited when treated with dihydrocoumarin at 1 g l-1, and its root and shoot growth were significantly inhibited at an application rate of 0.005 g l-1. Dihydrocoumarin showed highly efficient inhibition of root and shoot growth in barnyardgrass at low concentrations compared with coumarin.These findings show that weeds may be used as cover crops to provide allelochemicals for weed suppression and that dihydrocoumarin has the potential to be used as a bioherbicide.

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“…Bioconversion product was purified as described in [ 37 ] with minor changes. The biotransformation broth was treated with concentrated HCl aq.…”

Section: Methodsmentioning

confidence: 99%

The First Step of Biodegradation of 7-Hydroxycoumarin in Pseudomonas mandelii 7HK4 Depends on an Alcohol Dehydrogenase-Type Enzyme

Krikštaponis

Urbelis

Meškys

2021

IJMS

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Coumarins are well known secondary metabolites widely found in various plants. However, the degradation of these compounds in the environment has not been studied in detail, and, especially, the initial stages of the catabolic pathways of coumarins are not fully understood. A soil isolate Pseudomonas mandelii 7HK4 is able to degrade 7-hydroxycoumarin (umbelliferone) via the formation of 3-(2,4-dihydroxyphenyl)propionic acid, but the enzymes catalyzing the α-pyrone ring transformations have not been characterized. To elucidate an upper pathway of the catabolism of 7-hydroxycoumarin, 7-hydroxycoumarin-inducible genes hcdD, hcdE, hcdF, and hcdG were identified by RT-qPCR analysis. The DNA fragment encoding a putative alcohol dehydrogenase HcdE was cloned, and the recombinant protein catalyzed the NADPH-dependent reduction of 7-hydroxycoumarin both in vivo and in vitro. The reaction product was isolated and characterized as a 7-hydroxy-3,4-dihydrocoumarin based on HPLC-MS and NMR analyses. In addition, the HcdE was active towards 6,7-dihydroxycoumarin, 6-hydroxycoumarin, 6-methylcoumarin and coumarin. Thus, in contrast to the well-known fact that the ene-reductases usually participate in the reduction of the double bond, an alcohol dehydrogenase catalyzing such reaction has been identified, and, for P. mandelii 7HK4, 7-hydroxycoumarin degradation via a 7-hydroxy-3,4-dihydrocoumarin pathway has been proposed.

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Biocatalytic Synthesis of Natural Dihydrocoumarin by Microbial Reduction of Coumarin

Cited by 13 publications

References 37 publications

Two Biotechnological Approaches to the Preparative Synthesis of Natural Dihydrocoumarin

Two Biotechnological Approaches to the Preparative Synthesis of Natural Dihydrocoumarin

The potential of Myosoton aquaticum extracts and compounds to control barnyardgrass (Echinochloa crus‐galli)

The First Step of Biodegradation of 7-Hydroxycoumarin in Pseudomonas mandelii 7HK4 Depends on an Alcohol Dehydrogenase-Type Enzyme

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