Development of a method to measure laccase activity on methoxyphenolic food ingredients and isomers

Manzano-Nicolas, Jesus; Marín-Iniesta, Fulgencio; Taboada-Rodríguez, Amaury; Garcı́a-Cánovas, Francisco; Tudela, José; Muñoz‐Muñoz, José Luis

doi:10.1016/j.ijbiomac.2019.10.152

Cited by 3 publications

(4 citation statements)

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“…The products of the reaction of these three enzymes (laccase, peroxidase, and tyrosinase) when acting on OL and HT are semiquinones that evolve towards o‐ quinones, in the case of the first two (Manzano‐Nicolas, Marin‐Iniesta, et al., 2020; Manzano‐Nicolas, Taboada‐Rodriguez, et al., 2020; Rodríguez‐López, Gilabert, et al., 2000) and directly o‐ quinones in the case of tyrosinase (Espín et al., 2001; Muñoz et al., 2006; Rodríguez‐López, Fenoll, et al., 2000). It is known that o‐ quinones are unstable, especially at high pH values.…”

Section: Methodsmentioning

confidence: 99%

“…It is known that o‐ quinones are unstable, especially at high pH values. Thus, o‐ quinones absorb in the visible region of the spectrum and make it possible to measure the activity of these enzymes; however, their instability means that only laccase can be measured at pH = 4.0, at its optimal pH (Manzano‐Nicolas, Marin‐Iniesta, et al., 2020; Manzano‐Nicolas, Taboada‐Rodriguez, et al., 2020). The other two enzymes peroxidase (Rodríguez‐López, Gilabert, et al., 2000) and tyrosinase (Rodríguez‐López, Fenoll, et al., 2000), which have an optimal pH of 5.5 and 6.8, respectively, cannot be correctly measured, as shown below, in which case a spectrophotometric chronometric method is proposed, as it is described below.…”

Section: Methodsmentioning

confidence: 99%

“…The enzymatic activity of peroxidase and tyrosinase on OL and HT was followed spectrophotometrically in the visible zone, measuring the formation of the corresponding products after the consumption of a determined amount of AH 2 (micromolar) by the reaction with the different quinones and semiquinones generated by the enzymes. Since in all cases the product absorbs in the visible area, the classic chronometric method is used (Muñoz et al., 2006), since the AH 2 spectrum does not influence the measurement (Manzano‐Nicolas, Marin‐Iniesta, et al., 2020; Manzano‐Nicolas, Taboada‐Rodriguez, et al., 2020; Rodríguez‐López, Fenoll, et al., 2000; Rodríguez‐López, Gilabert, et al., 2000). Except where otherwise indicated, the experimental conditions were as follows: pH 5.5 and 50‐mM acetate buffer for peroxidase, while pH 6.8 and 50‐mM phosphate buffer for tyrosinase; temperature was maintained at 25°C.…”

Section: Methodsmentioning

confidence: 99%

“…In previous works, we have kinetically characterized the action of laccase, peroxidase, and tyrosinase on a series of substrates (Manzano‐Nicolas, Marin‐Iniesta, et al., 2020; Manzano‐Nicolas, Taboada‐Rodriguez, et al., 2020; Rodríguez‐López, Fenoll, et al., 2000; Rodríguez‐López, Gilabert, et al., 2000). Due to the instability of the products, the activity measurement method used consists of avoiding following these molecules, and this is achieved by using a small amount of ascorbic acid (AH 2 ) (µM), achieving the reversion of free radicals or o‐ quinones to their original substrate and ascorbic acid is oxidized to dehydroascorbic (Manzano‐Nicolas, Marin‐Iniesta, et al., 2020; Manzano‐Nicolas, Taboada‐Rodriguez, et al., 2020; Rodríguez‐López, Fenoll, et al., 2000; Rodríguez‐López, Gilabert, et al., 2000).…”

Section: Introductionmentioning

confidence: 99%

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Enzymatic oxidation of oleuropein and 3‐hydroxytyrosol by laccase, peroxidase, and tyrosinase

et al. 2021

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The oxidation of oleuropein and 3‐hydroxytyrosol by oxidases laccase, tyrosinase, and peroxidase has been studied. The use of a spectrophotometric method and another spectrophotometric chronometric method has made it possible to determine the kinetic parameters Vmax and KM for each enzyme. The highest binding affinity was shown by laccase. The antioxidant capacities of these two molecules have been characterized, finding a very similar primary antioxidant capacity between them. Docking studies revealed the optimal binding position, which was the same for the two molecules and was a catalytically active position. Practical applications One of the biggest environmental problems in the food industry comes from olive oil mill wastewater with a quantity of approximately 30 million tons per year worldwide. In addition, olive pomace, the solid residue obtained from the olive oil production, is rich in hydroxytyrosol and oleuropein and the action of enzymatic oxidases can give rise to products in their reactions that can lead to polymerization. This polymerization can have beneficial effects because it can increase the antioxidant capacity with potential application on new functional foods or as feed ingredients. Tyrosinase, peroxidase, and laccase are the enzymes degrading these important polyphenols. The application of a spectrophotometric method for laccase and a chronometric method, for tyrosinase and peroxidase, allowed us to obtain the kinetic information of their reactions on hydroxytyrosol and oleuropein. The kinetic information obtained could advance in the understanding of the mechanism of these important industrial enzymes.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Enzymatic oxidation of oleuropein and 3‐hydroxytyrosol by laccase, peroxidase, and tyrosinase

et al. 2021

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Kinetic characterization of the oxidation of catecolamines and related compounds by laccase

Manzano-Nicolas

Taboada-Rodríguez

Teruel-Puche

et al. 2020

International Journal of Biological Macromolecules

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The pathways of melanization and sclerotization of the cuticle in insects are carried out by the action of laccases on dopamine and related compounds. In this work, the laccase action of Trametes versicolor (TvL) on catecholamines and related compounds has been kinetically characterized. Among them, dopamine, L-dopa, Lepinephrine, L-norepinephrine, L-isoprenaline L-α-methyldopa and L-dopa methylester. A chronometric method has been used for this characterization, with the help of a small amount of ascorbic acid. The use of TvL has allowed docking studies of these molecules to be carried out at the active site of this enzyme. The hydrogen bridge interaction between the hydroxyl oxygen at C-4 with His-458, and with the acid group of Asp-206, would make it possible to transfer the electron to the copper centre of the enzyme. The presence of an isopropyl group bound to nitrogen (isoprenaline) makes it especially difficult to catalyse. The formation of the ester (L-dopa methyl ester) practically does not affect catalysis. The addition of a methyl group (α-methyl dopa) increases the rate but decreases the affinity for catalysis. L-epinephrine and Lnorepinephrine have a similar affinity to isoprenaline, but faster catalysis, probably due to the greater nucleophilic power of their phenolic hydroxyl.

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A Treatment for Rice Straw and Its Use for Mealworm (Tenebrio molitor L.) Feeding: Effect on Insect Performance and Diet Digestibility

Saura-Martínez,

Montalbán,

Manzano-Nicolás

et al. 2024

Insects

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The development of reuse processes for plant by-products for both animal and human food offers numerous possibilities for quality-of-life improvements that align with a circular economy model. For this reason, we divided this study into two experiments. First, we designed a combined treatment consisting of laccase, ultrasound, and ascorbic acid to hydrolyze rice straw plant fibers and used the resulting feed as the basis for T. molitor diets. Second, we formulated diets with different inclusion levels (0%, 25%, 50%, 75%, and 100%) of rice straw and treated rice straw to assess their impact on larvae growth and diet digestibility. For each treatment, six replicates were employed: four for the growth–performance–digestibility trial and two for complementary uric acid determination tests. The combined laccase enzyme, ultrasound, and ascorbic acid treatment hydrolyzed 13.2% of the vegetable fibers. The diets containing treated rice straw resulted in higher larvae weight and a better feed conversion ratio; however, reaching 100% by-product inclusion values led to similar results between both diets. In conclusion, these treatments improve the potential of low-nutritional-value vegetable by-products as part of a T. molitor diet, opening the possibility of new methodologies for the use of recalcitrant vegetable by-products for insect rearing.

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Development of a method to measure laccase activity on methoxyphenolic food ingredients and isomers

Cited by 3 publications

References 46 publications

Enzymatic oxidation of oleuropein and 3‐hydroxytyrosol by laccase, peroxidase, and tyrosinase

Enzymatic oxidation of oleuropein and 3‐hydroxytyrosol by laccase, peroxidase, and tyrosinase

Kinetic characterization of the oxidation of catecolamines and related compounds by laccase

A Treatment for Rice Straw and Its Use for Mealworm (Tenebrio molitor L.) Feeding: Effect on Insect Performance and Diet Digestibility

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