A rapid and continuous spectrophotometric method to measure ‐glucosidase activity based on p‐nitrophenyl ‐O‐D‐glucopyranoside hydrolysis

Arnaldos, Tomás López; Serrano, María del Carmen Martínez; Calderón, Antonio A.; Muñoz, Romualdo

doi:10.1002/(sici)1099-1565(199907/08)10:4<171::aid-pca452>3.3.co;2-w

Cited by 3 publications

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“…The first phase of this study sought to adapt and validate the assay developed by Arnaldos et al [25] for measuring β-glucosidase activity in a non-saliva matrix to whole Fig. 2 Comparison of almond β-glucosidase activity measured at 400 and 405 nm.…”

Section: Discussionmentioning

confidence: 99%

“…A method to measure β-glucosidase activity from solubilized strawberry callus protein developed by Arnaldos et al [25] was evaluated and modified for use in human saliva. The reagents were 6 mM p-nitrophenyl-β-O-Dglucopyranoside (Sigma Aldrich, St. Louis, MO, USA), 6 mM α-cyclodextrin (Sigma Aldrich, St. Louis, MO, USA), 0.025 units of almond β-glucosidase (Sigma Aldrich, St. Louis, MO, USA) and 0.1 M phosphate buffer pH of 6.5, used to correct to a total volume of 2 mL.…”

Section: Assay Development and Validationmentioning

confidence: 99%

“…The reagents were 6 mM p-nitrophenyl-β-O-Dglucopyranoside (Sigma Aldrich, St. Louis, MO, USA), 6 mM α-cyclodextrin (Sigma Aldrich, St. Louis, MO, USA), 0.025 units of almond β-glucosidase (Sigma Aldrich, St. Louis, MO, USA) and 0.1 M phosphate buffer pH of 6.5, used to correct to a total volume of 2 mL. α-Cyclodextrin was included as it is reported to increase the sensitivity of the assay [25]. One enzyme unit is defined as the protein that hydrolyzes 1 μmol of salicin per minute at pH 5 and 37°C.…”

Section: Assay Development and Validationmentioning

confidence: 99%

“…1). Due to different reported values for the p-nitrophenol wavelength maxima (405 nm, [25]; 400 nm, [34]), we measured almond β-glucosidase activity using 0.1 M phosphate buffer as well as 0.1 M phosphate buffer containing 25 μL of heat-inactivated human saliva at both 400 and 405 nm. There was no significant difference between the reaction rates observed for each wavelength for either matrix (Fig.…”

Section: Assay Development and Validationmentioning

confidence: 99%

“…Menguy et al [24] suggested that approximately one in five people possess β-glucosidase in their saliva; however, the sample size of their study was quite small. Later, Arnaldos et al [25] modified a spectrophotometric assay for strawberry callus protein for potential application in determining β-glucosidase in human saliva.…”

Section: Introductionmentioning

confidence: 99%

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Development and application of assay for determining β-glucosidase activity in human saliva

Stradwick

Inglis

Kelly

et al. 2017

Flavour

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Background: β-glucosidase is an enzyme important to flavour enhancement. It hydrolyzes glucosides to release aglycones-aroma precursors that are bound to a sugar molecule-thereby making them available to contribute to the flavour of foods and beverages. While there is strong interest within the food and beverage industry to optimizing flavour through the use of exogenous and endogenous glucosidase in production, little is known regarding the possible occurrence of these enzymes within the human oral cavity. This could be an important source of flavour release and/or account for some differences between individuals in flavour perception. In the present study, we determined whether β-glucosidase is present in human saliva. First, an existing spectrophotometric assay that uses p-nitrophenyl-β-O-D-glucopyranoside as a substrate was modified and optimized for use in human saliva. The following variables were evaluated and where necessary, optimized: linearity of the assay signal, possible matrix interference, the effect of heat inactivation of the saliva, absorbance wavelength maxima, substrate saturation concentration, maximum saliva volume and the inclusion of α-cyclodextrin. The modified assay was then used to screen for β-glucosidase activity in the saliva of 20 individuals. Of the 20 samples analyzed, four were tentatively identified as containing active β-glucosidase and were further investigated. Results: Significant differences (p < 0.05) in absorbance values (A400 nm ) between these saliva samples confirm low levels of β-glucosidase activity in approximately 20% of the population sampled. Conclusions: Inter-individual variability exists in β-glucosidase activity within the oral cavity. The described method can be applied to rapidly assay a large population of individuals, and further elucidate the extent and significance of salivary β-glucosidase activity within the context of human flavour perception and enhancement.

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Section: Discussionmentioning

confidence: 99%

Section: Assay Development and Validationmentioning

confidence: 99%

Section: Assay Development and Validationmentioning

confidence: 99%

Section: Assay Development and Validationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development and application of assay for determining β-glucosidase activity in human saliva

Stradwick

Inglis

Kelly

et al. 2017

Flavour

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Changes in phenol content during strawberry (Fragaria×ananassa, cv. Chandler) callus culture

Arnaldos

Muñoz

Ferrer

et al. 2001

Physiologia Plantarum

Self Cite

111

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Total soluble phenols, soluble flavanols, (+)-catechin, ferulic acid and 1-O-feruloyl-beta-d-glucose were analyzed during the development of a strawberry (Fragariaxananassa, cv. Chandler) callus culture. The time-course changes of the different phenols assayed were well correlated with callus growth and morphology. The changes in polyphenol oxidase (EC 1.10.3.1-2) and beta-glucosidase (EC 3.2.1.21) activities in the callus were also examined. The total phenol, soluble flavanols and (+)-catechin contents were high during the preexponential and exponential phases of growth. The subsequent decrease in (+)-catechin concentration coincided with high levels of polyphenol oxidase activity. The 1-O-feruloyl-beta-d-glucose content was highest as callus growth ceased, and its subsequent decrease was accompanied by the increased production of ferulic acid. This increase in ferulic acid was accompanied by an increase in beta-glucosidase activity. The ferulic acid content decreased at the end of culture, when callus growth had stopped and showed clear symptoms of senescence. This decrease in the ferulic acid concentration was accompanied by an increase in the levels of ferulic acid bound to cell wall components.

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Enzymes and wine quality

Canal-Llaubères¹

2010

Managing Wine Quality

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A rapid and continuous spectrophotometric method to measure ‐glucosidase activity based on p‐nitrophenyl ‐O‐D‐glucopyranoside hydrolysis

Cited by 3 publications

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Development and application of assay for determining β-glucosidase activity in human saliva

Development and application of assay for determining β-glucosidase activity in human saliva

Changes in phenol content during strawberry (Fragaria×ananassa, cv. Chandler) callus culture

Enzymes and wine quality

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