Practical enzyme kinetics: A biochemical laboratory experiment

Rowe, Helen A.

doi:10.1021/ed065p548

Cited by 10 publications

(5 citation statements)

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“…[16,28] Collagenase activity of the pear extract was tested with azocoll, a commercially available azo dye-labeled collagen, which is a commonly used substrate for collagenase assays. [29] According to a previous study, [17] azocoll stock suspensions were extensively pre-washed to reduce non-specific release of azo dye. Azocoll collagen solution was kept suspended by agitation during the assay time course since agitation has been shown to have a critical effect on Azocoll hydrolysis.…”

Section: Functional Caharcterization Of Pear Cysteine Proteasementioning

confidence: 99%

Identification and Functional Characterization of Cysteine Protease from Nine Pear Cultivars (Pyrus pyrifolia)

Nam

Walsh

Kim

et al. 2015

International Journal of Food Properties

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This study was performed to compare the total protein and protease content among the whole fruit, flesh, and peel of nine different pear cultivars. Pear proteases were functionally characterized with respect to three enzyme assays. Proteases from pears were further identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis, in gel activity staining, and matrix assisted laser desorption/ionization-time of flight mass spectrometry analysis. Flesh from Whasan, Nikita, and Hanareum cultivars contained relatively more total protein and protease and showed high enzyme activities, while Chuwhang contained the lowest amount of protein and protease activity. Protease content and enzyme activities found in the pear flesh or whole fruits were two to six times higher than those in the pear peel. Pear cultivars contained one or two protease bands with molecular weights of 36 kDa and/or 38k Da. The larger band was further identified as a cysteine proteinase with 70% homology to the pear cysteine protease from Pyrus pyrifolia.

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Section: Functional Caharcterization Of Pear Cysteine Proteasementioning

confidence: 99%

Identification and Functional Characterization of Cysteine Protease from Nine Pear Cultivars (Pyrus pyrifolia)

Nam

Walsh

Kim

et al. 2015

International Journal of Food Properties

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“…The source of the enzyme is dried papaya, which is available from Sigma. Although contact lens tablets may also be used as a source of papain for this investigation (11), the dried papaya is less costly. In addition, as contact lens wearers tend to prefer disposable lenses, enzyme tablets containing papain are less widely available.…”

Section: Reagents and Equipmentmentioning

confidence: 99%

“…This investigation also makes use of a chromogenic substrate (10). Other investigators have studied the kinetics of papain using the nonspecific substrate Azocoll (11). Here we use a more specific substrate, taking advantage of the fact that papain interacts with a phenylalanine residue two amino acids away from the peptide bond cleaved (12).…”

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confidence: 99%

Kinetics of Papain: An Introductory Biochemistry Laboratory Experiment

et al. 1999

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“…Although many undergraduate enzyme kinetics experiments for biochemistry laboratories have been published, − in most, the rate data are only analyzed using one, − , two, ,,,, or three types of plots and a single enzyme. ,,,,,, This procedure does not provide advanced undergraduate students an opportunity to explore the strengths and weaknesses of the Michaelis–Menten, Lineweaver–Burk, Eadie–Hofstee, and Hanes plots, which are commonly used to describe enzyme kinetics …”

Section: Introductionmentioning

confidence: 99%

Enzyme Kinetics Experiment with the Multienzyme Complex Viscozyme L and Two Substrates for the Accurate Determination of Michaelian Parameters

Guerra

2017

J. Chem. Educ.

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A laboratory experiment in which students study the kinetics of the Viscozyme-L-catalyzed hydrolysis of cellulose and starch comparatively was designed for an upper-division biochemistry laboratory. The main objective of this experiment was to provide an opportunity to perform enhanced enzyme kinetics data analysis using appropriate informatics and statistical tools. By fitting the Michaelis–Menten, Lineweaver–Burk, Eadie–Hofstee, and Hanes equations to the experimental data, students explore the strengths and weaknesses of each plot and determine whether the two Viscozyme-L-catalyzed reactions obey Michaelis–Menten kinetics. Comparing the values of the Michaelian constants (v max and K M) and the efficiencies obtained for each enzyme-catalyzed reaction allows advanced undergraduates to determine if the differences between these parameters are statistically significant.

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Practical enzyme kinetics: A biochemical laboratory experiment

Abstract: The authors share an experiment using papain, a substance found in papaya that has been used in meat tenderizer as the basis of a practical study of enzyme kinetics.

Cited by 10 publications

References 0 publications

Identification and Functional Characterization of Cysteine Protease from Nine Pear Cultivars (Pyrus pyrifolia)

Identification and Functional Characterization of Cysteine Protease from Nine Pear Cultivars (Pyrus pyrifolia)

Kinetics of Papain: An Introductory Biochemistry Laboratory Experiment

Enzyme Kinetics Experiment with the Multienzyme Complex Viscozyme L and Two Substrates for the Accurate Determination of Michaelian Parameters

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