Measurement of Enzyme Kinetics by Use of a Blood Glucometer: Hydrolysis of Sucrose and Lactose

Heinzerling, Peter; Schrader, Frank Charles; Schanze, Sascha

doi:10.1021/ed200735f

Cited by 32 publications

(33 citation statements)

References 15 publications

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“…21 Briefly, these analytical methods can be classified into two common approaches; (a) Spectrophotometry coupled to colorimetric based sugar detection assays, 11,13 and (b) Amperometry coupled to enzyme-based sugar detection assays. 22 Colorimetric based assays were first developed over 150 years ago to quantify glucose in urine and blood samples to diagnose diabetic patients and have been adapted over the years as part of several undergraduate-level teaching and clinical laboratories. 23 Bio-Rad Laboratories Incorporated developed a biofuel enzyme instructional kit for students to explore the Aspergillus derived cellobiase enzyme kinetics using model soluble cellulosic substrates tethered to chromophoric leaving groups (e.g., p-nitrophenyl based cellobiose).…”

Section: Figure 1 Overview Of Complex Carbohydrate Structures Found mentioning

confidence: 99%

Comparison of analytical methods for rapid & reliable quantification of plant-based carbohydrates for the quintessential bioenergy educator

Chundawat

2020

Preprint

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Biochemical conversion of plant-based insoluble carbohydrate polymers, such as starch from corn grains or cellulose-hemicellulose from corn stover, into soluble fermentable sugars (e.g., glucose and xylose) for bioenergy production has seen tremendous research activity and commercial-scale biorefineries deployment over the last three decades, particularly in regions around the world that have a dominant agricultural-based economy. Therefore, educators in schools and universities have developed various hands-on experimental activities to engage the general public and students either in outreach events or lab/classroom-based settings to instruct students on various inter-disciplinary concepts relevant to bioenergy and biochemicals production. One of the limitations of most available protocols is the lack of systematic and comprehensive comparison of educator-friendly analytical tools and protocols for quantitative analysis of water-soluble carbohydrates commonly encountered in a biorefinery backdrop during the biochemical conversion of lignocellulosic biomass to biofuels/biochemicals. Here, we systematically compare and validate findings from four leading analytical approaches for detection and quantification of lignocellulosic biomass derived soluble carbohydrates. We compare these assay methods based on the overall ease of use, detection accuracy/sensitivity, commercial availability, analytical cost per assay run, and suitability for use by instructors in biorefining specific hands-on activity protocols. Next, we provide a detailed instructional protocol that utilizes one of these validated soluble sugar assays as part of a ~90 min hands-on bioenergy focused activity (called 'Grass-to-Gas!') conducted at Rutgers University with pre-university high school students. 'Grass-to-Gas!' activity involves students running biochemical assays that helps them understand the various facets of cellulosic biomass hydrolysis by commercial cellulase enzymes and monitoring the total glucose product released using our validated sugar assays to finally estimate the fractional conversion of cellulose-to-glucose. Lastly, we further demonstrate how such carbohydrate-based analytical methods can be used by instructors to help university students explore and understand various chemistry, biochemistry, and chemical engineering concepts relevant to other advanced operations involved in lignocellulose biorefining. These activity protocols would greatly aid educators teaching interdisciplinary science and engineering concepts to students in the backdrop of lignocellulose biorefining.

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Section: Figure 1 Overview Of Complex Carbohydrate Structures Found mentioning

confidence: 99%

Comparison of analytical methods for rapid & reliable quantification of plant-based carbohydrates for the quintessential bioenergy educator

Chundawat

2020

Preprint

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“…Danach wurden die enzymatische Rohrzuckerhydrolyse mithilfe von Trockenhefe als Invertasequelle und die Lactosehydrolyse mithilfe von Lactase-Kapseln untersucht [11,12] …”

Section: Experimenteunclassified

“…Terumo-Spritze gearbeitet werden. In einer früheren Untersuchung wurde die Linearität der Reaktion getestet: Es kann zwischen 10 und 30 Minuten gemessen werden [12]. Nach 30 Minuten wird die Glucose-Konzentration gemessen.…”

Section: Lactosehydrolyse Mit Oligase 600unclassified

Enzymkinetische Untersuchungen mit Nahrungsergänzungsmitteln und moderner Medizindiagnostik

Heinzerling

Schanze

2014

Chemkon

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Zusammenfassung: Enzymkinetische Untersuchungen erfordern in der Regel einen hohen apparativen und analytischen Aufwand und führen in der Schule nicht immer zu optimalen Ergebnissen. In der Schulliteratur wird überwiegend mit Urease gearbeitet. Wir verwenden als Enzymquellen frei verkäufliche Nahrungsergän-zungsmittel aus der Apotheke. Analytisch werden die Resultate mit einem Glucometer erfasst. Diese Alltagsgeräte liefern in fünf Sekunden fertige Resultate. Die Experimente waren auch mit Inhibitoren erfolgreich. Stichworte: Enzymkinetik · Inhibition · Michaelis-Menten-KonstanteEnzyme Kinetics with dietary supplements and modern medical diagnostics Abstract: Experimental enzyme kinetics is of high technical and analytical complexity. The results under conditions of school laboratories are suboptimal in many cases. In textbooks experiments are mostly done with urease. We use as enzymes over the counter dietary supplements available at pharmacies. The analytical results were obtained without any transformation by a glucometer within five seconds. The experiments were successful with inhibitors, too.

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“…Educational protocols involving the analysis of lactase from over-the-counter formulations have been proposed, but these are best suited for entry-level undergraduate laboratory courses [14][15][16][17][18][19]. Protocols in which lactase Michaelis-Menten kinetic parameters are obtained typically rely upon the model substrate o-nitrophenyl-β-Dgalactopyranoside (oNPGal), since the color change during the reaction is easy to monitor.…”

Section: Introductionmentioning

confidence: 99%

Analysis of lactase in lactose intolerance supplements

Stourman

Moore

2018

Biochem Molecular Bio Educ

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Lactase is the enzyme responsible for the digestion of the disaccharide lactose, and deficiency in this enzyme causes the prevalent medical condition lactose intolerance. Management of lactose intolerance can be achieved through the administration of lactase supplements. Lactase is an appropriate platform for advanced enzymatic study because its medical application is a motivator for student learning. The following is an upper‐level biochemistry laboratory sequence that integrates student inquiry and exposure to advanced laboratory techniques. Students investigate three different lactase supplements through experimentation that includes the Bradford assay, SDS‐PAGE, continuous and discontinuous kinetic assays, and zymography. Upon completion of this project, students compile their results and conclusions in a scientifically formatted paper comparing supplement protein content and activity. This safe and inexpensive laboratory project enriches student understanding of key biochemical concepts while mirroring work performed in a realistic research setting. © 2018 International Union of Biochemistry and Molecular Biology, 46(6):652–662, 2018.

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Measurement of Enzyme Kinetics by Use of a Blood Glucometer: Hydrolysis of Sucrose and Lactose

Cited by 32 publications

References 15 publications

Comparison of analytical methods for rapid & reliable quantification of plant-based carbohydrates for the quintessential bioenergy educator

Comparison of analytical methods for rapid & reliable quantification of plant-based carbohydrates for the quintessential bioenergy educator

Enzymkinetische Untersuchungen mit Nahrungsergänzungsmitteln und moderner Medizindiagnostik

Analysis of lactase in lactose intolerance supplements

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