We created a novel laboratory experience where undergraduate students explore the techniques used to study protein misfolding, unfolding, and aggregation. Despite the importance of protein misfolding and aggregation diseases, protein unfolding is not typically explored in undergraduate biochemistry laboratory classes. Yeast alcohol dehydrogenase…

The inhibition of porcine pancreatic lipase by the drug orlistat was studied through a series of activities that combine experimental and computational procedures. The enzymatic activity of the lipase was determined by measuring enzyme kinetics and calculating parameters such as K[subscript m] and V[subscript max] from experimental data. Assays of…

This paper describes an undergraduate biochemistry laboratory module consisting of a set of experiments designed around a purification scheme for bovine serum albumin (BSA). Students purify BSA from cow plasma by a combination of salt and acetone precipitation, equilibrium dialysis, ion exchange, and size exclusion chromatography. Students use the…

Here, we present a 13-week research-based biochemistry laboratory curriculum designed to provide the students with the experience of engaging in original research while introducing foundational biochemistry laboratory techniques. The laboratory experience has been developed around the directed evolution of an enzyme chosen by the instructor, with…

The kinetic parameters for the conversion of alpha-D-glucose to beta-D-glucose were measured using a blood glucometer. The reaction order, rate constant, and Arrhenius activation energy are reported for the noncatalyzed reaction and turnover number and Michaelis constant are reported for the reaction catalyzed by porcine kidney mutarotase. The…

Developments in analytical chemistry were made using acoustically levitated small volumes of liquid to study enzyme reaction kinetics and by detecting volatile organic compounds in the gas phase using single-walled carbon nanotubes. Experience gained in engineering, electronics, automation, and software development from the design and…

A new method is presented for measuring the rate of the oxidation of glucose in the presence of glucose oxidase. The improved method employs luminescence measurements to directly determine the concentration of oxygen in real time, thus obviating complicated reaction schemes employed in previous methods. Our method has been used to determine…

Describes an undergraduate enzyme kinetics experiment which uses bovine erythrocyte carbonic anhydrase, a very stable enzyme commercially available in lyophilized form. Includes background information, reactions involved, procedures used, and the calculation of typical results obtained. (JN)

Background information, procedures used, typical results obtained, and calculations are provided for a short (4-6 hours) kinetic experiment which includes different methods both in performance and in analysis of the results. The experiment can be modified and extended depending on the level of students and the available laboratory time. (JN)

Describes a set of experiments (for senior-level biochemistry students) which permit evaluation and estimation of rate and equilibrium constants involving an intermediate in the alpha-chymotrypsin mediated hydrolysis of ortho-hydroxy-alpha-toluenesulfonic acid (I). The only equipment required for the experiments is a well-thermostated double beam…

Described is an advanced undergraduate kinetics experiment using buffer dilutions to determine spontaneous rate, catalyzed rate, and reaction order. The reaction utilized is hydrolysis of p-nitro-phenyl acetate in presence of imidazole, which has been shown to enhance rate of the reaction. (Author/JN)

Describes an experiment to examine the kinetics of carbamate decomposition and the effect of buffer catalysis on the reaction. Includes background information, laboratory procedures, evaluation of data, and teaching suggestions. (Author/JN)

Discusses an enzyme kinetics laboratory experiment involving a two substrate system for undergraduate biochemistry. Uses the enzyme gamma-glutamyl transpeptidase as this enzyme in blood serum is of clinical significance. Notes elevated levels are seen in liver disease, alcoholism, and epilepsy. Uses a spectrophotometer for the analysis. (MVL)

Examines the influence of enzyme and substrate concentrations, pH, temperature, and inhibitors on catalytic activity. Follows the influence of different phosphate concentrations in the growth medium on enzyme activity. Studies regulation of enzyme synthesis by repression. Includes methodology for six experiments. (MVL)