Rapid Microscale Isolation and Purification of Yeast Alcohol Dehydrogenase Using Cibacron Blue Affinity Chromatography

Morgan, Chad J.; Moir, Neil J.

doi:10.1021/ed073p1040

Cited by 8 publications

(5 citation statements)

References 3 publications

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“…It is commercially available in crystalline form, stable enough for student use, and the spectrophotometric YADH assay can be completed in less than two minutes, using simple equipment. Several experiments have been published using YADH, including its isolation from yeast and purification [5], kinetics [6,7], and pH stability [8]. Suelter developed an entire inquiry-based laboratory course around the study of YADH [9].…”

Section: Introductionmentioning

confidence: 99%

The competitive inhibition of yeast alcohol dehydrogenase by 2,2,2-trifluoroethanol

Taber

1998

Biochemical Education

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Section: Introductionmentioning

confidence: 99%

The competitive inhibition of yeast alcohol dehydrogenase by 2,2,2-trifluoroethanol

Taber

1998

Biochemical Education

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“…The dye-ligand chromatography step is still done by gravity feed chromatography, but it is very straightforward and can be completed in one or two lab periods. Similar purification procedures for dehydrogenases have been described previously for undergraduate biochemistry lab courses (26,27). Students then perform protein and activity assays and construct a protein purification regularly exceeds scheduled class time.…”

Section: Course Schedulementioning

confidence: 99%

A Project-Oriented Biochemistry Laboratory Course

Craig

1999

J. Chem. Educ.

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A project-oriented laboratory course has been designed to introduce students to the study of biochemistry as it is practiced. The course is designed to be a capstone experience for students enrolled in a variety of majors at the Rochester Institute of Technology, including those who enter our new B.S. Biochemistry program. The experiments in this course enable the students to explore the protein chemistry, enzymology, and molecular biology of a single enzyme, threonine dehydrogenase, in a series of integrated experiments. The laboratory incorporates both traditional methods (centrifugation, UV-vis spectroscopy, gel electrophoresis, and chromatography) and more recent developments in the field (polymerase chain reaction). Students use a small computer network to prepare for experiments (using simulation software developed at RIT), to evaluate data, to access sequence homology databases over the Internet, and to visualize and model proteins and nucleic acids. The change in the biochemistry teaching lab from a sequence of unrelated experiments to an integrated series of experiments is a model that can be readily adapted by other educators, who can change their courses to focus on a single enzyme with which they are most familiar.

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“…Alcohol dehydrogenase (ADH) is an ideal enzyme for the undergraduate biochemistry laboratory. − It is purified relatively easily from yeast − and liver, is commercially available in purified form, and has been extensively studied and reviewed. − The enzyme has two substrates, alcohol and NAD + , and is therefore more interesting to work with than the single-substrate or pseudo-first order enzymes that are typically studied in undergraduate laboratories . An added advantage is that NADH, the reduced product, is a UV chromophore that is easily assayed spectrophotometrically.…”

Section: Introductionmentioning

confidence: 99%

The Alcohol Dehydrogenase Kinetics Laboratory: Enhanced Data Analysis and Student-Designed Mini-Projects

Silverstein

2016

J. Chem. Educ.

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A highly instructive, wide-ranging laboratory project in which students study the effects of various parameters on the enzymatic activity of alcohol dehydrogenase has been adapted for the upper-division biochemistry and physical biochemistry laboratory. Our two main goals were to provide enhanced data analysis, featuring nonlinear regression, and also to give students experience in experimental design. Students use appropriate kinetic and thermodynamic equations to fit their data from Michaelis–Menten plots, enzyme activity pH profiles, inhibitor and denaturant concentration profiles, and temperature-dependence plots. Experiments at the end of this project are designed and implemented by student pairs, thus preparing them for independent research.

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Rapid Microscale Isolation and Purification of Yeast Alcohol Dehydrogenase Using Cibacron Blue Affinity Chromatography

Cited by 8 publications

References 3 publications

The competitive inhibition of yeast alcohol dehydrogenase by 2,2,2-trifluoroethanol

The competitive inhibition of yeast alcohol dehydrogenase by 2,2,2-trifluoroethanol

A Project-Oriented Biochemistry Laboratory Course

The Alcohol Dehydrogenase Kinetics Laboratory: Enhanced Data Analysis and Student-Designed Mini-Projects

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