Purification and Characterization of Secondary Metabolites: A Laboratory Manual for Analytical Biochemistry provides students with a working knowledge of the fundamental and advanced techniques of experimental biochemistry.
Presented in this manual is an overview of the microbiological and biochemical methods that are typically used for purification of metabolites. It discussed the biological significance of secondary metabolites that are secreted by three diverse species of bacteria. These molecules function in cell to cell signaling, acquisition of iron and phytotoxicity.
Included in this manual are instruction and experiments that involve purification and characterization of enzymes from various source materials giving students excellent experience kinetics analysis and data analysis opportunities. Additionally, this lab manual covers. The theory and practice of the most commonly-used techniques of analytical biochemistry, UV-vis and IR spectrophotometry, high-performance liquid chromatography, mass spectrometry, X-ray crystallography and nuclear magnetic resonance and how to evaluate and effectively use scientific data.
For each of the three metabolites, a set of exercises with detailed protocols, is included. In these exercises: 1) cultures of bacteria are grown, 2) the metabolite is purified and 3) the metabolite is analyzed with various physical and chemical techniques. Students are also given guidance in designing their own experiments to characterize secondary bacterial metabolites having features distinct from those of the metabolites addressed.
Instructors will find this book useful because the modular nature of the lab exercises allows them to apply the exercises.
Written in a logical, easy-to-understand manner, Purification and Characterization of Secondary Metabolites: A Laboratory Manual for Analytical Biochemistry is an indispensible resource for both students and instructors in the fields of biochemistry molecular biology, chemistry, pharmaceutical chemistry, and related molecular life sciences such as cell biology, neurosciences, and genetics.
- Offers project lab formats for students that closely simulate original research projects
- Provides instructional guidance for students to design their own experiments
- Includes advanced analytical techniques
- Includes access to a website with additional resources for instructors
1. The Structure and Function of Secondary Metabolites that are Secreted by Bacteria 2. The Reagents, Supplies and Equipment that are Necessary to Grow Cultures of Bacteria in the Laboratory and to Purify Secreted Metabolites 3. Overview of the Methods for Purification of Metabolites that are Secreted by Bacteria 4. Absorption Spectrophotometry: Ultraviolet-visible and Infrared 5. High-performance Liquid Chromatography 6. Mass Spectrometry 7. X-ray Crystallography 8. Nuclear Magnetic Resonance Spectroscopy 9. Exercises in Purifying and Characterizing a Quorum-sensing Signal 10. Exercises in Purifying and Characterizing Iron-chelating Molecules 11. Exercises in Purifying and Characterizing a Chloroplast-targeting Phytotoxin 12. Designing your Own Experiments
Thomas Crowley studied biochemistry as an undergraduate at the University of Illinois Urbana-Champaign. He then pursued graduate studies in molecular biology in the Division of Biology at the California Institute of Technology. As a graduate student and later during postdoctoral studies he used biochemical and genetic methods to examine the regulation of gene expression and the intracellular localization of proteins during animal development. He has taught courses covering a wide range of chemical and biological topics such as general chemistry, biochemistry , microbiology, cell biology and developmental biology. These courses were taught at Montclair State University in New Jersey, Columbia University in New York City, the University of California San Diego and National University in La Jolla, California. He has authored articles derived from his research in a variety of journals and articles derived from his teaching in Biochemistry and Molecular Biology Education (BAMBED). He is currently an adjunct faculty member in the Department of Mathematics and Natural Sciences at National University and a member of the American Chemical Society.