Purification and Characterization of Secondary Metabolites

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

Established scientists managing biochemical research laboratories at universities or specialized research institutes may use this lab manual as a reference when training new students or employees

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