BLBS102-c08 BLBS102-Simpson March 21, 2012 12:8 Trim: 276mm X 219mm Printer Name: Yet to Come
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Enzyme Activities
D. J. H. Shyu, J. T. C. Tzen, and C. L. Jeang
Introduction
Features of Enzymes
Most of the Enzymes are Proteins
Chemical Composition of Enzymes
Enzymes are Specific
Enzymes are Regulated
Enzymes are Powerful Catalysts
Enzymes and Activation Energy
Enzymes Lower the Activation Energy
How Does an Enzyme work?
Enzyme Kinetics and Mechanism
Regulatory Enzymes
Feedback Inhibition
Noncompetitive Inhibition
Competitive Inhibition
Enzyme Kinetics
Data Presentation
Untransformed Graphics
Lineweaver–Burk Plots
Eadie–Hofstee Plots
Hanes–Wolff Plots
Eisenthal–Cornish-Bowden Plots (the Direct Linear
Plots)
Hill Plot
Factors Affecting Enzyme Activity
Enzyme, Substrate, and Cofactor Concentrations
Effects of pH
Effects of Temperature
Methods Used in Enzyme Assays
General Considerations
Types of Assay Methods
Detection Methods
This manuscript was reviewed by Dr. Hsien-Yi Sung, Department of Biochemical
Science and Technology, National Taiwan University, Taipei, Taiwan, Republic
of China, and Dr. Wai-Kit Nip, Department of Molecular Biosciences and Bio-
engineering, University of Hawaii at Manoa, Honolulu, Hawaii, USA.
Spectrophotometric Methods
Spectrofluorometric Methods
Radiometric Methods
Chromatographic Methods
Electrophoretic Methods
Other Methods
Selection of an Appropriate Substrate
Unit of Enzyme Activity
References
INTRODUCTION
Long before human history, our ancestors, chimpanzees, might
have already experienced the mild drunk feeling of drinking
wine when they ate the fermented fruits that contained small
amounts of alcohol. Archaeologists have also found some sculp-
tured signs on 8000-year-old plates describing the beer-making
processes. Chinese historians wrote about the lavish life of Tsou,
a tyrant of Shang dynasty (around 1100bc), who lived in a castle
with wine-storing pools. All these indicated that people grasped
the wine fermentation technique thousands of years ago. In the
Chin dynasty (around 220bc) China, a spicy paste or sauce made
from fermentation of soybean and/or wheat was mentioned. It is
now called soybean sauce.
Although people applied fermentation techniques and ob-
served the changes from raw materials to special products, they
did not realize the mechanisms that caused the changes. This
mystery was uncovered by the development of biological sci-
ences. Louis Pasteur claimed the existence and function of or-
ganisms that were responsible for the changes. In the same era,
Justus Liebig observed the digestion of meat with pepsin, a
substance found in stomach fluid, and proposed that a whole
organism is not necessary for the process of fermentation.
In 1878, Kuhne was the first person to solve the conflict by in-
troducing the word “enzyme,” which means “in yeast” in Greek.
Food Biochemistry and Food Processing, Second Edition. Edited by Benjamin K. Simpson, Leo M.L. Nollet, Fidel Toldr ́a, Soottawat Benjakul, Gopinadhan Paliyath and Y.H. Hui.
©C2012 John Wiley & Sons, Inc. Published 2012 by John Wiley & Sons, Inc.
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