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Enzyme Location. Extracellular (secreted)IntracellularCytosolic (soluble)Membrane-associatedIntegralPeripheral. Definitions of Enzyme Activity. Unit: amount of enzyme that catalyzes the formation of 1 mmol of product per minSpecific activity: units per mg proteinTurnover number: moles of substrate converted to product per sec by 1 mol of enzyme.
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1. Enzymes
Enzymes are biological catalysts responsible for supporting almost all of the chemical reactions that maintain biological homeostasis. The macromolecular components of almost all enzymes are composed of protein, except for a class of RNA modifying catalysts known as ribozymes. Ribozymes are molecules of ribonucleic acid that catalyze reactions on the phosphodiester bond of other RNAs.
2. Enzyme Location Extracellular (secreted)
Intracellular
Cytosolic (soluble)
Membrane-associated
Integral
Peripheral
9. Definitions of Enzyme Activity Unit: amount of enzyme that catalyzes the formation of 1 mmol of product per min
Specific activity: units per mg protein
Turnover number: moles of substrate converted to product per sec by 1 mol of enzyme
10. Enzyme Measurement Chemical: analysis of protein
Activity: conversion of substrate to product
Loss of substrate
Formation of product
Change in cofactor
11. e.g., Lactate Dehydrogenase Pyruvate + NADH ? lactate + NAD
Measure decrease in absorbance at 340 nm and calculate decrease in NADH concentration using the extinction coefficient (?NADH = 6.22 x 103 M-1 cm-1)
Concentration = absorbance/extinction coefficient
20. Michaelis-Menton Kinetics E + S <---> ES <---> ES* <---> EP <---> E + P
21. Lineweaver-Burk Plot
34. Types of Inhibitors
36. Enzyme Kinetics and Food Quality Freezing and dehydration
(concentration of solutes)
Substrates
Cofactors
Enzymes
Inhibitors
Activators
37. Processing and Enzyme Kinetics E + S <---> ES <---> ES* <---> EP <---> E + P
39. Temperature and Food Quality Fish: Enzyme activity at low temperature
Bacteria: Enzyme activity at high temperature
Consider the source of enzyme
40. Lipases Triacylglycerol ? diacylglycerol + fatty acid
Diacylglycerol ? monoacylglycerol + fatty acid
Monoacylglycerol ? glycerol + fatty acid
Flavor generation, emulsifier, rancidity
41. Phospholipases Phospholipase A1
Phospholipid ? 2-lysophospholipid + fatty acid
Phospholipase A2
Phospholipid ? 1-lysophospholipid + fatty acid
Membrane disruption, flavor, rancidity, cell signaling
42. Phospholipases Phospholipase C
Phospholipid ? diacylglycerol + X-phosphate
Phospholipase D
Phospholipid ? phosphatidic acid + X
Membrane disruption, cell signaling
(loss of food quality)
43. Lipoxygenase Unsaturated fatty acid ? free radicals
(cis, cis- 1,4-pentadiene double bond system)
Lipid oxidation, rancidity, bleaching
(color, flavor, texture, nutrition)
44. Carboxyprotease
Aminoprotease
Acid protease
46. Cellulase Cellulose ? oligosaccharides, glucose
Softening, sweetener
47. Pectinases Pectin methylesterase
Pectin ? demethylated pectin
Pectin lyase
Pectin ? hydrolyzed pectin
Juice clarification, prevents gelling, softening
48. Amylases a-Amylase
Amylose (Amylopectin) ? oligosaccharides
b-Amylase
Amylose (Amylopectin) ? b-Maltose
Malting, baking, starch degradation
53. Deleterious Reactions Flavor: proteases, lipases, lipoxygenase
Color: phenolase, invertase, amylase, maltase
Texture: phospholipases, proteases, pectinase, cellulase
54. Enzymes and Food Processing Natural, nontoxic, and specific
Active at low concentrations
Active under mild conditions
Can control reactions by knowing enzyme properties
55. Analytical Tools Acetic acid (cheese, mayonnaise)
Ascorbic acid (potatoes, flour)
Cholesterol (noodles, egg liqueur)
Citric acid (bread, meat, cheese)
Lactose (cheese, ice cream)
Lecithin (egg, noodles)
Triacylglycerol (margarine, salad oil)
56. Biotechnology Immobilized enzymes
Batch process
Continuous process
Recombinant DNA Technology
Gene cloning
Enzyme expression
(flavor, color, texture, nutrition)