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E-S
E+P
E-S#
Change in environment
Lactose
Sucrose
The reason enzymes are able to speed up chemical reactions so much comes from their ability to bind to their substrates with very high specificity and significantly lower the activation energy (Ea) of the reaction converting a substrate to a product Speed up reactions by 103-1011 compared to other catalysts; 108-1020 compared to uncatalyzed reactions A
No enzyme
C C E EA
Free energy
Ea E-S E+S Ea
Enzyme
E+P
Progress of reaction
There are six main types/groups of enzymes classified based on their chemical reaction mechanism 1. Oxidoreductases Catalyze oxidations or reductions of substrates Some important food reaction examples: Lipid oxidation lipoxygenase (adds an oxygen on fatty acids) Browning polyphenol oxidase (oxidizes phenols in food) 2. Transferases Catalyze a shift of a chemical group from a donor to acceptor substrate Not so important in foods
Chemical kinetics
E + S E-S E + P
k-1
k1
k2
o This reaction curve is shared by most enzyme and gives us very useful information on the activity of the enzyme and the affinity for its substrate o Vmax gives us the maximum velocity that the enzyme can produce (under the conditions tested) the higher the faster o Km (determined as Vmax) tells us the affinity of the enzyme for its substrate o Vmax/Km = catalytic efficiency (higher number means more efficient)
Extremes of pH can affect the enzyme by denaturing it (remember it is a protein) or affecting the charge of critical amino acids in its active site (or charge on the substrate) For this reason pH control of foods with undesirable enzymes is important
Pepsin Activity Trypsin
pH
12
Enzyme 1
Activity
Enzyme 2
aw
2. Non-competitive inhibitors
Bind to enzyme at another site than active site Enzyme can bind to both S and I at the same time
3. Un-competitive inhibitors
Can only bind to the E-S complex (the intermediate state) Enzyme binds first to S and then can bind to I
These can be reversible or irreversible Some food use for these but many have flavor, odor, color and toxicity problems, plus can be very expensive
Glucoamylase Pullulunase
These enzymes are naturally present in the food or are found in microorganisms added to the food
Brewing
High level of amylases in barley malt (no need to add more) During mashing (milled barley malt and water at 50C) amylases hydrolyze starch to give maltose for yeast to utilize and produce CO2 and ethanol
CO O C H O O
CO O H
O O O
CO O C H
O
PME
O
O
O O O O O
CO O C H O O
CO O H
O O O
CO O C H
O
O O Ca
CO O H
CO O C H 3
C O O C H O O
C O O H
O O O
C O O C H
O
O O
O O
O
CO O HO
O O O CO O C H 3 O O
CO O C H 3
O O O O O O
O CO O H
O Ca
C O O C H O O
C O O H
O O O
C O O C H
O
O O O
O
O CO O H
O O
CO O C H 3 OO O O O O
O O Ca
CO O H
CO O C H 3
CO O H
O O
CO O C H 3
O O
Converts colloidal pectin to non-colloidal pectin We add this enzyme when clarity is desired (e.g. apple juice)
Most other proteases can initiate a milk clot like chymosin but they would continue the casein hydrolysis producing bitter peptides and eventually breaking the clot
Two classes
a) 1,3-lipases: preferentially hydrolyze ester bonds at SN1 and SN3 b) 2-lipases preferentially hydrolyze ester bonds at SN2
The enzyme is located in the water droplet of a water-in-oil emulsion and acts on the oil surrounding the water droplet
Product called high fructose corn syrup Made from corn syrup (which is made by amylase digestion of starch) Enzymes are immobilized in large columns where the reaction takes place can reuse them
42% (HFCS) 52%
Enzymes that catalyze the oxidation or reduction of substrates Found in a wide variety of plants (primarily legumes) and have also been identified in animal tissue (e.g. in the skin of fish) Specific for the oxidation of fatty acids that have a cis, cis penta-1,4diene unit, so there are three naturally occurring fatty acids that can be substrates
Linoleic acid (2 double bonds) Linolenic acid (3 double bonds) Arachidonic acid (4 double bonds)
A) Lipoxygenase
Undesirable
Lipid oxidation and reactions of its products
Breakdown products of hydroperoxides give off-flavors and odors Oxidation products (the free radicals or hydroperoxide) can bind and/or oxidize proteins to lead to textural problems Lipid oxidation also leads to nutritional loss of essential polyunsaturated fatty acids Vitamins may also be oxidized by the oxidation products Chlorophylls and carotenes can be bleached
Polymerizes Melanins Undesirable browning of apples, bananas, mushrooms, shrimp, lobster, human freckles? Up to 50% economic loss of tropical fruit due to PPO activity Desirable browning of tea, coffee, cocoa, raisins, prunes, tobacco, human tan, freckles?