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Angela Chen

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Angela Chen. Sweeteners from Starch…. Sweeteners from Starch…. Sweeteners from Starch…. Sweeteners from Starch…. Sweeteners from Starch…. Sweeteners from Starch…. Hydrocolloids. Binding water with carbohydrates. Starches- Our #1 Hydrocolloid?. - PowerPoint PPT Presentation

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Page 1: Angela Chen

Angela Chen

Page 2: Angela Chen

Sweeteners from Starch….

Page 3: Angela Chen

Sweeteners from Starch….

Page 4: Angela Chen

Sweeteners from Starch….

Page 5: Angela Chen

Sweeteners from Starch….

Page 6: Angela Chen

Sweeteners from Starch….

Page 7: Angela Chen

Sweeteners from Starch….

Page 8: Angela Chen

Hydrocolloids

Binding water with carbohydrates

Page 9: Angela Chen

Starches- Our #1 Hydrocolloid? Hydrocolloids are substances that will form a gel or

add viscosity on addition of water.

Most are polysaccharides and all form significant H-bonding with water with processing.

Size, structure, and charge are the most important factors relating to texture and physical features of foods

Page 10: Angela Chen

Small versus LargeSmall molecule sugars would create a high

osmotic pressure if stored in sufficient quantities to be useful.

Polymerized sugars reduce the number of molecules present and hence the osmotic effects.

Free polymers are too thick to allow cell to function

Thus, plants store energy into starch granules

Page 11: Angela Chen

AMYLOSELinear polymer of glucoseα 1 - 4 linkagesDigestable by humans (4 kcal/g)250-350 glucose units on average

Varies widely

Corn, wheat, and potato starch ~10-30% amylose

Page 12: Angela Chen

AMYLOPECTINBranched chain polymer of glucoseα 1 - 4 and α 1 - 6 glycosidic linkagesMostly digestible by humans1,000 glucose units is common

Branch points every ~15-25 units

Page 13: Angela Chen

StarchAmylose may have a few branched chains

Helical structure with a hydrophobic core Core may contain lipids, metals, etc.

Amylose to Amylopectin ratios ~ 1:4 Varies with the plant source

Waxy starches are ~100% amylopectinSugary “mutant” starches have more amylose

Page 14: Angela Chen

Straight-Chained Starch = AmyloseGlucose polymer linked α-1,4 and α-1,6

Page 15: Angela Chen

Starch

Page 16: Angela Chen

Birefringence When starch granules are viewed under the microscope by polarized light they exhibit a phenomenon known as birefringency - the refraction of polarized light by the intact crystalline regions to give a characteristic "Maltese cross" pattern on each granule. The cross disappears upon heating and gelatinization.

Page 17: Angela Chen

Modified Starches

Gelatinization is the easiest modification Heated in water then dried.

Acid and/heat will form “dextrins” α-Amylase

hydrolyzes α (1-4) linkage random attack to make shorter chains

β-Amylase Also attacks α (1 - 4) linkages Starts at the non-reducing end of the starch chain Gives short dextrins and maltose

Both enzymes have trouble with α (1 - 6) linkages

Page 18: Angela Chen

Gelatinization of Starch Native starch granules are insoluble in cold water, despite

some “swelling” Heated water increases kinetic energy, breaking some

intermolecular bonds, and allows water to penetrate The gelatinization point is where crystallinity is lost

GTR is the temperature range over which gelatinization occurs.

As water is bound, the viscosity increases. GTR is different from different starch types There must be enough water to break open and bind to

the starch hydrogen binding sites.

Page 19: Angela Chen

Starch grains swell when heated in water

Gelatinization

Page 20: Angela Chen

H-bonds break, amylose can spill from the grain

Page 21: Angela Chen

Gelatinization is done

Gains may loose integrity

Page 22: Angela Chen

During cooling, junction zones form Between amylose and amylopectin

Page 23: Angela Chen

water

water

water

waterwater

water

Water is trappedForming a gel.

Page 24: Angela Chen

WATER

As the gel dehydrates and/or junction zonesTighten, water is “squeezed” from the gel, in a syneresis process.

Page 25: Angela Chen

Starch ModificationsCross-linking (common modification)

Alkali treatment (pH 7.5-12) with salt Phosphorus oxychloride Sodium trimetaphosphate Adipic and acetic anhydride Starch phosphates formed after neutralization

Page 26: Angela Chen

Cross-LinkingResists viscosity breakdownResists prolonged heating effectsResists high shear ratesResists high acid environments Increased viscosity Increased texture

Page 27: Angela Chen

Starch ModificationsStarch Substitutions

Adding monofunctional groups “Blocking Groups” added to the starch Acetyl (2.5% max starch acetates) Hydroxypropyl, phosphates, ethers

Slows retrogradation (re-association of amylose) Lowers GTR, stabilizes the starch

Acetate + Starch

Page 28: Angela Chen

Starch ModificationsOxidation and Bleaching

Hydrogen peroxide Ammonium persulfate Na/Ca hypochlorite

0.0082 lbs chlorine/pound of starch

K-permanganate Na-chlorite

Whitens the starch Removes carotenes and other natural pigments

~25% of oxidizers break C-C linages ~75% of oxidizers will oxidize the hydroxyl groups Lowers viscosity, improves clarity of gels

Page 29: Angela Chen

Polysaccharide Breakdown Products

Page 30: Angela Chen

Hydrolytic Products Maltose Maltitol Maltodextrins Dextrins Dextrans

Maltose = glucose disaccharide Maltitol = example of a “polyol” Maltodextrins = enzyme converted starch fragments

DextrDextriinsns = starch fragments (α-1-4) linkages produced by hydrolysis of amylose

DextrDextraansns = polysaccharides made by bacteria and yeast metabolism, fragments with mostly α (1 - 6) linkages

Page 31: Angela Chen

Maltodextrins and enzyme-converted starch:

STARCHSTARCH fermentation SUGARS

ETHANOL

MODIFIED STARCHESMODIFIED STARCHES

GELATINIZED STARCHGELATINIZED STARCH alpha amylase Maltodextrins

Corn Syrups

Sugars

Page 32: Angela Chen

The smaller the size of the products in these reactions, the higher the dextrose equivalence (DE), and the sweeter they are

Starch DE = 0 Glucose (dextrose) DE = 100

Maltodextrin (MD) DE is <20

Corn syrup solids (CS) DE is >20

Low DE syrup alpha amylase MD beta amylase High DESyrup

Page 33: Angela Chen

DextrinizationA non-enzymatic method to product low-

molecular weight fragments High heat treatment of acidified starch “Pyro-conversion” of starch to dextrins

Both breaks and re-forms bonds Wide-range of products formed

Vary in viscosity Solubility Color (white, yellow) Reducing capacity Stability

Page 34: Angela Chen

Hydrocolloids

Binding water with carbohydrates

“Gums”

Page 35: Angela Chen

“Vegetable gum” polysaccharides are substances derived

from plants, including seaweed and various shrubs or trees, have the ability to hold water, and often act as thickeners, stabilizers, or gelling agents in various food products.

Plant gums - exudates, seeds (guar, xanthan, locust bean, etc)

Marine hydrocolloids - extracts from seaweeds(Carageenan, agar, alginates)

Microbiological polysaccharides - exocellular polysaccharides

Modified, natural polysaccharides

Page 36: Angela Chen

FUNCTIONS IN FOOD Gelation Viscosity Suspension Emulsification and stability Whipping Freeze thaw protection Fiber (dietary fiber)

Gut health Binds cholesterol

Page 37: Angela Chen

STRUCTURAL CONSIDERATIONS

Electrical charge, pH sensitive Interactions with

Oppositely charged molecules Salts Acids

Chain length Longer chains are more viscous

Linear vs Branched chains Inter-entangled, enter-woven molecules

Page 38: Angela Chen

“Structural” Polysaccharides

CellulosePolymer of glucose linked ß-1,4

HemicelluloseSimilar to celluloseConsist of glucose and other monosaccharides

Arabinose, xylose, other 5-carbon sugars

PectinPolymer of galacturonic acid

Page 39: Angela Chen

MODIFIED CELLULOSESChemically modified celluloseDo not occur naturally in plantsSimilar to starch, but β-(1,4) glycosidic bondsCarboxymethyl cellulose (CMC) most common

Acid treatment to add a methyl group Increases water solubility, thickening agent Sensitive to salts and low pH

Fruit fillings, custards, processed cheeses, high fiber filler

Page 40: Angela Chen

PECTINS Linear polymers of galacturonic acid

Gels form with degree of methylation of its carboxylic acid groups

Many natural sources

Susceptible to degrading enzymes Polygalacturonase (depolymerize) Pectin esterases (remove methyl groups)

Longer polymers, higher viscosity Lower methylation, lower viscosity Increase electrolytes (ie. metal cations), higher viscosity pH and soluble solids impact viscosity

Page 41: Angela Chen

PECTIC SUBSTANCES: cell cementing compound; fruits and vegetables; pectin will form gel with appropriate concentration, amount of sugar and pH.

Basic unit comprised of galacturonic acidgalacturonic acid.

Page 42: Angela Chen

BETA-GLUCANSExtracts from the bran of barley and oatsLong glucose chains with mixed ß-linkagesVery large (~250,000 glucose units)

Water soluble, but have a low viscosity Can be used as a fat replacer Responsible for the health claims (cholesterol) for

whole oat products Formulated to reduce the glycemic index of a food

Page 43: Angela Chen

Beta-Glucan

Beta-glucans occur in the bran of grains such as barley and oats, and they are recognized as being beneficial for reducing heart disease by lowering cholesterol and reducing the glycemic responseglycemic response.

They are used commercially to modify food texture. and as fat replacerfat replacer .

                                                                                                                                                      

         

Beta-Glucan

Page 44: Angela Chen

OthersCHITIN Polymer of N-Acetyl-D-glucosamine Found in the exoskeleton of insects and shellfish Many uses in industry, food and non-food.

INULIN Chains of fructose that end in a glucose molecule

Generally a sweet taste Isolated from Jerusalem artichokes and chicory Act as a dietary fiber Potentially a pre-biotic compound

Page 45: Angela Chen

Paper ReviewProducing fructo-oligosaccharides: For Tuesday

Page 46: Angela Chen

StarchStarch must be cooked to act as a thickening

agent Pre-gelatinized starch is made by quickly

cooking a starch and drying the product. Pre-gelatinized starch rapidly re-hydrates

without further cooking Useful thickening agent Can be used in dried sauces and salad dressings Used in products that do not require more cooking

Page 47: Angela Chen

StarchStarch suspensions are not stable to heating Swollen starch granules break down in hot,

stirred or acidic conditions Combinations (ie. heat and acid) will

depolymerizeCross-linking can help stabilize and slow or

maybe prevent breakdown

Page 48: Angela Chen

Starch Starch gels change their properties during storage Slow retrogradation of amylopectin is common The texture of a starch gel will change and show some

syneresis. Again, modified starch will resist changes during

storage Starch acetates or phosphates are common

modifications, altering the helical arrangements, and slow or inhibit retrogradation.

All stabilized starches must also be labeled as “modified starch” on an ingredient list.

Page 49: Angela Chen

“Vegetable gum” polysaccharides are substances derived

from plants, including seaweed and various shrubs or trees, have the ability to hold water, and often act as thickeners, stabilizers, or gelling agents in various food products.

Plant gums - exudates, seeds (guar, xanthan, locust bean, etc)

Marine hydrocolloids - extracts from seaweeds(Carageenan, agar, alginates)

Microbiological polysaccharides - exocellular polysaccharides

Modified, natural polysaccharides

Page 50: Angela Chen

FUNCTIONS IN FOOD Gelation Viscosity Suspension Emulsification and stability Whipping Freeze thaw protection Fiber (dietary fiber)

Gut health Binds cholesterol

Page 51: Angela Chen

STRUCTURAL CONSIDERATIONS

Electrical charge, pH sensitive Interactions with

Oppositely charged molecules Salts Acids

Chain length Longer chains are more viscous

Linear vs Branched chains Inter-entangled, enter-woven molecules

Page 52: Angela Chen

“Structural” Polysaccharides

CellulosePolymer of glucose linked ß-1,4

HemicelluloseSimilar to celluloseConsist of glucose and other monosaccharides

Arabinose, xylose, other 5-carbon sugars

PectinPolymer of galacturonic acid

Page 53: Angela Chen

MODIFIED CELLULOSESChemically modified celluloseDo not occur naturally in plantsSimilar to starch, but β-(1,4) glycosidic bondsCarboxymethyl cellulose (CMC) most common

Acid treatment to add a methyl group Increases water solubility, thickening agent Sensitive to salts and low pH

Fruit fillings, custards, processed cheeses, high fiber filler

Page 54: Angela Chen

PECTINS Linear polymers of galacturonic acid

Gels form with degree of methylation of its carboxylic acid groups

Many natural sources

Susceptible to degrading enzymes Polygalacturonase (depolymerize) Pectin esterases (remove methyl groups)

Longer polymers, higher viscosity Lower methylation, lower viscosity Increase electrolytes (ie. metal cations), higher viscosity pH and soluble solids impact viscosity

Page 55: Angela Chen

PECTIC SUBSTANCES: cell cementing compound; fruits and vegetables; pectin will form gel with appropriate concentration, amount of sugar and pH.

Basic unit comprised of galacturonic acidgalacturonic acid.

Page 56: Angela Chen

BETA-GLUCANSExtracts from the bran of barley and oatsLong glucose chains with mixed ß-linkagesVery large (~250,000 glucose units)

Water soluble, but have a low viscosity Can be used as a fat replacer Responsible for the health claims (cholesterol) for

whole oat products Formulated to reduce the glycemic index of a food

Page 57: Angela Chen

Beta-Glucan

Beta-glucans occur in the bran of grains such as barley and oats, and they are recognized as being beneficial for reducing heart disease by lowering cholesterol and reducing the glycemic responseglycemic response.

They are used commercially to modify food texture. and as fat replacerfat replacer .

                                                                                                                                                      

         

Beta-Glucan

Page 58: Angela Chen
Page 59: Angela Chen
Page 60: Angela Chen

Yeast ß-Glucan Isolation

Page 61: Angela Chen
Page 62: Angela Chen

Sugar Reactions

(Gluconic acid)(Glucuronic acid)

Page 63: Angela Chen

Properties of GlucoseC1 of glucose is the carbonyl carbonGlucose has 4 chiral centers

Non-super-imposable on its mirror imageCarbons 2, 3, 4, 5 are chiral carbons

The carbonyl carbon (C1) is also the site of many reactions involving glucose They have two enantiomeric forms, D and

L, depending on the location of the hydroxyl group at the chiral carbons.

Page 64: Angela Chen

SugarsThey have two enantiomeric forms, D and L,

depending on the location of the hydroxyl group at the chiral carbons. An enantiomer is one of two stereoisomers that are

mirror images of each other, non-superposable.

Isomerism in which two isomers are mirror images of each other. (D vs L).

Vary in their 3-D space

Page 65: Angela Chen

AnomersAn anomer is one of a special pair of

diastereomeric (isomer) aldoses or ketoses A stereoisomer that is not an enantiomer

They differ only in configuration about the carbonyl carbon (C1 for aldoses and C2 for ketoses)

Page 66: Angela Chen

Carbonyl CarbonsCarbonyl carbons are subject to nucleophilic

attack, since it is electron deficient. Electrons are drawn to this site

-OH groups on the sugar act as the nucleophile, and add to the carbonyl carbon to recreate the ring form

Page 67: Angela Chen

Carbonyl Carbons

Anomers α-anomer (~36%) β- anomer (~64%)

Page 68: Angela Chen

Sugar Anomers => Mutarotation Interconversion of α- and β- anomers The α- and β- anomers of carbohydrates are typically

stable. In solution, a single molecule can interchange between

straight and ring form different ring sizes α and β anomeric isomers

The process is dynamic equilibrium due to reversibility of reaction

All isomers can potentially exist in solution energy/stability of different forms vary

Page 69: Angela Chen

Mutarotation α- and β- anomers

Page 70: Angela Chen

IsomerizationKeto-Enol Tautomerism (equilibration)

Hydrogen migration; switch from SB to DB

Enol is predominant in aldose sugarKeto is predominant in ketose sugarKeto and Enol forms are tautomers of each other

Page 71: Angela Chen

IsomerizationGlucose and mannose are enantiomers, but

with dramatically different propertiesGlucose and fructose are isomers

Page 72: Angela Chen

Pectins in Foods

Page 73: Angela Chen

Plant Cell Wall

Middlelamella

Primary wall

Plasmalemma

Cytoplasm

Vacuole

Nucleolus

Nucleus

Water-Filled

Page 74: Angela Chen

PECTINS Linear polymers of galacturonic acid

Gels form with degree of methylation of its carboxylic acid groups

Many natural sources

Susceptible to degrading enzymes Polygalacturonase (depolymerize); PG Pectin esterases (remove methyl groups), PME

Longer polymers, higher viscosity Lower methylation, lower viscosity Increase electrolytes (ie. metal cations), higher viscosity pH and soluble solids impact viscosity

Page 75: Angela Chen

Composition: polymer of galacturonic acids; may be partially esterifiedesterified.

Pectic Acid

                                                                                                  

                                                                                                                 

Pectin Molecule

Page 76: Angela Chen

Pectins Pectins are important because they form gelsgels

Mechanism of gel formation differs by the degree of esterification (DE) of the pectin molecules DE refers to that percentage of pectin units with a methyl group attached

Free COOH groups can crosslink with divalent cationsdivalent cations

Sugar and acid under certain conditions can contribute to gel structure and formation

LM pectin “low methoxyl pectin”LM pectin “low methoxyl pectin” has DE < 50% ; gelatin is controlled by adding cations (like Ca++ and controlling the pH)

HM pectinHM pectin “high methoxyl pectin” has DE >50% and forms a gel under acidic conditions by hydrophobic interactions and H-bonding with dissolved solids (i.e. sugar)

Page 77: Angela Chen

Hydrophobic attractions between neighboring pectin polymer chainspromote gelation

Ca++ Ca++

Page 78: Angela Chen

ProteinsMany important functions

Functional Nutritional Biological

EnzymesStructurally complex and large compoundsMajor source of nitrogen in the diet

By weight, proteins are about 16% nitrogen

Page 79: Angela Chen

Properties of Amino AcidsAliphatic chains: Gly, Ala, Val, Leucine, IleHydroxy or sulfur side chains: Ser, Thr, Cys, MetAromatic: Phe, Trp, TryBasic: His, Lys, ArgAcidic and their amides: Asp, Asn, Glu, Gln

Page 80: Angela Chen

Properties of Amino Acids:

Aliphatic Side Chains

Aromatic Side Chains

Acidic Side Chains

SulfurSideChains

Page 81: Angela Chen

Properties of Amino Acids:Zwitterions are electrically neutral, but carry a

“formal” positive or negative charge.

Page 82: Angela Chen

The Zwitterion Nature Zwitterions make amino acids good acid-base buffers.

Accepting H+ is acidic environments; donating H+ in basic environments

For proteins and amino acids, the pH at which they have no net charge in solution is called the Isoelectric Point of pI (i.e. IEP).

The solubility of a protein depends on the pH of the solution.

Similar to amino acids, proteins can be either positively or negatively charged due to the terminal amine -NH2 and carboxyl (-COOH) groups.

Proteins are positively charged at low pH and negatively charged at high pH. When the net charge is zero, we are at the IEP.

A charged protein helps interactions with water and increases its solubility.

As a result, protein is the least soluble when the pH of the solution is at its isoelectric point.

Page 83: Angela Chen

Physical Nature of Proteins

Page 84: Angela Chen

Secondary protein structureThe spatial structure the protein assumes along

its axis (its “native conformation” or min. free energy)

This gives a protein functional properties such as flexibility and strength

Page 85: Angela Chen

Tertiary Structure of Proteins3-D organization of a polypeptide chainCompacts proteins Interior is mostly devoid of water or charge groups

3-D folding of chain

Page 86: Angela Chen

Quaternary Structure of ProteinsNon-covalent associations of protein units

Page 87: Angela Chen

ProteinsChanges in structure Denaturation

Breaking of any structure except primary Examples:

Heat Salt/Ions Alcohol pH extremes Shear Enzymes

Page 88: Angela Chen

Emulsoids and Suspensiods

Proteins should be thought of as solids Not in true solution, but bond to a lot of water

Can be described in 2 ways:

Emulsoids- have close to the same surface charge, with many shells of bound water

Suspensoids- colloidal particles that are suspended by charge alone

Page 89: Angela Chen

Functional Properties of Proteins3 major categories Hydration properties

Protein to water interactions Dispersibility, solubility, adhesion, Water holding capacity, viscosity

Structure formation Protein to protein interactions Gel formation, precipitation, Aggregation

Surface properties Protein to interface interactions Foaming, emulsification

Page 90: Angela Chen

1. Hydration Properties (hydration)

Proteins are important hydrocolloids As ingredients, many are sold as dry powders Hydrating and processing w/o denaturation

Solubility- Mostly, denatured proteins are less soluble than native proteins

Many (but not all) proteins (particularly suspensoids) aggregate or precipitate at their isoelectric point (IEP)

Protein viscosity is influenced by amount, size, shape, pH, water content, and solubility of the proteins

Page 91: Angela Chen

2. Structure Formation (protein interactions)

Gels – a 3-D network of protein and water. Attractive and repulsive forces between adjacent polypeptides

Gelation- when denatured proteins aggregate and form an ordered protein matrix Water absorption and thickening Formation of viscous, solid, or visco-elastic gels

For many proteins, heated followed by cooling forms the gel

Texturization – Proteins are responsible for the structure and texture of many foods Meat, bread dough, gelatin Texturized proteins are modified with with salts, acid/alkali,

oxidants/reductants “Pink Slime” Can also be processed to mimic other proteins (i.e. surimi)

Page 92: Angela Chen

3. Surface Properties (interfaces)

Emulsions- Exposure of protein hydrophobic regions to lipids (ie. tertiary structures) Not all proteins make good emulsifiers Can strengthen a normal emulsion system

Foams- trapping gas bubbles in a viscous medium Protein is usually soluble Air bubble size is critical (nebulized air) Duration and shear rate Temperature and physical kinetics Food ingredient interactions (i.e. salt, acid, and lipids)..bad. Metal ions, hydrocolloids, and sugar can increase stability

Page 93: Angela Chen

Enzymes

Page 94: Angela Chen

Enzyme Influencing Factors

Temperature-dependence of enzymesEvery enzyme has an optimal temperature for

maximal activityThe effectiveness of an enzyme: Enzyme activityFor most enzymes, it is 30-40°CMany enzymes denature >45°CEach enzyme is different, and vary by isozymes Often an enzyme is at is maximal activity just

before it denatures at its maximum temperature

Page 95: Angela Chen

pHLike temp, enzymes have an optimal pH where

they are maximally activeGenerally between 4 and 8

with many exceptions

Most have a very narrow pH range where they show activity.

This influences their selectivity and activity.

Page 96: Angela Chen

Water ActivityEnzymes need “free” water to operateLow Aw foods have slower enzyme reactions

Ionic StrengthSome ions may be needed by active sites on the

protein (salting in) Ions may be a link between the enzyme and substrate Ions change the surface charge on the enzyme Ions may block, inhibit, or remove an inhibitor Others, enzyme-specific

Page 97: Angela Chen

Common Enzymes in FoodsPolyphenol oxidasePlant cell wall degrading enzymesProteasesLipasesPeroxidase/CatalaseAmylaseAscorbic acid oxidaseLipoxygenase

Page 98: Angela Chen

Enzyme Influencing Factors Enzymes are proteins that act as biological catalysts They are influenced in foods by:

Temperature pH Water activity Ionic strength (ie. Salt concentrations) Presence of other agents in solution

Metal chelators Reducing agents Other inhibitors

Also factors forInhibition, including:

Oxygen exclusionand

Sulfites

Page 99: Angela Chen

The “Raw Foods” Movement Enzymes present in raw foods help in digesting the foods we eat

But they have to enter the digestive system.

Cooking destroys food enzymes forcing the body to produce more of its own digestive enzymes Eating these enzymes saves your both the work.

Our body has a finite amount of enzyme producing potential The more enyzmes we eat, the more we preserve health and longevity Our digesting enzyme potential can be exhausted.

Enzymes in raw food also carry our "life force" When our ability to produce digestive enzymes is exausted, we die.

Page 100: Angela Chen

Enzymes Before a chemical reaction can occur, the activation energy (Ea)

barrier must be overcome Enzymes are biological catalysts, so they increase the rate of a

reaction by lowering Ea

Page 101: Angela Chen

Enzymes

The effect of temperature is two-fold From about 20, to 35-40°C (for enzymes) From about 5-35°C for other reactions

Q10-Principal: For every 10°C increase in temperature, the reaction rate will double

Not an absolute “law” in science, but a general “rule of thumb”

At higher temperatures, some enzymes are much more stable than other enzymes

Page 102: Angela Chen

Enzymes Enzymes are sensitive to pH – most enzymes active only within a pH range of

3-4 units (catalase has max. activity between pH 3 & 10)

The optimum pH depends on the nature of the enzyme and reflects the environmental conditions in which enzyme is normally active: Pepsin pH 2; Trypsin pH 8; Peroxidase pH 6

pH dependence is usually due to the presence of one or more charged AA at the active site.

Page 103: Angela Chen

Worthington Enzyme Manual

http://www.worthington-biochem.com/index/manual.html

Page 104: Angela Chen

Nomenclature

Each enzyme can be described in 3 ways: Trivial name: -amylase Systematic name: -1,4-glucan-glucono-hydrolase

substrate reaction

Number of the Enzyme Commission: E.C. 3.2.1.1 3- hydrolases (class) 2- glucosidase (sub-class) 1- hydrolyzing O-glycosidic bond (sub-sub-class) 1- specific enzyme

Page 105: Angela Chen

Enzyme Class Characterizations

1. OxidoreductaseOxidation/reduction reactions

2. TransferaseTransfer of one molecule to another (i.e. functional groups)

3. HydrolaseCatalyze bond breaking using water (ie. protease, lipase)

4. LyaseCatalyze the formation of double bonds, often in dehydration reations

5. IsomeraseCatalyze intramolecular rearrangement of molecules

6. LigaseCatalyze covalent attachment of two substrate molecules

Page 106: Angela Chen

Enzyme CommissionEnzyme Nomenclature

International Union of Biochemistry and Molecular Biology (IUBMB)

International Union of Pure and Applied Chemistry (IUPAC)

Joint Commission on Biochemical Nomenclature (JCBN)

IUPAC-IUBMB-JCBNhttp://www.chem.qmul.ac.uk/iubmb/enzyme/

Page 107: Angela Chen

1. OXIDOREDUCTASES

OxidationIsLosing electrons

ReductionIsGaining electrons

Xm+ Xm2+

e-

oxidizedreduced e-

Electron acceptor

Electron donor

Redox active (Transition) metals (copper/ iron containing proteins)

Page 108: Angela Chen

1. Oxidoreductases: GLUCOSE OXIDASE -D-glucose: oxygen oxidoreductase Catalyzes oxidation of glucose to glucono- -lactone

-D-glucose Glucose oxidase D glucono--lactone

FAD FADH2 +H2O

H2O2 O2 D Gluconic acidCatalase

H2O + ½ O2

Oxidation of glucose to gluconic acid

Page 109: Angela Chen

How Glucose Oxidase + Catalase Works:

Reaction 1: Glucose + O2 Gluconic acid + H2O2

Reaction 2: H2O2 H2O + 1/2 O2

Reaction 3: Glucose + 1/2 O2 Gluconic acid

GO

CAT

GO/CAT

Page 110: Angela Chen

1. Oxidoreductases: PEROXIDASE (POD)

Donor: Hydrogen peroxide oxidoreductase

Iron-containing enzyme. Has a heme prosthetic group

Thermo-resistant – denaturation at ~ 85oC

Since is thermoresistant - indicator of proper blanching (no POD activity in properly blanched vegetables)

N N

NN

Fe

Page 111: Angela Chen

1. Oxidoreductases: Catalase

Hydrogen peroxide oxidoreductase Catalyzes conversion of 2 molecules of H2O2 into

water and O2:

Uses H2O2 When coupled with glucose oxidase the net result is

uptake of ½ O2 per molecule of glucose Occurs in MO, plants, animals

H2O2 ------------------- H2O +1/2 O2

Page 112: Angela Chen

1. Oxidoreductases: LIPOXYGENASE

OOH

HH

HC

C

H

H

CC

C

cistrans

HH

H

CC

H

H

CC

C

cis cis

+ O2

……..………

……..

Oxidation of lipids with cis, cis groups into conjugated cis, trans hydroperoxides.

Page 113: Angela Chen

Enzymatic Determination of Starch

or other simple sugars

PRINCIPLE Starch is hydrolyzed

into glucose units by enzymatic conversion

D-glucose can then be quantified by enzymatic methods

Page 114: Angela Chen

1. Oxidoreductases: POLYPHENOLOXIDASES (PPO)

Phenolases, PPO Copper-containing enzyme Oxidizes phenolic compounds to o-quinones: Catalyze conversion of mono-phenols to o-diphenols In all plants; high level in potato, mushrooms, apples, peaches,

bananas, tea leaves, coffee beans

Tea leaf tannins

CatechinsProcyanidins PPO o-Quinone + H2OGallocatechins O2

Catechin gallates

Colored products

Page 115: Angela Chen

Worthington Enzyme Manual

http://www.worthington-biochem.com/index/manual.html

Page 116: Angela Chen

Functional Proteins

Page 117: Angela Chen

Protein FunctionalityHydrodynamic-Aggregation

Viscosity, Elasticity, Viscoelasticity Solubility, Water holding capacity

Hydrophobic- Surface Active Emulsion and foam stabilization Flavor binding

Page 118: Angela Chen

Concentration

Vis

cosi

ty

Dilute

Sem

i-di

lute

Con

cent

rate

d

Hydrodynamic Functionality

Page 119: Angela Chen

Viscosity A property of liquids Viscosity is the resistance to flow. The amount of

energy you need to expend to get a given flow rate.

Stress (force per unit area) is proportional to rate of strain (i.e., flow rate)

Particles of any type in a fluid will increase its viscosity

Large, well hydrated polymers contribute most to viscosity

Page 120: Angela Chen

Elasticity A property of solids Elasticity is the force to achieve a given

percentage change in length Stress (force per unit area) is proportional to strain

(fractional deformation) An elastic material must have some solid-like

network throughout the structure The more load bearing structures the more elastic The more inter-structure links the more elastic

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ViscoelasticityMany materials simultaneously show solid

and liquid like properties If they are stretched they will partly and

slowly return to their original shape Elastic solids would completely recover Viscous liquids would retain their shape

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Water BindingWater Binding Gels contain pores Water can flow out of

the pores If the gel contracts it

may expel liquid SYNERESIS

Due to closer association of protein with protein

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pH

1 2 3 4 5 6 7 8

Sol

ubili

ty /%

0

20

40

60

80

100

SolubilityEmulsoid

Suspensoid

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Whey vs. Casein Dense, ordered

globular proteins

2D Gel

Loose, disordered, flexible chains

Loop-train-tail model

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Practical Applications

A quick stroll through the literature…

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WH= whole hydrolysate

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Story Behind the Story Amy-Acrylamide

Andrea-Maillard ingredients

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Effect of Citric Acid and Glycine Addition on Acrylamide andFlavor in a Potato Model System

Class discussion;

Bianca and Cassie

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A quick review

Protein Analysis Methods

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