tg cheese and yogurt 012710
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Dairy Applications for Dairy Applications for TransglutaminaseTransglutaminase
Ajinomoto Food Ingredients, LLC
TransglutaminaseTransglutaminaseEnzyme BasicsEnzyme Basics
TransglutaminaseTransglutaminase
TG is an enzyme that covalently crossTG is an enzyme that covalently cross--links links proteinprotein
Found throughout nature (from Found throughout nature (from microorganisms to mammals)microorganisms to mammals)
ACTIVAACTIVA®® TransglutaminaseTransglutaminase
TG TG enzyme used by Ajinomoto is produced enzyme used by Ajinomoto is produced via fermentation by a GMOvia fermentation by a GMO--free free microorganismmicroorganism
Marketed as Marketed as TG preparationsTG preparations by Ajinomoto by Ajinomoto Food Ingredients (AFI) under the Food Ingredients (AFI) under the ACTIVAACTIVA®® brand namebrand name
Linking ReactionLinking Reaction
TGTG
Transglutaminase: Cross-linking Mechanism
GLN-C-NH2 + H-N-LYSGLN-C-NH2 + H-N-LYS||
O ||
O
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TGTG
NH3GLN-C-N-LYSGLN-C-N-LYS|
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O H|| |
O H
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H |
H
γ Carbonyl Group of Glutamine ε Amino group of Lysine
Forms permanent covalent bonds unaffectedby freezing or cooking conditions
ACTIVAACTIVA®® TG Temperature ImpactTG Temperature Impact
Activity and Stability over Temperature Range
0
20
40
60
80
100
0 10 20 30 40 50 60 70
Temperature (C)
Rel
ativ
e A
ctiv
ity (%
)
RelativeActivityStability
Temperature range: 32 to ~150FOptimal Temperature range: 115-130F (45-55C)
(Specific denaturation temp depends on system composition)
ACTIVAACTIVA®® TG pH ImpactTG pH Impact
pH Stability
0
20
40
60
80
100
120
3 4 5 6 7 8 9 10
pH
Rel
ativ
e A
ctiv
ity (%
pH Range: 4-9Optimal pH: 6-7
Protein substrate specificity of TGProtein substrate specificity of TG
MilkCasein
a-Lactoalbuminb-Lactoglobulin
EggEgg whiteEgg yolk
Wheat
Meat
CollagenGelatinMyosinActin
Soybean
Food Protein Reaction Strength+++
++
+++
++++
++++++
-+++
+++ reacts very well
+ reacts depending on conditions
++ reacts well
- mostly does not react
Reaction time and temperature:Reaction time and temperature:speed of reactionspeed of reaction
Temperature Time5℃ 240min15℃ 105min20℃ 70min30℃ 35min40℃ 20min
Required time for the reactions at each temperature to the same extent as the reaction at pH6.0, 50℃ 10min
Functions & BenefitsFunctions & Benefits in Food Processingin Food Processing
Cross-linking protein molecules
Increase gel strength
Provide food bond
Modify protein property
Reaction Function Benefit-Texture improvement of gel based foods, such as sausage, surimi, yogurt, cheese and etc.-Cost reduction by reducing necessary protein levels
-Bonding meat pieces into desired size or shape. (portion control, ham)
-Reduced syneresis by improved water retention
-Smooth, thick texture
-Pasta texture improvement
ACTIVA® STG-MS
Super CurdACTIVA® TI ACTIVA® YG
ACTIVA® TI
Surimi Noodle / PastaBonding
Ham Sausage Tofu
Dairy
Specific ACTIVASpecific ACTIVA®® Preparation ApplicationsPreparation Applications
ACTIVA® TI
ACTIVA® TI
ACTIVA® RM/GS
ACTIVA® BFACTIVA® TI
TransglutaminaseTransglutaminaseUse in dairy productsUse in dairy products
Use of TG in Dairy ProductsUse of TG in Dairy Products
Casein protein has a high concentration of Casein protein has a high concentration of Glutamine and Lysine, making it a great Glutamine and Lysine, making it a great substrate for TG activitysubstrate for TG activity
ACTIVAACTIVA®® YG YG –– for use in situations where raw for use in situations where raw milk is being used in the processmilk is being used in the processACTIVAACTIVA®® TI TI –– used when processed milk is being used when processed milk is being usedused
Can develop enhanced structure and handling Can develop enhanced structure and handling characteristicscharacteristics
Use of Use of TransglutaminaseTransglutaminase in Cheese in Cheese ProductsProducts
TG in Natural Cheese ApplicationsTG in Natural Cheese Applications
Good potential functionality in cheese Good potential functionality in cheese manufacturemanufactureUse of low levels of TG in formulation can offer Use of low levels of TG in formulation can offer enhanced yield and cost savingsenhanced yield and cost savings
Retain more whey proteinRetain more whey proteinRetain more moistureRetain more moisture
Mozzarella Cheese StudyMozzarella Cheese StudyExamine use of TG incorporated into industrial Examine use of TG incorporated into industrial processprocess
TG either denatured, or active in final productTG either denatured, or active in final product
ResultsResultsBest use: 0.0625 units TG/gram proteinBest use: 0.0625 units TG/gram protein
UndenaturedUndenatured processprocess~$1.68/ton cheese produced~$1.68/ton cheese produced0.46% increase in final yield0.46% increase in final yield
9.2 lb increase per ton cheese produced9.2 lb increase per ton cheese produced~$17.00 savings/ton~$17.00 savings/ton
OrganolepticOrganoleptic and physical quality same or better than and physical quality same or better than control over study time (4 weeks)control over study time (4 weeks)
TG in Cheese Sauce ApplicationsTG in Cheese Sauce Applications
The addition of TG to a cheese sauce The addition of TG to a cheese sauce application can give distinct advantagesapplication can give distinct advantages
Melt RestrictionMelt RestrictionReduction of Reduction of ““oiling offoiling off”” upon meltingupon meltingEnhanced elasticityEnhanced elasticityImproved shredImproved shredReplacement of Replacement of texturizingtexturizing ingredientsingredients
Milk protein: Whey powder, whey isolateMilk protein: Whey powder, whey isolate
Study Study –– TG use in Cheese SauceTG use in Cheese Sauce
Enhanced Melt restriction with dramatic Enhanced Melt restriction with dramatic reduction in reduction in ““oiling outoiling out””
Control - Without TG
Avg: 0.5”high x2.25”x axis x2.5”y axis
#101 - With TG (0.041% TI)
Avg: 0.75”high x1.75”x axis x1.5”y axis
Melt test: 1” cubes heated covered at 400F for 7 minutes
Study Study -- TG use in Cheese Sauce TG use in Cheese Sauce Melt Restriction and Moisture DataMelt Restriction and Moisture Data
pH Fat MoistureTotal
ProteinSalt
Total Protein
FatTotal
ProteinFat
3902 - 2001Pasteurized Process CheeseSauce
Control 0.0000 5.28 24.72 45.40 --- 2.61 --- --- --- --- 50 x 11
3902 - 2002Pasteurized Process CheeseSauce - 100% texturants
0.2500 0.0410 5.30 25.09 43.74 --- 2.87 --- --- --- --- 42 x 13
3902 - 2003Pasteurized Process CheeseSauce - 50% texturants
0.2500 0.041 5.32 24.51 46.10 --- 2.56 --- --- --- --- 45 x 12
3902 - 2004Pasteurized Process CheeseSauce - 25% texturants
0.2500 0.0410 5.30 25.14 46.88 --- 2.50 --- --- --- --- 46 x 10
3902 - 2005Pasteurized Process CheeseSauce - 0% texturants
0.2500 0.0410 5.31 26.84 45.39 --- 2.56 --- --- --- --- 53 x 12
3902 - 5101 Cream Cheese Control 0.0000 4.57 40.30 48.29 --- --- 2.62 16.26 --- --- ---3902 - 5102 Cream Cheese 0.0200 0.0030 4.55 42.03 45.96 --- --- 1.28 3.32 --- --- ---3902 - 5103 Cream Cheese 0.0400 0.0050 4.55 42.26 46.62 --- --- 1.40 4.91 --- --- ---3902 - 5104 Cream Cheese 0.0500 0.0060 4.57 40.61 47.12 --- --- 1.47 4.39 --- --- ---3902 - 5105 Cream Cheese Control 0.0000 4.70 42.02 49.34 --- --- --- --- --- --- ---3902 - 5106 Cream Cheese 0.0200 0.0030 4.65 44.29 46.29 --- --- 1.28 0.68 --- --- ---3902 - 5107 Cream Cheese 0.0400 0.0050 4.60 40.58 47.38 --- --- 1.47 2.06 --- --- ---3902 - 5108 Cream Cheese 0.0500 0.0060 4.53 40.09 46.30 --- --- 1.47 2.20 --- --- ---
Kraft Philadelphia Cream Chease 4.65 31.42 53.95 6.25Kraft Philly Chease Spread 4.83 26.84 59.39 4.98Best Choice Cream Cheese Spread 4.82 26.90 58.92 5.30Best Choice Cream Cheese 4.95 32.56 53.66 5.17
TI Amount(units/gram)
Formulation Number
Ajinomoto Analytical Testing Results v5
CheeseWhey Analysis Milk AnalysisCheese Analysis
TI Amount(percentage)
Melt Test(mm)
A positive melt restriction is seen when TG is A positive melt restriction is seen when TG is used in combination with other used in combination with other texturantstexturants
Melt test measurements are width vs. heightMelt test measurements are width vs. height
103
101
102
Control
104
Enhanced moisture holding characteristics while Enhanced moisture holding characteristics while maintaining comparable textural attributesmaintaining comparable textural attributes
46% moisture in #102 vs. 43% moisture in #10146% moisture in #102 vs. 43% moisture in #101Studies indicate similar cost savings benefits Studies indicate similar cost savings benefits seen when used in spreadable/pourable cheese seen when used in spreadable/pourable cheese sauces sauces ACTIVAACTIVA®® TG has the potential for delivery of TG has the potential for delivery of cost reduced cheese saucecost reduced cheese sauce--type products with type products with enhanced physical characteristicsenhanced physical characteristics
Study Study –– TG use in Cheese SauceTG use in Cheese Sauce
Use of Use of TransglutaminaseTransglutaminase in Yogurt in Yogurt ProductsProducts
Increased gel strengIncreased gel strengthth/viscosity/viscosityImproved creaminess Improved creaminess Reduced syneresisReduced syneresisReduction of added dry matter and/or Reduction of added dry matter and/or stabilizersstabilizersImproved gelling capacity of reduced fat Improved gelling capacity of reduced fat yogurtyogurt
TG use in Yogurt TG use in Yogurt -- BenefitsBenefits
ACTIVAACTIVA®® Preparations for Dairy Preparations for Dairy ProductsProducts
ACTIVAACTIVA®® TITIFor use in preFor use in pre--processed milkprocessed milk
ACTIVAACTIVA®® YGYGFor use in raw milkFor use in raw milk
Raw milk contains TG inhibitor. YG has reducing agent Raw milk contains TG inhibitor. YG has reducing agent to eliminate inhibitionto eliminate inhibition
Yogurt Yogurt –– Enhanced Gel StrengthEnhanced Gel Strength
0
500
1000
1500
2000
2500
-TG -TG 0.5 U/g p 1 U/g p
Visco
sity(
mPa
・s
) 10% NFDM11% NFDM
Powder milk(10Powder milk(10%%, 11, 11%% NFDM, milk fat 2.7NFDM, milk fat 2.7%)%)↓↓ ACTIVA TI ACTIVA TI (0(0,,0.5, 1U/g protein)0.5, 1U/g protein)
Incubation Incubation ((2525℃℃, 2 hours), 2 hours)
↓↓Cooling after enzyme deactivation Cooling after enzyme deactivation ((7070℃℃))↓↓Starter additionStarter addition
↓↓Cooling after fermentationCooling after fermentation
↓↓StirringStirring
↓↓Stirred yogurt sampleStirred yogurt sample
↓↓Cooling O/NCooling O/N→→Viscosity measurementViscosity measurement
Viscosity is increased with TG, achieving equivalence at 1U per gram protein when treating 10% SNF addition to 11% SNFWith ACTIVA® YG, dosage should be 1/3 that of ACTIVA® TI when using raw milk
Study Study –– Reduction of NFDMReduction of NFDM
Yogurt - NFDM replacement by TG-YG
0
200
400
600
800
1000
1200
Vis
cosi
ty (c
ps) 5% NFDM
4% NFDM4% NFDM/0.3% YG4% NFDM/0.6% YG3% NFDM/0% YG3% NFDM/0.6% YG
Addition of ACTIVA® YG allows reduction of NFDM levels while achieving desired viscosification
Potential Savings through NFDM Potential Savings through NFDM ReductionReduction
ACTIVA YG cost reduction calculationNFDM WPC80
5.45$
Added Reduced cost Total costACTIVA NFDM WPC80 Total ACTIVA YG unit/g milk protein
Dosage reduction
Reduced cost / 1t milk
Reduced cost / 1t milk
Reduced cost / 1 ton milk
0.05 0.2% 9.08$ 0.2% -$ (7.62)$ 0.1 0.3% 18.17$ 0.3% -$ (15.24)$ 0.2 0.7% 36.33$ 0.7% -$ (30.48)$ 0.3 1.0% 54.50$ 1.0% -$ (45.73)$ 0.6 2.0% 109.00$ 2.0% -$ (91.45)$
Shelf Life Textural StabilityShelf Life Textural Stability
Yogurt Viscosity
0
500
1000
1500
2000
2500
3000
3.6%Protein, 0 U
TG
3.5%Protein, 0 U
TG
3.5%Protein, 0.1
U TG
3.5%Protein, 0.2
U TG
3.4%Protein, 0 U
TG
3.4%Protein, 0.2
U TG
3.4%Protein, 0.3
U TG
3.3%Protein, 0.4
U TG
cps
1 day7 days14 days21 days42 days
Addition of ACTIVA® YG allows a higher final viscosity through the end of shelf life.
Enhanced Gelling with Lower MFEnhanced Gelling with Lower MF
0
5
10
15
20
25
30
35
-TG -TG 1 u/g p 2 u/g p
Brea
king s
treng
th(
g/cm
2)
MF 3.6% MF 1.0%Powder milkPowder milk((8.28.2%% NFDM, 3.6% or 1.0NFDM, 3.6% or 1.0%% milk fatmilk fat))↓↓ ACTIVA TIACTIVA TI (0, 1, 2U/g protein)(0, 1, 2U/g protein)
Incubation Incubation ((2525℃℃, 2 hours), 2 hours)
↓↓Cooling after enzyme deactivation Cooling after enzyme deactivation ((7070℃℃))↓↓Starter additionStarter addition
↓↓StuffingStuffing
↓↓FermentationFermentation
↓↓Set type yogurtSet type yogurt
↓↓Cooling O/N Cooling O/N →→ Breaking strength measurementBreaking strength measurement
An increase in gelling character and creaminess can be obtained in low fat yogurt products through the use of TG without the necessity of thickening agents. With ACTIVA® YG, dosage should be 1/3 of ACTIVA® TI when using raw milk.
TransglutaminaseTransglutaminaseRegulationsRegulations
TG: Use Levels Affirmed as GRASTG: Use Levels Affirmed as GRAS
FOOD CATEGORY USE Level as TG
Proc Meat & Poultry Non-Standard & select Std Product Chicken Breast Products
65 ppm
100 ppm Processed Seafood GMP*
Meat Analogs/Substitutes GMP*
Natural Cheese/Processed Cheese GMP*
Cream Cheese GMP*
Refrigerated Yogurt GMP*
Frozen Desserts GMP*
* GRAS Notice 0095
CanadaCanada
Canada Gazette Part II Canada Gazette Part II vol. 138, No. 9vol. 138, No. 9
Apr 22, 2004Apr 22, 2004
NonNon--standardized prepared fish productsstandardized prepared fish products
Simulated meat productsSimulated meat products
NonNon--standardized cheesestandardized cheese
Processed cheese and cream cheese productsProcessed cheese and cream cheese products
Yogurt and NonYogurt and Non--standardized frozen dairy dessertsstandardized frozen dairy desserts
at GMP levelat GMP level
TransglutaminaseTransglutaminaseIngredient StatementsIngredient Statements
Labeling of TG by itself:Labeling of TG by itself:Current RegulationsCurrent Regulations
Area Category Remarks
Japan Food additive If enzyme is active, state "enzyme"
EU Processing aid No statement required
USA GRAS State "enzyme", "TG enzyme" or "TGP enzyme"
Canada Food additive State "transglutaminase"
TransglutaminaseTransglutaminase preparations preparations seriesseries
ACTIVA® RM BindingSodium caseinate, Maltodextrin, Transglutaminase
Sodium caseinate, Maltodextrin, Enzyme Contains Milk
ACTIVA® GS
Binding (strong bind, stable solution)
Sodium Chloride, Gelatin, Trisodium Phosphate, Maltodextrin, Transglutaminase, Safflower Oil
Salt, Gelatin, Trisodium Phosphate, Enzyme None
ACTIVA® TI Sausage, Ham Maltodextrin, Transglutaminase Maltodextrin, Enzyme None
ACTIVA® BF HamAmmonium glutamate, Maltodextrin, Transglutaminase, Safflower oil
Ammonium glutamate, Enzyme None
ACTIVA® YG Yogurt, Cheese
Lactose, Yeast extract, Maltodextrin, Safflower oil, Transglutaminase
Lactose, Yeast Extract, Enzyme None
AllergenPreparation Typical usage
Ingredients (as shown on the label)
Ingredient statement in final products
USAUSA
*Note: Information is for reference only. Product labeling must be determined by manufacturer.
Lactose not considered allergen
Gelatin not considered allergen
TransglutaminaseTransglutaminase preparations preparations seriesseries
ACTIVA® RM Binding Sodium caseinate, Maltodextrin, Transglutaminase
Sodium caseinate, Maltodextrin, Transglutaminase
Contains Milk
ACTIVA® GS
Binding (strong bind, stable solution)
Sodium Chloride, Gelatin, Trisodium Phosphate, Maltodextrin, Transglutaminase, Safflower Oil
Salt, Gelatin, Trisodium Phosphate, Transglutaminase
None
ACTIVA® TI Sausage, Ham
Maltodextrin, Transglutaminase Maltodextrin, Transglutaminase None
ACTIVA® BF HamAmmonium glutamate, Maltodextrin, Transglutaminase, Safflower oil
Ammonium glutamate, Transglutaminase None
ACTIVA® YG Yogurt, Cheese
Lactose, Yeast extract, Maltodextrin, Safflower oil, Transglutaminase
Lactose, Yeast Extract, Transglutaminase None
AllergenPreparation Typical usage
Ingredients (as shown on the label)
Ingredient statement in final products
CanadaCanada
*Note: Information is for reference only. Product labeling must be determined by manufacturer.
Lactose not considered allergen
Gelatin not considered allergen
Determining whether “fish” must be on label
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