yieldmax customer jan2010
TRANSCRIPT
YieldMAX™ PL - enhancing Fat Recovery in Mozzarella / Pizza Cheese
2010 YieldMAX in pasta filata2
Background
Cost reduction by enhancing fat and protein retention an ongoing focus in the cheese industryMuch effort has concentrated on designing optimal cheese making equipment and process parameters, e.g. cutting timeDespite these efforts, fat and protein losses are observed especially after cutting, during early stage of stirring and during stretchCheese yield may be increased by protein concentration. However, this approach may lead to impaired flavour, texture and melting propertiesThe aim of this overview is to explain about the novel enzyme technology using a phospholipase YieldMAX™ PL to increase cheese yield and thereby production economy in the manufacturing of Pasta Filata type cheese
Business case on YieldMAX in Mozzarella/Pizza cheese
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Where does the value of YieldMAX come from
YieldMAX modifies part of the fat globule membraneby acting on the phospholipidsAs a result more fat is retained in the cheeseRecovery of protein also increasesWater binding capacity is improved Altogether yield increases about 2%
YieldMAX results in more cheese from thesame amount of milk
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What are the economic benefits of increasing fat recovery in pizza cheese manufacturing
Higher cheese yieldMore cheese from the same milk
Higher capacity of plant
Milk fat has the highest value when used in cheeseHigher recovery of fat Option to take more sweet cream out of cheese milk
Sweet cream has higher value than whey cream
Lower volume of whey creamRecovery of whey fat often lower than milk fat in cheeseUsing whey cream increases risk of bacteriophagesHigh usage may influence cheese quality negatively
Much less fat in stretching waterFat from stretching water is non-kosher
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Example of economic benefits of using YieldMAX in 1m lbs milk/day pizza cheese production
Benefit of using YieldMAX360 days/year
Element Q ValueExtra cheese 792,000# 1,1m$
YieldMAX cost 570,000$
Gross earnings 539,000$
Gross return on investment 95%
Gross earning at cheese price of 1,6$/lbs
697,000$
Cheese price wheregross RoI is 50%
1.08$/lbs
Constants used
Cheese price 1.4$/lbs
Standard cheese yield
11%
Fat in cheese milk
2,6%
Yield benefit 2%
Milk volume/day
1 million lbs
Pilot plant and industrial validation of concept value
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How to use YieldMAX™ PL
Simple additionAdd to cheese milk 20 min before renneting
While filling vatDosage (LMPS)
5 units per g of fat1.5 l per 25,000 kg milk≈45 fl oz per 55,000# milk
Up to 10x dilution gives no loss in activity YieldMAX™ does not affect the coagulation.
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Pilot plant cheese trial design
Cheese type: Pizza 40+Variable: Addition of YieldMAX™ PL, 20 min
before rennetingAddition rate: 5 u per g fatVats per day: 4 vats: 2 control and 2 experimentalNumbers of days: 4
Cheese yield std. deviation between control vats = 56 g from 150 kg of milk
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Effect on cheese - Pilot scale Mozzarella trial (LMPS)
VariableUntreated
(n = 8)With YieldMAX
Difference %
Fresh curds yield, % 10.80 + 1.1
Cheese yield a, % 10.35 + 3.2
Fat in whey, % 0.132 - 4.3
Fat in cooking water, % 0.300 - 2.1
Moisture in cheese, % 47.12 + 1.9
DM yield b, % 46.43 + 1.5
Fat recovery, % 87.20 + 0.4
Protein recovery, % 76.69 + 1.7a Cheese weight divided by milk weight. b Cheese dry matter divided by milk dry matter.
Lilbæk H.M .. ... .2006. J. D.... S... 89 .11. 4114.4125
P...... ........... .........
U......... ......... .. ....
.......
1H.NMR ..... ......
........ ..... .......
..... ..........
L... ... .... .. .... ...
.......... .....
Statistically significant
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Cheese Functionality - (Confocal Laser Scanning Microscopy)
Fresh cheese, prior to stretching
3 days
14 days
28 days
Control YieldMAX™
No significant difference in image analysis with respect to
size and numbers of fat globule unitsdegree of coalescencecoarseness of protein network
Microstructure shows swelling of casein network and diminishing of whey pockets during storage
Meltability, stretchability and browning similar in control and treated cheeses (Ref: Lilbaek et al. 2006 Journal Dairy Science
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Cheese Control YieldMAX SEM Difference P-value
Phospholipid retention, % 45.4 79.3 3.2 +75 % <0.001
Whey Control YieldMAX
Total phospholipids, µMSphingomyelin, µM
183.052.4
68.446.4
Lysophospholipids preferentially retained
Correlation between PL content and water content !
H.... .. ... 1985 J D.... S.. 68. 1603.1607
D.... .. ... 1988 J F... S.. 63. 1018.1023
T..... .. ... 2001 L... 81. 429.442
G...........L.... .. ... 2006 J D.... S.. 89. 454.467
Retention of phospholipids in cheese increased
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Standard whey fat flow diagram
Milk in Cheese out
Mozzarella without YieldMAX™ PL
Cheese vat Stretching
Whey, 0.2-0.3% fat
Cooking water,2-4% fat
Whey, 0.05% fatWhey cream(25-30% fat)
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YieldMAX whey fat flow diagram
Less whey cream to handle – Less fat in defatted whey
Milk in Cheese out
Mozzarella with YieldMAX™ PL
Cheese vat Stretching
Whey, (<0.2-0.3% fat)
Cooking water,(1-3% fat)
Whey, < 0.05% fatWhey cream(25-30% fat)Less volume
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Improved whey fat separation
YieldMAX PL treatment
% Fat in whey after centrifugation
95 % Confidence interval
+ 0,031 (0,028;0,033)
- 0,040 (0,038;0,042)
Impact of YieldMax PL treatment of cheese milk on separability of whey from small scale (150 L cheese milk) mozzarella cheese production. Values in brackets are 95% confidence intervals.
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Lower recovery of whey fatEffect of recirculation of whey cream
Whey cream fat is to a lesser extend in globular form
More susceptible to oxidationNon globular fat will form large pods of on surface
Less whey fat is retained in cheese lowering the overall fat retentionQuality issues
Recirculation of phageMoisture retention Flavour defects at higher dosage levels
% whey fat recovered
% total fat recovered
% FDM % yield
93.0 93,0 55,2 10,54
80,0 92,4 55,0 10,51
60,0 91,4 54,8 10,44
40,0 90,5 54,6 10,38
Based on whey cream fat as 5% of total fat: Milk 3.88% fat, 2.49% caseinCheddar with 38% moisture
Uni. of Wisconsin, October 1997
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Industrial validation – effect on cheese
Yield data from industrial trials (≥500 cheese vats)
% yieldincrease
95%interval
Recipe 1 1,4 (0,3; 2,4)
Recipe 2 0,7 (-0,5; 1,9)
Recipe 3 2,1 (0,1; 4,0)
Recipe 4 (high fat) 3,8 (1,6; 6,0)
Significant increases seen infat in dry mattermoisture in cheesefat retentionprotein retention in two recipes
H...... E .. ... .2006. T.. A......... J. .. D.... T.... 61 .2. 179.182
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Industrial validation - effects on whey fat
Significant decrease in whey fatTypical pilot plant data shows decrease from 0.22 to 0.19%9-10% decrease in industrial trialsSame effects seen on all tested recipes
Whe
y fa
t %LS
Mea
ns0,2
0,25
0,3
0,35
0,4
0,45
Recipe 1Recipe 2Recipe 3Recipe 4
Control pLipase
Treatment
LS Means Plot
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Industrial validation - Effects on mixer/molder water
Fat recovery in cheese increased – industrial dataLess fat/protein loss in cooking water
Treated vats
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Effects on process - foaming
YieldMAX addition may lead to some foam formation
To be addressed depending on individual circumstancesSpraying of water dropletsMinimizing air incorporation during processing (plant design)Ultrasound treatmentOther anti foaming methods
Untreated Treated
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Summarizing the current knowledge -
A suggested mechanism for cheese yield increase
MilkC.....
W...
+ YieldMAX™ PL
. 1.3.
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Process step Effect of YieldMAX™ PL
Incubation of milk with phospholipase
• Conversion of phospholipids to lysophospholipids• Release of lysophospholipids from fat globules
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Process step Effect of YieldMAX™ PL
Incubation of milk with phospholipase
• Conversion of phospholipids to lysophospholipids• Release of lysophospholipids from fat globules
Coagulation • Complexation of lysophospholipids with casein
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Process step Effect of YieldMAX™ PL
Incubation of milk with phospholipase
• Conversion of phospholipids to lysophospholipids• Release of lysophospholipids from fat globules
Coagulation • Complexation of lysophospholipids with casein
Curd cutting, stirring, scalding
• Lysophospholipids act as an emulsifier- protect fat globules from rupture - re-emulsify fat “leaked out” from damaged globules
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Process step Effect of YieldMAX™ PL
Incubation of milk with phospholipase
• Conversion of phospholipids to lysophospholipids• Release of lysophospholipids from fat globules
Coagulation • Complexation of lysophospholipids with casein
Curd cutting, stirring, scalding
• Lysophospholipids act as an emulsifier- protect fat globules from rupture - re-emulsify fat “leaked out” from damaged globules
Whey drainage • Lysophospholipid – casein complexes retained in the curd
• Less un-emulsified liquid fat is lost in the whey
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Process step Effect of YieldMAX™ PL
Incubation of milk with phospholipase
• Conversion of phospholipids to lysophospholipids• Release of lysophospholipids from fat globules
Coagulation • Complexation of lysophospholipids with casein
Curd cutting, stirring, scalding
• Lysophospholipids act as an emulsifier- protect fat globules from rupture - re-emulsify fat “leaked out” from damaged globules
Whey drainage • Lysophospholipid – casein complexes retained in the curd
• Less un-emulsified liquid fat is lost in the whey
Curd stretching • Lysophospholipid acts as an emulsifier- protect fat globules from breaking- re-emulsify fat “leaked out” from damaged globules- reduce syneresis more water is retained in cheese
Activity of YieldMAX in the milk
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Phopsholipids in milk products
0
0,2
0,4
0,6
0,8
1
1,2
1,4
1,6
Milk
Skimmed
milk
Cream 40
%Butt
er
Butterm
ilk
Anhyd
rous m
ilk fat
% w
/w % in product% in solids
Based upon data fra Walstra et.al 2006. Dairy Science and Technology CRC Press
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Total amount in milk 0.02 – 0.04 % w/wIn fresh raw milk ∼80% is on the fat globule surfaceLipoprotein aggregates/vesicles in milk plasma
50 – 400 ..
1 – 10 µm
Phospholipids in milk
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Phospholipids in milk
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Phospholipids in milk (mol-%)
√ Phosphatidyl ethanolamine (PE) 22-49 %
√ Phosphatidyl choline PC) 21-40 %
√ Phosphatidyl inositol (PI) 4-11 %
√ Phosphatidyl serine (PS) 2-12 %
÷ Sphingomyelin (SM) 18-32 %
Note! Sphingomyelin is not hydrolyzed.
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Hydrolysis of milk phospholipids:catalyzed by YieldMAX™ Phospholipase A1
Example of a substrate: Phosphatidyl choline
P........... L............... . F....
....
Y....MAX PL
H
2
0
YieldMAX PL(PLA1)
OO
O
O
OPO
O-
ON
+
Hydrolyzes ester bonds in long-chain fatty acids
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Phospholipids can be extracted from milk and analyzed by HPLC
Almost complete depletion of phosphatidylethanolamine and phosphatidylcholine
Development of long chain free fatty acids can be observed
Lysophospholipids are detected in whey, amount corresponds to approx. 5 % of content in hydrolyzed whole milk - 95% remains in cheese matrix
Sphingomyelin is not hydrolyzed by YieldMAX™ PL
0
25000
50000
75000
100000
125000
150000
0 5 10 15 20Time, min
mAU
PE
PCSph
0
25000
50000
75000
100000
125000
150000
0 5 10 15 20
Time, min
mA
U
PE PC
LPE
LPC
Sph
Untreated control
Phospholipase treated
The YieldMAX™ PL action
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DH% in whole milk vs. incubation time YieldMAX PL dose 5 or 50 LEU/g fat
Max. Degree of hydrolysis 65-80%
0
10
20
30
40
50
60
70
80
0 5 10 15 20Time (h)
DH
% PE (5 Units /g fat)
PC (5 Units /g fat)
PE (50 Units /g fat)
PC (50 Units /g fat)
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Phospholipids are in the fat globules -Lysophospholipids are in serum phase
∼ 50.60 . PC.PE
W.... ....
S... ....
∼ 50.60 . .. ............ PC.PE∼ 100 . ................. LPC.LPE
.Y....MAX
PC LPC
PE LPE
Lilbæk H.M .. ... .2007. J. A..... F... C.... I. .....
C.......... .......... C..........
..........
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Fat globule stability unchanged
Whole milk particle size distribution
Particle diameter [µm]
0,01 0,1 1 10 100 1000
Rel
ativ
e di
strib
utio
n [%
]
0
2
4
6
8
10
12
14
16
B.
A.U........
Y....MAX
Phospholipase Control
Fat globules in cheese curd
Documentation on whey streamand casein
Positive effect on whey processing
No effect seen on whey protein fractions or their ratio
No effect seen on casein
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Improved processing capabilities of YieldMAX whey
Whey contains less fat due to better separation properties of YieldMAX whey
Data from processing confirm thisLower fat in whey means easier processingLower fat means easier membrane filtration for WPI
Improved functionality of WPIsLower means less risk of oxidative flavor changesReplace fat with low value lactose in WPC
YieldMAX reduces surface tension of the wheyTheoretical explanation on functionality of lyso-phospholipidsProven in laboratory model Customer references
Longer runs between CIP on UFLess fouling
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Method:Wilhelmy plate
Sample: Skim milk, separated from treated whole milk
General phenomenon:Surface tension is also reduced upon phospholipase treatment of:
-Cream
-Skim milk
-WPC-solution
-Milk phospholipid
suspension
-In whey from treated
cheese milk
Surface tension is reduced by YieldMAX
Control, no enzymePLA1 F. venenatum (YieldMAX)PLA2 Porcine pancreasPLC B. cereus
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YieldMAX has high specificity
Enzyme heat labile - ≥90% of the enzyme is destroyed at low pasteurization of the whey(72°C/160°F, 20 sec)
Side activities removedAmylase and glucoamylases absent according to specsConformity checks also for ß-galactosidase, lipase and proteolytic activity
No effect on whey protein fractions
K-sorbate
Na-benzoateEnzyme
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Products incubated with YieldMAX™ PL
3 different commercial whey protein concentrates (WPC)2 WPC from Mozzarella trials with or without use of YieldMAX™ PL4 different commercial whey protein isolates
Sample Whey protein (powder products)
123456789
Whey Protein Concentrate I (34%)Whey Protein Isolate WPC Trial (from Mozzarella production using YieldMAX™ PL)WPC Control (from Mozzarella production)Bovine Lactoferrin ILactoferrin / LactoperoxidaseBioactive Whey ConcentrateBovine Lactoferrin IIWhey Protein Concentrate II
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Experimental setup 0.4 g Whey Protein
40 mL Buffer* 200 µL 5% Na-Azid
20 mL A
10 mL B
10 mL C
Mixing WPS
Incubation at 35°C for 21 hr
+ 5 µL YieldMAX PL
+ 5 µL HT YieldMAX PL
SEC SDS-Page
Detect proteins between 2.5 to 200 kDa
Detect proteinsless than 2000 kDa(optimal <200 kDa)
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SDS-gel on WPC80
No appearance of new bands
No reduction of bands intensity
No effect observed of YieldMAX™ PL incubation25
Samples: WPC80 with and without YieldMAX™
45
10 ul 20 ul
kDa
116
66,2
35
18,414,4
Trea
ted
13-3
-06
Con
trol
14-3
-06
Trea
ted
21-3
-06
Con
trol
22-3
-06
Trea
ted
13-3
-06
Con
trol
14-3
-06
Trea
ted
21-3
-06
Con
trol
22-3
-06
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Bovine Lactoferrin I & Lactoferrin/Lactoperoxidase
SECNo new peaks observedSame level of peak areasNo differences
1 % solutions of products
Bovine Lactoferrin I
-0,5
0
0,5
1
1,5
2
2,5
3
0 20 40 60 80
Time (min)
Abs
orba
nce
220
nm
WPS5
+YieldMAX
+HT YieldMAX
MwSTD
-0,5
0
0,5
1
1,5
2
2,5
3
0 20 40 60 80
Time (min)
Abs
orba
nce
220
nm
WPS7
+YieldMAX
+HT YieldMAX
MwSTD
Lactoferrin/Lactoperoxidase
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Effect on casein
1. Sodium caseinate (from Sigma, C-8654), αs1-casein and β-casein were solubilised in 50 mM sodium phosphate buffer, pH 6.4, containing 0.025 % sodium azide. Protein concentration of the solutions were 5 mg/ml2. YieldMAX™ PL (Batch No. 2663736) was added to the solutions at a concentration of 0.05 % (v/v), and samples were incubated for 21 hours at 35ºC.3. Control reactions without YieldMAX™ PL addition were also included.4. Samples were analyzed using SDS-PAGE
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SDS-PAGEcaseins incubated with YieldMAX™ PL
1: Molecular weight standards2: Sodium caseinate standard3: Sodium caseinate incubated without YiedMAX PL4: Sodium caseinate incubated with YieldMAX PL5: as1-casein standard6: as1-casein incubated without YieldMAX PL7: as1-casein incubated with YieldMAX PL8: b-casein standard9: b-casein incubated without YieldMAX PL10: b-casein incubated with YieldMAX PL
kDa
116.0
66.2
45.0
35.0
25.0
18.4
14.4
1 32 54 76 8 109
=> No hydrolysis of the caseins by YieldMAX™ PL
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Summary
YieldMAX™ PL incubated with different whey protein and casein products under optimal conditions
No differences observed fromSDS-Page Gel ElectrophoresisSize Exclusion Chromatography
No effect of YieldMAX™ PL detected towards different whey protein or casein fractions
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Regulatory overviewUSA
FDA notified GRAS accepted June 2004YieldMAX™ PL is a GRAS substance,covered by GRAS Notice GRN000142
(Canada – Health Canada Approval)
The Interim Marketing Authorization (IMA) for YieldMax™ PL was published in Gazette I on August 5, 2006. YieldMAX™ PL is legal for sales in Canada from that date.
Allergen statusNo allergens as defined by regulatory authorities in EU and USA
Dietary statusYieldMAX™ PL is certified Kosher Pareve
GMO StatusYieldMAX™ PL is an enzyme from Fusarium venenatum produced with a strain of Aspergillus oryzae. By current EU rules, YieldMAX™ PL should not be GM labelled
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Conclusion
YieldMAX™ PL successfully applied in Pasta Filata cheese makingeasy to implement
Significant economic benefits through yield increasesignificant increased fat retentionincreased protein retentionhigher fat in dry matter (at unchanged milk fat content)higher moisture content (at unchanged parameters)
No detrimental effects found on whey products
No functional changes in cheese