Transglutaminase for Bonding and Texture

Transcription

1 Transglutaminase for Bonding and Texture

2 History of Transglutaminase New TG Discovered A new transglutaminase from microorganism was discovered in 1987 AJINOMOTO CO.,LTD. has succeeded in developing a method for the mass production and commercialization of this enzyme in 1993

3 Basic Properties of TG TG

4 Basic Properties of TG Primary Structure ACTIVA TG is a single polypeptide chain consisting of 331 amino acids (AA) with a Cysteine residue in the active center. Ca2+ - independent Activity ACTIVA TG is Ca 2+ -independent enzyme,unlike the other Ca 2+ -dependent TGs derived from mammals. Therefore ACTIVA TG is easy to use in food processing.

5 Basic Properties of TG How Does It Work? Catalyses the formation of a covalent bond between free amine group and form cross-linked protein polymers TG

6 Mechanism of Enzyme Reaction Crosslinking between Glutamine and Lysine residues are formed through TG reaction Amino acids Protein chains Inter-molecular cross-link Lys Gln Gln Lys Gln Lys Intra-molecular cross-link

7 Mechanism of Reaction Reaction Catalyzed by Transglutaminase Glutamine Lysine Forms a covalent bond unaffected by freezing or cooking temperatures

8 Transglutaminase TG enzyme used by Ajinomoto is produced via fermentation by a GMO-free microorganism (specifically Streptomyces Mobaraensis), then isolated from the culture medium and purified. No microbial contamination Marketed worldwide by Ajinomoto as TG preparations under the Activa brand name

9 How is TG Used? Texture Enhancement and Food Bonding

10 Factors to Consider Meat Quality Age of Meat As meat wet/dry age proteolysis increases and those enzymes can interfere with the bonding process ph This relates to the age of meat as well as potentially the marinades used to enhance the meat. Ex. Pineapple juice due to its low ph will denature the TG enzyme Thawing State Frozen meat cannot be bonded together. The meat must first be thawed before bonding can be done. Reaction Time & Temperature TG needs time to create the protein cross-links and a conducive temperature to speed up or slow down the reaction Application Activa has been designed to give you either fast action (Activa RM) or processing time (Activa GS) from point of application. Air Removal TG does not bond air, and any air pockets that are in the seam between pieces of meat will be areas of weak bonding. You can use perforated packaging or vacuumized systems to help remove air and improve the quality of the product. Set Time Minimum reaction time is process dependent, but an over night hold (>12hr) will give a nice bond

11 Optimal Conditions for TG Reaction (The enzymatic activities were measured at 37 and/or ph 6)

12 ph and Thermal-Stability of TG ph Temp. [ºC] (holding time: 10 minutes at ph 7). ph-stability between 5 and 9 Heat-stability up to a temperature of ~ 40. Gradually inactivated after ~ 50. Quick inactivation after 60

13 Substrate Specificity (Food Proteins) AJI- TG reacts with various proteins

14 Relationship Between Reaction Temperature and Time Temperature Time 5 C 240 min 15 C 105 min 20 C 70 min 30 C 35 min 40 C 20 min the same extent as the reaction at ph 6.0 and 50 C for 10 minutes Important : Reaction time and Temperature

15 Inactivation of Transglutaminase Temperature Time 65 C 2 hours or more 70 C Within 15 min 75 C Within 5 min 80 C Within 1 min Time required for inactivation in sausage (3cm diameter) by heating. The time required for deactivation depends on the internal temperature.

16 Functions & Benefits in Food Processing Cross-linking protein molecules Increase gel strength Provide food bond Modify protein property -Texture improvement of foods, such as sausage, surimi, yogurt, cheese and etc. - Cost reduction by reducing necessary protein levels - Bonding meat pieces into desired size or shape (Tenderloins, scallops) - Change protein gelling properties - Change in protein solubility - Improve processing of ingredient

17 Reference Guide to Activa Preparations

18 Reference Guide to Activa Preparations

19 Processing Methods

20 Methods of Use Dry Addition (0.50% % Activa RM, GS) Depends on application, raw material, formula and conditions Can be dipped, sprinkled on to substrate Add directly to a mixer/tumbler Liquid Slurry Addition (0.60% 1.20% Activa GS) Depends on application, raw material, formulation and conditions 1:4 ratio Activa TG : water Can be dipped, sprayed or brushed onto large intact muscles Can be used in mixers and tumblers *** May not be used with meat pretreated with tenderizer***

21 Dip Method Step 1 - Coat product Dip product into powder so that all surfaces to be bonded are coated with powder. Shake off excess Step 2 Align surfaces Meat to meat give best results. Bonding skins sides together or fat sides will not work. Step 3 Form or mold Vacuum packaging, Casings, Molds or Plastic Wrap Step 4 Reaction Chill product overnight Step 5 Portion

22 Sprinkle Method Step 1 - Coat Product Sprinkle product onto substrate so that all surfaces to be bonded are coated with powder. Shake off excess Step 2 Align surfaces Step 3 Form or mold Vacuum package within about 5-10 minutes, or wrap with plastic wrap Step 4 - Reaction Chill product overnight Step 5 Portion *Note Dip method distributes Activa preparations more evenly than does sprinkle method.

23 Dry Addition Mixer / Tumbler Step 1 Prepared Ingredients Weigh ingredients and prepare meat Step 2 Mix Ingredients Add Activa while mixing to avoid clumps Step 3 Form and mold Vacuum stuff into casing within 1 hour Step 4 Reaction Allow product to rest for minimum 8 hours or overnight for best results Step 5 Portion From fresh or frozen

24 Slurry Preparation Step 1 - Prepare Ingredients Weigh ingredients and prepare meat. Step 2 Mix Ingredients Slurry is typically made at a 1 part Activa GS to 4 parts warm water Mix using a whisk or high speed mixer until watery in consistency Avoid foaming as this will have to break before slurry can be used Note High shear mixers can be used, but will generate an enormous amount of foam and will take >1 hour for foam to break

25 Slurry Addition Step 1 Add slurry to meat Make sure slurry is well distributed, but not absorbed, product should be glossy in appearance Step 2 Form or mold Vacuum stuff into casing or mold within one hour Step 3 Reaction Allow product to rest for minimum 8 hours or overnight for best results Step 4 Portion From fresh or frozen

26 Benefits Comparison Activa GS Increased handling time Stays in slurry form until applied to product Can be used in mixer or tumbler Activa RM Quick bonding Can be used as a dry powder Short ingredient statement Can be used in mixer

27 Bonding Applications

28 Texture Enhancement Slicing Yield Improvement Sodium Reduction

29 Bonding Demonstration

30 Improving Slice Yield and Texture of Deli Meats

31 Sodium Reduction / Improved Slice Yield Slice Yield Study - Ham Protocol Carriage slicer 0.5mm thick slice Compare acceptable (whole pieces, uniform, minimal breakage) to unacceptable (broken) slices Calculate on weight basis Results Sample Mg Na/100g Slice Yield. Full Sodium % 75% Reduced Sodium % 75% Red Sodium +0.03% Activa BF %

32 Sodium Reduced / Texture Improvement Ham - Textural Improvement 46% Sodium Reduction, 25% Inject Brine Force (g) Distance (mm) Con 0.01 BF 0.02 BF 0.03 BF 0.00 Force Distance

33 Slice Yield Improvement Reduction of Defects During Slicing Slicing Yield of Activa TI = 99.65% Slice Yield Without Activa TI = 93.16% 8.00% 7.00% 6.00% 5.00% 4.00% 3.00% 2.00% 1.00% % of Non-Conforming Slices 0.00% Cooked Ham With Activa TI Without Activa TI

34 Slice Yield Results Improvement of slice stability/connectivity in cooked ham Confirmed benefits of Activa TI usage: 1. Improvement of inter- and intramuscular connectivity especially between connective tissue (fat and tendons). 2. Improved peelability especially with wafer-thin slice thicknesses. 3. Reduced variance and reduction of slicing loss 6.49 % in internal trial. Typical problem in cooked ham slice stability without Activa TI Cooked ham slice with Activa TI

35 Delivered Cost Savings Calculation includes meat and brine costs, 18% pump Cost of standard ham before cooking: $4.77/kg $2.17/lb Cost increase by cooking loss: (11.1% loss) + $0.48/kg $0.22/lb Cost increase by slicing loss (6.84%) + $0.35/kg $0.16/lb Total cost (after cooking and slicing) $5.60/kg $2.55/lb Cost of Activa TI ham before cooking $4.84/kg $2.20/lb Cost increase by cooking loss (12.0% loss): + $0.50/kg $0.23/lb Cost increase by slicing loss (0.35%) + $0.02/kg $0.01/lb Total cost (after cooking and slicing) $5.36/kg $2.43/lb Cost savings $0.24/kg $0.11/lb Dramatic cost benefits may be seen through the use of Activa TI in injected low extension ham products.

36 Slice Yield Improvement Conclusions Activa improves slice stability and texture of processed ham and deli meats Through tensile strength improvement, Activa can also improve quality of PSE meat Activa allows the production of ham and deli meats with reduced sodium levels that still exhibit desirable texture Cost savings delivered through the allowance of enhanced pump rates as well as less slice waste Activa BF can also be used for injections containing functional proteins Soy proteins Whey proteins Other proteins

37 Sodium Reduction of Sausage Type Products

38 Reduced Sodium Franks Control (2% salt) 960 mg sodium/100 g product Low sodium control (0.4% salt) 180 mg sodium/100 g product Low sodium % Activa TI 180 mg sodium/100 g product Low sodium % Activa TI 180 mg sodium/100 g product Low Sodium % Activa TI 180 mg sodium/100 g product

39 Firmness Results Overall firmness Force (g) % Control 0.40% Control 0.025% TI 0.05% TI 0.075% TI Force

40 Sodium Reduction / Improved Texture Conclusion TG can be used to build back structure to reduced sodium sausage/franks without addition of binders Similar results can be seen for low sodium ham textural improvement TG can be used in conjunction with sodium replacers to deliver desired flavor Level of sodium reduction can be optimized for individual applications

41 Transglutaminase Use in Seafood Products

42 Improved Texture Activa Use in Seafood Products Transglutaminase has a long history of use in seafood products Surimi Other seafood uses Use of TG depends on surimi formulation Grade and source of fish protein Cost reduction through use of lower grade fish + TG Additional texturizing materials in formulation Egg white and soy protein Can be partially replaced by TG Important to optimize dosage according to specific customer formulations

43 Other Seafood Uses Bonding of other seafood products and/or byproducts Scallops, shrimp, mussels, other shellfish Fish Roe Surimi Novel applications Bonding of bacon to scallops Applying skin onto fish filet Unique/colorful fish block

44 Transglutaminase Regulations for Use

45 USDA Regulations TG: Use Levels Affirmed as GRAS FOOD CATEGORY Proc Meat & Poultry Non-Standard & select Std Product Chicken Breast Products Processed Seafood Meat Analogs/Substitutes Natural Cheese/Processed Cheese Cream Cheese Refrigerated Yogurt Frozen Desserts * GRAS Notice 0095 USE Level as TG 65 ppm 100 ppm GMP* GMP* GMP* GMP* GMP* GMP*

46 Safety & Regulatory Activa TG is the Only TG Approved Generally Recognized As Safe (GRAS) status in the U.S., authorized its use in meat, dairy, bread, and fish products Official approval in Canada Official approval in Europe* Many other approvals around the world, ACTIVA TG has been safely used in various food applications in the world for the last 2 decades. *France and Denmark have their own enzyme regulation. Besides these 2 countries, transglutaminase is regarded as processing aid in EU countries under Framework Directive (89/107/EEC), same as other food enzyme. Safety Standards Guideline (Guideline for food enzyme, Scientific Committee on Food, 27th series) Acute toxicity (rats) Sub-acute toxicity (rats, dogs) Mutagenicity Antibiotic residues Allergenicity Pathogenicity on producing microorganism

47 USDA Regulations U.S. Meat Approvals Standardized Products (Fed Reg. 66:211, 31-OCT-01) Fabricated Steaks Formed or Reformed Formed Beef Tenderloin* Roast Beef Parboiled and Steam Roasted Cured Pork Products Ham water added, Ham and water product-x%, Ham with Natural juices Sausages Frankfurter, frank, furter, hotdog, weiner, vienna, bologna and similar products Fabricated Poultry Reformed and Shaped Chicken Breast Poultry Rolls

48 Canadian Regulations Canada Gazette Part II vol. 138, No. 9 Apr 22, 2004 Non-standardized prepared fish products Simulated meat products Non-standardized cheese Processed cheese and cream cheese products Yogurt and Non-standardized frozen dairy desserts at GMP level

49 Canadian Regulations Interim Marketing Authorization (IMA) Volume 141 #47 Nov 24, 2007 Immediate use approved (at GMP level) for the following standardized meat products: Prepared meat and by products Prepared poultry meat and by-products Solid cut meat with or without added phosphate salts Solid cut poultry meat with or without added phosphate salts Pumping pickle and cover pickle Preserved meat & poultry and by-products Sausage Meat loaf

50 Activa Ingredient Statement

51 Government Labeling Labeling of TG by Itself Area Category Remarks Japan Food additive If enzyme is active, state enzyme EU Processing Aid No statement required (as of January 2008) USA GRAS State enzyme Canada Food additive State transglutaminase Mexico GRAS State enzyme

52 FDA / USDA Product Labeling Active Preps RM GS TI BF YG Product Usage FDA Label USDA Label* Allergen Meat / Poultry / Seafood / Fish Bonding Meat / Poultry / Seafood / Fish Bonding Texture in all food categories Texture and injections of meats Dairy Sodium caseinate, maltodextrin, transglutaminase, safflower oil Salt, fish gelatin (cod, Pollack, haddock, hake), trisodium phosphate, transglutaminase, safflower oil Maltodextrin, transglutaminase, safflower oil Ammonium glutamate, maltodextrin, transglutaminase, safflower oil Lactose, yeast extract, maltodextrin, transglutaminase, safflower oil Sodium caseinate, maltodextrin, enzyme Salt, fish gelatin, sodium phosphate, enzyme Maltodextrin, enzyme Ammonium glutamate, maltodextrin, enzyme Lactose, yeast extract, maltodextrin, enzyme Milk Fish None None None *Note: Information is for reference only. Product labeling must be determined by manufacturer.

53 FDA / USDA Product Labeling Active Preps STG-MS KS-TS-MH Product Usage Pasta Surimi FDA Label USDA Label* Allergen Maltodextrin, glucose oxidase, transglutaminase, safflower oil Sodium phosphate, potato starch, transglutaminase, safflower oil Maltodextrin, enzymes Sodium phosphate, potato starch, transglutaminase, safflower oil Wheat None Super Shrimp Crustaceans Sodium carbonate, sodium bicarbonate, glucose oxidase, glucose, calcium oxide, maltodextrin, yeast extract, transglutaminase, safflower oil Sodium carbonate, sodium bicarbonate, dextrose, calcium oxide, maltodextrin, yeast extract, enzymes, safflower oil None *Note: Information is for reference only. Product labeling must be determined by manufacturer.

54 Thank You!