PROTEIN BLENDS Dr. Laurice Pouvreau Senior Scientist Protein Functionality laurice.pouvreau@nizo.com May 23-24, 2017
NIZO FOOD RESEARCH FOR BETTER FOOD & HEALTH Independent, private contract research company for food and health Proteins Bacteria Processing HQ in The Netherlands (Food Valley) 140 professionals From lab to practice Food-grade pilot plant Processing Centre Research Center 2
PROTEIN FUNCTIONALITY UNLOCKING THE FULL POTENTIAL OF PROTEINS Dairy, Plant, Animal, Algae, Insect Proteins and Hydrolysates
AGENDA What do we mean by protein blends? Same source blends: how functionality is driven by protein-protein interactions. Multi-source blends: can animal-plant blends solve the new source challenges?
INGREDIENT FLEXIBILITY Protein blends: Nutritional value, sustainability and price Choice or necessity? Not dependent on one supplier Textural properties One-to-one protein or ingredient replacement is not always possible: Ingredient compatibility or Smart blending of ingredient Nutritional quality Proteins from different sources have different amino acid and digestibility Technical functionality Solubility, gelling, emulsifying, foaming Interaction with hydrocolloids Taste and smell Understanding ingredient functionality and ingredient compatibility 5
PROTEIN BLENDS Protein blends: From the same source: milk or soy proteins From different sources: mixes of protein from different origin Mixes of intact and hydrolyzed proteins Ingredient compatibility: Effect of the processing conditions (ph, T, and ratio between the different proteins) on the final product properties, Relation between the powder/protein properties and the final product properties, TO ENABLE USING/CONTROLLING VARIOUS INGREDIENTS
FRACTIONATION OF MILK FROM MILK TO SERUM MF 200nm Micellar casein Native whey UF 10 kda Whey proteins Milk permeate Cream UF 10 kda Milk protein concentrate
SOY PROTEIN EXTRACTION 40% Protein 70% glycinin β-conglycinin Gel hardness Elasticity 8 Research into single-protein functionality enables better understanding of the blend
AGENDA What do we mean by protein blends? Same source blends: how functionality is driven by protein-protein interactions. Multi-source blends: can animal-plant blends solve the new source challenges?
EGG PROTEIN INGREDIENTS A WIDE RANGE OF APPLICATIONS Egg yolk Phospholipids, proteins Mayonnaise, sauces, bakery products Egg white powder Ovalbumin is major protein Processing controls functionality/application Egg white powder / Crystallised egg white High foam / High gel Foaming, gelling, water binding Lysozyme Protein with high iso-electric point Preservative against gram-positive bacteria in many food and beverage products 10 www.bouwhuis-enthoven.com
EGG WHITE COMPOSITION Ovalbumin is the main protein in egg white and also the most studied. Not available commercially (too expensive process) Lysozyme is extracted on commercial scale 11 Abeyrathne et al, Poultry Science 92 (2013), 3292 3299
OVALBUMIN VERSUS EGG WHITE GELLING PROPERTIES Ovalbumin Egg white Ovalbumin Egg white Increasing salt concentration Water holding (%) Pouvreau et al, JFS 81 (2016), N982 N990 Protein purity directly influences their functional properties
CASEIN AND WHEY PROTEINS YOGURT Pre-heating treatment of yoghurt mix required to: Inactivate bacteria Induce whey protein denaturation to create desirable texture Fractionation & analysis Whey protein-casein micel Aggregated whey protein Native whey protein 13
CASEIN AND WHEY PROTEINS SOLUBLE, DENATURED WHEY PROTEINS ESSENTIAL FOR FIRMNESS Interactions of whey with casein micelle highly sensitive to ph Optimised pre-heating step: soluble, denatured whey proteins essential for firmness 14
CASEIN AND WHEY PROTEINS CASEIN AND WHEY RATIO V isc o sity (P o sth u m u s se c ) 200 WPC35* WPC35 100 30 2 4 6 WPC35 (closed symbol) WPC35* (open symbol) casein m icelle/w hey prot Increased % denaturation upon increased amount of whey proteins To obtain a viscosity enhancing effect, an optimal amount of whey proteins (β-lactoglobulin) should be present, as a non-optimal amount decreased the viscosity.
INGREDIENT COMPATIBILITY STABILITY OF INFANT FORMULA Infant formula: Typically contains both milk powder and WPC as ingredients Heat-load during infant formula manufacture can result in protein instability, leading to: Insolubility White flecks
INGREDIENT COMPATIBILITY STABILITY OF INFANT FORMULA Skim milk powder Skim milk powder 1: ~50% whey protein denaturation (medium-heat) Skim milk powder 2: ~95% whey protein denaturation (high-heat) Whey protein concentrate Whey protein concentrate 1: ~20% whey protein denaturation Whey protein concentrate 2: ~50% whey protein denaturation Behavior of blends vs. individual ingredients in relation to heat stability
INGREDIENT COMPATIBILITY INFANT FORMULA Heat coagulation time at 110 C (s) 2500 2000 1500 1000 500 0 SMP 1 SMP 2 6.3 6.5 6.7 6.9 7.1 ph Heat coagulation time at 95 C (s) SMP1+WPC1 SMP1+WPC2 15 10 5 0 SMP2+WPC2 SMP2+WPC1 6.3 6.5 6.7 6.9 7.1 ph Heat stable SMP works does not give highest stability in combination with WPC Test in appropriate systems when selecting ingredients Combine ingredient functionality with ingredient compatibility
PROTEIN INGREDIENTS FOR HIGH-PROTEIN BARS High-protein bars have high protein content, but also high carbohydrate content, and low fat and low moisture content. 19
MODEL PROTEIN BAR SYSTEMS Ingredients: 45% (m/m) carbohydrate syrup (mixture of 50% fructose and 50% glucose at 70 g solids/100 g solution); 45% (m/m) milk protein ingredient; 10% (m/m) glycerol. NaCas CaCas1 CaCas2 Storage time at 37 C 1 d 8 d 15 d 28 d Casein-based powders WPI1 WPI2 WPI3 WPC80 Storage time at 37 C 1 d 8 d 15 d 28 d Whey protein-based powders Protein bars stored in sealed containers at 37 C. Analyzed for: Water activity; Texture analysis; Color changes. CaCas3 MPC90 MCC80 Acid casein Rennet casein CH1 CH2 CPP WPH1 WPH2 Protein hydrolysates 20
BLENDS OF PROTEINS AND PEPTIDES Protein bar hardness (N) 1000 100 10 1 0.1 0.01 1 Day 8 Days 0 20 40 60 80 100 NaCas (% in NaCas : CH mixture) Protein bar hardness (N) 1000 100 10 1 0.1 1 Day 8 Days 0 20 40 60 80 100 WPI (% in WPI : WPH) Model system protein bars prepared with blends of sodium caseinate and casein hydrolysate, and whey protein isolate and whey protein hydrolysate, respectively. Striking difference between NaCas and WPI: very high contribution to hardness by NaCas at similar ratios Differences in hardness of the blends due to powder properties or protein properties 21
AGENDA What do we mean by protein blends? Same source blends: how functionality is driven by protein-protein interactions. Multi-source blends: can animal-plant blends solve the new source challenges?
PROTEIN BLENDS WITH PLANT PROTEINS Plant proteins are used in a wide range of products ranging from: Protein bars, processed meat, Infant formula, elderly and clinical food, Heat stability and ingredient solubility Viscosity, Ingredient compatibility = smart blending. BUT a broader and more extensive use of plant proteins is often limited by lack of dispersibility Increase plant protein use by blending with animal proteins, Protein-protein interactions in various applications, Soluble versus commercial plant proteins.
HEAT STABILITY PROTEIN BLENDS WHEY AND PEA PROTEINS 3.5 % w/w protein Heating: 120 C between 1 and 10 min Increasing heating time decreases the amount of insoluble materials.
HEAT STABILITY PROTEIN BLENDS WHEY AND PEA PROTEINS 3.5 % w/w protein Heating: 120 C between 1 and 10 min ph > 7.0 is crucial for pea protein heat stability Increasing ph increases heat stability of pea protein
ACIDIFIED (GDL) GELS SODIUM CASEINATE AND NIZO EXTRACTED SOY PROTEINS 10% m/m protein 100% NaCas No heat treatment 70/30 NaCas/SP Heat treated soy proteins Addition of soy proteins resulted in changes in microstructure, Decrease in mechanical property Martin, Pouvreau et al. (2016) Food hydrocolloids, 58
TARGETED PRE-TREATMENT FOR RECOVERING TEXTURE ACIDIFIED GEL OF NaCas AND NIZO EXTRACTED SP 10% m/m protein 100% NaCas 70/30 NaCas/SP No treatment Treated NaCas/SP Smart mixing and processing allows texture recovery Martin, Pouvreau et al. (2016) Food hydrocolloids, 58
TARGETED PRE-TREATMENT FOR RECOVERING TEXTURE ACIDIFIED GEL OF NACAS AND NIZO EXTRACTED SP Smart mixing and processing: The ph at pre-treatment The ratio NaCas and SP during pre-treatment Understanding protein-protein interaction
HEAT SET GELS WHEY PROTEINS AND NIZO EXTRACTED SOY PROTEINS 10% m/m protein 100mM NaCl, ph 7.0 WP:SP=10:0 9:1 7:3 5:5 The substitution of 1% WP by 1% SP has a large impact on the microstructure of the heat set gels of WP and SP Jose, Pouvreau, Martin (2016) Food Hydrocolloids, 60 29
HEAT SET GELS WHEY PROTEINS AND SELF EXTRACTED SOY PROTEINS 10% m/m protein 100mM NaCl, ph 7.0 Partial replacement change in textural properties Increase in water holding Mouth feel while chewing may be affected Jose, Pouvreau, Martin (2016) Food Hydrocolloids, 60 30
HEAT SET GELS WHEY PROTEINS AND SELF EXTRACTED SOY PROTEINS 10% m/m protein 100mM NaCl, ph 7.0 Heat induced gelation of WP and SP leads to a gel in which both protein contribute to the gel strength Synergistic effect appears when higher concentration of SP is added Jose, Pouvreau, Martin (2016) Food Hydrocolloids, 60 31
HEAT SET GELS WHEY PROTEINS AND COMMERCIAL SOY PROTEINS WP:SP=10:0 9:1 7:3 10% m/m protein 150mM NaCl, ph 7.0 120 70 Young modulus (Kpa) 100 80 60 40 20 0 Commercial soy NIZO extracted soy 0 1 2 3 SP concentration (% m/m) Fracture stress (KPa) 60 50 40 30 20 10 0 Commercial soy NIZO extracted soy 0 1 2 3 SP concentration (% m/m) 32
HEAT SET GELS WHEY PROTEINS AND COMMERCIAL SOY PROTEINS 10% m/m protein 150mM NaCl, ph 7.0 WP:SP=9:1 No treatment SP heated Homogenized 250/50 400/50 Heating SP prior to gelling has a positive effect on protein dispersibility but not a clear effect on microstructure Homogenization is having a clear effect on gel microstructure 33
120 HEAT SET GELS WHEY PROTEINS AND COMMERCIAL SOY PROTEINS 60 10% m/m protein 150mM NaCl, ph 7.0 Young modulus (Kpa) 100 80 60 40 20 0 Commercial soy Self extracted soy Heated commercial soy Homegenized 250/50 com soy Homogenized 400/50 com soy 0 1 2 SP concentration (% m/m) 3 Fracture stress (KPa) 50 40 30 20 10 0 0 1 2 3 SP concentration (% m/m) Minor effect of the pre-treatment on the stiffness of the gels, Effect of homogenization more significant on the firmness of the gels 34
EMULSION CAPACITY/STABILITY NA-CAS AND PEA PROTEINS Na-Cas:PPI=0.5:0.5 Na-Cas:PPI=0:1 Na-Cas:HF-PPI=0.5:0.5 Composition Clarification rate (µs/s) Cream layer (%) Na-Cas:PPI = 1:0 1.20 32.0 Na-Cas:PPI = 0.75:0.25 1.39 36.0 Na-Cas:PPI = 0.50:0.50 1.65 38.5 Na-Cas:PPI = 0.25:0.75 1.98 42.5 Na-Cas:PPI = 0:1 2.55 45.3 Na-Cas:HF-PPI = 0.50:0.50 1.40 32.5 Increasing creaming layer Lack of soluble protein increases emulsion instability Pouvreau & van de Velde (2015), World of Food Ingredients 35
TAKE HOME MESSAGES The same two proteins can either be synergistic or antagonistic, depending on their individual processing history, the precise conditions when adding/blending the two together, their shared processing history. Smart blending can address common obstacles to use of new source proteins, namely a complete nutritional profile, an excellent texture, using protein efficiently, a clean taste profile. Functionality starts with dispersibility. 36
THANKS Laurice Pouvreau Fred van de Velde Thom Huppertz Marja Kanning 37