Micellar Casein Concentrate Research Articles

Impact of high-pressure treatments on physicochemical and structural changes of reconstituted micellar casein concentrates from bovine and caprine milk: A comparative study

This study investigated the impact of high-pressure processing (HPP) on the physicochemical and structural changes in reconstituted micellar casein concentrates (MCC) sourced from bovine (B) and caprine (C) milk. Applying pressures ranging from 100 to 500 MPa for 30 min at 25 ± 2°C, it was observed that BMCC exhibited a notable increase in soluble minerals and solubility percentages with a simultaneous reduction in particle size and zeta potential compared with CMCC. In BMCC, calcium (Ca) and phosphorus (P) solubility rose from 27.71% and 33.02% to 90.33% and 92.01%, respectively. In CMCC, the solubility of Ca and P increased from 36.12% and 38.29% at 100 MPa to 52.57% and 56.77% at 500 MPa. Additionally, the reduction in particle size was more pronounced in BMCC, with a 53% decrease at 500 MPa, compared with a 19.90% reduction in CMCC. Furthermore, fluorescence intensity, indicative of surface hydrophobicity, decreased in BMCC samples (from 3478 to 2571 a.u.), whereas it increased in CMCC samples (from 2388 to 4058 a.u.) across the same pressure range. Comparative analysis revealed that HP-treated CMCC samples exhibited a decline in α-helix content until the treatment at 300 MPa, followed by an increase post-treatment at 500 MPa. Additionally, a rise in β-sheet structure content was observed, except for the sample treated at 500 MPa, where β-sheet content significantly decreased. These findings suggest that HPP holds promise as a viable technological approach in the dairy sector for enhancing the functional characteristics of CMCC.

Transglutaminase Crosslinked Milk Protein Concentrate and Micellar Casein Concentrate: Impact on the Functionality of Imitation Mozzarella Cheese Manufactured on a Small Scale Using a Rapid Visco Analyzer.

In dairy-based imitation mozzarella cheese (IMC) formulations, intact casein is critical and imparts IMC with a firm and elastic, stringy, melted texture. Rennet casein (RCN) is the desired ingredient to provide intact casein in IMC and is preferred over milk protein concentrate (MPC) and micellar casein concentrate (MCC). Transglutaminase (TGase), a crosslinking enzyme, alters the physical properties of MPC or MCC and may change IMC functionality. The objective of this study was to determine the effect of TGase-crosslinked MPC and MCC powders on the functionality of IMCs. The TGase treatment included TGase at 0.3 (L) and 3.0 (H) units/g of protein and a control (C) with no TGase addition. Each IMC formulation was balanced for constituents and was produced in a Rapid Visco Analyzer (RVA). The MCC or MPC powder with high TGase enzyme in IMC formulation did not form an emulsion. The IMC containing TGase-treated powders had a significantly (p ≤ 0.05) higher RVA-viscosity during manufacture and transition temperature (TT), and a significantly (p ≤ 0.05) lower Schreiber melt test area. The IMC made from MPC (with or without TGase) had lower TT values and Schreiber melt test area as compared with that made from MCC. The TGase-treated MPC and MCC, when used for IMC manufacture, were comparable to IMC manufactured with RCN in texture and some measured melted characteristics. In conclusion, TGase treatment alters the melt characteristics of MCC and MPC in IMC applications.

Oil–water interfacial behaviour of different caseins and stability of emulsions: Effect of micelle content and caseins concentrations

This study aimed to investigate the interfacial behaviour of caseins in different micelle content and its effect on the stability of emulsions, including micellar casein concentrate (MCN), calcium caseinate (CaC) and sodium caseinate (NaC). Results revealed that at high protein concentrations (0.5 %–2.5 %), MCN, CaC and NaC exhibited similar interfacial behaviour as well as unfolding rate constants (k1) of 3.11–3.41 × 10−4 (s−1), 2.96–3.35 × 10−4 (s−1) and 2.75–3.27 × 10−4 (s−1), respectively. The interfacial layer formed was dominated by non-micelles, and microscopic images revealed the thickness of the interfacial layer to be 10–20 nm. By contrast, at low concentrations, the differences in the slope of E–π curves and k1 indicated that the micelle content of casein affects protein interfacial behaviour and properties and that micellar casein is involved in the formation of the interfacial layer. The formation of large numbers of droplets during emulsion preparation results in a similar low concentration environment. Cryo-TEM showed adsorption of micellar casein in all three casein-stabilised emulsions, and the amount of adsorption was proportional to the micelle content. NaC has faster adsorption and rearrangement rates due to fewer micelles and more non-micelles, so that NaC forms smaller droplets and more stable emulsions than those formed by MCN and CaC within the range of 0.5 % to 2.0 %.

Technological Parameters of Microfiltration in the Production of Micellar Casein Concentrate

Background:: There are many different factors that influence the process of microfiltration, including membrane pore size, pressure, temperature, and membrane origin. Studying the joint influence of technological parameters in order to justify rational modes of microfiltration is relevant. Objective:: This research aimed to define the rational technological parameters of microfiltration, which allow to achieve the target parameters of retentate in the process of obtaining micellar casein concentrate. Methods:: Standard and modified methods and techniques have been used in the process of experimental research. The mathematical processing was carried out by methods of mathematical statistics according to several trials data at three stage sequence. Results:: As a result of experimental investigations, the applicability of ceramic membranes for microfiltration of skim milk has been shown. The rational pore size has been established as 0,1 μm. To obtain the micellar casein concentrate, it is advisably to carry out the microfiltration of skim milk to the mass fraction of dry solids in concentrate (13 ± 3) %, followed by two-phase diafiltration with a double volume of reverse osmosis water. The rational operating conditions, allowing to get the target value of protein fractions correlation, have been established, as described as follows: input pressure as 0,2 – 0,3 MPa, and temperature as 10 – 15°С. Conclusion:: The analysis of microfiltration concentrates and permeates composition of raw milk, including vitamins and minerals content, showed both their high nutritional and biological value and the viability of further processing into food ingredients, including the ingredients for the specialized food products.

Development of a premix based on micellar casein for fortification of meat systems with vitamin A

A novel approach to the protection of unstable fat-soluble vitamins, using retinol as an example, is presented in this work. This method is based on introducing vitamin A molecules into casein micelles. Protective properties of micellar casein towards different forms of retinol (native vitamin and palmitic acid ester) in vitro and in emulsion-type meat products are investigated. A technology of the introduction using micellar casein concentrate (MCC) has been developed. Conditions similar to those in which vitamin molecules can be in meat emulsions during heat treatment are simulated in vitro. The optimal time of “encapsulation” (2 hours) and the need for additional surfactant (tween-80) are identified. The use of the casein micelles protection made it possible to increase the number of retinol molecules that did not undergo decomposition under model conditions (in vitro) from ~30% to ~80%. Using the vitamin premix the degree of degradation of vitamin molecules does not exceed 4% after heat treatment. Data received allowed us to determine the efficiency of the protection properties of casein micelles for unstable vitamin A molecules.

Digestive properties of β-CN and α-La in different milk protein ingredients

The digestion behavior of proteins not only differs between protein types but also between ion saturation and protein profile. This study aimed to investigate the digestive properties of α-La and β-CN in different dairy protein ingredients, including β-CN concentrate (βCC), micellar casein concentrate (MCC), α-La concentrate (α-LaC), and whey protein concentrate (WPC80), using an in-vitro infant static digestion model. α-La digested much faster in WPC than in α-LaC in the gastric phase, whereas β-CN digested faster in βCC than in MCC during gastric digestion. In the intestinal phase, α-La released more amino acids than β-CN. The released essential amino acids index (EAAIS) was higher in α-LaC than WPC80 and higher in βCC than MCC. This study facilitates better understanding of the effect of protein profile, protein structure, and calcium binding on the digestion behavior of β-CN and α-La, which provide reference for selection of dairy ingredient for infant formula.

Structure and digestibility of bovine casein complexes formed by enriching κ- and β-caseins in micellar casein concentrate together with minerals adjustment

Colloidal structure and infant in vitro gastrointestinal digestibility of casein complexes simulating caseins and minerals compositions of human casein micelles were studied. κ- and β-Caseins were fractionated from bovine micellar casein concentrate (MCC), and mixed with MCC to increase their ratios to 20% and 68%, followed by adding citrate (Cit), calcium (Ca), and inorganic phosphate (Pi) to obtain the casein complexes. The complexes enriched with κ- and β-caseins showed higher percentages of serum Ca, Pi and caseins, larger particle size, and looser internal structure. During gastric digestion, the complexes enriched with κ- and β-caseins formed smaller and looser coagula, which promoted degradation of intact caseins and formation of free amino groups and small peptides. During initial intestinal digestion, the complexes enriched with κ- and β-caseins formed more free amino groups and smaller peptides. These results offered a potential strategy to form human casein micelles analogues for use in infant formula.

Ultrasonication of Micellar Casein Concentrate to Reduce Viscosity-Role of Undissolved Material.

This research reveals the underlying mechanisms that make high-intensity ultrasound an effective tool to reduce the viscosity of micellar casein concentrates and to enhance the solubility of the respective powders. Micellar casein concentrates (MCC) gained great importance in the production of valuable food products with high protein content, but the processing properties of the reconstituted solutions are deficient. Even though several presumptions were established, the reasons why ultrasound is able to reduce the product viscosity and what limitations occur when using sonication technology are still not clear yet. Our study aims to investigate those reasons by combining analyses of viscosity measurements, particle size distributions, solubility, and hydration. The data presented demonstrate that undissolved, highly hydrated particles play an important role in micellar casein concentrates showing a high viscosity. We conclude on the high voluminosity of those particles, since improved solubility and decreased viscosity are accompanying effects. The determined voluminosities of those particles are 35-40% higher than for colloidal dissolved micelles. Hence, the viscosity reduction of up to 50% can be only obtained by sonicating micellar casein concentrates derived from powder reconstitution, whereas ultrasonication of freshly prepared membrane-filtrated MCC does not reduce viscosity.

The physicochemical properties and heat stability of milk fortified with blackberry juice and different casein-dominant ingredients

The influence of blackberry juice (BBJ) on the physicochemical properties and heat stability of skimmed milk with and without sodium caseinate or micellar casein concentrate (MCC) addition was investigated. This was performed by measuring the ionic calcium content, heat coagulation time, viscosity, particle size distribution and analytical centrifugation profiles of the different samples. BBJ decreased the ionic calcium of the samples, which resulted in a higher heat coagulation time at pH < 6.5. Its addition also prevented the increase in viscosity observed in skimmed milk and skimmed milk with MCC at pH 6.2 after heating. The particle size distribution, turbidity and analytical centrifugation profiles indicated this was due to a prevention of protein aggregation. In conclusion, the blackberry juice improved the heat stability of different types of milk systems in the pH range between 6.2 and 6.4, which was likely due to binding of calcium by blackberry polyphenols.

Effect of enzymatic dephosphorylation on caprine casein micelle structure

The effect of enzymatic dephosphorylation, using intestinal alkaline phosphatase, on the structure of casein micelles in caprine micellar casein concentrate (MCC) was studied. An optimal condition, involving preheating the MCC dispersion, pH 6.4, 2.5 mg casein/mL and 0.4 U phosphatase/mL, was established and used to prepare MCC with 0–49% dephosphorylation by incubation at 37 °C for 0–180 min. β-Casein showed marked dephosphorylation and formed multi-phosphorylated isoforms depending on dephosphorylation degree, whereas αs- and κ-caseins showed limited and fast dephosphorylation, respectively. With increasing dephosphorylation, both the colloidal calcium and the calcium sensitive micellar caseins, especially β-casein, were gradually dissociated, and the calcium insensitive serum κ-casein was gradually associated with the micelles. The dissociated β-casein fraction was predominated by isoforms with lower number of phosphate groups. For micelles with increasing dephosphorylation, the molar mass decreased, the gyration and hydrodynamic radii decreased, the ratio of gyration to hydrodynamic radii and micellar hydration increased, the spherical morphology was generally retained, and the internal protein inhomogeneity disappeared progressively. These results suggest that after dephosphorylation, the caprine micelle framework underwent an intra-micellar mass redistribution, and become more loose and homogeneous.

Effect of dipotassium phosphate addition and heat on proteins and minerals in milk protein beverages

Our objective was to determine the effects of dipotassium phosphate (DKP) addition, heat treatments (no heat, high temperature, short time [HTST]: 72°C for 15 s, and direct steam injection UHT: 142°C for 2.3 s), and storage time on the soluble protein composition and mineral (P, Ca, K) concentration of the aqueous phase around casein micelles in 7.5% milk protein-based beverages made with liquid skim milk protein concentrate (MPC) and micellar casein concentrate (MCC). Milk protein concentrate was produced using a spiral wound polymeric membrane, and MCC was produced using a 0.1-µm ceramic membrane by filtration at 50°C. Two DKP concentrations were used (0% and 0.15% wt/wt) within each of the 3 heat treatments. All beverages had no other additives and ran through heat treatment without coagulation. Ultracentrifugation (2-h run at 4°C) supernatants of the beverages were collected at 1, 5, 8, 12, and 15-d storage at 4°C. Phosphorus, Ca, and K concentrations in the beverages and supernatants were measured using inductively coupled plasma spectrometry. Protein composition of supernatants was measured using Kjeldahl and sodium dodecyl sulfate-PAGE. Micellar casein concentrate and MPC beverages with 0.15% DKP had higher concentrations of supernatant protein, Ca, and P than beverages without DKP. Protein, Ca, and P concentrations were higher in MCC supernatant than in MPC supernatant when DKP was added, and these concentrations increased over storage time, especially when lower heat treatments (HTST or no heat treatment) had been applied. Dipotassium phosphate addition caused the dissociation of αS-, β-, and κ-casein, and casein proteolysis products out of the casein micelles, and DKP addition explained over 70% of the increase in supernatant protein, P, and Ca concentrations. Dipotassium phosphate could be removed from 7.5% of protein beverages made with fresh liquid MCC and MPC (containing a residual lactose concentration of 0.6% to 0.7% and the proportional amount of soluble milk minerals), as these beverages maintain heat-processing stability without DKP addition.

Effect of temperature and protein concentration on the protein types within the ultracentrifugation supernatant of liquid micellar casein concentrate

Liquid micellar casein concentrate (MCC) is an ideal milk-based protein ingredient for neutral-pH ready-to-drink beverages. The texture and mouthfeel of liquid MCC-based beverages depend on the beverage protein content, as well as the composition of soluble proteins in the aqueous phase around the casein micelle. The objective of this study was to determine the composition of soluble proteins in the aqueous phase around the casein micelles in skim milk and liquid MCC containing 7.0% and 11.6% protein content. Skim milk was pasteurized and concentrated to 7% protein content by microfiltration and then to 18% protein content by ultrafiltration. The 18% MCC was then serially diluted with distilled water to produce 11.6% and 7.0% protein MCC. Skim milk, 7.0% MCC, and 11.6% MCC representing starting materials with different protein concentrations were each ultracentrifuged at 100,605 × g for 2 h. The ultracentrifugation for each of the starting materials was performed at 3 different temperatures: 4°C, 20°C, and 37°C. The ultracentrifugation supernatants were collected to represent the aqueous phase around the casein micelle in MCC solutions. The supernatants were analyzed by Kjeldahl to determine the crude protein, casein, and casein as a percentage of crude protein content, and by sodium dodecyl sulfate PAGE to determine the composition of the individual proteins. Most of the proteins in MCC supernatant (about 45%) were casein proteolysis products. The remaining proteins in the MCC supernatant consisted of a combination of intact αS-, β-, and κ-caseins (about 40%) and serum proteins (14-18%). Concentrations of αS-casein and β-casein in the supernatant increased with decreasing temperature, especially at higher protein concentrations. Temperature and interaction between temperature and protein explained about 80% of the variation in concentration of supernatant αS- and β-caseins. Concentration of supernatant κ-casein, casein proteolysis products, and serum protein increased with increasing MCC protein concentration, and MCC protein concentration explained most of the variation in supernatant κ-casein, casein proteolysis products, and serum protein concentrations. Predicted MCC apparent viscosity was positively associated with the dissociation of αS- and β-caseins. Optimal beverage viscosity could be achieved by controlling the dissociation of these proteins in MCC.

Microstructural Characterization of High-Protein Dairy Powders

Dairy proteins are potential and multipurpose ingredients used to prepare various food products. It exhibits many beneficial functionalities and bioactivities in the processing of food products. All the functionalities of different dairy proteins depend on their peculiar structural characteristics. So, the present study aimed to characterize high-protein powders with different matrices (milk protein concentrate (MPC), rennet casein (RCN), micellar casein concentrate (MCC), whey protein isolate (WPI), and native whey concentrate (NWC)) for their composition, handling, and reconstitution properties, and microstructure and compared them with whole milk powder (WMP) and skim milk powder (SMP). The bulk density of the high-protein powders was significantly (p &lt; 0.05) lower than WMP and NFDM. Due to the low bulk density of the high-protein powders, the wettability of these powders in the water was very high. Microstructural analysis of powders by scanning electron microscopy reveals that the high-protein powder had tetrahedron-to-polyhedron-shaped particles depending on the protein level. The idea regarding the powder’s morphology might be helpful for further improvement in the production processes to modify the functional properties of high-protein milk powders.

Gastrointestinal digestibility of micellar casein dispersions: Effects of caprine vs bovine origin, and partial colloidal calcium depletion using in vitro digestion models for the adults and elderly.

In vitro coagulation and digestion of caprine and bovine micellar casein concentrate (MCC) with or without partial colloidal calcium depletion (deCa) were studied under simulated adult and elderly conditions. Gastric clots were smaller and looser for caprine than bovine MCC, and were further looser with deCa and under elderly condition for both caprine and bovine MCC. Casein hydrolysis and concomitant formation of large peptides was faster for caprine than bovine MCC, and with deCa and under adult condition for caprine and bovine MCC. Formation of free amino groups and small peptides were faster for caprine MCC, and with deCa and under adult condition. Upon intestinal digestion, proteolysis occurred rapidly, and was faster under adult condition, but showed less differences with increasing digestion between caprine and bovine MCC, and with and without deCa. These results suggested weakened coagulation and greater digestibility for caprine MCC and MCC with deCa under both conditions.

Manufacture of loaf‐type processed cheese products using transglutaminase crosslinked micellar casein concentrate and milk protein concentrate

The aim of this research was to determine the effect of transglutaminase (TGase; 0, 0.3 and 3.0 units/g of protein) crosslinked micellar casein concentrate (MCC) and milk protein concentrate (MPC) on the functionality of process cheese product (PCP) loaf‐type formulation. The results showed that TGase significantly (P ≤ 0.05) increased viscosity after manufacture. Higher TGase levels also significantly (P ≤ 0.05) affected post‐manufacture melt characteristics such as melt viscosity and melt area measured using a rapid visco analyser and Schreiber melt test respectively. The study shows that the TGase treatment alters meltability of PCP.

Acid gelation functionality of casein micelles in concentrated state: Influence of calcium supplementation or chelation combined with enzymatic stabilization

The main structure-building component in gelled dairy products is the casein micelle (CM). It was previously shown that in concentrated state of CMs, the changes of their compositional and colloidal properties in response to the addition or chelation of calcium divers from the reactions at milk-like casein concentration. This study aimed to evaluate the relationship between these specifically induced properties of CMs and the acid gelation functionality of a micellar casein concentrate (MCC). A covalent stabilization of the structures of pre-modified and native CMs by treatment with transglutaminase (Tgase) prior to acidification with glucono-δ-lactone was additionally tested. During acidification, structure formation was monitored in a rheometer to gain insights on mechanistic changes in the course of structural changes during acidification compared to gelation of unmodified MCC. Gelation pH, gel firmness, and serum binding capacity were analyzed to characterize gel structures at a final pH of 4.6. The structure formation process was highly affected by prior changes in composition, integrity, and structural stabilization of CMs. Gelation pH and acid gel firmness were reduced by ionic strength. Thermal and Tgase treatment induced opposing effects, leading to compensation when combined with prior salt supplementation. A synergistic increasing effect on serum binding capacity was found for trisodium-citrate supplementation and subsequent Tgase treatment. New mechanistic insights on acid gel structure formation progression were gained. This study provides a better insight into the relationship between structural CM properties prior to acid gelation and the resulting acidification progress, as well as the final acid gel properties.

Characteristics of imitation Mozzarella cheese manufactured without emulsifying salts using a combination of culture-based acid curd and micellar casein concentrate.

The objectives of this study were to develop a process to produce acid curd from micellar casein concentrate (MCC) using starter cultures and to manufacture imitation Mozzarella cheese (IMC) using a combination of acid curd and MCC that would confer emulsification ability to the caseins without the use of emulsifying salts (ES). The formulations were targeted to produce IMC with 49.0% moisture, 20.0% fat, 18.0% protein, and 1.5% salt. In the IMC formulation made without ES (FR-2:1), the acid curd was blended with MCC so that the formula contained a 2:1 ratio of protein from acid curd relative to MCC. IMC with ES was also produced as a control. The melt and stretch characteristics of IMC made from FR-2:1 were similar to those of control IMC. We conclude that IMC can be made without ES using a 2:1 ratio of protein from acid curd relative to MCC.

Effect of dipotassium phosphate and heat on milk protein beverage viscosity and color

Our objective was to determine the effect of addition of dipotassium phosphate (DKP) at 3 different thermal treatments on color, viscosity, and sensory properties of 7.5% milk protein-based beverages during 15 d of storage at 4°C. Micellar casein concentrate (MCC) and milk protein concentrate (MPC) containing about 7.5% protein were produced from pasteurized skim milk using a 3×, 3-stage ceramic microfiltration process and a 3×, 3-stage polymeric ultrafiltration membrane process, respectively. The MCC and MPC were each split into 6 batches, based on thermal process and addition of DKP. The 6 batches were no postfiltration heat treatment with added DKP (0.15%), no postfiltration heat without added DKP (0%), postfiltration high-temperature, short time (HTST) with DKP, postfiltration HTST without DKP, postfiltration direct steam injection with DKP, and postfiltration direct steam injection without DKP. The 6 MCC milk-based beverages and the 6 MPC milk-based beverages were stored at 4°C. Viscosity, color, and sensory properties were determined over 15 d of refrigerated storage. MCC- and MPC-based beverages at 7.5% protein with and without 0.15% added dipotassium phosphate were successfully run through an HTST and direct steam injection thermal process. The 7.5% protein MCC-based beverage contained a higher calcium and phosphorus content (2,425 and 1,583 mg/L, respectively) than the 7.5% protein MPC-based beverages (2,141 and 1,338 mg/L, respectively). Pasteurization (HTST) had very little effect on beverage particle size distribution, whereas direct steam injection thermal processing produced protein aggregates with medians in the range of 10 and 175 μm for MPC beverages. A population of casein micelles at about 0.15 μm was found in both MCC- and MPC-based beverages. Larger particles in the 175-μm range were not detected in the MCC beverages. In general, the apparent viscosity (AV) of MCC beverages was higher than MPC beverages. Added DKP increased the AV of both MCC- and MPC-based beverages, while increasing heat treatment decreased AV. The AV of beverages with DKP increased during 15 d of 4°C of storage for both MCC and MPC, whereas there was very little change in AV during storage without DKP and a similar effect was observed for sensory viscosity scores. The L value of beverages was higher with higher heat treatment, but DKP addition decreased L value and sensory opacity greatly. Sulfur-eggy flavors were detected in MPC beverages, but not MCC-based beverages.

Fractionation of κ-casein from caprine micellar caseins using differential precipitation

The effects of potentially important processing conditions at each step of differential precipitation on the yield and purity of κ-casein fractionated from caprine micellar casein concentrate (MCC) were investigated. The optimal conditions were: rehydrating (3% casein) MCC using homogenisation followed by dissociating micelles at pH 11.0; complexing caseins using 45 mm added Ca at 25 °C for 60 min, followed by precipitating β-/αS-caseins using 2 m acetic acid at pH 7.0 again at 25 °C for 60 min. Two fractionation cycles were used with 50 mm added Ca for the second cycle, followed by precipitating κ-casein at pH 3.8 and 50 °C. To increase processing capacity for scale-up, increased casein concentrations of 4–7% along with 60 mm added Ca resulted in comparable yields and purity of the κ-casein. Maximum yield and purity of 84.6% and 84.4%, respectively, were achieved for κ-casein from the MCC dispersions at 4% casein.

Manufacture of a novel cultured micellar casein concentrate ingredient for emulsifying salt–free process cheese products applications

Micellar casein concentrate (MCC) is a high protein ingredient that is typically produced using 3 stages of microfiltration with a 3× concentration factor and diafiltration. Acid curd is an acid protein concentrate, which can be obtained by precipitating the casein at pH 4.6 (isoelectric point) using starter cultures or direct acids without the use of rennet. Process cheese product (PCP) is a dairy food prepared by blending dairy ingredients with nondairy ingredients and then heating the mixture to get a product with an extended shelf-life. Emulsifying salts are critical for the desired functional characteristics of PCP because of their role in calcium sequestration and pH adjustment. The objectives of this study were to develop a process to produce a novel cultured micellar casein concentrate ingredient (cMCC; culture-based acid curd) and to produce PCP without emulsifying salts using different combinations of protein from cMCC and MCC in the formulations (2.0:1.0, 1.9:1.1, and 1.8:1.2). Skim milk was pasteurized at 76°C for 16 s and then microfiltered in 3 microfiltration stages using graded permeability ceramic membranes to produce liquid MCC (11.15% total protein; TPr and 14.06% total solids; TS). Part of the liquid MCC was spray dried to produce MCC powder (75.77% TPr and 97.84% TS). The rest of the MCC was used to produce cMCC (86.9% TPr and 96.4% TS). Three PCP treatments were formulated with different ratios of cMCC:MCC, including 2.0:1.0, 1.9:1.1, and 1.8:1.2 on the protein basis. The composition of PCP was targeted to 19.0% protein, 45.0% moisture, 30.0% fat, and 2.4% salt. This trial was repeated 3 times using different batches of cMCC and MCC powders. All PCP were evaluated for their final functional properties. No significant differences were detected in the composition of PCP made with different ratios of cMCC and MCC except for the pH. The pH was expected to increase slightly with elevating the MCC amount in the PCP formulations. The end apparent viscosity was significantly higher in 2.0:1.0 formulation (4,305 cP) compared with 1.9:1.1 (2,408 cP) and 1.8:1.2 (2,499 cP). The hardness ranged from 407 to 512 g with no significant differences within the formulations. However, the melting temperature showed significant differences with 2.0:1.0 having the highest melting temperature (54.0°C), whereas 1.9:1.1 and 1.8:1.2 showed 43.0 and 42.0°C melting temperature, respectively. The melting diameter (38.8 to 43.9 mm) and melt area (1,183.9 to 1,538.6 mm2) did not show any differences in different PCP formulations. The PCP made with a 2.0:1.0 ratio of protein from cMCC and MCC showed better functional properties compared with other formulations.