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-, ß-, 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-, ß-, and κ-caseins (about 40%) and serum proteins (14-18%). Concentrations of αS-casein and ß-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 ß-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 ß-caseins. Optimal beverage viscosity could be achieved by controlling the dissociation of these proteins in 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.

Efficiency of removal of whey protein from sweet whey using polymeric microfiltration membranes.

Our objective was to measure whey protein removal percentage from separated sweet whey using spiral-wound (SW) polymeric microfiltration (MF) membranes using a 3-stage, 3× process at 50°C and to compare the performance of polymeric membranes with ceramic membranes. Pasteurized, separated Cheddar cheese whey (1,080 kg) was microfiltered using a polymeric 0.3-µm polyvinylidene (PVDF) fluoride SW membrane and a 3×, 3-stage MF process. Cheese making and whey processing were replicated 3 times. There was no detectable level of lactoferrin and no intact α- or ß-casein detected in the MF permeate from the 0.3-µm SW PVDF membranes used in this study. We found BSA and IgG in both the retentate and permeate. The ß-lactoglobulin (ß-LG) and α-lactalbumin (α-LA) partitioned between retentate and permeate, but ß-LG passage through the membrane was retarded more than α-LA because the ratio of ß-LG to α-LA was higher in the MF retentate than either in the sweet whey feed or the MF permeate. About 69% of the crude protein present in the pasteurized separated sweet whey was removed using a 3×, 3-stage, 0.3-µm SW PVDF MF process at 50°C compared with 0.1-µm ceramic graded permeability MF that removed about 85% of crude protein from sweet whey. The polymeric SW membranes used in this study achieve approximately 20% lower yield of whey protein isolate (WPI) and a 50% higher yield of whey protein phospholipid concentrate (WPPC) under the same MF processing conditions as ceramic MF membranes used in the comparison study. Total gross revenue from the sale of WPI plus WPPC produced with polymeric versus ceramic membranes is influenced by both the absolute market price for each product and the ratio of market price of these 2 products. The combination of the market price of WPPC versus WPI and the influence of difference in yield of WPPC and WPI produced with polymeric versus ceramic membranes yielded a price ratio of WPPC versus WPI of 0.556 as the cross over point that determined which membrane type achieves higher total gross revenue return from production of these 2 products from separated sweet whey. A complete economic engineering study comparison of the WPI and WPPC manufacturing costs for polymeric versus ceramic MF membranes is needed to determine the effect of membrane material selection on long-term processing costs, which will affect net revenue and profit when the same quantity of sweet whey is processed under various market price conditions.

Measurement of casein in milk by Kjeldahl and sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Our objectives were to determine if milk casein as a percentage of true protein (CN%TP) estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is equivalent to CN%TP estimated by Kjeldahl, and to determine the proportion of casein (CN), casein proteolysis products (CNPP), and serum protein (SP) from milk true protein (TP) that goes into the Kjeldahl noncasein nitrogen (NCN) filtrate and the proportion that stays in the NCN precipitate using SDS-PAGE. Raw milk samples were collected from 16 mid-lactation Holstein cows twice a week for 2 wk. These milks were analyzed for Kjeldahl total nitrogen, nonprotein nitrogen, and NCN content in duplicate, and by SDS-PAGE. The CN%TP determined by Kjeldahl was compared with the CN%TP estimated by SDS-PAGE calculated in 2 ways: as a percentage of only intact caseins divided by TP and as a percentage of both intact caseins and CNPP divided by TP. Three milks varying in fat, lactose, TP, CN, and SP content were formulated. These milks were analyzed in duplicate for Kjeldahl total nitrogen, nonprotein nitrogen, and NCN content, and each of the NCN filtrate and NCN precipitate were analyzed in duplicate by SDS-PAGE for relative quantity (%) of CN, CNPP, and SP. We found that the estimate of CN%TP by Kjeldahl was higher than the estimate of CN%TP by SDS-PAGE that was calculated as only intact CN divided by the total of all protein bands. However, no difference was detected in the estimate of CN%TP by Kjeldahl compared with CN%TP by SDS-PAGE when CNPP were included as CN in the calculation of SDS-PAGE results. Based on SDS-PAGE results, we found that a majority (89%) of the CNPP from the milk (approximately 10.13 out of 11.41% TP) were retained in the Kjeldahl NCN precipitate. Thus, CN%TP measured by Kjeldahl underestimates the amount of proteolytic damage that has been done to CN in milk. It is important for the dairy industry to correctly and rapidly measure the extent of proteolytic damage to milk protein to correctly value milk from a product quality and yield point of view. A rapid and quantitative measure of proteolytic damage to milk protein is needed.

Determination of the efficiency of removal of whey protein from sweet whey with ceramic microfiltration membranes.

Our research objective was to measure percent removal of whey protein from separated sweet whey using 0.1-µm uniform transmembrane pressure ceramic microfiltration (MF) membranes in a sequential batch 3-stage, 3× process at 50°C. Cheddar cheese whey was centrifugally separated to remove fat at 72°C and pasteurized (72°C for 15 s), cooled to 4°C, and held overnight. Separated whey (375 kg) was heated to 50°C with a plate heat exchanger and microfiltered using a pilot-scale ceramic 0.1-µm uniform transmembrane pressure MF system in bleed-and-feed mode at 50°C in a sequential batch 3-stage (2 diafiltration stages) process to produce a 3× MF retentate and MF permeate. Feed, retentate, and permeate samples were analyzed for total nitrogen, noncasein nitrogen, and nonprotein nitrogen using the Kjeldahl method. Sodium dodecyl sulfate-PAGE analysis was also performed on the whey feeds, retentates, and permeates from each stage. A flux of 54 kg/m2 per hour was achieved with 0.1-µm ceramic uniform transmembrane pressure microfiltration membranes at 50°C. About 85% of the total nitrogen in the whey feed passed though the membrane into the permeate. No passage of lactoferrin from the sweet whey feed of the MF into the MF permeate was detected. There was some passage of IgG, bovine serum albumen, glycomacropeptide, and casein proteolysis products into the permeate. ß-Lactoglobulin was in higher concentration in the retentate than the permeate, indicating that it was partially blocked from passage through the ceramic MF membrane.

Effect of uncertainty in composition and weight measures in control of cheese yield and fat loss in large cheese factories.

Our objective was to develop a computer-based cheese yield, fat recovery, and composition control performance measurement system to provide quantitative performance records for a Cheddar and mozzarella cheese factory. The system can be used to track trends in performance of starter cultures and vats, as well as systematically calculate theoretical yield. Yield equations were built into the spreadsheet to evaluate cheese yield performance and fat losses in a cheese factory. Based on observations in commercial cheese factories, sensitivity analysis was done to demonstrate the sensitivity of cheese factory performance to analytical uncertainty of data used in the evaluation. Analytical uncertainty in the accuracy of milk weight and milk and cheese composition were identified as important factors that influence the ability to manage consistency of cheese quality and profitability. It was demonstrated that an uncertainty of ±0.1% milk fat or milk protein in the vat causes a range of theoretical Cheddar cheese yield from 10.05 to 10.37% and an uncertainty of yield efficiency of ±1.5%. This equates to ±1,451 kg (3,199 lb) of cheese per day in a factory processing 907,185 kg (2 million pounds) of milk per day. The same is true for uncertainty in cheese composition, where the effect of being 0.5% low on moisture or fat is about 484 kg (1,067 lb) of missed revenue opportunity from cheese for the day. Missing the moisture target causes other targets such as fat on a dry basis and salt in moisture to be missed. Similar impacts were demonstrated for mozzarella cheese. In analytical performance evaluations of commercial cheese quality assurance laboratories, we found that analytical uncertainty was typically a bias that was as large as 0.5% on fat and moisture. The effect of having a high bias of 0.5% moisture or fat will produce a missed opportunity of 484 kg of cheese per day for each component. More accurate rapid methods for determination of moisture, fat, and salt contents of cheese in large cheese factories will improve the accuracy of yield performance evaluation and control of consistency of cheese composition and quality.