The rising demand for sustainable foods has accelerated interest in plant-based cheese; however, replacing casein with plant proteins often compromises texture, meltability, and overall quality. This study investigated lentil-gluten protein systems in dairy-free cheese formulations and evaluated the effects of protein, modified tapioca starch, chitosan, maltodextrin, and oil using response surface methodology (RSM). A central composite design with 50 formulations was used to model effects on moisture, protein, fat, ash, color, meltability, and texture properties. Functional characterization showed that modified tapioca starch exhibited the highest swelling capacity (9.21 mL/g) and water retention capacity (WRC) (10.03 g/g), while lentil protein showed the greatest water solubility index (WSI) (9.47%), and oil-holding capacity (OHC) (0.61 g/g). Linear models accurately described trends in moisture, ash, fat, and protein contents (R2 = 0.95-0.99). Protein (p < 0.01), chitosan (p < 0.05), and starch (p < 0.05) significantly increased hardness, cohesiveness, and springiness, whereas oil enhanced meltability (p < 0.01). Higher protein levels decreased lightness (L*) but increased a* and b* values, reflecting natural pigments and Maillard reaction products. The optimized formulation-28.31% protein powder, 20% oil, 1.5% chitosan, 9.99% starch, and 2.03% maltodextrin-achieved high overall desirability and closely matched predicted values, confirming model validity. Overall, this study provides the first integrated quantitative assessment of lentil-gluten systems in plant-based cheese and identifies ingredient interactions that govern structure and melting behavior. The findings establish a data-driven formulation framework capable of producing high-protein, texturally stable dairy-free cheese with improved functional and sensory potential.

The rennet and acid coagulation properties of reconstituted micellar casein concentrate prepared using cold or warm microfiltration (MF), at similar casein contents, were investigated, with low-heat skim milk powder (LHSMP) as a control. The MF retentates had higher casein content (as % of total protein) compared with LHSMP, and heat-induced whey protein-casein aggregates were only present in LHSMP. All MF retentates showed shorter rennet coagulation times and higher gel strengths than LHSMP, which may be linked to lower levels of whey protein (either native or denatured). At similar casein contents, longer rennet coagulation times were evident for cold MF retentates compared with warm MF retentates, as the ratio of κ-CN as a function of total increased with the depletion of β-CN. In terms of acid-induced coagulation, all MF retentates coagulated at a pH >5, higher than the gelation pH of LHSMP (4.7-4.9), which was confirmed by microscopic and textural analysis. An inflection point (increase, followed by a decrease) in the storage modulus value was seen during the acidification of warm MF retentates, but not cold MF retentates; this may be related to structural rearrangements of the gel initiated by release of colloidal calcium phosphate and compacting of the structure of warm MF retentate gels as pH decreased. Both warm and cold MF retentates exhibited shorter rennet coagulation times, stronger rennet-induced gels and higher acid-induced gelation pH compared with LHSMP, which might influence their use for the manufacture of cheese or yogurt with tailored functionalities.

Recently, with the efforts of food specialists, protein sources in the diet is rapidly changing. Consequently, novel products based on plant proteins or lab-grown proteins are increasingly being introduced to consumers. However, the food industry is facing new challenges in creating natural, innovative, accessible, and nutritious dairy-free alternatives. Designing dairy-free cheese alternatives with beneficial nutritional profiles, bioavailability, and digestibility requires a thorough understanding of the molecular characteristics of plant-derived ingredients, their structural assembly, and their role in physicochemical and sensory properties of the final product. In other words, with a deep knowledge of the nature of plant-based ingredients and the intelligent control of intermolecular interactions, these ingredients can be used to produce dairy-free cheese alternatives with the same characteristics as dairy products. Therefore, this review aims to synthesize current knowledge on production techniques and key factors influencing the design of dairy-free cheese alternatives. By examining the interactions between carbohydrates, proteins, and other ingredients, as well as their ability to replicate the functional and textural properties of traditional dairy cheeses, this review provides valuable insights for the development of high-quality dairy-free cheese alternatives. In the design of these alternatives, fractionation and tissue disruption routes are employed. The difference in the number and distribution of their surface groups has a significant effect on ability of plant polysaccharides and proteins to interact with other molecules. Gel strength decreases with decreasing plant biopolymer concentration, bond strength, and cross-linking density. While heat treatment can unfold proteins and minimize their net charge, the hydrophobic attraction becomes the primary driver of protein-protein interactions in plant-based cheeses. In contrast, the 3D-structure of dairy cheeses is stabilized through a combination of hydrophobic interactions, hydrogen bonds, and calcium bridges. (c) 2026 The authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

In response to population growth, ethical considerations, and the environmental impacts of animal proteins, researchers are intensifying efforts to find alternative protein sources that replicate the functionality and nutritional profile of animal proteins. In this regard, plant-based cheese alternatives are becoming increasingly common in the marketplace, as one of the emerging dairy-free products. However, the dairy industry faces challenges in developing dairy-free products alternatives that meet the demands of customers with specific lifestyles or diets, ensure sustainability, and retain traditional customers. These challenges include food neophobia, the need to mimic the physicochemical, sensory, functional, and nutritional properties of dairy products, the inefficient conversion factor of plant-based proteins into animal proteins, and high production expenses. Given the distinct nature of plant-based milks, understanding their differences from cow's milk is crucial for formulating alternatives with comparable properties. Designing dairy-free cheese analogs requires overcoming electrostatic repulsion energy barriers among plant proteins to induce gelation and curd formation. Innovative approaches have substantially enhanced the physicochemical and sensory properties of these alternatives. Researchers are exploring the application of microalgae as a plant protein source and investigating new microbial fermentation methods to increase protein content in dairy-free products.

Proteolytic activities and specificities of three types of fermentation-produced chymosin were investigated in cheese and in sodium caseinate (NaCN) digests made using these coagulants. The highest level of enzyme required to hydrolyse all intact aS1-CN in NaCN solution at pH 5.2 over 24 h was for a modified camel chymosin (mCC; 13.34 IMCU mL-1), followed by camel chymosin (4 IMCU mL-1) and bovine chymosin (0.4 IMCU mL-1). Many peptides were identified using liquid chromatography-mass spectrometry from both Cheddar cheese and NaCN digests produced using each chymosin. Besides previously reported casein-derived peptides produced by bovine and camel chymosins and corresponding cleavage sites, several new cleavage sites were identified. The proteolytic specificity of mCC on casein was determined. Most of the peptides observed in Cheddar cheese samples were also identified in NaCN digests. The overall proteolytic activity of mCC was the lowest, which may have implications for ripening and functionality of cheese.(c) 2023 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

This study evaluated protein transmission, permeate flux and energy consumption during MF of skim milk at 7 [[EQUATION]] C using two discrete 800 kDa polymeric MF membranes. Filtration trials determined optimum process parameters (i.e., transmembrane pressure and volume concentration factor) and membrane configuration (in-series or in-parallel) to maximise serum protein permeation. This study demonstrated that a combination of higher VCF (3) and lower TMP (75 kPa), with an in-parallel membrane configuration resulted in the most efficient rate of permeation of serum protein per kg of permeate produced. However, from an energy perspective, an in-parallel configuration with a TMP of 75 kPa and a lower VCF2 was the most efficient process, consuming between 1.28 – 1.57 kW h kg -1 of crude protein permeated. Additionally, the permeation of serum β-casein at low temperature was governed by the uniformity of the pore size distribution in discrete MF membranes with the same nominal molecular weight cut-off.

The effect of I3 -casein reduction (LB) and high heat treatment (HHT, 120 degrees C x 15 s) of whey protein reduced milk on the ripening properties of Emmental cheese was investigated. Cheeses were manufactured from low-heat skim milk powder (LH), micellar casein concentrate powder (MCC) and I3 -casein reduced MCC powder (LB-or HHT LB) respectively, then ripened for 120 days. There was no significant difference between the ripening properties of LB and MCC cheeses. Compared with the other treatments, HHT LB cheese had significantly lower plasmin activity (day 120 of ripening) and flowability (days 70 and 120 of ripening), higher level of a-value (redness), and similar numbers of peptides and levels of casein hydrolysis (days 1 and 120 of ripening). Overall, reduction of I3 -casein by 4.25% in MCC did not influence the maturation properties of Emmental cheese; however, high heat treatment of whey protein-reduced milk impaired cheese functionality and appearance.(c) 2022 Elsevier Ltd. All rights reserved.

The principles of the manufacture and ripening of rennet- and acid-coagulated cheese are discussed in this chapter. Rennet or acid coagulation is essential for the manufacture of cheese, and various methods can be applied to study the coagulation properties of milk. In cheese manufacture, many processes are used to concentrate the casein and fat content of milk in the form of cheese. Membrane filtration processes are a relatively novel technology that alters the cheesemaking properties and composition of milk; cheese manufacture with membrane filtration processes has the potential to allow manufacture of cheese with tailored functionalities.

Membrane performance during microfiltration (MF) of skim milk carried at 7 degrees C (cold MF) and 50 degrees C (hot MF) using 800 kDa spiral wound polymeric membranes was assessed. Permeate flux, protein and mineral partitioning, fouling resistance (R-f) and energy consumption as a function of temperature are reported. MF performance at 50 degrees C resulted in significantly (p < 0.05) higher flux than at 7 degrees C, with the energy requirements (in kW h kg(-1) protein removed) significantly (p < 0.05) lower than cold MF. Overall R-f was significantly (p < 0.05) higher for the cold MF process, with reversible fouling accounting for 97 and 71% of the Rf in the cold and hot MF processes, respectively. On a dry matter basis, cumulative serum protein permeation was similar at both temperatures at 60.4 and 62.1% for the cold and hot MF processes, respectively, with higher beta-casein and calcium permeation observed for the cold MF process. (c) 2023 Published by Elsevier Ltd.

Removal of 40–95% of native serum proteins from skimmed milk may be achieved by microfiltration (MF) to generate micellar casein concentrate (MCC). The MCC may be used to prepare cheese milk of the target composition and resultant cheeses of the desired quality and functionality. Herein, factors affecting the composition of MCC, different methods to preserve MCC as well as the application of MF in tailoring the composition and quality of cheese milk and resultant cheeses are reviewed.

Efficient separation of serum from colloidal proteins in bovine milk in their native form can be achieved by microfiltration (MF). This study assessed partitioning of serum proteins (SP) in skim milk by a MF process using 0.1 mu m graded permeability ceramic membranes with two diafiltration (DF) steps, from a mass balance and energy utilisation perspective. A mass balance focused on dry matter and true protein yielded recoveries of 99.5 and 95.3%, respectively. However, an accurate mass balance relative to colloidal and SP contents in the retentate and permeate streams was not achieved, linked to errors surrounding quantification of nitrogen fractions in processed streams using standard methodologies designed for raw milk. Additionally the energy required for each MF/DF step was 13.1, 13.7 and 20.6 kW h kg(-1) of SP removed, respectively, demonstrating the dynamic relationship between SP partition relative to diafiltrant utilisation and its impact on energy consumption. (C) 2022 Elsevier Ltd. All rights reserved.

Micellar casein concentrate (MCC) of high casein content (93.64% of total protein), was produced by microfiltration of pasteurised skim milk. Cheesemilk of typical (1 x ) or high (1.5 x ) casein content were formulated from MCC which received either: no further heat treatment; pasteurisation (72 degrees C x 15 s) or high heat treatment (90 degrees C x 15 s), prior to combination with other membrane streams and cream. Cheddar cheeses were manufactured in triplicate and ripened for 180 days. Cheeses made from pasteurised or high heat treated MCC had similar pH as well as comparable flowability, hardness and volatile profiles over 180 days of ripening. On increasing cheesemilk casein content, levels of primary proteolysis in resultant cheeses decreased, and pH and hardness levels increased. Overall, increasing the heat treatment temperature from 72 to 90 degrees C for extension of the microbial shelf life of MCC does not impair the texture, functionality and volatile profile of resultant Cheddar cheeses. (C) 2021 Elsevier Ltd. All rights reserved.

Cheddar-type cheese was manufactured using fermentation-produced bovine chymosin (BC), fermentation-produced camel chymosin (CC) and a modified fermentation-produced camel chymosin (mCC) and ripened for 180 days. Only minor differences were found in cheese composition and pH between the cheeses made with any of the chymosins studied. Proteolysis in cheese made with mCC was reduced compared with cheese made with BC or CC. Significantly higher instrumental and sensory hardness and significantly lower meltability were found in cheeses made using CC or mCC compared with cheese BC after 180 days of ripening. Descriptive sensory analysis results showed that cheese made with CC or mCC had less sulphur and barny flavour; the brothy flavour and bitter taste of cheese made with mCC were also lowest. In conclusion, the modified camel chymosin appears to be suitable for the manufacture of Cheddar cheese with modified functionalities.

To study the effect of removing beta-casein of cheesemilk on cheese manufacture, cheesemilk was formulated from low heat skim milk powder (LH CM), micellar casein concentrate with 1.83% beta-casein reduction (MCC CM) and beta-casein-reduced micellar casein concentrate with 4.25% beta-casein reduction (LB CM) or high heat treatment (HHT; 120 degrees C, 15 s) LB CM to manufacture Emmental cheeses. MCC- and LB CM had similar rennet coagulation properties and similar compositions and yield in the resultant cheeses. Compared with LB CM, HHT LB CM had an increased level of denatured whey protein and a reduced, yet acceptable, curd firming rate. Higher fat losses, but lower protein losses, from cheesemilk to whey in HHT LB cheeses resulted in a similar yield compared with LB CM cheese. Overall, it was possible to remove b-casein from cheesemilk or apply high heat treatment to whey protein reduced milk without adversely affecting cheese composition and yield. (C) 2021 Published by Elsevier Ltd.

Microfiltration at 0.10 mu m removed 70.29% of serum proteins from milk and the resultant micellar casein concentrates (MCC) were subjected to no heat treatment (control), pasteurisation (72 degrees C x 15s) and high heat treatment (HHT; 90 degrees C x 15s) before formulation of cheese milk for Cheddar cheese manufacture. MCC showed good heat stability due to low serum protein content. For cheese milk of typical casein content, both pasteurisation and HHT did not significantly influence pH, calcium distribution and rennet coagulability, or subsequent cheese composition and yield; although HHT elongated cheese make time significantly. On increasing casein content from 3.09% to 4.31%, there was no significant difference for rennet to cut time between cheeses made from milk with different thermal histories and casein contents. Overall, HHT of MCC had no significant impact on cheese make properties, cheese composition and yield of Cheddar cheese. (C) 2020 Elsevier Ltd. All rights reserved.

The pH of cheese is determined by the amount of lactose fermented and the buffering capacity of the cheese. The buffering capacity of cheese is largely determined by the protein contents of milk and cheese and the amount of insoluble calcium phosphate in the curd, which is related to the rate of acidification. The objective of this study was to standardize both the lactose and casein contents of milk to better control final pH and prevent the development of excessive acidity in Cheddar cheese. This approach involved the use of low-concentration factor ultrafiltration of milk to increase the casein content (∼5%), followed by the addition of water, ultrafiltration permeate, or both to the retentate to adjust the lactose content. We evaluated milks with 4 different lactose-to-casein ratios (L:CN): 1.8 (control milk), 1.4, 1.1, and 0.9. All cheesemilks had similar total casein (2.3%) and fat (3.4%) contents. These milks were used to make milled-curd Cheddar cheese, and we evaluated cheese composition, texture, functionality, and sensory properties over 9 mo of ripening. Cheeses made from milks with varying levels of L:CN had similar moisture, protein, fat, and salt contents, due to slight modifications during manufacture (i.e., cutting the gel at a smaller size than control) as well as control of acid development at critical steps (i.e., cutting the gel, whey drainage, salting). As expected, decreasing the L:CN led to cheeses with lower lactic acid, residual lactose, and insoluble Ca contents, as well as a substantial pH increase during cheese ripening in cheeses. The L:CN ratio had no significant effect on the levels of primary and secondary proteolysis. Texture profile analysis showed no significant differences in hardness values during ripening. Maximum loss tangent, an index of cheese meltability, was lower until 45 d for the L:CN 1.4 and 0.9 treatments, but after 45 d, all reduced L:CN cheeses had higher maximum loss tangent values than the control cheese (L:CN 1.8). Sensory analyses showed that cheeses made from milks with reduced L:CN contents had lower acidity, sourness, sulfury notes, and chewdown cohesiveness. Standardization of milk to a specific L:CN ratio, while maintaining a constant casein level in the milk, would allow Cheddar cheese manufacturers to have tighter control of pH and acidity.

Pink discoloration defects in cheese manifests as the appearance of pink patches within cheese blocks and has recently been associated with Thermus thermophilus . Swiss‐type cheeses were prepared at pilot scale using thermophilic starter cultures, Propionibacterium freudenreichii , and one of Thermus thermophilus HB27, Thermus scotoductus SE1 or Thermus thermophilus DPC6866 or a control without Thermus . Significantly, a higher level of redness was observed in cheeses with Thermus thermophilus HB27 relative to the other cheeses and suggests that the development of the pink defect is dependent on the strain of Thermus present and on other, as yet unknown, factors that require further study.

Control of acidity is critical for cheese quality, as high acidity can be associated with poor flavor and textural attributes. We investigated an alternative method to control cheese acidity, specifically in low-fat (LF) and reduced-fat (RF) milled curd, direct-salted Gouda cheese, which involved altering the initial lactose content of cheesemilk. In traditional Gouda cheese manufacture, a critical technique to control acidity is whey dilution (WD); that is, partial removal of whey and its replacement with water. Direct standardization of the lactose content of milk during the ultrafiltration process could be a simpler and more effective technique to control cheese acidity. This study compared the effect of traditional WD at 2 different levels, 15 and 30% (WD15 and WD30), with the alternative approach of adjustment of the lactose content of milk using low-concentration-factor ultrafiltration (LCF-UF). The composition, texture, functionality, and sensory properties of these LF and RF Gouda cheeses were evaluated. A milled curd, direct-salted cheese manufacturing protocol was used. Milks used for cheesemaking had a lactose-to-casein (L:CN) ratio of approximately 1.8, which is the typical ratio found in milk, whereas milks prepared with lactose standardization (LS) were made from UF concentrated milks with water added during filtration to achieve a L:CN ratio of approximately 1.1. Cheeses made with LS exhibited lower lactose and lactic acid contents than WD30 and WD15, leading to significantly higher pH values in the cheese. Dynamic small-amplitude oscillatory rheology indicated that use of LS led to cheeses with a lower crossover temperature (melting point) than the cheeses made with WD. Cheeses made with LS had lower insoluble Ca contents, likely caused by the addition of water required to achieve the lower L:CN ratio in these milks. Sensory analysis also indicated that LS cheeses had lower acidity and softer texture. These results suggest that standardization of the L:CN ratio of milk could be a useful alternative to WD (or a curd rinse step) to reduce acidity in cheeses. In addition, LS could be used to help soften texture and increase meltability, if desired in lower-fat cheese types.

The production of Cheddar cheese using micellar casein concentrate (MCC), a novel milk ingredient powder with a high casein content (∼92%), was evaluated. Four types of Cheddar cheese were manufactured and ripened for 180 days from the following starting materials: standardised control milk (control), skim milk with cream (SC), reconstituted MCC with cream (MC) and reconstituted low-heat skim milk powder with cream (PC). Only minor differences were found in composition between treatments, but MC cheese showed higher levels of proteolysis compared with other treatments, linked to significantly higher plasmin and chymosin activities. No differences were observed in hardness between treatments (60, 120 and 180 days), but the springiness and cohesiveness of MC and PC cheeses were significantly higher than that of the control and SC cheeses at 60, 120 and 180 days. The use of casein-dominant dairy streams thus has the potential for production of Cheddar cheese with tailored functionality.