Fresh white cheeses from buttermilk with polymerized whey protein: texture, color, gloss, cheese yield

doi:10.21203/rs.3.rs-3226862/v1

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Fresh white cheeses from buttermilk with polymerized whey protein: texture, color, gloss, cheese yield

https://doi.org/10.21203/rs.3.rs-3226862/v1

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Buttermilk and whey, despite their documented health and technological potential, are still not sufficiently utilized for the development of new products. In this research, the effect of heat treatment on whey protein’s texture, color, and gloss was analyzed using fresh white cheeses from buttermilk with polymerized whey protein. Additionally, the influence of the polymerization process on cheese yield and composition was examined. Four fresh white cheese samples were prepared: without whey protein concentrate (FWC); with whey protein concentrate (FWC/WPC); with single-heated polymerized whey protein (FWC/SPWP); and with double-heated polymerized whey protein (FWC/DPWP). The introduction of whey protein in buttermilk cheese production increased cheese yield over 2-fold. There were no differences in color and gloss between FWC/SPWP and FWC/DPWP samples.

Earth and environmental sciences/Ecology

Physical sciences/Engineering

Buttermilk cheese

Fresh white cheese

Polymerized whey protein

Texture

Color

Cheese Yield

Buttermilk and skim milk are characterized by a similar composition in terms of their content of lactose, casein, whey proteins, and minerals^1,2. However, they differ in phospholipids content. Buttermilk contains up to 10 times more phospholipids than skimmed milk¹. The source of phospholipids in buttermilk is the milk fat globule membrane (MFGM), which surrounds and stabilizes the fat globules, which is released during the churning process³. Hence, buttermilk is classified as a functional food⁴, and publications have cited the health benefits of MFGM, e.g., infection prevention, cognitive improvement and brain system development, immunity and protection metabolism⁵, reduction in cholesterol levels, and anticancer².

Due to its structure-forming, emulsifying, and water-binding properties, buttermilk is used in the production of yogurts^6–8; ice cream^9,10; as a component in mixtures for chocolate production¹¹; meat marinating¹²; and improve flavor and texture of bakery products¹³.

The cheesemaking sector is a large part of the dairy industry¹⁴, where innovative concepts for the enhancement of cheese quality are of concern, e.g., implementation of probiotic bacteria, non-fat cheese production, but also sensory properties. Sensory properties, texture, viscosity, and moisture retention of cheeses can be improved using buttermilk¹⁵ and/or the introduction of whey proteins to its composition¹⁶. During the cheesemaking process, it is common to increase the protein content to increase cheese yield^14,17. In the production of fresh and hard cheese, a high concentration of casein is recommended, while in the production of quark cheese, a high concentration of total protein and a high degree of whey protein denaturation is also desirable. The protein’s composition can be modulated using membrane processes or the introduction of protein concentrates into the processed milk¹⁸. The texture of cheeses can be affected by moisture, fat content, pH and, importantly, the structure of the protein matrix¹⁴. The structure of the protein’s matrix can be modified, e.g., conducting the polymerization process of whey proteins induced by heat treatment^19–21. The obtained whey protein aggregates are used as a thickening agent for the formulation of fermented milk²²; for food texture design²³; for microencapsulation of Lactobacillus acidophilus, which improves their survivability in yogurts²⁴; to increase the water holding capacity and reduce the syneresis of fermented milk²⁵; as a fat replacement in the development of low-fat yogurts²⁶; and in determining the behavior of water in cheese from buttermilk¹⁶.

Buttermilk and whey are considered the main by-products in the dairy industry, which, despite their documented health and technological potential, are still not sufficiently utilized for the development of new products¹³. In cheesemaking, milk is primarily employed as the basic raw material, and buttermilk is usually an additive aimed at improving rheological or sensory properties^27,28. The replacement of milk with buttermilk in cheese production can successfully lead to a new product with improved functions and nutritional value¹⁵.

The aim of our research was to directly use buttermilk as a by-product of butter production and indirectly use whey as a whey protein concentrate obtained from buttermilk, allowing for the development of an innovative dairy product: fresh white cheese from buttermilk with polymerized whey protein. The effect of heat treatment of whey proteins on the texture, color, and gloss of buttermilk cheese was analyzed for the parameters to adhere to consumer expectations. We also examined how the polymerization process impacted cheese yield and composition, which will provide the necessary knowledge for future cheesemaking.

Fresh white cheese from buttermilk preparation

Buttermilk leftovers were obtained from the industrial production of butter from cream (Great Poland, Poland). The fresh white cheese from buttermilk was prepared with the addition of whey protein concentrate powder (5.62%, w/v) and whey protein concentrate after the polymerization process (28%, w/v). Single-heated polymerized whey protein (SPWP, 28%, w/v) and double-heated polymerized whey protein (DPWP, 28%, w/v) were prepared in accordance with the methodology described by Bielska et al.²¹. The buttermilk (20 L) with whey proteins (ratio of 0.23 L of solution PWP for every 1 L of buttermilk) was mixed in double coat cheese kettle type SKM50 (Plevnik, Dobrova, Slovenia) with an automatic processor GPC 145, equipped with an automatic propeller stirrer at 15°C for 10 min, 36 rpm. Four fresh white cheese samples from buttermilk were prepared without the addition of whey protein concentrate (FWC); addition of whey protein concentrate (FWC/WPC); addition of single-heated polymerized whey protein (FWC/SPWP); and addition of double-heated polymerized whey protein (FWC/DPWP) (Fig. 1). Technical and technological conditions for the production of fresh white cheese from buttermilk were in accordance with the methodology described by Cais-Sokolińska et al.¹⁶. The cheeses were stored under refrigerated conditions at 4 ± 0.3°C and analyzed within the first 24 h after production.

Composition and cheese yield

Moisture in the prepared fresh white cheese from buttermilk was determined by standard methods AOAC 926.08 ²⁹. Fat content was determined using standard methods ISO 1735 ³⁰. The content of total nitrogen (TN), casein nitrogen (CN), non-casein nitrogen (NCN), and non-protein nitrogen (NPN) were determined according to methods described by Svanborg et al.³¹. The content of total protein (TP) and whey protein (WP) was calculated according to the following equations³²:

TP = (TN – NPN) × 6.38 (1)

WP = (NCN – NPN) × 6.38 (2)

where 6.38 represents the factor indicated for protein derived from milk

Titratable acidity was determined by standard methods AOAC 920.124 ³³. pH was measured using a CP-402 pH-meter (Elmetron, Zabrze, Poland) equipped with a IONODE IJ44A electrode (Ionode Pty. Ltd., Tennyson, Australia). The cheese yield (CY) was calculated according to Hanafy et al.¹⁴ methods with modification:

CY % = amount of cheese (kg) / amount of buttermilk (kg) × 100 (3)

Texture

Texture parameters were measured using a texturometer (Stable Micro Systems Ltd., Surrey, UK) equipped with an attachment: hardness and brittleness were measured using A/WEG attachment: pre-test speed 1.0 mm/s, test speed 2.0 mm/s, post-test speed 10.0 mm/s, and distance 10.0 mm; firmness and stickiness were measured using P/1S attachment: pre-test speed 1.5 mm/s, test speed 2.0 mm/s, post-test speed 10.0 mm/s, and distance 5 mm. Results were recorded using Texture Exponent E32 version 4.0.9.0 software (Godalming, Surrey, UK).

Color and gloss

Measurements were using an X-Rite SP-60 camera (X-Rite, Grandville, MI, USA). The whiteness index (WI), yellowness index (YI), chroma (C*), and browning index (BI) were calculated using the following equation:

WI = 100 − [(100 − L*)² + a*² + b*²]^0.5 (4)

YI = 142.86b* · L*⁻¹ (5)

C* = (a*² + b*²)^0.5 (6)

BI = [100(x − 0.31)] · 5.88 (7)

where x = (a* + 1.750 · L*) · (5.645 · L* + a* − 3.012 · b*)^-1

The calculations assumed: L = 100, a* = 0, and b* = 0.

The gloss was measured using the gloss meter (DT 268, TestAn, Gdańsk, Poland), measurement geometry 60°.

Statistical evaluation

Verification of statistical hypotheses was achieved using a level of significance of α = 0.05 by two-way analysis of variance followed by Tukey’s HSD post hoc test for multiple comparisons. Data were analyzed using Statistica data analysis software, version 13 (TIBCO Software Inc., Palo Alto, California, USA).

Composition and cheese yield

The obtained results showed no differences in moisture, fat content, titratable acidity and pH among all analyzed samples (Table 1; p > 0.05). The introduction of whey proteins to buttermilk in the production of FWC increased TN in the analyzed samples by 7.4% in FWC/WPC and by 10.6% in FWC/SPWP compared to fresh white cheese with no additive (p < 0.05). However, no TN differences were found between the samples of FWC/SPWP and FWC/DPWP) (p > 0.05). TP in the control sample was 167.4 g/kg and approx. 4% lower than in the other analyzed samples (p < 0.05). TP was the same among the samples with the addition of whey protein concentrate and whey protein concentrate after the polymerization process (p > 0.05). No differences were observed between samples in CN and NPN (p > 0.05), while NCN after the addition of whey protein (regardless of the polymerization process) increased (p < 0.05). According to Masotti et al.³⁴, during cheese production, unused whey should be recycled, therefore, the introduction of whey protein in cheese production is an innovative approach responding to consumer’s expectations of a product with a new texture and functionality. Mileriene et al.³⁵ reported that curd cheese supplemented with thermo-coagulated whey protein is a response to the global interest in sustainable dairy foods.

Table 1

Composition and cheese yield of fresh white cheeses from buttermilk with polymerized whey protein.
Parameters	FWC	FWC/WPC	FWC/SPWP	FWC/DPWP
Moisture (g/kg)	783.7 ± 1.52 ^a	784.5 ± 1.36 ^a	784.3 ± 0.95 ^a	783.6 ± 3.03 ^a
Fat (g/kg)	21.4 ± 0.20 ^a	21.2 ± 0.23 ^a	21.1 ± 0.11 ^a	21.2 ± 0.66 ^a
TN (g/kg)	28.4 ± 0.11 ^a	30.5 ± 0.06 ^b	31.4 ± 0.26 ^c	31.1 ± 0.06 ^c
CN (g/kg)	142.1 ± 0.15 ^a	142.0 ± 0.40 ^a	141.9 ± 0.66 ^a	142.3 ± 0.26 ^a
NCN (g/kg)	0.1 ± 0.06 ^a	4.6 ± 0.10 ^b	4.5 ± 0.23 ^b	4.5 ± 0.00 ^b
NPN (g/kg)	3.1 ± 0.06 ^a	3.0 ± 0.00 ^a	3.0 ± 0.06 ^a	3.0 ± 0.06 ^a
TP (g/kg)	167.4 ± 1.42 ^a	173.7 ± 0.53 ^b	174.4 ± 0.64 ^b	174.9 ± 0.30 ^b
WP (g/kg)	167.4 ± 1.42 ^a	173.7 ± 0.53 ^b	174.4 ± 0.64 ^b	174.9 ± 0.30 ^b
Titratable acidity (%)	1.6 ± 0.10 ^a	1.5 ± 0.10 ^a	1.6 ± 0.10 ^a	1.6 ± 0.10 ^a
pH	4.61 ± 0.01 ^a	4.60 ± 0.01 ^a	4.59 ± 0.03 ^a	4.58 ± 0.01 ^a
Cheese yield (%)	9.75 ± 0.05 ^a	21.97 ± 0.00 ^d	20.34 ± 0.09 ^b	21.34 ± 0.06 ^c
FWC/WPC, fresh white cheese from buttermilk with whey protein concentrate; FWC/SPWP, fresh white cheese from buttermilk with single-heated polymerized whey protein; FWC/DPWP, fresh white cheese from buttermilk with double-heated polymerized whey protein; TN, total nitrogen; CN, casein nitrogen; NCN, non-casein nitrogen; NPN, non-protein nitrogen; TP, total protein; WP, whey protein. ^a−c different small letters in superscript in the rows indicate between parameters statistically significant differences (p = 0.05).

Studies have shown that the introduction of whey proteins in buttermilk cheese production increases CY over 2-fold (Table 1; p < 0.05). CY can be increased by increasing the fat and protein content; stopping or re-adding whey proteins; and introducing other milk components (e.g., lactose; ash; moisture)¹⁴. Elbarbary and Saad³⁶ described the quality of buffalo’s milk soft cheese using the addition of camel’s whey protein concentrate, and showed that the higher the amount of added WPC, the higher CY. The authors suggested that the increase in CY may be related to the addition of WPC powder and/or denatured WP, which resulted from heat treatment. This enhanced the integration of whey protein into cheese or higher retention of serum in cheese matrix while increasing CY. As shown by Stankey et al.³⁷, denatured whey protein has a better ability to bind water, thus increasing the moisture content in cheese and CY. The same observations are made by Perreault et al.³⁸, where denatured whey protein concentrate significantly increased CY (from 12.5–15%, compared to the control) as a result of the higher cheese moisture. Giroux et al.³⁹ evaluated the cheesemaking properties of milk, where denaturation of whey proteins resulted in higher CY.

Texture

A 3.7-fold reduction in the hardness of FWC/WPC (Table 2; Fig. 2) was observed compared to FWC (p < 0.05). According to Hanafy et al.¹⁴, the introduction of 2–4% of WPC in cheese production increased the hardness, but the addition of 6% significantly reduced the hardness. The same authors suggested that the characteristics that determine cheese texture were moisture, fat, pH, and the structure of the protein matrix. Similar results were presented by Mohamed⁴⁰, where the impact of the addition of 1–3% WPC in the production of Kariesh cheese was analyzed. The results showed that hardness, cohesiveness, springiness, gumminess and chewiness values decreased after the addition of inulin or WPC. The addition of single-heated polymerized whey protein to buttermilk reduced the cheese hardness by 40% (FWC/SPWP; p < 0.05) compared to the control sample, but promoted a 2-fold increase in hardness compared to FWC/WPC (p < 0.05). Sołowiej et al.⁴¹ showed that the addition of polymerized whey protein thickened the cheese mass due to the ability to emulsify fat and increase water absorption. Casein could crosslink with whey protein in cheesemaking. This accumulates in the cheese possessing a higher hardness⁴². However, in our research, further heating of the whey protein and the addition of a double-heated polymerized whey protein significantly decreased the hardness. The addition to buttermilk whey protein (regardless of type) decreased firmness (compared to FWC; p < 0.05). Hence, in accordance with the results presented by Cais-Sokolińska et al.¹⁶, the addition of whey protein concentrates in powder and after the polymerization process to buttermilk in the production of buttermilk cheese caused a significant decrease in firmness (despite the use of other firmness measurement conditions). There were no differences observed in brittleness in all cheese samples (p > 0.05), and the stickiness increased 3-fold only for FWC/WPC compared to the control sample. The obtained results suggested that the texture of fresh white cheese from buttermilk could be modified via the modification of the composition of the main raw material with simultaneous thermal modification of whey protein concentrates. This was previously confirmed by Domingos et al.⁴³, where the texture problem caused by fat reduction could be improved through the addition of WPC. Tashakori et al.⁴⁴ found that the addition of 20% WPC improved the overall texture of spread cheese. However, Mohamed⁴⁰ reported that the ability of whey proteins to form gels that could hold water, lipids, and provide the appropriate texture, was important for consumer acceptability. After 10 days of cold storage, significant changes were found in the hardness, firmness, and stickiness parameters, mainly for FWC/SPWP (Table 2; Fig. 1). Hardness and firmness increased 2-fold, and stickiness showed a 2-fold decrease (FWC/SPWP; p < 0.05). Refrigerated storage had no effect on the brittleness of all samples analyzed (p > 0.05). According to Gamlath et al.⁴⁵, denatured aggregates of whey proteins could avoid impairment of rennet gelation caused by native whey protein, and the combination of heat treatment with ultrasonication of whey proteins could potentially improve cheese texture.

Table 2

Texture of fresh white cheeses from buttermilk with polymerized whey protein.
Parameters	Storage (d)	FWC	FWC/WPC	FWC/SPWP	FWC/DPWP
Hardness (g)	0	254.71 ± 14.47 ^cB	69.48 ± 7.57 ^aA	152.55 ± 21.86 ^bA	84.86 ± 6.71 ^aA
Hardness (g)	10	178.74 ± 24.89 ^bA	62.69 ± 4.66 ^aA	360.64 ± 24.25 ^cB	156.69 ± 14.81 ^bB
Brittleness (mm)	0	9.46 ± 0.63 ^aA	9.79 ± 0.17 ^aA	9.68 ± 0.31 ^aA	7.59 ± 1.70 ^aA
Brittleness (mm)	10	8.28 ± 1.19 ^aA	9.88 ± 0.10 ^aA	9.99 ± 0.00 ^aA	9.02 ± 1.35 ^aA
Firmness (g)	0	1027.55 ± 72.68 ^cB	222.14 ± 23.80 ^aB	378.60 ± 8.23 ^bA	335.64 ± 12.57 ^bA
Firmness (g)	10	295.69 ± 65.28 ^bA	128.95 ± 43.14 ^aA	756.10 ± 66.06 ^cB	327.36 ± 43.47 ^bA
Stickiness (g)	0	7.12 ± 2.00 ^aA	22.05 ± 6.25 ^bB	14.52 ± 2.58 ^aB	9.60 ± 0.98 ^aA
Stickiness (g)	10	6.31 ± 0.58 ^aA	8.52 ± 2.52 ^aA	7.67 ± 0.97 ^aA	9.55 ± 1.16 ^aA
d, day; FWC/WPC, fresh white cheese from buttermilk with whey protein concentrate; FWC/SPWP, fresh white cheese from buttermilk with single-heated polymerized whey protein; FWC/DPWP, fresh white cheese from buttermilk with double-heated polymerized whey protein. ^{a−c, A−B} different small letters in superscript in the rows and different capital letters in columns indicate between parameters statistically significant differences (p = 0.05).

Color

No differences in color were found between samples of FWC/SPWP and FWC/DPWP (Table 3; p > 0.05). The least lightness was cheese with WPC, which was approx. 6% lower compared to the control sample (FWC; p < 0.05) and 5% lower compared to samples with polymerized whey protein (p < 0.05). The cheese sample with WPC was also characterized by the smallest whiteness index. This was consistent with the results reported by Cais-Sokolińska et al.¹⁶, where buttermilk cheese with added WPC powder was darker than the samples of cheese with added polymerized whey proteins, and the calculated value of the total color difference (ΔE) was above 3.3. Such a result may negatively impact consumer ratings of overall desirability. Brodziak et al.⁴⁶ also showed that the introduction of whey protein powder to yogurt caused the final product to be further from the ideal whiteness standard and thus less light.

Table 3

Color of fresh white cheeses from buttermilk with polymerized whey protein.
Parameters	Storage (d)	FWC	FWC/WPC	FWC/SPWP	FWC/DPWP
L	0	94.91 ± 0.05 ^cA	89.31 ± 0.20 ^aB	94.10 ± 0.15 ^bA	94.35 ± 0.21 ^bA
L	10	95.08 ± 0.38 ^bA	88.49 ± 0.40 ^aA	94.70 ± 0.12 ^bB	94.78 ± 0.21 ^bA
WI	0	89.28 ± 0.10 ^cA	84.19 ± 0.10 ^aA	87.86 ± 0.01 ^bA	87.93 ± 0.16 ^bA
WI	10	89.64 ± 0.33 ^bA	84.52 ± 0.31 ^aA	89.04 ± 0.11 ^bB	88.97 ± 0.30 ^bB
YI	0	14.12 ± 0.14 ^aA	18.58 ± 0.07 ^cB	16.05 ± 0.08 ^bB	16.07 ± 0.15 ^bB
YI	10	13.62 ± 0.31 ^aA	16.64 ± 0.15 ^cA	14.44 ± 0.12 ^bA	14.63 ± 0.38 ^bA
C	0	9.44 ± 0.09 ^aB	11.65 ± 0.06 ^cB	10.61 ± 0.07 ^bB	10.66 ± 0.08 ^bB
C	10	9.11 ± 0.17 ^aA	10.35 ± 0.07 ^cA	9.60 ± 0.07 ^bA	9.72 ± 0.23 ^bA
BI	0	9.34 ± 0.09 ^aA	12.82 ± 0.05 ^cB	10.92 ± 0.06 ^bB	10.85 ± 0.08 ^bB
BI	10	9.04 ± 0.22 ^aA	11.31 ± 0.12 ^cA	9.89 ± 0.05 ^bA	10.12 ± 0.27 ^bA
GU	0	2.2 ± 0.10 ^aA	2.8 ± 0.06 ^bA	2.9 ± 0.15 ^bB	2.6 ± 0.10 ^bA
GU	10	2.3 ± 0.06 ^aA	3.1 ± 0.06 ^bB	2.2 ± 0.26 ^aA	3.4 ± 0.00 ^bB
d, day; L, lightness; WI, whiteness index; YI, yellowness index; C, chroma; BI, browning index; FWC, fresh white cheese from buttermilk; FWC/WPC, fresh white cheese from buttermilk with whey protein concentrate; FWC/SPWP, fresh white cheese from buttermilk with single-heated polymerized whey protein; FWC/DPWP, fresh white cheese from buttermilk with double-heated polymerized whey protein; GU, gloss. ^{a−c, A−B} different small letters in superscript in the rows and different capital letters in columns indicate between parameters statistically significant differences (p = 0.05).

The addition of whey proteins after the polymerization process in the production of fresh white cheese from buttermilk resulted in increased whiteness index and reduced yellowness index by 4.4% and 13.6%, respectively, compared to FWC/WPC. Cheese with WPC also had the highest browning index and chroma (Table 3; p < 0.05). The addition of whey proteins increased the gloss of the analyzed cheese samples compared to the control sample (p < 0.05). The gloss between the cheeses with whey protein concentrate and polymerized whey protein was the same (p > 0.05). After 10 days of cold storage, the lightness decreased in FWC/WPC and increased in FWC/SPWP, but the differences were less than 1% (p < 0.05). In FWC/SPWP and FWC/DPWP cheeses, the whiteness index after 10 days of storage increased slightly above 1% (p < 0.05). In all samples, after 10 days of refrigerated storage, the chroma parameter decreased (p < 0.05). Color is the first sense that registers with the consumer and uses to accept or reject the product^47,48. Whey protein may undergo slight color changes when heated⁴⁹. According to Ortega et al.⁵⁰ color of whey proteins after heat treatment correlated with antioxidant activity due to the Maillard reaction.

In traditional cheesemaking, whey proteins are lost in the whey. Therefore, it is considered reasonable to increase the technological quality of buttermilk cheese through the introduction of whey protein concentrates to buttermilk. The use of buttermilk after the production of butter and whey proteins forms cheese whey, which is part of the global trend of sustainable food production and closed-loop food production. The obtained results can significantly influence the development of dairy products and improve technologist in dairy industry, how to use whey protein aggregates to modify the texture, color and improve the yield of fresh white cheese.

Funding

This study was funded by Poznań University of Life Sciences (Poland) as the research program “First grant”, no. 11/2022 and by the Ministry of Education and Science (Poznań, Poland), MEN-UPP 506.784.03.00/KMIP.

Author contributions

PB and DCSconceptualization, methodology, formal analysis and writing-original draft preparation, DCS investigation, PB visualization and supervision. All authors read and approved the final manuscript.

Declaration of competing interest

The authors declare no conflict of interest.

Data availability

The datasets used during the current study available from the corresponding author on reasonable request.

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No competing interests reported.

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Fresh white cheeses from buttermilk with polymerized whey protein: texture, color, gloss, cheese yield

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Abstract

Figures

Introduction

Materials and Methods

Fresh white cheese from buttermilk preparation

Composition and cheese yield

TP = (TN – NPN) × 6.38 (1)

WP = (NCN – NPN) × 6.38 (2)

Texture

Color and gloss

Statistical evaluation

Results and Discussion

Composition and cheese yield

Texture

Color

Conclusions

Declarations

References

Additional Declarations

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