incorporation of whey proteins in cheese

International Dairy Journal 11 (2001) 495–503

Incorporation of whey proteins in cheese

J. Hinrichs*

Institute for Food Technology, Animal Foodstuff Technology, University of Hohenheim, Garbenstr. 54, D-70599 Stuttgart, Germany

Abstract

In traditional cheese-making casein forms the curd structure while whey proteins are lost in the whey. When whey proteins areintegrated into fresh, soft, semi-hard and hard cheese, this not only improves the nutrient value and yield but also causes changes tofunctional properties. There are several technologies for re-integrating whey proteins into cheese during processing. The application

and process adaptations depend mainly on the type of cheese and the desired texture. Whey proteins may be retained by applyinghigh heat treatment in order to affix the whey proteins to caseins or by using membrane technology to reduce the aqueous phase.Alternatively, whey proteins may be removed from drained whey by ultrafiltration and then added to curd after special heat

treatment or by recycling into cheese milk. r 2001 Elsevier Science Ltd. All rights reserved.

Keywords: Cheese; Whey protein; Ultrafiltration; Nanofiltration; High heating; Yield

1. Introduction

Cheese is a dairy product which has played a key rolein human nutrition for centuries. The broad range ofdifferent cheeses available is based mainly on regionalconditions and production technology, which has beenrepeatedly adapted and optimized. The main objectivehas always been and still is to convert milk, which isperishable, into a product with a longer shelf-life whilstpreserving its nutrients. In order to preserve a productwithout thermal treatment and appropriate packaging,it is necessary to lower its water activity (aw-value).Rennet coagulation of milk in combination withfermentation is an effective means of dehydrating theresulting curd that forms at the expense of losingvaluable whey proteins. New technologies have enabledthe integration of the whey proteins and different milkconstituents into the cheese matrix, in order to improveits nutrient value as well as the economic effectiveness ofcheese production (e.g. Lawrence, 1989; Pedersen, 1991;Kessler, 1996).

In traditional rennet cheese manufacturing only 6–30 kg milk constituents in form of curd is recoveredfrom 100 kg milk depending upon cheese type whilst theremainder is represented by whey. The essentialstructure of the cheese matrix is formed by the caseins,

which constitute about 80% of the milk proteins. Theremaining 20%, the whey proteins, are lost to a largeextent in the whey. In the meantime, different technol-ogies have been tried and tested, and some are now well-established to integrate whey proteins into cheese, andthus, improve the use of the raw material, milk. Wheyproteins may be incorporated in principle both in anative form and in a denatured state into cheese(Lawrence, 1989, 1993a, b, c). In their native form, theymay be retained during processing (Fig. 1) whenultrafiltration membranes are used for the ‘‘partial’’ or‘‘full’’ concentration of the milk. In a denatured formdifferent technologies are available: (1) high heattreatment of the milk in order to affix the whey proteinsto the caseins so that they are retained in the cheesematrix; (2) combining high temperature heating andmembrane technology to retain the denatured andaggregated whey of the serum phase; (3) recyclingthermally modified, particulated whey proteins viaprotein incorporation into cheese milk or (4) addingthese to the cheese matrix (Fig. 1).

Yield is of special interest in cheese manufacturing forprofitability reasons and may be calculated from

Yield ¼mcheese

mmilk� 100: ð1Þ

The yield of cheese is influenced by different factors, e.g.,it is increased (i) as the fat and protein contents of milkincrease; (ii) by retaining or re-incorporating wheyproteins, and (iii) by integrating other milk constituents

*Tel.: +49-711-4593792; fax: +49-711-4593443.

E-mail address: [email protected] (J. Hinrichs).

0958-6946/01/$ - see front matter r 2001 Elsevier Science Ltd. All rights reserved.

PII: S 0 9 5 8 - 6 9 4 6 ( 0 1 ) 0 0 0 7 1 - 1

such as lactose or ash, as well as water. In order tocalculate the incorporation of whey proteins from milkinto the cheese matrix (yield), the mass of whey proteinsin both milk and cheese should be included in Eq. (1).

2. Fresh cheese

Fresh cheese is a very popular dairy product inEurope. Fig. 2 shows the individual steps involved in the

production process and the improvements in technologyfor the incorporation of whey proteins. Using the moretraditional method (a), skim milk is HTST (hightemperature, short-time heated to 741C for 40 s) treated,fermented and whey separated by a centrifugal separa-tor. The final product is firm and it is possible to achievea high total solids content. The maximum yield ofintegrated whey proteins of milk is approx. 15%.

In an improved process (b), to obtain what is knownas thermoquarg, milk is heated to high temperatures inorder to denature whey proteins and induce self-aggregation and interaction with caseins. The amountof integrated whey proteins may be increased to between50% and 70% depending on the degree of denaturation.Appropriate heating conditions within a range of82–951C for about 360–80 s ensure that the degree ofwhey protein denaturation exceeds more than 90% inskim milk (Kessler, 1996). High preheat temperatureimproves yield, stability of the product and the texturewhich is smoother than that achieved with process (a)due to increased water binding of the denatured wheyproteins.

In order to extend whey protein yield in fresh cheese,a retentate produced by means of ultrafiltration of theacid whey (c) is added to the curd. However, if too muchretentate is added, a strange whey taste often develops in

Fig. 1. Possibilities for the incorporation of native and denatured

whey proteins into cheese.

Fig. 2. Incorporation of whey proteins into fresh cheese processing.

J. Hinrichs / International Dairy Journal 11 (2001) 495–503496

the final fresh cheese during storage, so the amount ofwhey added must be limited.

Further attempts have been made to retain the wheyproteins by introducing ultrafiltration during freshcheese processing (d) (Pedersen & Ottesen, 1992;Kessler, 1996). This technology allows the integrationof all whey proteins, but a bitter taste develops duringstorage, which is attributed to (i) the high bufferingcapacity of the retentate, (ii) increased starter growthand proteolytic activity, and (iii) the higher amount ofcalcium compared with the fresh cheeses produced bythe processes (a)–(c) (Puhan & Gallmann, 1981). Theseproblems were overcome by exchanging the order ofunit operations (B.auerle, Walenta, & Kessler, 1984) sothat fermentation takes place first followed by concen-tration (e). The prior-acidification of milk facilitates therelease of casein-bound calcium into the permeateduring concentration. High temperature heating (as inb), which is responsible for the well-known smooth andcreamy curd structure of the UF fresh cheese comparedwith thermoquarg (Fig. 3), have helped establish thistechnology on an industrial scale.

Process (f) constitutes yet a further improvement. Themilk is pre-concentrated up to a volume concentrationratio (VCR) of 2, which does not alter the fermentation(Schkoda & Kessler, 1996; Schkoda & Kessler, 1997;Schkoda, Stumpf, & Kessler, 1998). Ultrafiltration aswell as nanofiltration may be applied for the firstconcentration step. Up to 100% of the whey proteins ofthe milk are integrated depending on whether the secondstep is carried out by separator or ultrafiltration. Inaddition, the amount of lactose and calcium is increasedcompared to process (e). Despite the high amount ofcalcium, a bitter flavor does not develop, which indicatesthat the proteolytic activity of the starter culture and notthe calcium content is the major factor associated withbitter flavor in fresh cheese (Schkoda & Kessler, 1997).

Rheological properties are also influenced by theprocess method and the amount of integrated wheyproteins and other milk constituents. Fig. 4 shows thedifferences between thermoquarg, UF fresh cheese andfresh cheese produced by the new FML process(Schkoda & Kessler, 1997). A cylinder penetrates thesample twice and force is shown as a function of themeasuring time. In comparison, the curves demonstratethat the thermoquarg structure is firmer than that of theFML fresh cheese and UF fresh cheese.

In summary, modifications of process technology inorder to improve the integration of valuable wheyproteins or other milk constituents are accompanied bytextural changes. Thus, it may be necessary to adaptprocess conditions as well as the starter culture to obtaina high quality product and consumer acceptance.

3. Cheeses produced by UF technology

Ultrafiltration technology has meanwhile been estab-lished on an industrial scale in modern dairies enablingwhey proteins and other milk constituents to be retainedin the cheese. Among the examples are fresh cheese, asalready shown, fresh cream cheese, soft cheese (i.e.Camembert), Feta cheese, Pasta Filata (i.e. Mozzarella),Cheddar cheese, cheese base, cottage cheese and buttercheese (King, 1986; Hansen, 1987; Rao & Renner, 1988;Hansen, 1989)

Ultrafiltration may be integrated into the cheese-making process either for ‘‘partial’’ or ‘‘full’’ concentra-tion. In the latter application, curd cutting and wheydrainage are entirely eliminated and 100% of the wheyproteins of milk are retained in the cheese matrix.However, this technology may not be applied success-fully to produce cheeses with a high total solid contentsuch as semi-hard and hard cheese because the viscosity

Fig. 3. Comparison of the rheological properties of thermoquarg and

UF fresh cheese.

Fig. 4. Texture profile of samples of thermoquarg, UF fresh cheese

and FML fresh cheese at 51C (Schkoda & Kessler, 1996).

J. Hinrichs / International Dairy Journal 11 (2001) 495–503 497

of the product as well as permeation flux limit the degreeof concentration (Fig. 5). At >5� concentration, only10% of typical skim milk flux is attained due to the highviscosity of the retentate and fouling, which rendersultrafiltration ineffective. Typical and very successfulproducts of this technology are UF soft cheese types,e.g. Camembert and Feta, which are produced fromabout fivefold concentrated milk. However, one mustbear in mind that the changes to the composition andtechnology influence the texture of the product andadaptations are necessary to produce a high qualitycheese.

‘‘Partial’’ concentration may be applied also in themanufacturing process to produce cheeses with a higheramount of total solids, e.g. SiroCurd process (Radford,Freeman, Jameson, Leeuwen, & Sutherland, 1988) oryellow cast cheese (APV, 1992; Hansen, 1989; Dybing,1995; Guinee et al., 1995). The milk is concentrated upto a volume concentration ratio of 2 to 3, followed bythe customary unit operations of cheese manufacturingwith adaptations. As the amount of drained whey isreduced, this technology allows the integration of morewhey proteins compared to the traditional method. Thehigher the volume concentration ratio, the higher theamount of retained whey proteins and fat (Gernedel,1980). Further efforts to increase yield require thatcareful attention must be paid to the final productbecause the texture, ripening and also melting condi-tions may change. It is necessary to adapt the processingconditions depending on the degree of concentration.

When milk is ultrafiltered the fat globules are alsorecovered in addition to the caseins and whey proteins.This enables the concentration of whole milk and cream

and the standardization of the cheese milk. Fig. 6 showsthe limitations (called maximum operation lines) for thecombined concentration of protein and fat in homo-genized and non-homogenized milk. The technicallimitations identified by Grimm, Huss, and Kessler(1995) are a result of the combined effects of viscosityincrease and deposit formation, which is in agreementwith the experiments carried out by Gernedel (1980). Inparticular, the viscosity increase induced by homogeni-zation of cream results in a lower limit line forhomogenized milk and cream in contrast to the non-homogenized samples (Fig. 6). An example shall serve toexplain the diagram with the objective of being able toproduce a cream fresh cheese with 30% fat and 7%protein by means of ‘‘full’’ concentration (see Fig. 6):according to the fat and protein ratio the starting pointis located on the separator operation line at about 15%fat, which represents the composition of the milk uponleaving the centrifugal separator. Therefore, cheese milkrequires standardization to 15% fat by the separatorbefore ultrafiltration is applied. The same proceduremay be used for other cheese products.

Fig. 6 may be useful also for applied situationsinvolving ‘‘partial’’ concentration (2x23x) as alreadymentioned. On the one hand, gel strength of thecoagulum increases so that processing has to be adaptedbut whey proteins are still lost in the whey. In freshcheese technology, high temperature heating of the milk

Fig. 5. Permeation flux as a function of the degree of concentration in

a tubular UF module (Gernedel & Kessler, 1980 cit. in Kessler (1996)).

Fig. 6. Composition of UF retentate depending on the fat concentra-

tion of the milk (adopted from Grimm et al. (1995)).


may be used to increase whey protein recovery, but thequestion arises about its applicability to cheese milkintended for semi-hard or hard cheese.

Rennet-induced gelation was hindered when morethan 60% whey proteins of cheese milk were denatured(Steffl, 1999). In addition, gelation time was prolonged.Therefore, high heat treatment of milk should be ruledout as a possible application for the production of semi-hard and hard cheese production.

Schreiber (2000) studied the limitations of highheating ultrafiltration concentrates on gel formation inorder to affix whey proteins to the caseins and increasetheir retention in the cheese matrix. The inset diagram(Fig. 7) illustrates schematically the effect that blockingof the caseins by denatured whey proteins and caseincontent have on gel formation and resulting gel strength.The first row demonstrates that ultrafiltration withoutsubsequent heating and denaturation of the wheyproteins leads to an increase in casein content andconnections in the network with an increase of the F-60-value (defined as the gel strength after 60min of rennetaction). In the next row, caseins are blocked by heat-denatured whey proteins. Rennet-induced aggregation isimpeded and reduces the amount of connections in thecasein network. The more caseins that are blocked, thelower the F-60-value. But concentration of milkcounteracts the reduction in gel strength because morecaseins for network formation are available (Maubois,Mocquot, & Vassal, 1975; McMahon, Savello, Brown,& Kalab, 1991; McMahon, Yousif, & Kalab, 1993;Guinee, O’Callaghan, Pudja, & O’Brien, 1996; Smith &McMahon, 1996; Waungana, Singh, & Bennet, 1996;Schreiber, 2000).

In summary, gel strength decreases due to blockage ofcaseins and increases with milk concentration. Adiagonal representing a line of equal gel strength maybe imagined corresponding to the counter-effects of

blocking the casein and the concentration of the caseinon the network formation (Fig. 7).

The relationship between the degree of whey proteindenaturation and casein concentration which is neces-sary to reach the gel strength of pasteurized skim milk isdescribed in Fig. 7. Greater denaturation of milk ispermitted when casein content is increased. Further-more, above a protein concentration of about 8% (VCRabout 3), all whey proteins may be denatured becausethen the gel strength attained is higher than that of skimmilk (Schreiber, 2000).

This means that high heating and denaturation ofwhey proteins is possible, in order to integrate them intothe cheese, if the milk is concentrated. Pilot-scale testsfor semi-hard cheese type Edamer, resulted in anincreased yield and the possibility of heating theconcentrate in order to inactivate spore formers(Schreiber, 2000) and eliminate the addition of bacter-iocides. However, ripening as well as the functionalproperties may be influenced by high heating ofconcentrated milk and the altered composition of theresulting cheese (Berg van den & Exterkate, 1993;Guinee et al., 1995; Enright & Kelly, 1999 cit. inSchreiber, 2000; Steffl, 1999).

4. Addition of whey protein particles

The incorporation of whey proteins into the matrix ofcheese with a high total solids content may be increasedby ‘‘partial’’ concentration of milk, although retentiondoes not reach 100%. Bachmann, Schaub, and Rolle(1975) showed that added denatured whey proteins incheese milk are mechanically retained in the rennet-induced gel network. The consequences of adding wheyprotein concentrates in terms of yield and characteristicsof different cheeses are illustrated in Table 1.

Whey protein addition leads to increased yield butmay result in a slightly poorer quality cheese flavor andtexture. Improved yield is attributed to both anincreased retention of serum in the cheese matrix, andthe incorporation of whey proteins. In particular,denatured and highly hydrated whey proteins obstructsyneresis, so that less water drains off during the cheese-making process (Lucey & Gorry, 1994). By applying theusual technological countermeasures, such as intensifiedcurd working, water content may be reduced withbeneficial influences on quality-relevant parameters.

Valuable whey proteins are retained preferentially in adenatured and more aggregated (particulated) state in agel network (Fig. 8). Particulation is a technology bywhich whey proteins are denatured and aggregated byheating with simultaneous shearing into particles in awhey concentrateFthe efficiency depends on composi-tion and the process conditions, e.g. Simplesss (Singer,Yamamoto, & Latella, 1988; Fang, 1991) Dairy-LoTM

Fig. 7. Line of equal effect for rennet-induced gels depending on the

casein content and the degree of whey protein denaturation (adopted

from Schreiber (2000)).


(Asher, Mollard, Thomson, Maurice, & Caldwell, 1992)or produced by other means (Queguiner, Dumay, Salou-Cavalier, & Cheftel, 1992; Paquin, Lebeuf, Richard, &Kalab, 1993; Huss & Spiegel, 1999; Spiegel, 1999).

The effect of degree of denaturation on the retentionof whey proteins in soft and semi-hard cheese wasstudied using two whey protein concentrates (WPC 1

and WPC 2) and a whey protein isolate (WPI) whichwere heated and shear-treated in order to particulate thewhey proteins (Fig. 8). The amount of WPC or WPIadded was varied between 0.3% to 1.25% protein. Therewas a significant increase in retention as a result ofincreasing degree of denaturation for WPC and WPI.The maximum retention of 70% for semi-hard cheesewas achieved for >90% denaturation. During softcheese production a higher recovery of whey protein ispossible, especially when WPI was particulated.

Whey protein particles (WPP) are inserted inertly intothe pores of the casein network like fat globules (Steffl,1999), while added undenatured whey proteins as well asnative whey proteins of milk are lost in the whey. Thepore size of the network is indicated as approx. 10 mm(Walstra & van Vliet, 1986 cit. in Steffl, 1999), whichmeans that this is the critical diameter for addedparticles. WPP between 1 and 10 mm are integratedinertly into the structure; however, larger particlesdisturb the homogeneity of the network, and result ina reduction of the firmness (Hinrichs & Steffl, 1998).There is a further advantage of WPP: due to their sizeand structure they act similarly to integrated fat globulesdo, so that the quality of cheese, particularly with a lowfat content, is improved (Steffl, 1999).

However, adjustments in cheese processing must bemade when recycling whey proteins in the form ofparticles in order to lower the water content on the one

Table 1

Influence of the addition of whey protein concentrates on the yield and properties of various cheese types (adopted from Steffl, 1999)

Cheese type Whey concentrate Added

amount (%)

Additional

yield (%)

Flavor Texture Reference

Camembert Whey-UF

(851C, 20 s)

5, 10 Yes Bitter,

atypical

Soft, uneven

ripened

Birkkjaer (1976)

Semi-hard cheese Whey UF 5.56 12 Sour Less holes Birkkjaer (1976)

(851C, 20 s)

Gouda Centri-whey

processt (pH 5)

Not given Yes Less Crumbly Berg van den (1979)

Saint-Paulin Centri-whey

processt (UF)

0–10 11–17 Sour,

atypical

Soft, crumbly Abrahamsen (1979)

Hard cheese

(type Emmentaler)

Centri-whey

processt (UF)

0–9 14 Less Less Abrahamsen (1979)

Cheddar Whey UF

(751C, 30min)

3.5 4 Sour Soft Brown and Ernstrom

(1982)

Cheddar centri-whey processt

(pH 4.5)

5–6 12–15 Normal Minimal softer Banks and Muir (1985)

Cheddar Whey protein

concentrate

5–10 Max. 7 Atypical Sticky Baldwin et al. (1986)

Gouda Whey protein

concentrate

Not given Yes Acceptable Soft Zoon and Hols (1994)

Red. fat Gouda Simplesses 2 15 Sour Minimal softer Lucey and Gorry (1994)

Semi-hard cheese Whey protein

concentrate

(901C, 5min)

10 3 Normal Not given Santoro and Faccia (1996)

Cheddar Dairy-Lot 1 Not given Less Less Fenelon (1997)

Soft cheese

(type Camembert)

Whey protein

particle

1 Max. 30 Normal Soft Steffl (1999)

Fig. 8. Effect of the degree of denaturation of whey protein particles

on their retention in soft cheese and semi-hard cheese (adopted from

Hinrichs and Steffl (1998)).


hand and to remove milk constituents from the cheeseon the other hand, which are additionally recycled withWPP such as lactose. The basic steps in the productionprocess of soft, semi-hard and hard cheese-making areshown in Fig. 9. When adding whey protein particles,renneting properties may to be improved by raisingfermentation and renneting temperature by 2–31C, as isusually applied for high fat cheeses. Further processingof the cheese curdFwhey removal and addition ofwashing waterFcontributes to a mild cheese byreducing lactose content. Lactose is also increased whenadding whey protein particles produced from WPCwhich means that washing should be intensified.Additionally, moisture of the cheese increases. Certainmeasures may be used to reduce water content, e.g.cutting curd into smaller cubes and/or increasing theprocessing temperature after cutting to improve thesyneresis. These adaptations lead to pH-values within anormal range at the end of the ripening phase. Saltingtime may be reduced depending on moisture content(Schreiber, Neuhauser, Schindler, & Kessler, 1998a;Schreiber, Post, & Kessler, 1998b).

There are limits to the adaptations that may be madeto the cheese-making process. Therefore, the extent ofrecycling of whey protein particles is restricted depend-ing, e.g. on the moisture specification of individualcheese types and on the texture. In this manner, highquality cheeses may be produced with fortified wheyproteins. The maximum amount of added whey protein

particles and the resulting recovery of whey proteins inthe cheese matrix is determined by the dry mattercontent of the cheese (Table 2). The higher the requiredtotal solids content, the lower the amount of addedWPP and, thus, the lower the integration of wheyprotein. In order to benefit from whey protein integra-tion and satisfy high quality standards, adaptations ofeither the particulation process or cheese-making shouldbe carried out.

5. Conclusions

In traditional cheese-making, casein forms the curdstructure while the whey proteins are lost in the whey. Inorder to use the nutritionally valuable whey proteinsand increase yield several attempts have been made torecover or to re-integrate them into the cheese matrix.Industrial applications and the required adaptations tothe cheese-making process depend mainly on therequirements of cheese type and the desired texture.Whey proteins are retained by either applying hightemperatures in order to interact with the caseins orrecovering by membranes in order to reduce the aqueousphase. Alternatively, whey proteins may be restrainedand recovered from drained whey by ultrafiltration sothat they may be added after special treatment (i.e.particulation), added to the curd or recycled into thecheese milk. Meanwhile, several advanced processes,depending on the type of cheese, are being established indairies.

The advantages of incorporating whey protein intocheese are higher nutritional value, increased cheeseyield and, especially in the case of low fat cheese, sensoryimprovement. Besides, it puts whey to good use.Furthermore, when fermentation takes place in theretentate, not only the amount of retained whey proteinincreases, rennet and starter cultures are also saved. Butin order to benefit from the incorporation of the wheyprotein, cheese-making conditions require adaptationsto suit the composition of the cheese milk and therequirements of the product in order to produce a high

Fig. 9. Process modification in cheese manufacture of soft, semi-hard

and hard cheese when adding whey protein particles.

Table 2

Maximum addition of whey protein particles in the FML-process for

cheese fortified with whey protein (adopted from Schreiber et al.,

1998a; Schreiber et al., 1998b; Steffl, 1999)a

Cheese type Addition of particulated

WPCb in g protein/100 g

milk

Integrated whey

proteins of milk

in %

Soft cheese Max. 0.7 g Max. 100

Semi-hard cheese Max. 0.5 g Max. 70

Hard cheese Max. 0.3 g Max. 50

aMax., maximum amount.bDegree of whey protein denaturation of about 90%.


quality cheese. The textural changes are not negativeand may even present an opportunity to createinnovative cheese products with a creamy and softstructure and a high amount of valuable whey proteins.

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