Abstract
Acid whey, a byproduct of dairy processing, particularly from Greek yogurt and cottage cheese production, presents significant environmental and economic challenges due to its high organic load and disposal restrictions. However, the unique composition of acid whey, containing proteins, lactose, minerals, and bioactive compounds, presents promising opportunities for the development of functional, value-added products. This review explores innovative approaches to acid whey valorization, emphasizing biotechnological methods, fermentation techniques, and advanced membrane filtration processes. Comparative analysis of sweet and acid whey compositions underscores specific challenges and advantages in acid whey utilization, various valorization strategies, such as membrane filtration and ultrafiltration, osmosis, enzymatic, and microbial processing, highlighting their effectiveness in ingredient recovery and product development. It also identifies suitable microorganisms capable of efficiently metabolizing acid whey and enhancing the nutritional and functional profiles of derived products. Special attention is given to fermented beverages and other functional products developed from acid whey, incorporating novel strategies to optimize fermentation processes. Moreover, this review details recent advancements in probiotic microencapsulation technologies, demonstrating their effectiveness in maintaining and enhancing probiotic viability in acid whey-based functional beverages. The success of these functional products significantly depends on selecting appropriate probiotic strains, encapsulation materials, and innovative microencapsulation methods. Finally, the article addresses current limitations and outlines future research perspectives, highlighting the potential applications of acid whey-derived products in food, beverages, animal feed, and bioenergy sectors.
1 Acid whey utilization challenges
Whey is a residue after cheese production and rich in carbon and nitrogen, that contains high concentration of lactose, proteins and minerals (). Depending on the type of milk coagulation, there are 2 types of whey: rennet whey, which remains after enzymatic milk coagulation, and acid whey, a byproduct of milk coagulation by acidification ().
According to Solieri et al. (
), there are 2 main ways to use whey:
Filtration to obtain useful whey components such as whey permeate, whey protein isolate, whey protein concentrate (WPC), whey powder, or lactose.
Biotechnological treatment uses enzymes and microorganisms to convert lactose and whey proteins into useful biochemical.
Also, in addition to biotechnological methods for processing acid and sweet whey due to the high lactose content; chemical methods are also available. For example, Brönsted acid catalysts (hydrochloric acid, nitric acid, sulfuric acid, and phosphoric acid) were used in the acid catalyst experiment ().
The biotechnological approach to utilize sweet and acid whey is preferable due to cost-effectiveness, and research on whey processing using microorganisms remains easier and more beneficial.
But while sweet whey has commercial uses in products such as nutritional supplements, a major proportion of acid whey from the dairy industry is discarded as effluent, constituting a form of pollution (). Kazakhstan also has many small dairy industries where acid whey is a co-product after production; however, because of its liquid consistency, it is less suitable for processing, making its utilization more challenging compared to rennet whey.
In the book of Rocha-Mendoza et al. () acid whey is described as a waste produced during the production of cottage cheese, and authors Rocha-Mendoza et al. () described acid whey after Greek yogurt, and in both productions it has less proteins, contains high salt and acid concentration in comparison with sweet whey, and this cause the problem with further utilization of acidic whey.
There are many types of research devoted to the utilization of acid whey as shown in review of Rocha-Mendoza et al. (), that concentrated on the industrial trends, applications and health benefits of acid whey products.
For utilization of acid whey usually are used biotechnological way with the cultivation of various cultures of microorganisms and their combinations. For example authors described commercial microorganisms combinations such as Streptococcus thermophilus, Bifidobacterium spp., Lactobacillus acidophilus, Bifidobacterium lactis, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus, and Lactococcus lactis (commercial); 2 combination of microorganisms of home using Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus, and Bifidobacterium spp., Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus paracasei, and Streptococcus thermophilus. At the same time, acid whey, regardless of which combinations of microorganism cultures were used, has approximately the same content of fats, proteins, lactose and medium pH, which indicates metabolic products obtained during fermentation (). Acid whey can be converted into functional products, but due to its too-low pH and lactose content, products based on it are mostly liquid. In addition, acid whey’s acidic pH optimum and severe product inhibition limits its application for lactose hydrolysis in milk ().
Therefore, despite of rich content, the acid whey is a by-product that remains largely underutilized and represents an environmental problem by contributing to water and soil pollution (, ). Although numerous works discuss acid whey utilization, its processing remains a significant challenge.
What are the problems with acid whey utilization? Firstly, manufacturers consider acid whey as waste and send it to wastewater, which becomes too expensive to treat due to its high Biochemical Oxygen Demand (BOD) ().
Secondly, acid whey is high in lactose and low in protein.
Thirdly: Due to the high content of lactic acid, it is difficult to dry acid whey or extract lactose from it (, ). But it should be noted that despite the difficulties in processing acid whey, there are advantages in its use in comparison with sweet whey. Although acid whey contains a lower protein content than sweet whey, the presence of approximately double the content of calcium as compared to sweet whey increases the value of acid whey as a source of nutraceuticals ().
Therefore, there is a trend in utilizing acid whey to produce the useful products and high-value components.
2 Valorization of acid whey: challenges and approaches
Acid whey, which is the byproduct of acid coagulation in cheese production, has traditionally been considered a waste product with limited value. However, recent research has shown that acid whey, a rich source of nutrients, including carbohydrates, proteins, isoflavones, and micronutrients, can be utilized in a variety of high-end products, ranging from beverages to cosmetics (). There is biotechnological approach for utilization acidic whey, and biotransformation of whey can be achieved by biotechnological tools, including microbial fermentation and enzymatic treatment () (Figure 1).
Figure 1
For developing value-added uses of dairy co-products such as acid whey waste it is very important to know its content (
In addition, acid whey composition has some differences with sweet whey composition, and consists mostly of water, with lactose, high acidity and mineral content, which influence the strategies for their processing (
Table 1
| Composition | Acid whey | Sweet whey | Reference |
|---|---|---|---|
| Total solid, % wb | 4.27–5.92 | 60.64 ± 1.15 g/L | ( |
| Minerals g/L | 3.49–5.04 | 1.2 | ( |
| Ca2+, wt, % | 1.98 wt, % 1.91 ± 0.01b g/L | 0.69 wt, % 0.29 ± 0.00 g/L | ( |
| Phosphate ions | 0.8–0.92 g/L | 0.34–0.4 g/L | ( |
| Total Proteins % wb | 0.16–0.48 7.5 + 0.2 wt% | 5.52 ± 0.47 g/L 9.2 + 0.3 wt% | ( |
| Lactose % wt | 67% Wt 2.1–3.5 | 72–73% wt 49.44 ± 0.55 g/L | ( |
| Lactic acid | 7.19–13.07 g/L 14.4 + 1.5 wt% | 1.2 + 0.1 wt% | ( |
| pH | 4.21–4.7 | 5.9–6.4 | ( |
| COD mg/L | 31.9–62.400 | 50–102 g/L | ( |
| BOD mg/L | 32.7–50.5 | 35–60 g/L 35.000–55.000 mg l−1 | ( |
| Ash % wt | 0.6–0.73% wt | 0.5% wt 4.41 g/L | ( |
Comparison of sweet and acid whey composition.
Because of its rich content in protein and excellent amino acid profile production of high-quality protein powders is one of the potential uses of acid whey (
At the same time the whey powder production from Acid whey is more problematic compared with Sweet whey because AW has a more acidic environment due to the high content of organic acids, and lower content of lactose and proteins (
However, as the authors note (
Whey concentrate produced as a product of acid whey utilization is an ingredient of nonfat yogurt (
Some works also considered the potential use of acid whey for the production of dairy products such as kefir and yogurt (
Another problem of acid whey is the high content of lactose that can be utilized by probiotic bacteria to produce lactic acid, which helps to lower the pH of the product and give it a tangy flavor (
Finally, acid whey can be utilized in the production of cosmetics and personal care products (
Some biotechnological approaches are devoted to using acid whey as substrate for converting lactose into galactooligosaccharides (GOS) with prebiotic activity using commercial β-galactosidases or microorganisms with high β-galactosidase activity, and products produced during valorization can be used for people with lactose intolerance (
Some works described that treatment of whey leads to the release of nutraceuticals such as bioactive peptides, prebiotics, exopolysaccharides, organic acids, bacteriocins, isoflavone aglycones, and to the production of industrially important enzymes including β-galactosidase, protease, and amylase, and development of novel functional foods with health beneficial effects (
The mechanism of producing the β-galactosidase enzyme from lactic acid bacteria using AW as a nutrient medium substrate is considered by Kolev et al. (
For people who are lactose intolerant, it is important to eat lactose-free foods. Because acid whey is rich in carbohydrates, the selection of microorganisms capable of utilizing it as a substrate for the bioconversion of lactose into galactooligosaccharides is an important direction for the whey valorization and functional products development.
Besides, biotransformation of whey carbohydrates into prebiotics by recombinant enzymes has proven effective for the valorization of whey (
Some authors considered attempts to convert AW to biogas, ethanol, or lactose food products and they concluded that these attempts have all been hindered by the presence of lactic acid and high capital investment requirements, which restrict AW conversion in small-scale dairies. Using whey lactose and lactate for polyhydroxyalkanoates (PHAs) production is an opportunity that would not only provide low-cost renewable feedstock without competing with edible products but also simultaneously solve an environmental problem (
Table 2
| Source | Research | Product | Method | Strain/enzyme | Key findings |
|---|---|---|---|---|---|
| ( | Acid whey metabolomics | Metabolic profile | Analytical | - | Acid whey has a rich metabolic composition |
| (120) | Formulating ranch dressing | Formulating ranch dressing by replacing buttermilk with Yogurt Acid Whey (YAW). No lactic acid was added (lactic acid is naturally present in YAW) | Food technology | - | Up to 60% of buttermilk can be replaced by acid whey |
| ( | GOS synthesis | GOS | Microbial/enzymatic | Cryptococcus laurentii, Aspergillus oryzae | Acid whey as substrate for GOS synthesis |
| (121) | prebiotic GOS | Prebiotic galacto-oligosaccharides production from acid whey | Enzymatic | β-galactosidases (from Kluyvero-myces lactis and Aspergillus oryzae) and one novel, in-house produced (from Thermothielavioi-des terrestris), | Efficient, cost-effective production of valuable prebiotics from acid whey. The maximum GOS yield was 25.7% when using concentrated acid whey with 20% lactose content and the enzyme from T. terrestris. |
| ( | Production of Polyhydroxybu-tyrate (PHB) by use of Recombinant Escherichia coli LSBJ | Polyhydroxybuty-rate (bioplastic) | Microbio-logical | Recombinant Escherichia coli LSBJ | Bioconversion of acid whey (AW) to PHB |
| ( | β-galactosidase | Biocatalysis | Enzymatic | LAB, Bifidobacterium spp. | LAB strains assessed for enzyme activity in acid whey |
| ( | Fermented beverages based on processing of acid whey. | Fermented probiotic beverages The production process included combining pasteurized acid whey with UHT milk, unsweetenedcondensed milk or skim milk powder-introduced milk to enrich casein content and obtain a product with characteristics similar to that of fermented milk drinks. | Microbiolo-gical | Lactobacillus acidophilus LA-5 or Bifidobacterium animalis ssp. lactis BB-12 | Acid whey is a suitable base for functional beverages. L. acidophilus provides more acidity of beverages in comparison with B. animalis. |
| ( | Functional ingredients for yogurt | Acid whey protein concentrate after neutralization by food-grade ammonium hydroxide, and then ultrafiltration and diafiltration. | Technolo-gical | — | Whey protein concentrate is used for yogurt making, stable sensory and physicoche-mical properties |
| ( | GOS | GOS from acid whey | Enzymatic | β-galactosidase from Aspergillus oryzae and Kluyveromyces lactis Trichoderma terrestris | Enzymes from Aspergillus oryzae and Kluyveromyces lactis, Trichoderma terrestris have thermostable and acid pH-stable β-galactosidase. |
| ( | Bioenergy | Medium-chain carboxylic acid | Microbio-logical | Microbial composition | Bioconversion of acid whey |
| ( | Probiotic beverage | Whey and pineapple juice drink | Microbio-logical | Lactobacillus acidophilus, Bifidobacterium bifidum | Probiotic beverage with good sensory properties |
| ( | Functional beverage | Fermented beverage | Microbio-logical | LAB | Probiotic potential and antioxidant properties |
| ( | β-galactosidase activity | Hydrolysis of acid whey | Microbiolo-gical/Enzy-matic | LAB strains | Acid whey valorization using LAB enzyme potential |
| ( | Fermented probiotic beverage | Fermented probiotic beverage | Microbio-logical | LAB and yeast | Enhanced antimicrobial and antioxidant properties |
| ( | Functional beverage | Beverage based on acid whey | Microbio-logical | Brettanomyces claussenii | Lactose conversion |
| ( | Review of acid whey benefits | Multiple functional foods | Review | - | Acid whey has health potential and industrial uses |
| ( | Bioactive peptides | Functional peptides | Microbio-logical | Streptococcus thermophilus | Biopeptides |
| ( | β-galactosidase optimization | Milk and whey processing | Enzymatic | β-galactosidase (acid- and cold-tolerant) | Enzyme suitable for acidic whey conditions |
| ( | Whey spirit production | Fermented acid whey spirits | Microbio-logical/ Distillation | Fermenting yeast strains | Characterized volatile compounds during distillation stages |
| ( | Membrane processing | Treated acid whey | Technolo-gical | — | Electrodialysis and nanofiltration alter whey composition |
| (119) | Mineral removal from acid whey | Calcium and lactate extraction | Technolo-gical | — | Electrodialysis effective in adjusting mineral content |
| ( | Chemical conversion | 5-HMF and levulinic acid | Chemical / Thermoche-mical | — | Optimized synthesis from acid whey and lactose |
| ( | Exopolysaccharide production in whey | Iron-complexing EPS | Microbio-logical | Lactic acid bacteria | LAB produce EPS in acid whey with iron-binding properties |
| (122) | Whey fermentation promoters | Genetic regulation insights | Microbio-logical | — | New regulatory promoters discovered in acid whey fermentation |
| ( | Value of acid whey stream | Functional ingredients | Review | — | Acid whey is becoming more valuable than the main dairy product |
| ( | Microbial and enzymatic valorization | Nutraceuticals: bioactive peptides, prebiotics, exopolysaccharides, organic acids, bacteriocins, isoflavone aglycones; and industrially important enzymes: β-galactosidase, protease, and amylase. Whey enriched with bioactive compounds can be utilized for the functional and nutritional enhancement of foods and the development of novel functional foods with health beneficial effects. | Microbial and enzymatic | LAB, enzymes | Acid whey enriched with bioactive compounds can be utilized for the functional and nutritional enhancement of foods and the development of novel functional foods with health beneficial effects. |
| ( | GOS synthesis from whey | Galactooligosaccharides | Enzymatic | lactases from Aspergillus oryzae and Kluyveromyces lactis | Both acid and sweet whey are suitable for GOS production |
| ( | Cosmetic biotechnology | Innovative cosmetic product for hair based on lactoserum | Technolo-gical | — | Whey based concentrates in cosmetics |
| ( | Composition analysis | Acid whey from Greek yogurt (GAW), acid whey from cottage cheese (CAW), and milk permeate (MP). | Analytical | - | Composition data of coproduct streams: acid whey from Greek yogurt (GAW), acid whey from cottage cheese (CAW), and milk permeate (MP). |
| ( | Membrane technology applications | Dairy coproducts | Techno-logical | — | Membrane processes used for treating acid whey |
| ( | Historical and future outlook | Acid whey | Review | — | Trends and possibilities for whey valorization |
| ( | Engineering the optimum pH galactosidase | Glycoside hydrolase | Engineering | Aspergillus oryzae | Y138F and Y364F mutants exhibited better hydrolytic ability than lacA in milk lactose hydrolysis. |
| ( | GOS | GOS | Enzymatic/microbio-logical | Aspergillus oryzae galactosidase, Kluyveromyces marxianus and Saccharomyces cerevisiae cells | Reacted medium without nutrient supplementation (raw GOS) was fermented with Kluyveromyces marxianus cells obtaining GOS of 95% purity containing mostly tri- and tetrasaccharides with total recovery of GOS after 24 h. |
Strategies to valorize of acid whey.
3 Valorization strategies using membrane filtration and ultrafiltration
Advanced filtration techniques can separate valuable components such as proteins and minerals from acid whey. Ultrafiltration and reverse osmosis enhance protein recovery while reducing waste volume.
Advanced filtration techniques, such as ultrafiltration (UF), nanofiltration (NF), and reverse osmosis (RO), play a crucial role in separating valuable components from acid whey, reducing waste, and improving its functional applications. These membrane-based technologies enable the selective recovery of proteins, minerals, and lactose, transforming acid whey from an environmental burden into a resource for functional food and ingredient development.
3.1 Ultrafiltration for protein recovery
Ultrafiltration is commonly employed to concentrate and isolate proteins from acid whey. The membrane pore size (typically 1–100 nm) allows the retention of high-molecular-weight proteins, such as α-lactalbumin and β-lactoglobulin, while permitting the passage of smaller molecules like lactose, minerals, and organic acids (
3.2 Nanofiltration for partial desalination
Nanofiltration is used after UF to remove excess minerals from the acid whey stream while retaining valuable peptides and carbohydrates. The partial removal of minerals improves the sensory properties of acid whey-derived ingredients, making them more suitable for incorporation into food formulations (
3.3 Reverse osmosis for concentration and waste reduction
Reverse osmosis, with its tighter membrane structure, effectively concentrates acid whey by removing water, reducing transportation and storage costs. This process enhances the sustainability of acid whey valorization by minimizing waste volume and facilitating further processing into powdered whey ingredients (
3.4 Emerging membrane technologies
Innovative membrane-based techniques such as electrodialysis (ED) and forward osmosis (FO) are gaining attention for selective separation and demineralization of acid whey, enabling its application in beverages, dairy alternatives, and nutraceuticals (
Advanced filtration techniques provide a sustainable and economically viable solution for acid whey valorization. The integration of UF, NF, and RO, alongside emerging membrane technologies, enhances the functional properties of acid whey by concentrating valuable proteins and reducing waste. Continued research and optimization of membrane processes will further improve the efficiency and commercial feasibility of acid whey-based functional product development.
4 Microorganisms for acid whey utilization
Lactic Acid Bacteria (LAB) for Fermentation.
Lactic acid bacteria are naturally present in acid whey and can be employed for fermentation-based valorization. LAB such as Lactobacillus delbrueckii, Lactobacillus plantarum, and Streptococcus thermophilus efficiently utilize lactose and lactic acid from acid whey, converting them into lactic acid which is a precursor for biodegradable plastics and food preservatives (
4.1 Yeasts for ethanol and biopolymer production
Certain yeast strains can metabolize lactose and organic acids in acid whey, leading to bioethanol production, an important biofuel alternative. Kluyveromyces marxianus is widely studied due to its ability to ferment lactose directly into ethanol (
4.2 Filamentous fungi for protein and enzyme production
Filamentous fungi such as Aspergillus oryzae and Rhizopus oligosporus can utilize acid whey components to produce Single-cell proteins (SCPs) for animal feed and food applications (
4.3 Microbial consortia for biogas and biohydrogen production
Mixed microbial communities have been successfully employed to convert acid whey into biogas (methane) and biohydrogen: Anaerobic digestion of acid whey by methanogenic consortia yields biomethane, a renewable energy source (
Microorganisms play a pivotal role in acid whey valorization, offering diverse biotechnological applications such as fermentation for functional food ingredients, biofuel production, enzyme synthesis, and biogas generation. Future research should focus on genetic and metabolic engineering of microbial strains to enhance their efficiency in acid whey bioconversion while optimizing industrial-scale fermentation processes.
Several types of microorganisms can utilize acid whey for various purposes, such as producing value-added products or treating waste.
Among microorganisms’ lactic acid bacteria are most used because of their ability to ferment lactose. LAB are commonly used to ferment acid whey into products such as kefir, yogurt, and sour cream. These bacteria convert lactose, a major component of acid whey, into lactic acid, which lowers the pH and gives the product a tangy flavor. LAB can also produce other compounds such as exopolysaccharides, which can enhance the texture and stability of dairy products (
Lactobacillus bulgaricus and Streptococcus thermophilus are two species of lactic acid bacteria commonly used in the production of yogurt and other fermented dairy products. They can utilize lactose in acid whey to produce lactic acid, which contributes to the characteristic tangy flavor of yogurt.
Propionibacterium freudenreichii is a species of propionic acid bacteria that can be used to produce Swiss cheese. It can utilize the lactose and other components of acid whey to produce propionic acid and carbon dioxide, which contribute to the characteristic flavor and texture of Swiss cheese.
Yeasts - Some yeast strains can use lactose in acid whey as a carbon source to produce ethanol and other organic compounds (
Saccharomyces cerevisiaes commonly used in the production of beer, wine, and other alcoholic beverages. It can ferment the lactose in acid whey to produce ethanol and other organic compounds, which can be used to produce premium alcoholic beverages (
Methanogenic archaea - These microorganisms can convert the organic matter in acid whey into biogas, which is mainly composed of methane and carbon dioxide (
Fungi - Certain filamentous fungal strains can use acid whey as a substrate to produce enzymes such as proteases, amylases, and lipases. These enzymes can have various industrial applications such as in the food, textile, and detergent industries. Aspergillus oryzae is a filamentous fungi and Cryptococcus laurentii used in the production of functional sugars and related value-added compounds (
In summary, acid whey can be a valuable substrate for several types of microorganisms, which can be converted it into useful products or treated it as waste. Harnessing the potential of these microorganisms can lead to the development of more sustainable and efficient processes for utilizing acid whey (Figure 2).
Figure 2

Graphical representation of acid whey utilization pathways. Reproduced from (118), with permission from The Royal Society of Chemistry.
Kaur et al. works devoted to utilizing whey with using of microorganisms such as proteolytic LAB strains, yeasts, Bacillus spp., fungi, and algae, and treatment of whey by enzymes including β-galactosidase, protease, dextransucrase, levansucrase, and β-glucosidase that can produce several bioactive compounds with functional properties, and novel functional food products (
For example, enzymes from Aspergillus oryzae and Kluyveromyces lactis have been used to convert lactose using acid whey. Zerva et al. noted that A thermostable β-galactosidase (TtbGal1) from Thermothielavioides terrestris can be used for the valorization of acid whey, with minimal preparation of the starting material, because this microorganism has a thermostable and acid pH-stable β-galactosidase, and GOS yields reached up 14.8% (
Besides LAB, β-galactosidase synthesizing microorganisms such as yeast strains Kluyveromyces marxianus, Kluyveromyces lactis, fungi Aspergillus oryzae, bacteria species Bacillus circulans can be used for the production of GOS from whey (
Wherry et al. (
5 Acid whey fermented beverages and functional products -new ways to increase acid whey fermentation
Many papers are devoted to producing beverages based on sweet whey. In the paper authors (
Kaziullayeva et al. (
Jitpakdee et al. (
Very interesting way to use the fermented whey described by authors of work (
As an Skryplonek et al. (
Functionality, Formulations, Health Benefits, and Applications of whey-based beverages are described in the work of Chavan (
Nielsen et al. (
The work found new ways to increase whey fermentation by using orange juice, orange peel and molasses to increase activity.
It has been found that the use of whey in mixtures with low-cost raw materials and the application of a continuous process promote effectively its fermentation by kefir, leading to the potential production of novel alcoholic drinks. The authors reveals new findings concerning the substantial promotional activity of orange juice, peel, and molasses on whey fermentation using single-cell culture of kefir microorganisms, to increase the capacity of whey exploitation and to create a new research concept.
Durpekova et al.(
Rocha-Mendoza et al. (
Authors (
The authors concluded that acid whey was a good medium for probiotic bacteria and grew very well and their concentration was higher than the therapeutic dose.
Islam et al. (
6 Ways to increase probiotic viability of functional beverages by use of microencapsulation
Functional beverages are beverages fortified with bioactive substances having health-promoting or disease-preventing properties. Incorporation of bioactive compounds into beverages is a technologically demanding process concerning maintaining their bioactivity and adequate delivery to the organism. The significant bioactive substances are probiotic bacteria which are beneficial to the host only when administered in adequate amounts (
Krunić et al. (
Encapsulation is an advanced technology and the most effective way of incorporating probiotics into functional food and drink (
The composition of encapsulating material is the main determinant of microcapsule functional properties and effective use of the bioactive ingredient (
Various biopolymers such as polysaccharides and proteins, or a mixture of both, are regarded as suitable materials that retain their structure during their pass through the highly acidic environment of the stomach and deliver them in the intestine (
Among polysaccharides used for encapsulation (alginate, chitosan, agar, carrageenan, gum arabic, dextrans, xanthan and cellulose (ethyl-cellulose, acetyl-cellulose, methyl-cellulose, carboxymethyl-cellu-lose, nitrocellulose) alginate is commonly used to encapsulate probiotics by various methods such as extrusion, emulsion and spray drying (especially freeze drying) (
The advantages of alginate as a matrix are its ability to form gels with relatively good mechanical properties, high porosity and biodegradability, easy manipulation as well as good biocompatibility. At very low pH values, the use of the alginate encapsulation matrix alone is limited due to fast degradation and rapid release of encapsulated agents (84). These are the reasons why alginate microparticles are often not sufficient for the encapsulation of bioactive. The functionality of alginate particles and their ability to preserve probiotics bioavailability and assure minimum effective concentration can be adjusted by using alginate-based composites, i.e., by mixing alginate gels with other biopolymers (hydrocolloids, proteins, or starches) in correct proportions (85, 86).
Coating of alginate microspheres with oppositely charged polyelectrolyte, like chitosan, increases microparticle chemical and mechanical stability improving the effectiveness of encapsulation, reducing porosity, limiting the adverse effect of functional food constituents (for example the transition of acids and flavonoids from the fruit juice into microcapsules), reduced release of the encapsulated bacteria, and increased stability at various pH ranges (87, 88). Some examples of improved survival of probiotics bacteria encapsulated in alginate-chitosan microcapsules compared to alginate microparticles are (i) Lactobacillus casei, Bifidobacterium longum, Bifidobacterium longum subsp. infantis and Bifidobacterium breve in digestive juices (83), (ii) Lactobacillus casei and Lactobacillus acidophilus in yogurt and milk (89), (iii) Lactobacillus acidophilus in simulated gastric solution (90), (iv) Bifidobacterium breve in simulated gastric solution (91), (v) Bifidobacterium longum in gastrointestinal fluids and at elevated temperature conditions (92).
Examples of the increased survival in simulated gastric solution and bile salts of probiotics encapsulated in alginate-composite compared to alginate microparticles only are (i) Bifidobacterium bifidum loaded in alginate-coated poly-L-lysine microcapsules (93), (ii) alginate-pectin composite with encapsulated Lactobacillus casei (94), (iii) Lactobacillus acidophilus and Bifidobacterium animals subsp. lactis encapsulated in alginate-modified starch composite (95), (iv) Lactobacillus delbrucekii encapsulated into succinylated alginate (96), (v) Lactobacillus plantarum encapsulated in alginate coated with whey protein (97, 98), (vi) Lactobacillus delbrueckii subsp. bulgaricus and Lacticaseibacillus paracasei encapsulated in alginate-whey protein isolate microcapsules (99).
Common materials for protein-based probiotics encapsulating materials are casein, gluten, albumin, and whey proteins. Whey proteins are often the preferred source for ready-to-drink protein beverages because of their excellent nutritional qualities, bland flavor, ease of digestibility, and unique functionality in beverage systems (
Casein used as a matrix for protecting Lacticaseibacillus. paracasei and Bifidobacterium lactis during gastric transit improved cell viability in pH 2.5 buffer by 20% relative to the free cells (102). Lactobacillus acidophilus and Bifidobacterium lactis encapsulated in spray-dried coacervates of casein with pectin resulted in an excellent increase in cell survival at very low pH values (103), Lactobacillus rhamnosus GG encapsulated in microparticles with whey protein isolate enabled very good cell survival in ex vivo porcine gastric contents (104). It was shown that during incubation in the stomach loss of probiotic content was minimal but in the contents of the intestine a release over 30 min occurred.
The addition of proteins to the alginate matrix increases cell viability and encapsulation efficiency. Streptococcus thermophilus and a probiotic strain Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophilus and Bifidobacterium bifidum were encapsulated in whey protein-alginate and whey protein hydrolysate-alginate (
Many health benefits of probiotics are associated with their survivability and stability in carrier food and gastrointestinal conditions. Among many beneficial health effects, probiotic bacteria also present significant antioxidant abilities. In vitro and in vivo studies indicate that probiotics may reduce oxidative damage, and free radical scavenging rate, and modify the activities of key antioxidant enzymes in human cells (105). Various strains of lactobacilli and bifidobacteria exhibit remarkable antioxidant activity in the host intestine and promote the production of antioxidant enzymes helping to remove reactive oxygen species (106, 107). In a review paper Wang et al. (106) summarized the mode of action of probiotic bacteria in antioxidation (probiotics may modulate the redox status of the host via their metal ion chelating ability, regulate signaling pathways, enzyme-producing reactive oxygen species, and intestinal microbiota).
Besides numerous investigations on encapsulated probiotics’ health benefits, the antioxidant properties of encapsulated probiotics have been investigated to a lesser extent. Krunić et al. showed that enriching the alginate matrix with whey protein and whey protein hydrolysates can preserve the antioxidant capacity and stability of whey-based beverages (
A relatively novel trend in the functional beverage design is the use of microcapsules containing both probiotics and prebiotics (known as synbiotics). Prebiotics are food ingredients that selectively promote the growth and activity of the beneficial bacteria in the gut over detrimental bacteria. There has been little published data on this topic, but some of the examples are: (i) optimization of Lactobacillus casei, Lactobacillus acidophilus, Bifidobacterium longum and Bifidobacterium bifidum encapsulated in alginate with prebiotics revealed improved protection and survival (108); (ii) presence of prebiotic, oligosaccharides, stimulates the growth of Lactobacillus acidophilus 5 encapsulated in alginate-chitosan microcapsules incorporated in orange juice (89); (iii) Lactobacillus casei 359 microencapsulated in whey protein concentrate with prebiotic inulin revealed very high survivability in litchi juice. During the encapsulation process, inulin protects the cells from drying providing tolerance to a higher temperature (109). In a review paper, Rovinaru and Pasarin presented the synbiotic concept, challenges for synbiotic formulation in fruit drinks and future perspectives (110). They concluded that the encapsulation of synbiotics is a successful improvement of the viability and stability of probiotics in fruit juices.
In vivo evaluation of probiotics encapsulation efficacy performed mainly on rats and mice revealed a direct and easy delivery of probiotic cells in the intestinal region (111, 112). The viability of microencapsulated probiotic cells after oral administration may be followed by molecular biological approaches real-time PCR (113) and fluorescence in situ hybridization (114). However, to obtain relevant results, detailed in vivo studies and models of the digestive system more similar to humans are needed (115).
Various research works revealed that microcapsules loaded with probiotics (microcapsule formulations) increase the viability of the encapsulated ingredient during processing and in the final product, a functional beverage. Besides enhancing probiotics viability during processing and storage, microcapsule formulations also provide targeted delivery ensuring desirable health-promoting effects of probiotics on the host. The supply of probiotics to the site of action not only depends on the amount of the probiotic in a beverage but also on its bioavailability. Bioavailability is the amount of a probiotic that enters the circulation when introduced into the body to have an active effect. The bioavailability of a probiotic after ingestion depends on its absorption and in part on the changes that occur during passage through the gastrointestinal tract (116).
The selection of the matrix materials, probiotics to be encapsulated, methods of microencapsulation and application are interdependent (117). To obtain a well-designed microcapsule formulation for a specific application, it is important to optimize parameters during microcapsule preparation bearing in mind the properties of probiotic and matrix components, the method used for encapsulation as well as regulatory requirements, cost, industrial scalability, consumer needs, and economic issues. Although different techniques and wall materials have been reported for the effective encapsulation of probiotics, maintaining the viability of probiotics remains a major challenge and further research is needed in this regard (115). Proper selection of microcapsule formulation variables helps in designing microcapsules with the desirable release. Designing optimal formulations of microcapsules that ensure sufficient survival of probiotics in a functional product, prevent the release of probiotics and reactions with other compounds of the base drink, and enable the action of probiotics only at the point of use requires an interdisciplinary approach as well as a good understanding of the molecular interactions of all present ingredients. Any probiotic delivery system should be designed so that the probiotics are released within the colon for their health-promoting effect. Microencapsulation is a complex area that includes scientific research on colloidal, physical and surface chemistry and microbiology. A better understanding of the complex interactions of probiotics with complex beverage matrices and the physiological mechanism involved when beverages pass through the gastrointestinal tract is needed to ensure the optimal viability of probiotics under all circumstances.
7 Conclusion
Acid whey, a byproduct of dairy processing, presents both challenges and opportunities due to its high organic load and valuable nutrient composition. This review explored innovative strategies for acid whey valorization, including biotechnological processing, fermentation, and ingredient recovery. Potential applications span food, beverages, animal feed, bioenergy, and even cosmetics. Despite challenges such as its high acidity, lactose content, and mineral concentration, acid whey can be effectively utilized to create value-added products, including protein powders, fermented dairy products, alcoholic beverages, and nutraceuticals. Biotechnological approaches, such as enzymatic conversion of lactose into galactooligosaccharides, further enhance its potential for use in functional foods. Additionally, recent research highlights ways to increase probiotic viability in functional beverages through microencapsulation. Functional beverages, fortified with bioactive compounds, including probiotics, require technological advancements to maintain probiotic viability.
Encapsulation techniques, such as using an alginate matrix enriched with whey, have been shown to improve the antioxidant capacity and stability of whey-based beverages. This approach not only enhances probiotic survival during storage and gastrointestinal transit but also contributes to the sustainability of whey utilization by addressing environmental concerns related to whey disposal. Encapsulation materials, including biopolymers such as polysaccharides and proteins, play a crucial role in protecting probiotics and ensuring their controlled release in the intestine. Various encapsulation methods, such as spray drying, extrusion, and coacervation, offer promising solutions for developing stable probiotic-enriched functional beverages.
Overall, acid whey valorization presents a sustainable solution to reduce dairy industry waste while creating high-value functional products. Future research should focus on optimizing processing techniques, improving probiotic stability, and expanding applications to maximize the potential of acid whey as a functional ingredient. By addressing the current challenges and leveraging biotechnological advancements, acid whey can be transformed into a resource that benefits both industry and consumers.
A promising method for improving the viability and targeted distribution of probiotics in functional foods and beverages is microencapsulation. Achieving the intended functional qualities and making efficient use of bioactive compounds depend on the choice of suitable microorganisms, encapsulating materials, and microencapsulation techniques.
Microorganisms: Because of their probiotic and antioxidant qualities, strains of Lactobacillus and Bifidobacterium are commonly employed.
7.1 Encapsulation systems
Commonly used matrices include alginate, chitosan, whey protein, and casein. Whey proteins provide superior nutritional characteristics, while alginate is used extensively since it is inexpensive and biocompatible. Probiotic survival can be further increased by adding prebiotics (synbiotics) or combining ingredients like alginate and chitosan.
7.2 Procedures
Spray drying, extrusion, emulsion, coacervation, and lyophilization are examples of common procedures. The best methods for preserving probiotic viability are gentle ones.
The optimization of microcapsule preparation parameters, including the properties of probiotic and matrix components, regulatory requirements, cost, and consumer needs, is essential for designing effective microcapsules. Future research should focus on biopolymer research for encapsulating probiotics with various beneficial properties and on gaining a better understanding of probiotic interactions within complex beverage matrices to ensure optimal viability and health-promoting effects. In vivo studies and models that closely mimic the human digestive system are also needed for relevant results.
Statements
Author contributions
BM: Methodology, Validation, Software, Formal analysis, Data curation, Visualization, Investigation, Writing – review & editing, Conceptualization, Resources, Writing – original draft. ET: Visualization, Writing – review & editing, Conceptualization, Investigation. GM: Project administration, Writing – review & editing, Supervision, Investigation, Funding acquisition, Resources, Conceptualization. LŽ: Writing – review & editing. VT: Writing – review & editing. AI: Data curation, Formal analysis, Writing – review & editing. MV: Investigation, Conceptualization, Methodology, Writing – review & editing, Writing – original draft.
Funding
The author(s) declare that financial support was received for the research and/or publication of this article. This research is funded by the Science Committee of the Ministry of Science and Higher Education of the Republic of Kazakhstan (Grant no. AP19679879).
Acknowledgments
The authors gratefully acknowledge the Bolashak International Program “500 Scientists” for facilitating academic networking opportunities at the University of Westminster, London, United Kingdom.
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Generative AI statement
The authors declare that Gen AI was used in the creation of this manuscript. The authors used OpenAI’s ChatGPT to assist with the suggestion of a tabular format structure for Table 2.
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Summary
Keywords
acid whey, strategies for acid whey valorization, functional products, fermented beverages, microencapsulation, probiotic viability, acid whey fermentation
Citation
Mutaliyeva B, Turkeyeva E, Madybekova G, Živković L, Talluri VSSLP, Issayeva A and Vinceković M (2025) Acid whey valorization: novel approaches for probiotic and functional products development. Front. Nutr. 12:1630925. doi: 10.3389/fnut.2025.1630925
Received
18 May 2025
Accepted
01 September 2025
Published
19 September 2025
Volume
12 - 2025
Edited by
Photis Papademas, Cyprus University of Technology, Cyprus
Reviewed by
Maria Aspri, Cyprus University of Technology, Cyprus
Hannah Araujo, Federal University of Sergipe, Brazil
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© 2025 Mutaliyeva, Turkeyeva, Madybekova, Živković, Talluri, Issayeva and Vinceković.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: G. Madybekova, galiyamadybekova56@list.ru; E. Turkeyeva, e.turkeeva@iuth.edu.kz
†Present address: A. Issayeva, Scientific and Production Enterprise “Antigen” LLP”, Almaty, Kazakhstan
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