Interest in plant-based dairy imitations has consistently been growing over the years, with well-documented positive health effects causing consumers to demand these products and so markets are filling up (Cichońska et al., 2024). The global market for plant-based dairy is predicted to grow at a compounded annual rate of 12.5% and reach US$52.58 billion by 2028 Grand View Research. (2021); Plamada et al., (2023). These replacements have several favorable attributes compared to traditional dairy-based products. They are naturally free of lactose and cholesterol (Hartmann et al., 2018) as well as allergy-causing dairy proteins such as casein. On the other hand, they respond to increasing consumer worries concerning hormones and antibiotic residues in animal-based products and meet the conditions for vegan labeling (Cichońska and Ziarno, 2021).

Plant-based cheeses have come into focus over the last decade, in part because natural plant fats tend to be viewed as better-for-you options than those from animals. Plant-based cheese also serves as an essential alternative for people with dairy sensitivities. Nevertheless, food allergogenicity is still a major concern on plant proteins .allergic manifestations to dairy proteins vary from mild to life-threatening, necessitating the development of safe alternatives. As noted by Mäkinen et al.  (2016), grains, legumes and seeds constitute the fundamental matrix required to form the complex network that maintains the structural integrity and shape of plant-based cheese. Furthermore, organic acids, including lactic or citric acid, are also used for the pH control (Ismail et al., 2021) and consequently improving protein solubility. Addition of these acidulants not only enriches the flavour profile, it also extends secondary lipid oxidation, which contributes to maintaining sensory properties and overall quality of the final product (Kovačević et al., 2024). Therefore, the study aimed to develop a sustainable plant- based cheese analogue that can be locally produced in Iraq using potato as a carbohydrate source and peanut as a protein source.

Raw materials used
 
High-quality potatoes (Solanum tuberosum) representing commonly used culinary varieties, were obtained; peanuts (Arachis hypogaea), of Turkish origin; xanthan gum (E415) mesh 80, of Chinese origin, was purchased from Fooding Company (ISO 22000); citric acid; table salt (Sodium chloride, NaCl); artificial cheddar cheese flavoring (chemically safe for food use) and edible orange fooding grade color (E160b) were purchased from local markets in Baghdad. The probiotic strain, Lactobacillus plantarum (Standardized to 10 billion CFU per capsule), was obtained from Swanson Health Products (Fargo, ND, USA). The freeze- dried culture was maintained under sterile conditions and activated according to the manufacturer’s instructions before being incorporated into the cheese analogue formulations.
 
Manufacturing procedure of plant-based treatments
 
First
 
Preparation of the potato paste: The potatoes were boiled until tender, then peeled and mashed to obtain mashed potato.
 
Second
 
Preparation of the peanut emulsion: This was done using the method described by (9). The emulsion was prepared by adding 100 ml of water to 125 g of ground peanuts and treating it at 60°C for 15-20 minutes with continuous stirring.
 
Production of imitation plant-based (Vegan) cheese
 
A series of plant-based cheese formulations were developed by blending potato paste and peanut emulsion at a fixed ratio of 18:80 (w/w). This particular formulation was chosen to assess the best textural quality and organoleptic properties of the final product. Other ingredients (skim milk, xanthan gum, citric acid, sodium chloride, synthetic cheddar copper tone type NFFP-1 and orange colorant (E160b) were added to the base at the levels listed in (Table 1). After the thermal process, the blend was cooled down to an inoculation temperature of 35°C and a starter culture was added. The mixture was pipetted into moulds and placed in cold storage (4°C) under standard conditions for 21 days to mature and stabilise. Throughout shelf-life, the treatments were periodically analyzed in order to check quality and longevity.


 
Chemical composition analysis
 
The chemical qualities of the processed cheese were determined following (2) standard methods, including moisture, crude protein using the Kjeldahl method, fat employing the soxhlet method and ash and total carbohydrate content. The pH was measured potentiometrically by a Crison pH meter. the fat-in-dry-matter percentage (FDM%) was calculated as show in equation 1.
 

 

 
Microbiological analysis: Assessment of probiotic viability
 
The viable cell counts of the probiotic enhancer, Lactiplantibacillus plantarum (L. plantarum), were determined in accordance with the protocols established by the (3).
 
Sensory evaluation
 
Organoleptic properties of the plant cheese samples, was assessed by a trained panel of experts from Department of Food Science at College and agricultural Engineering Sciences University of Baghdad using a standard score sheet based on the method of (APHA, 2004).

Chemical composition analysis of raw materials used in analogue plant-based cheese production
 
Table (2) show the chemical composition percentages of analogues plant cheese, potatoes and peanuts. The raw potato materials were analyzed, which showed that the protein content was 2.9% prior to boiling and it increased to 9.5% after boiling of potatoes. Boiling and peeling increased protein concentration and the moisture content which was reduced from 74 % to about 18.7 % was doubled in other nutrients. Mandatory information: carbohydrates also increased largely to 61.59% having boiled and dried (Table 2). Meanwhile studies for protein levels 2-3%, fat at 0.5%, carbohydrates at 17-20%, fiber at 2-2.5% and ash content from (1) to (1.5%), while moisture content was in the range of between sevntieths to eightieth percent (%). These results are also consistent with those reported in Eco-physiological studies by Friedman (2014), although slight differences exist which may be attributed to variations in cultivars and environmental conditions.


 
Peanut chemical composition
 
The peanut protein (20.25%) was lower than that reported by 22 to 28%. The fat level was 45.26%, corresponding to the ranges of 45-52%. Carbohydrate was 21.34%, standard range from 18 to 25%. Fiber levels were low (2.3% vs the standard 7-10%) and may be associated with the species of peanut or processing method used. The ash content was 3.32% which falls slightly above the average of 2-3%. Lastly, moisture was 4.5%, which is within the limits (3-6%) established by standards. Upon processing into peanut paste, it was discovered to be of high fat (49.81%) and protein about (16.94%). Vegetable fats, potato starch and oats were also utilized to produce another style of vegetable cheese and other raw materials like chickpea flour and almonds. A study by (El-Abd et al., 2003) confirmed that peanuts are rich in protein (25-28%), monounsaturated fats (50%) and dietary fibres (8-10%).
 
Study of the effect of refrigerated storage on the chemical composition of prepared plant-based cheese
 
The changes in the chemical composition of the imitation plant-based cheese treatments were monitored during a21-days refrigerated storage period at 4°C. The results showed the following:
 
pH
 
The results in Table (3) indicate that the pH value of the control sample A1 was 6.5 on the first day and decreased to 5.02 on the 21^st day of refrigerated storage. Meanwhile, the pH value of the sample A2, to which the probiotic was added at a rate of 5%, was 5.8 at day 1 and decreased with increasing storage time, reaching 5.2 and 4.6 at days 7 and 15, respectively and decreasing further on day 21 to 4.04. The study conducted by (El-Abd et al., 2003) confirmed that the pH of plant-based cheeses ranged between 4.5 and 4.7 after 17 hours of fermentation. The reason for the extended fermentation time was explained by the fact that the amount of water-soluble carbohydrates was low in the legume protein used, which reduced the availability of fast-digesting sugars to the fermenting bacteria and thus slowed the rate of acid production compared to a medium rich in soluble sugars.



The statistical analysis results showed significant differences between treatments A1 and A2 during the 21-day refrigerated storage period. Significant differences were also found between the two treatments for days 1, 7 and 21.
 
Protein
 
The results show significant different in protein ratio over the storage period. A significant reduction in protein content was noted for all treatments ranging from day 1 to day 21. In the control (A1), protein content decreased from 11.22% on day 1 to 3.5% on day 21. Likewise, during treatment (A2), the proportion declined from 7.72% on do 1 to 3.6% on day 21. This decrease is suggested to be due to an increased rate of proteolysis. Proteases and peptidases-originating from the plant material of feed ingredients, or secreted by microorganisms (in our case probiotics in treatment A2) catalyze the hydrolysis of complex proteins like those in peanuts and potatoes into low-MW peptides that free amino acids. Furthermore, (El-Abd et al., 2003) stated that not only Lactobacillus spp. and Bifidobacterium spp. also produce extracellular protease that subsequently further degrades protein In fact, these levels of protein exceed that of other plant-based cheese analogs which contained much lower values varying between 0.1-1.7 g/100 g. These percentages are also particularly close to the recorded 12.9-18.2 g/100 g for processed dairy cheeses as reported by (Ali et al., 2025; Ouyang et al., 2020).

Fat
 
Fat content of both cheese treatments decreased throughout the storage period. The fat content decreased from 14.29% at day 1 to 11% by day 21 of storage in treatment A1. This decrease in fat percentage is mainly due to lipolysis, where the lipase enzymes formed by spoilage microorganisms or introduced probiotics induce hydrolysis of fats by degradation of triglycerides into free fatty acids. Furthermore, some of the fat could escape from the food matrix via oil leakage during storage (Reid and Yen, 2004; Lertpimonpan et al., 2019). Interestingly, Lactobacillus spp. in treatment A2 acted to retard this process of oxidation by both generating antioxidants like glutathione and superoxide dismutase.
 
Lipid oxidation and stability
 
The auto-oxidation of unsaturated fatty acid-rich vegetable fats from peanuts results in a loss in fat content during storage. However, the existence of probiotics has served to ameliorate this loss, through spawning secretion of endogenous antioxidants including superoxide dismutase. This is in conformity with the results of (Gupta et al., 2021) who mentioned that the presence of probiotics in vegetable-based cheese decreases lipid oxidation at refrigerated condition. Also, potato-based and xanthan-gum cheeses made from plants have a high rate of fat slumping. This is due to a protein network that encapsulates and entraps fats within the matrix.
 
Ash
 
The ash content is the total mineral content of a sample. The ash content was higher in the control treatment (A1) over time of storage. In contrast, ash content in treatment (A2) was relatively stable and fluctuated slightly between 1.4% and 2.5%. This stability indicates that mineral composition did not vary over the storage periodTex Parameteri-default pandoc ASH significant change are seldomly observed for this parameter and the reason is that ash materials are found to predominantly comprise of stable inorganic ones such as sodium (Na), potassium (K) and calcium (Ca). The constant characteristic of the ash content suggests that a stable mineral composition is maintained and also implies no solubility loss of salts during storage.
 
Carbohydrate 
 
The results show that the values of the percentage content of carbohydrates also showed a significant decrease at the beginning (after 7 days) for both treatments (A1 and A2). This decrease results in large part from the metabolism of lactose or starch by microorganisms. In particular, probiotics are reported to metabolize lactose, which is also present in the skim milk powder added to the composition, generating lactic acid and decreasing the digestible carbohydrate content. This was then followed by a notable and gradual increase in carbohydrate content until day 21. The high-treatment effect on A2 was appreciably reduced and consistently more moderate than the increase observed for A1. This increase is influenced by the decrease in dry matter content of fats and proteins caused by proteolysis and lipolysis, as with reduced proportions of fat and protein these are calculated for percent carbohydrates too. Moreover, the increase in moisture content also involved these relative values.
 
Moisture 
 
The experimental results revealed a progressive increase in moisture content for both treatments (A1 and A2) throughout the storage period, culminating in a maximum value of 68.77% by the end of the 21^st day. This increase is mainly due to the uptake of atmospheric water from the surrounding environment through the hygroscopicity of xanthan gum, a powerful water-retaining agent in the cheese microstructure. In addition, the increase in moisture is also associated with syneresis (liquid phase separating from solid ingredients (Table 4). It was also noted that the plant-based product absorbed more moisture during storage compared to animal cheeses, probably due to its weaker protein network structure; such a less dense and well-organized influence, determining its inferior mechanics of water migration, have previously been established by (Ismail et al., 2021). The results of the statistical analysis showed significant differences in the chemical composition of the imitation vegetable cheese during the 21-day refrigerated storage period, with significant differences in the percentage of fat, protein, nitrogen, carbohydrates and moisture.


 
Fat in dry matter (FDM%)
 
The fat in dry matter (FDM%) represents the lipid content remaining after the removal of moisture. It is considered the most critical indicator for classifying the quality, texture and flavor profile of plant-based cheese analogs. At the beginning of the storage period, the FDM values for treatments A1 and A2 were 33.79% and 28.27%, respectively. These percentages increased gradually as the storage progressed, reaching 31.38% and 34.90% by day 21 for both treatments. This upward trend in FDM can be attributed to the relatively minor changes in absolute fat content compared to the shifts in other solid components. These values are considered acceptable for low-fat plant-based cheeses, which typically range between (30-35%) (Table 5). The results of the current study differ from those reported by Ismail et al., (2021), who observed a decrease in FDM during the storage period of vegan cheese, attributing this decline to increased moisture content and fat loss.


 
The effect of refrigerated storage on microbial stability and the quality of plant-based cheese
 
Deterioration was observed in treatment A1 after 15 days of storage, while treatment A2, to which the probiotic enhancer was added, showed better stability until the end of the storage period, i.e., day 21, which confirms the role of the probiotic enhancer in extending the storage life through the production of organic acids and compounds that inhibit the growth of harmful bacteria such as bacteriocins. This can be explained by the biological activity of the probiotic leading to a reduction in the pH of the manufactured imitation plant-based cheese and inhibition of aerobic spoilage bacteria through the production of organic acids and bacteriocins (Table 6). While no mold or yeast growth was observed in either treatment during the storage period, it appeared only in treatment A1 on day 21. No growth was observed in treatment A2, indicating that the environment created by the probiotic was unfavorable to fungi due to the low pH and the production of acids and peroxides. Muthusamy et al., (2020) and Liu et al., (2018) reported that Lactobacillus plantarum and L. rhamnosus secrete short-chain fatty acids that inhibit fungal growth and no growth of Staphylococcus spp. was observed in either treatment A1 or A2 during the storage period.


 
Sensory evaluation
 
Sensory evaluation remains one of the most reliable methodologies for determining the fundamental characteristics of cheese and other food products, as well as gauging consumer acceptability of these attributes. Furthermore, sensory analysis reflects the extent to which the manufacturing and production objectives have been achieved. Table 7 illustrates the sensory evaluation results for the imitation plant-based cheese samples: the control (Treatment A1) and the probiotic-enriched imitation plant-based cheese (Treatment A2), fortified with L. plantarum during the storage period. The data indicates a significant superiority of Treatment A2 across all investigated sensory parameters. This reflects the pivotal role of the probiotic enhancer in improving flavor, texture and appearance. Additionally, the incorporation of 5% skim milk significantly synergized with the probiotic activity, effectively maintaining these sensory characteristics throughout the 21-day refrigerated storage period.

This study demonstrated the feasibility of producing plant-based cheeses from locally available plant materials and improving their sensory properties by adding xanthan gum and cheddar flavoring. It was further enriched with a probiotic to enhance texture, flavor and consistency, thereby increasing consumer acceptance. The study also concluded that fortifying these cheese alternatives with vitamins and essential nutrients is crucial for boosting their popularity and consumption.

The present study was supported Department of Food science, College of Agricultural Engineering Sciences, University of Baghdad.
 
Disclaimers
 
The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.

The authors declare that there are no conflicts of interest regarding the publication of this article. No. funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.