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Article

Development of Functional Bread: Exploring the Nutritional, Bioactive and Microbial Potential of Carob (Ceratonia siliqua L.) Pulp Powder

by
Jana Zahorec
1,
Dragana Šoronja-Simović
1,
Jovana Petrović
1,*,
Zita Šereš
1,
Branimir Pavlić
1,
Danica Božović
1,
Lidija Perović
2,
Nikola Martić
3,
Sandra Bulut
1 and
Sunčica Kocić-Tanackov
1
1
Faculty of Technology, University of Novi Sad, Bulevar Cara Lazara 1, 21000 Novi Sad, Serbia
2
Institute of Food Technology, University of Novi Sad, Bulevar Cara Lazara 1, 21000 Novi Sad, Serbia
3
Department of Pharmacology, Toxicology, and Clinical Pharmacology, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia
*
Author to whom correspondence should be addressed.
Processes 2024, 12(12), 2882; https://doi.org/10.3390/pr12122882
Submission received: 25 October 2024 / Revised: 11 December 2024 / Accepted: 12 December 2024 / Published: 16 December 2024
(This article belongs to the Special Issue Design of Food Processing and Technologies: Studies on Physical Properties, Thermodynamics, Rheology and Emulsification)
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Abstract

For the development of a high-quality functional bread, it is of crucial importance to find a compromise between improving the functional value of the product and impairing its quality. Therefore, the primary aim of this work was to define the effects of carob pulp powder (CP; 10–20% on flour basis) on the rheological behavior of dough and quality parameters of bread. The introduction of CP caused a major increase in dough resistance to extension while significantly decreasing dough extensibility, resulting in a lower volume with a denser crumb structure of the obtained bread samples. The negative effects of the CP on the dough and bread characteristics were partially compensated by the addition of dry sourdough (DS) as an improver at the level of 5–10% (flour basis). It was determined that by combining 15% CP and 10% DS, bread of very good sensory quality can be produced. The content of total dietary fiber (6.48 g), Mn (0.76 mg) and Cu (0.21 mg) per 100 g of the mentioned bread was sufficient for the nutritional claims “high in fiber”, “high in manganese” and “source of copper”. The obtained bread had a content of total phenols 10 times higher than the control and 5.5, 12 and 26 times higher antioxidant activity determined by FRAP, DPPH and ABTS tests, respectively. Moreover, microbiological tests confirmed the durability of the bread with 15% CP and 10% DS of up to seven days. Therefore, carob pulp powder can be successfully applied in the production of bread whose characteristics are adapted to modern trends in nutrition.

1. Introduction

By studying the health and physiological effects of certain food ingredients, many research studies have shown a direct correlation between diet rich in foods with bioactive components and a reduced risk of chronic non-communicable diseases [1]. For this reason, the importance of plant foods has been emphasized more and more in recommendations for proper nutrition. Thus, lesser-known plant varieties are gaining popularity among scientists, since they are a source of vitamins, minerals, dietary fiber and other beneficial content, also known as functional components [2].
The greatest effects of the mentioned functional components can be achieved if they are included in the composition of foods that are consumed on a daily basis. Cereal-based products, primarily bread, as the most common foods in a proper and balanced diet, are therefore justifiably the focus of such research [3,4]. However, any change in bread formulation or production processes can affect the acceptability of the enriched product due to changes occurring in taste, texture and other attributes [5]. Since functional food can achieve the desired effects only when products with a modified composition meet the majority of consumers’ expectations [6], it is clear that the bakery industry is faced with a difficult and challenging task when it comes to the production of functional bakery products.
Due to the abundance of functional compounds present in the composition of carob (Ceratonia siliqua L.) pulp powder, this raw material, which is a by-product of the industrial processing of carob fruit for obtaining locust bean gum (LBG) from seed endosperm, rightly attracts the attention of the scientific community [7]. The presence of crude dietary fiber (up to 47%), the largest part of which are insoluble fibers, as well as minerals and polyphenolic compounds, makes carob pulp powder interesting from the point of view of research and manufacturers, as it provides an opportunity to obtain innovative products, which can largely satisfy new requirements of the consumers [8]. The beneficial health effects of phenolics are considered in the light of their potent antioxidant, anti-inflammatory, antimicrobial and immunomodulatory properties, and the consequent reduction in the risk of developing mass non-communicable diseases, as well as their potential therapeutic effect [9].
In terms of appearance and taste, carob pulp powder is similar to cocoa, so it is often used as its substitute [10]. Carob pulp powder is used in various beverages, such as liqueurs [11], kvass [12], Moroccan traditional carob drink [13] and carob juice [14], as well as in dairy products, such as yogurt [15], kefir [16] and carob-based milk beverage [17]. Carob jams and liqueurs are produced in Turkey, Malta, Portugal, Spain and Sicily. In Libya, syrup is extracted from carob and used to prepare a traditional dessert [7].
However, in scientific research, the most commonly investigated aspect is the possibility of adding carob pulp powder to various cereal-based products: muffins [18], cookies [19], biscuits [20], cake [21], tarhana [22] and pasta [23].
Although the inclusion of powder obtained from carob pulp in the composition of wheat bread has only been examined in a few scientific works [24,25,26,27], inevitable changes in the rheological properties of the formed dough were recorded, which caused a reduction in the quality and acceptability of the obtained product. For this reason, great attention must be paid to the preparation of raw materials and optimization of the raw material composition of functional bread with carob.
Literature data indicate that pretreatment of fiber-rich raw materials, such as prehydration, can reduce the structural effects of gluten dehydration caused by the presence of fibers during kneading and the processing of dough to a certain extent [28,29]. However, in some cases, to achieve the stated goal the use of additives is inevitable. Considering that it is a functional product, it is very important that the additive used is in the clean label domain. Therefore, the modern bakery industry resorts to the use of different types of sourdough, given that they have been shown to be effective in improving the properties and acceptability of rye flour bread or bread with the addition of non-traditional raw materials [30].
Based on all of the above, the aim of this research was the utilization of carob pulp powder for the production of functional bread with improved nutritional quality and microbiological stability. In order to develop bread with the addition of carob pulp powder, it was necessary to define its effects on dough rheology and bread quality. In order to overcome the impaired physical and sensory properties of bread with carob powder, dry sourdough was introduced to the bread formulation and the optimal amount of carob pulp powder and dry sourdough was determined. Finally, for bread samples of optimal quality, the main indicators of its functionality were examined, i.e., nutritional profile, mineral composition, content of polyphenols, antioxidant activity, microbial load and stability during storage.

2. Materials and Methods

2.1. Raw Materials

The carob pulp powder (CP) used in this work was a commercial product available on the market in Serbia, supplied by Vega ADM (Senta, Serbia). The ingredients used for the preparation of bread were wheat flour (moisture 11.8%, ash 0.55% d.w., protein 12.7% d.w. and wet gluten content 27%) supplied by the local mill (Kikindski mlin AD, Kikinda, Serbia), salt (Solana d.d., Tuzla, BiH) and bakers’ yeast (Lesaffre, Budapest, Hungary). As a baking additive, a commercial concentrated mixture with dry sourdough for special types of bread and pastries, “Pecipan soft” (DS), was used (Kvasac d.o.o., Prigorje Brdovečko, Croatia).

2.2. Chemicals

Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) was purchased from Sigma-Aldrich (Milano, Italy). The following chemicals were purchased from Sigma-Aldrich Chem (Steinheim, Germany): Folin–Ciocalteu reagent, gallic acid, (±)-catechin, 2,2-diphenyl-1-picrylhydrazyl (DPPH), TPZT (2,4,6-tris (2-pyridil)-s triazine), ABTS (2,2′-azino-bis-(-3-ethylbenzothiazoline-6-sulfonic acid) diammoniumsalt), iron (III)-chloride, iron (II)-sulfatheptahydrate and potassium persulfate. All the other chemicals used in this study were of an analytical grade.

2.3. Experimental Plan

The full factorial experimental design 32 was used in order to investigate the dependence of the relevant parameters of the dough and bread quality as a function of two independent variables (CP, 10–20% and DS, 0–10%) at three levels (Table 1). CP was introduced as a functional ingredient into the dough while a part of the wheat flour was substituted with a corresponding amount of CP. On the other hand, DS was introduced due to its known ability to improve bread quality, reflected in better technological and sensory characteristics of the bread [31]. In this context, DS had the role of an improver and was added to the total flour weight in corresponding amounts (Table 1).

2.4. Preparation of Carob Powder

Before its incorporation into the dough, the CP was pre-hydrated, meaning it was hydrated with a precisely defined volume of water for a defined time period. The aim of the prehydration was to reduce the negative effects of the fibers from the CP on the gluten hydration during mixing and was a part of the investigation within another study. The amount of water for prehydration was defined by determining the water-holding capacity (WHC) of the CP, according to the method used in [32]. The volume of water required for the prehydration of the CP was calculated as the product of the CP weight and WHC (2.65 g/g). The time required for the complete hydration of the CP was defined by monitoring the WHC of the CP in time periods from 15 to 90 min. Since for the examined time period there was no significant difference in the obtained WHC values, the shortest time (15 min) was used for the prehydration of the CP before mixing it with the wheat flour. A representation of the pre-hydrated and untreated CP is shown in Figure 1.

2.5. Rheological Assesment

The mixing properties of the dough samples with CP and DS, as well as those of the control dough (without CP and DS), were determined using a Brabender farinograph (Duisburg, Germany), while the extensional behavior of the mentioned samples was determined using a Brabender extensograph (Duisburg, Germany) according to the standard methods [33,34]. In both methods, of the total amount of water required for kneading, part of the water was used for the prehydration of the CP.

2.6. Bread Making

The raw material composition of the control sample was wheat flour (900 g), salt (2%, on flour basis) and yeast (2.5%, on flour basis). Other bread samples were formulated according to the previously presented experimental plan (Table 1). The amount of water for kneading was calculated based on the farinograph water absorption determined for each sample and reduced by 1.4% (correction for water content in yeast). In samples with CP, of the total amount of water required for kneading the dough, part of the water was used for the prehydration of the CP. The temperature of the water for kneading was 32 °C. The dough was mixed in a high-speed spiral mixer MS-6 for 4 min, and the prepared dough was left in a fermentation chamber at 30 °C for 60 min, with one kneading after 45 min. The fermented dough was then divided into 3 dough pieces of 450 g and was rounded by hand. The dough was left to relax for 10 min, after which it was shaped into loaves and inserted into Teflon molds, followed by final proofing at 30 °C and 80% RH. The optimally fermented dough was baked in a laboratory oven (Chopin, Villeneuve-la-Garenne, France) for 25–30 min at 220 °C, i.e., until a 10% weight loss was achieved. After cooling for 2 h, the obtained bread samples were packaged in polyethylene bags and stored in ambient conditions.

2.7. Specific Volume

The bread samples were weighed and the volume was measured by the rapeseed displacement method [35]. Specific volume was calculated as the ratio of the bread volume to its weight, and it was expressed in cm3/g.

2.8. Sensory Analysis

The evaluation of the sensory properties of the bread was performed according to method described by Šoronja-Simović et al., 2017 [36] with some modifications, and was carried out 24 h after baking by a six-member panel of trained evaluators in an adequate space [37,38,39]. The evaluated quality attributes were external appearance, crumb structure, smell and taste. Given that different attributes contribute differently to the overall sensory quality of bread, for each attribute an importance factor (IF) was assigned as follows: external appearance 4.0, crumb structure 6.0, smell 4.0 and taste grade 6.0. Individual sensory attributes were scored with grades (G) on a scale from 1 to 5 and, by multiplication of the obtained grade and the corresponding importance factor for the observed attribute, the number of points (P) for the particular attribute was obtained. Additionally, the sum of the points for all the examined sensory attributes of each bread sample was calculated to give the total score (TS). Based on the total score, the bread samples were classified into five quality categories: excellent (89.1–100.0), very good (78.1–89.0), good (67.1–78.0), acceptable (56.1–67.0) and unacceptable (≤56.0).

2.9. Chemical Characterization

2.9.1. Nutritional Value

The chemical composition of the bread was determined according to the methods described by the Association of Analytical Chemists (AOAC): moisture (No. 926.5), ash (No. 930.22), fat (No. 935.38), protein (No. 950.36) and total, soluble and insoluble dietary fiber (No. 958.29) [40]. The factor 6.25 was used for the conversion of nitrogen to protein content. The content of total and available carbohydrates was determined by calculation.
The energy value of the bread (EV) was calculated based on the results of the chemical composition (protein—P, carbohydrates—CH, fat—F, dietary fiber—DF) and data on the energy value of individual nutrients using the following equation [41]:
E V k J / 100   g = a v a i l a b l e   C H · 17 + P · 17 + F · 37 + D F · 8

2.9.2. Mineral Content

The content of calcium, copper, iron, magnesium, manganese, potassium and zinc in bread was determined according to the ISO standard method [42]. The method is based on dissolving the sample in hydrochloric acid after incineration at 550 ± 15 °C. A suitably diluted solution was sprayed in a mixture of air and acetylene gas onto an atomic absorption spectrometer (Varian spectra AA 10, Varian techtron pty limited, Melbourne, Australia). For irradiation, corresponding cathode lamps were used for the determination of Ca, Cu, Fe, Mg, Mn, K and Zn, and measurements were performed at sensitive wavelengths: 766.5 nm for K, 422.6 nm for Ca, 285.2 nm for Mg, 324.8 nm for Cu, 248.3 nm for Fe, 279.5 nm for Mn and 213.8 nm for Zn. The measured absorbance of each element was compared with the absorbance of calibration solutions for the same metal [43]. The result was given as a mean value of three measurements.

2.9.3. Bioactive Components

In order to determine the content of polyphenols and the antioxidant activity, active compounds were extracted from the bread according to the procedure described by Gumul et al. [44]. First, 3 g of the sample (bread crumb only/crust and crumb) was weighed into a glass cuvette and mixed with 15 mL of 80% (w/w) ethanol. The cuvettes were placed in a rotary thermostat at room temperature for 3 h. The samples were centrifuged for 10 min at 4000 rpm. The supernatant was separated by decantation, collected in dark vials and stored at 4 °C until analysis. The procedures for determination of the total phenols content (TP), total flavonoids content (TF) and antioxidant activity by DPPH, FRAP and ABTS assays were described in detail in our previous study [45]. All the measurements were conducted in triplicate and the results were expressed per 100 g of bread.

2.10. Microbiological Quality

2.10.1. Microbiological Stability

The microbiological stability of the bread was assessed by monitoring the microbial load of the investigated samples for 14 days. The samples were stored in ambient conditions (~23 °C and 40% RH) in polyethylene bags. Sampling was performed on 4 ± 1 days, during which 20 g of bread crumb was taken and homogenized with 180 mL of sterile saline for 15 min at 200 rpm (Unimax 1010, Heidolph, Schwabach, Germany). The prepared stock solution was analyzed by the determination of mesophilic bacteria on plate count agar (Merck, Darmstadt, Germany) incubated for 3 days at 30 °C [46]; Enterobacteriaceae on violet red bile glucose agar (Merck, Darmstadt, Germany) incubated for 1 day at 37 °C [47]; yeast and molds on Sabouraud maltose agar (Merck, Darmstadt, Germany) incubated for 5 days at 25 °C [48]; and Bacillus cereus on mannitol egg yolk polymyxin (Merck, Darmstadt, Germany) incubated for 1 day at 30 °C [49]. In order to determine the total number of sporogenic aerobic bacteria, 10 mL was taken from the basic solution and transferred into sterile test tubes. The tubes were placed in a water bath and the contents were heated to 95 °C for 5 min. From the treated stock solution, 1 mL was transferred to an empty Petri plate, and then the plate count agar was poured. The plates were incubated for 3 days at 30 °C [50]. After incubation, the colonies of mesophilic bacteria, Enterobacteriaceae, yeast and molds were counted and presented as a log CFU/g of the bread, while the results for B. cereus were presented as positive (when typical growth had been observed) or negative.

2.10.2. Inoculation Test

An inoculation test was conducted to assess the antimicrobial potential of CP as a natural preservative. This test is crucial for developing new products with raw materials that have potential antimicrobial properties since it allows monitoring of product changes during its shelf life after intentional inoculation with the target microorganism. Aspergillus flavus (isolated from corn flour) and Penicillium aurantiogriseum (isolated from whole wheat flour), both products of the Serbian market, were used for the preparation of spore suspensions. The total number of mold spores was 1.8 × 108 CFU/mL for A. flavus and 1.0 × 108 CFU/mL for P. aurantiogriseum. Then, two slices of approximately 30 g were cut from each bread sample under sterile conditions and placed in large sterile Petri dishes. Next, 20 µL of the prepared mold suspension was applied to the middle of the bread slice. The prepared samples were stored in ambient conditions, and mold growth was monitored for 14 days by measuring the mycelia diameter [51].

2.11. Statistical Analysis

Statistical analysis was performed using the STATISTICA 14.0.0.15 software (TIBCO Statistica™, Palo Alto, CA, USA). In the analysis of variance (one-way ANOVA), Duncan’s test was used for estimation of the significance of the differences between the mean values at p < 0.05 significance level.

3. Results and Discussion

3.1. Rheological Properties of Dough

The results of the farinographic parameters of the dough with the addition of CP and DS are shown in Table 2. Substitution of part of the wheat flour with CP did not have a statistically significant effect on the water absorption (p > 0.05). The registered differences were less than 1% and remained at the level of the control sample. According to the literature, the substitution of 20% of wheat flour with CP can cause both an increase and a decrease in water absorption by about 2.5% [24,25], which can be explained by the different composition of the carob powder used in the aforementioned research, and therefore its different hydration characteristics [52].
On the other hand, a significant increase in water absorption, by about 10%, was caused by the addition of DS. The mentioned results were expected, given that DS consists mostly of wheat flour and contains 10.4% protein and 75.9% carbohydrates (mainly starch), which are responsible for the absorption of water. In addition, higher water absorption values with the addition of DS can also be explained by the increase in gluten solubility due to the increase in dough acidity and the reduction in the disulfide bonds of gluten proteins by heterofermentative lactic acid bacteria (LAB) [53]. The addition of DS (based on total flour weight) requires a larger amount of water to achieve optimal consistency, which contributes to a higher yield of dough, and further to a higher yield of bread.
It can also be seen that the substitution of wheat flour with CP up to 15% did not drastically affect the development time compared to the control sample. However, in the samples with the maximum amount of CP, the dough development time was extended 3.6 times compared with the control (Table 2). Data from the literature show that the addition of CP can prolong the dough development time to a much greater extent, from 8 to 11 min for samples with 10–20% CP [25,26]. It is assumed that the development time for the dough with CP in our study is lower compared to the literature data due to the applied prehydration of the carob powder. The above results are in agreement with the results of Nawrocka et al. [54] that prehydration of the fiber from carob powder shortens dough development and reduces the interaction between proteins and fibers, which makes it more resistant to mechanical damage during mixing. The mentioned research indicated that the substitution of part of the flour with prehydrated carob fibers caused mainly the formation of hydrated/extended β-sheets in the gluten structure, in contrast to the aggregates and pseudo-β-sheets that were present if the pretreatment was omitted. Therefore, the obtained results along with the literature suggest that the prehydration of CP can reduce the negative effects of carob powder constituents on the formation of the gluten network. Nevertheless, a statistically significant increase in development time (p < 0.05) in the sample with the maximum amount of CP flour can be explained by the effect of “gluten dilution” that occurred as a result of replacing a larger amount of wheat flour with CP [55], so it is assumed that with a higher proportion of carob powder rich in insoluble fibers, primarily cellulose, structural and conformational changes occur in the gluten network. In addition, CP is rich in polyphenols, which are able to interact with gluten by forming covalent or hydrogen bonds between protein SH groups and polyphenolic OH groups and thus can affect the redistribution of water and dehydration of gluten during mixing [56]. Also, the presented results (Table 2) indicate that the addition of DS caused a significant increase in the dough development time. The mentioned effect is more pronounced in dough samples with 10 and 15% CP, where the development time is 4.5 to 5.0 times longer compared to the samples without DS.
When it comes to the stability of the dough, it can generally be said that the addition of CP had a positive effect on stability. Compared to the stability of the control dough (0.5 min), in all samples there is an increase in stability by a factor of two to four. However, considering that viscoelastic properties are evaluated based on both the stability and the degree of softening, for a more realistic picture of the contribution of the mentioned increase in stability, the values of the degree of softening must also be considered. The results shown in Table 2 indicate that the addition of CP causes an increase in the degree of dough softening by about 80%. In practice, the higher stability and lower degree of softening indicate an improvement in the technological characteristics of the dough [55]. Therefore, the obtained results suggest that the dough with CP would not be able to withstand intensive mechanical processing during production. However, in this aspect, the use of DS in the production of bread with CP was justified, since there was a significant increase in the dough stability by 50% and 100%, but also a simultaneous decrease in the degree of softening by 40 to 50% when DS was added to the dough mixture with up to 15% CP (Table 2). A positive effect of DS on the stability of samples with 20% CP was absent, but the degree of softening with 5 and 10% DS was lower by about 50% compared to samples with a corresponding amount of CP without DS.
Based on the results obtained by extensographic measurements, it is evident that substitution of a part of the wheat flour with CP has a significant negative effect on all the parameters. The dough with 10% CP had a resistance two times higher than the control sample (315 EU), while increasing the amount of carob powder from 10 to 15 and 20% increased the resistance by an additional 25% (Figure 2). The sample with 20% CP had a resistance value almost three times higher than the control sample, which was also the highest registered value of the mentioned parameter. The observed effect is in agreement with the literature, although the level of recorded values for the resistance of dough with carob powder differs among the published data [25,26,57,58]. The mentioned effect can be explained by the interactions that occur between wheat proteins and the structural components of CP, primarily proteins and fibers, which lead to gluten strengthening [59,60]. However, the addition of DS to the dough with CP caused the opposite effect, where a significant decrease in the initially increased dough resistance was observed. Regardless of the amount of CP in the dough, the addition of 10% DS led to a decrease in resistance to an approximate value of 540 EU.
Even though the extensibility of the control sample was slightly above the value for standard quality flour, the addition of the CP resulted in a significant decrease in the mentioned parameter (Figure 2). The highest investigated amount of CP led to a decrease in extensibility by up to two times compared to the control. The obtained values confirmed the cause-and-effect relationship between dough resistance and extensibility. Based on the experimental results, as well as the literature data, it can be said that the incorporation of CP into wheat dough does not lead to gluten strengthening as much as to an increase in the stiffness of its structure, which results in less flexible dough and would inevitably cause a deterioration in the quality of the finished product [26]. In addition, when substituting wheat flour with raw materials rich in dietary fiber, the decrease in extensibility is also a consequence of the decrease in gluten content, which enables the dough to stretch during deformation [61].
On the other hand, the addition of DS increased the extensibility of the dough, but only to a small extent (Figure 2). The smallest changes were observed in the samples with 10% CP, while the mentioned effect was more pronounced in samples with 15 and 20% CP. The positive effect of DS addition on dough extensibility may be due to a decrease in pH value during sourdough fermentation, which contributes to an increase in gluten elasticity [62]. The mentioned effect coupled with the influence of sourdough on proteolysis leads to a “loosening” of the gluten structure, which was also observed visually during the analysis (Figure 3).
The energy needed for dough stretching is interpreted as an indicator of flour quality, i.e., the dough quality and the qualitative characteristics of the bread. It is considered that optimal rheological properties, good dough workability and consequently a large, well-shaped product can be achieved when energy values range between 80 and 110 cm2 [63]. In Figure 4, it is shown that the samples with 10 and 15% CP without DS have higher energy compared to the control dough. Although high energy of dough is highly desirable in breadmaking, the ratio of resistance to extensibility (R/E ratio) is also very important. The R/E ratio of the control sample was in the range of optimal values for bakery needs from 1.5 to 2.5 [55,64]. A significant increase in this parameter was recorded in all the other samples. The dough with 20% CP had the highest R/E ratio, six times higher than the control sample.
From the aspect of the energy value and the R/E ratio, a significant impact of the addition of DS was recorded. With an increase in the proportion of DS, there was a decrease in both the energy and R/E ratio of the dough with CP (Figure 4). The greatest impact of DS on the reduction in energy and the R/E ratio was registered in the samples with 15% CP. The results closest to the optimal values were obtained in the samples with 10 and 15% CP and with 5 and 10% DS, and it is predicted that those formulations could ensure the appropriate stability of the dough during fermentation with the ability to retain formed gases, and thus achieve the appropriate quality of bread.

3.2. Physical and Sensory Quality of Bread

Among the physical properties, the most important quality indicator for bread is its volume. A prerequisite for obtaining large-volume bread is suitable rheological properties of the bread dough. The obtained results for the specific volume of the bread with CP are shown in Figure 5. By comparing the specific volume of the samples with 10% CP and the control, it is noticeable that there is no statistically significant difference in the values of the mentioned parameter (p < 0.05). However, replacing wheat flour with 15 and 20% CP significantly reduced the specific volume of the bread compared to the control bread. Also, in the samples with 15 and 20% CP, there is a noticeable increase in the specific volume of the bread with the addition of DS, whereby the stated effect is statistically significant only in the samples with 15% CP (Figure 5). The presented results confirmed the predictions made on the basis of the results obtained by the rheological measurements. Namely, increasing the amount of CP reduced the extensibility and increased the resistance of the dough, prevented the expansion of gas bubbles during fermentation and ultimately resulted in a smaller product volume. The obtained results are also consistent with the registered positive effect of DS on the behavior of dough with CP, reaffirming the justification of its use in this specific bread formulation.
The sensory quality of the product is of crucial importance for the consumer when choosing and deciding on a certain type of product. For the development of functional products, whose characteristics often deviate from traditional products, it is extremely important that the changes in the parameters that define the sensory quality are within acceptable limits. This ensures its acceptability in the wider population and increases the intake of functional components in the daily diet.
The sensory quality of bread was tested using the scoring method, where the basic attributes of the product were evaluated; the obtained results are shown in Table 3 and the bread samples are presented in Figure 6. From the presented results, it can be seen that the only parameter on which the addition of CP and DS had no effect is the smell. Smell referred to the presence or absence of a scent characteristic for a given product and/or used raw materials. The smell of all the tested samples was described as characteristic, pleasant and rounded.
The external appearance of the control bread had the maximum score, while in the samples with the addition of 10% and 15% CP there was a slight decrease in the number of points for the external appearance of the bread. The addition of the maximum tested amount of CP had a negative effect on the external appearance, which resulted in a significant decrease in the total score (Table 3, Figure 6). In general, the addition of DS to the bread with CP did not improve the external appearance of the product.
The greatest importance factor among the tested sensory attributes was assigned to the taste and crumb structure of the bread and unequivocal trends were observed. Namely, for both parameters, there was a decrease in the obtained points with an increase in the amount of CP, while the reverse trend was achieved by including DS in the bread formulation (Table 3). When it comes to the taste of bread, half of the observed samples (with 10 and 15% CP) had the same or a higher number of points than the control bread (24.0). The exception is sample CP15DS0, where the number of points for taste was 25–40% lower compared to the previously mentioned samples. However, among the samples with 15% CP, another exception was the sample with 10% DS, which was characterized by good melting in the mouth, and therefore was easy to chew. A more pronounced negative influence of the carob powder on the taste of the bread was evident in the samples with 20% CP. Although the addition of DS to the bread with 20% CP had a positive effect on the taste of the product (increasing the number of points from 12.0 to 18.0), the sample had an undesirable doughy taste and smearing in the mouth during chewing was present.
As for the crumb structure, the samples with 10, 15 and 20% CP and without DS obtained significantly lower grades, which resulted in a decrease in points by 12, 25 and 37% (Table 3, Figure 6). Furthermore, the presented results indicate that the structure of bread with CP can be significantly improved by adding only a minimum amount of DS. It is important to point out that the compact, rather crumbly, underdeveloped structure of the bread with 20% CP was visibly improved by the addition of DS, which confirms the increase in the score for the structure by 20 and 33% (Table 3).
Based on the results for total score, it can be seen that with the increase in the proportion of CP, the quality of bread deteriorates, while the addition of DS, regardless of the applied amount, contributes to a positive change in the quality category compared to samples without sourdough. Thus, the samples with 10 and 15% CP with DS are in the excellent and very good category, which is at the level of the quality of the control sample or one category higher. The sample with 20% CP without DS had the worst sensory quality and belonged to the acceptable quality category.
Finally, the optimal raw material composition for the bread was chosen with the aim of achieving a compromise between the addition of the maximum possible amount of functional ingredients and the deterioration of the physical and sensory characteristics of the product. A sample that would fit within the specified limits was obtained by replacing a part of the wheat flour with carob powder at the level of 15% and adding the maximum amount of dry sourdough (10%).

3.3. Nutrients and Bioactive Compounds in Bread

The bread sample with optimal raw material composition (CP15DS10) was further characterized and compared to the control sample (CP0DS0). First, the nutritional composition and mineral content was analyzed; the obtained results are presented in Table 4. From the results shown, it is evident that the energy value of the examined samples was similar, since the composition of the main nutrients was quite uniform. The moisture content of the carob bread is 1.2% lower compared to the white bread. Also, only small differences were observed in the fat content. Although bread belongs to products with a low fat content, in the carob bread, the value of this parameter was even lower than in the white bread. On the other hand, the protein content in the carob bread was slightly higher compared to the control sample. Carbohydrates make up the largest share in the composition of bread, which is expected given that starch is dominantly present in wheat flour [65]. The difference in the content of total carbohydrates in the examined samples is negligible and amounts to about 55%; however, significant differences occur when looking at the content of available carbohydrates (starch, sugars) and dietary fiber. In the carob bread, there was a significant increase in the content of total dietary fiber from 1.27 to 6.48%, where 50% was soluble and 50% insoluble fiber (Table 4). According to the results shown and the Regulation of the European Parliament and of the council on nutrition and health claims made about foods [66], the bread made with 15% carob powder can carry the nutritional claim “rich in fiber”, which can be applied to products containing at least 6 g of dietary fiber per 100 g of product.
In addition, it can be observed that in the carob bread there was an increase in ash content compared to the white bread, which indicates an increase in the content of minerals. The detailed mineral composition of the carob and white bread is shown in Table 5. Based on the obtained results, the content of all the tested macro minerals (K, Ca and Mg) in carob bread was two times higher than the control sample. When looking at the content of micro minerals (Fe, Zn, Mn and Cu), the differences between the samples were not so obvious; however, they were significant. Namely, in the carob bread, the content of Fe, Mn and Cu was higher compared to the white bread. The obtained results were expected considering that carob powder is rich in minerals, which was previously shown in several scientific works [67,68,69]. However, the determination of mineral content in bread with carob powder was not found in the literature. Babiker et al. [70] examined the mineral composition of biscuits with different proportions of carob powder obtained by grinding whole carob fruit. Their results confirmed that the addition of carob increases the content of most of the investigated minerals in biscuits, which is in agreement with the results of our study.
Furthermore, Table 5 shows the Nutritional Reference Values (NRVs) and the value 15% NRV, which is defined as a Significant Amount of minerals. Those are a set of numerical values that are used for the purposes of nutrition labelling and relevant claims. Accordingly, the nutritional claim “source of minerals” is allowed when the mineral content in a food product contains at least a Significant Amount of minerals (15% NRV per 100 g of bread), while for the nutritional claim “high in minerals”, twice the Significant Amount of these nutrients is required [66]. The obtained amounts of minerals in the carob bread were sufficient for the use of the nutritional claim “rich in manganese” and “source of copper”. Provided that the legal conditions prescribed by the European Parliament and council [66] are met, in addition to the mentioned nutritional claims, carob bread can be a potential holder of eight health claims related to copper and four health claims related to manganese, which certainly confirms its functional characteristics.
Furthermore, the polyphenol content and antioxidant activity of the carob and white bread were determined and compared. According to the results shown in Table 6, the content of total phenols (TPs) in the carob bread was about 10 times higher than the control sample. An even more pronounced difference between the tested bread samples was confirmed by antioxidant tests. The carob bread had about a 5.5 times greater ability to reduce Fe3+ ions (FRAP test) and about a 12 times greater ability to neutralize DPPH radicals, while the ability to neutralize ABTS radicals in the mentioned bread was 26 times higher than the corresponding antioxidant activities of the white bread.
However, when it comes to the content of total flavonoids (TFs), it is evident that the obtained results are unrealistic. Namely, the registered content of TFs is higher than the content of TPs, which is illogical considering that flavonoids are a subgroup of phenols [71]. In addition, the values obtained for the content of total flavonoids in the control sample are initially high and relatively close to the values obtained for the sample with carob powder, which indicates that when performing the mentioned spectrophotometric analysis of bread samples, regardless of the type of additive or raw material composition, certain interferences occur. Since the TF content was not examined in any of the scientific publications dealing with the topic of bread with carob powder, it was not possible to compare the experimentally obtained values with other research. There are some studies investigating bread with different raw materials that also showed a higher content of TFs compared to TPs [72,73,74]; however, this phenomenon was not clarified or discussed.
It is important to point out that a higher content of phenolic compounds and a higher antioxidant activity were registered in the whole bread (crust and crumb) compared to the bread crumb only, especially in the case of the carob bread (Table 6). The obtained results are in agreement with the literature data, where it was shown that the mentioned increase in antioxidant activity in the bread crust occurs due to the formation of Maillard reaction products [75]. Although in the past the Maillard reaction products were characterized as anti-nutritive and toxic, the above results confirm that they can also be potential carriers of antioxidant activity in food products [76].

3.4. Microbiological Quality of Bread

In order to examine the effects of the addition of carob powder on the microbiological quality of bread, the total number of bacteria, spore-forming bacteria, Enterobacteriaceae, molds and yeasts, as well as the presence of Bacillus cereus, was determined. The microbiological quality of the examined samples was monitored for 14 days, and the results are shown in Table 7. During the 14 days of testing, the presence of B. cereus was not detected and the total number of Enterobacteriaceae and yeasts was <1.0 log CFU/g for both samples.
It can be observed that on the fourth day, the total bacterial count (TBC) in the control sample was >5 log CFU/g, while in the carob bread there was no change in the TBC (Table 7). During the first seven days, the difference between the samples was the most pronounced—carob bread had about a 4 and 3 log CFU/g lower TBC compared to the white bread. In the second week of monitoring, the mentioned differences decreased and were around 1 log CFU/g (day 11) and <1 log CFU/g (day 14).
The total spore count (TSC) in the white bread from the fourth day ranged from 2.3 to 2.9 log CFU/g. The changes in the total spore count (TSC) during the sampling period were not consistent. However, the obtained results mainly indicate that the carob bread had lower TSC than the control bread until day eleven. The presence of molds in the carob bread was not detected during the fourteen days of monitoring, while in the control bread their presence was detected on the first and fourth day (Table 7).
According to guidelines for the microbiological quality of certain groups of food products, it is required that the total count of aerobic bacteria in bakery products should not exceed <105 CFU/g, and total yeast and molds count should be <104 CFU/g [77,78]. Based on the mentioned prescribed values, the microbiological quality of the white bread after four days was unsatisfactory, while the quality of carob bread was within the acceptable limits for eleven days.
In order to obtain a more complete picture of the antimicrobial potential of carob powder and to examine the effects of its addition on mold growth and development in bread, an inoculation test was performed. The application of this microbiological test is especially important when developing new products with the addition of raw materials that have a certain antimicrobial potential [79]. The molds Aspergillus flavus and Penicillium aurantiogriseum, which were used to perform this test, are among the most common contaminants of basic ingredients for the production of bakery products and are the main cause of their spoiling [80].
Figure 7 shows the results of the inoculation test, which indicate that there was no visible growth of A. flavus during the first three days of testing. From day four to day eleven, the mycelia diameter of A. flavus in the carob bread was lower compared to the white bread. From the eighth to the thirteenth day, the growth stagnation of A. flavus on the white bread sample was observed, with a diameter of 50 mm. Similarly, in the carob bread, the mycelia diameter was constant, but only for three days, after which there was an increase in the observed parameter, and the registered values were higher compared to the control sample.
The effect of carob powder addition to the bread on the growth of P. aurantiogriseum was less pronounced compared to the growth of A. flavus (Figure 8). The first visible colony growth, as in the case of A. flavus inoculation, was recorded only on the fourth day. However, during the first six days of monitoring, there was no difference in the mycelia diameter between the examined bread samples. During the next six days, the mycelia diameter of P. aurantiogriseum on the carob bread was smaller compared to the mycelia on the white bread. Stagnation of the mycelia growth was observed from day nine to day twelve for both samples. After that, there was a sudden increase in the observed parameter, where, unlike A. flavus, there was no difference in the mycelia diameter of P. aurantiogriseum between the observed samples.
Based on the obtained results, it can be concluded that carob powder contributes to the improvement in the microbiological quality of the bread and due to the presence of antimicrobial components prolongs the durability of the product up to 7 days.

4. Conclusions

The addition of carob pulp powder (CP) negatively affects dough rheology, impairing bread’s physical and sensory properties. Prehydrating the CP for 15 min overcame these effects in samples with 10–15% CP. Dry sourdough (DS) addition improved dough resistance and extensibility, resulting in higher specific volume, well-developed crumb structure and enhanced sensory properties of the samples with CP. The best bread quality was achieved by combining 15% CP and 10% DS in the bread formulation, showing a specific volume of 2.82 cm3/g and a sensory score of 86.0 (max. 100), placing it into the quality class “very good”.
The supplementation of wheat flour with 15% CP also improved the nutritional value by increasing the fiber content to 6.48 g/100 g, supporting the “rich in fiber” claim. It also increased the mineral content, especially manganese and copper, and significantly increased antioxidant activity compared to the white bread. Microbiological tests confirmed the CP extended the bread’s freshness and durability for up to seven days, so that it can be labeled as bread with an extended shelf life. Therefore, CP can be successfully used in functional bread development, aligning with modern dietary trends.

Author Contributions

Conceptualization, D.Š.-S. and S.K.-T.; methodology, B.P. and S.B.; validation, J.P. and Z.Š.; formal analysis, L.P., D.B. and N.M.; investigation, J.Z.; data curation, B.P.; writing—original draft preparation, J.P. and J.Z.; writing—review and editing, D.Š.-S., S.K.-T. and B.P.; visualization, J.Z.; supervision, Z.Š. and D.Š.-S. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Ministry of Science, Technological Development and Innovation of the Republic Serbia, No. 451-03-66/2024-03/200134 and No. 451-03-65/2024-03/200134.

Data Availability Statement

Data are contained within the article.

Acknowledgments

The authors would like to thank Peter Raspor and Sonja Smole Možina (University of Ljubljana, Slovenia) for the original idea from which this research derived and therefore acknowledge their contribution.

Conflicts of Interest

The authors declare no conflicts of interest.

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Figure 1. Carob pulp powder (a) untreated and (b) pre-hydrated.
Figure 1. Carob pulp powder (a) untreated and (b) pre-hydrated.
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Figure 2. Influence of carob pulp powder (CP) and dry sourdough (DS) addition on (a) resistance and (b) extensibility.
Figure 2. Influence of carob pulp powder (CP) and dry sourdough (DS) addition on (a) resistance and (b) extensibility.
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Figure 3. Dough sample with 15% carob pulp powder for extensographic testing after 45 min rest period: (a) with 0% of dry sourdough; (b) with 10% of dry sourdough.
Figure 3. Dough sample with 15% carob pulp powder for extensographic testing after 45 min rest period: (a) with 0% of dry sourdough; (b) with 10% of dry sourdough.
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Figure 4. Influence of carob pulp powder (CP) and dry sourdough (DS) on energy and the ratio of resistance to extensibility (R/E ratio).
Figure 4. Influence of carob pulp powder (CP) and dry sourdough (DS) on energy and the ratio of resistance to extensibility (R/E ratio).
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Figure 5. Specific volume of bread as a function of the addition of carob pulp powder (CP) and dry sourdough (DS); results in bars marked with different lowercase letters are significantly different (p < 0.05).
Figure 5. Specific volume of bread as a function of the addition of carob pulp powder (CP) and dry sourdough (DS); results in bars marked with different lowercase letters are significantly different (p < 0.05).
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Figure 6. External appearance and crumb structure of bread samples with carob pulp powder (CP) and dry sourdough (DS).
Figure 6. External appearance and crumb structure of bread samples with carob pulp powder (CP) and dry sourdough (DS).
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Figure 7. Dependence of radial growth of Aspergillus flavus on bread composition.
Figure 7. Dependence of radial growth of Aspergillus flavus on bread composition.
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Figure 8. Dependence of radial growth of Penicillium aurantiogriseum on bread composition.
Figure 8. Dependence of radial growth of Penicillium aurantiogriseum on bread composition.
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Table 1. Experimental plan with two independent variables at three levels.
Table 1. Experimental plan with two independent variables at three levels.
Independent Variables 1Levels
−101
CP (%, g/100 g of wheat flour)101520
DS (%, g/100 g of wheat flour)0510
1 CP—carob pulp powder; DS—dry sourdough.
Table 2. The influence of independent variables on the farinographic parameters of dough.
Table 2. The influence of independent variables on the farinographic parameters of dough.
Sample 1Water Absorption (%)Development
Time (min)		Stability
(min)		Degree of Softening (FU)
CP0DS0 258.3 ± 0.20 a1.5 ± 0.17 a0.5 ± 0.30 a60 ± 1.0 e
CP10DS058.7 ± 0.25 a1.8 ± 0.16 a1.0 ± 0.05 b110 ± 0.0 g
CP10DS563.8 ± 0.17 b8.5 ± 0.14 c2.0 ± 0.01 d55 ± 1.5 d
CP10DS1068.3 ± 0.43 d9.0 ± 0.17 d2.0 ± 0.00 d45 ± 1.0 b
CP15DS058.5 ± 0.14 a2.0 ± 0.15 a1.0 ± 0.01 b105 ± 0.5 f
CP15DS565.1 ± 0.17 c9.0 ± 0.10 d1.5 ± 0.07 c55 ± 0.5 d
CP15DS1069.0 ± 0.19 d8.5 ± 0.12 c2.0 ± 0.15 d50 ± 0.5 c
CP20DS059.2 ± 0.16 a5.5 ± 0.11 b1.5 ± 0.02 c110 ± 0.5 g
CP20DS564.9 ± 0.15 b9.0 ± 0.12 d1.5 ± 0.07 c40 ± 1.0 a
CP20DS1069.7 ± 0.11 e9.2 ± 0.16 d1.5 ± 0.05 c55 ± 1.5 d
1 CP—carob pulp powder; DS—dry sourdough; 2 control sample. Results represent mean values of repeated measurements ± SD (n = 3); values in columns marked with different letters are significantly different (p < 0.05).
Table 3. Results of sensory analysis of bread with the addition of carob pulp powder and dry sourdough.
Table 3. Results of sensory analysis of bread with the addition of carob pulp powder and dry sourdough.
Sample 1External AppearanceCrumb StructureSmellTasteTotal ScoreQuality Category
CP0DS0 220.0 ± 0.0 a24.0 ± 0.6 a20.0 ± 0.0 a24.0 ± 0.6 a88.0Very good
CP10DS018.0 ± 0.5 b21.0 ± 0.5 b20.0 ± 0.0 a24.0 ± 0.0 a83.0Very good
CP10DS520.0 ± 0.0 a27.0 ± 0.5 c20.0 ± 0.0 a30.0 ± 0.0 b97.0Excellent
CP10DS1018.0 ± 0.5 b24.0 ± 0.6 a20.0 ± 0.0 a30.0 ± 0.0 b92.0Excellent
CP15DS018.0 ± 0.0 b18.0 ± 0.6 d20.0 ± 0.0 a18.0 ± 0.6 c74.0Good
CP15DS518.0 ± 0.6 b21.0 ± 0.5 b20.0 ± 0.0 a24.0 ± 0.6 a83.0Very good
CP15DS1018.0 ± 0.6 b21.0 ± 0.8 b20.0 ± 0.0 a27.0 ± 0.5 d86.0Very good
CP20DS012.0 ± 0.0 c15.0 ± 0.5 e20.0 ± 0.0 a12.0 ± 0.0 e59.0Acceptable
CP20DS512.0 ± 0.0 c18.0 ± 0.0 d20.0 ± 0.0 a18.0 ± 0.0 c68.0Good
CP20DS1012.0 ± 0.0 c20.0 ± 0.5 b20.0 ± 0.0 a18.0 ± 0.6 c71.0Good
1 CP—carob pulp powder; DS—dry sourdough; 2 control sample. External appearance and smell—max 20.0; Crumb structure and taste—max 30.0; total score—max 100.0. Results represent mean values of repeated measurements ± SD (n = 6); values in column marked with different letters are significantly different (p < 0.05).
Table 4. Nutritional composition and mineral content of bread with 15% carob pulp powder and 10% dry sourdough compared to the control sample.
Table 4. Nutritional composition and mineral content of bread with 15% carob pulp powder and 10% dry sourdough compared to the control sample.
Nutrients (g/100 g)CP0DS0CP15DS10
Moisture34.60 ± 0.05 b33.80 ± 0.07 a
Ash0.51 ± 0.01 a0.76 ± 0.01 b
Fat0.25 ± 0.12 b0.18 ± 0.09 a
Proteins9.47 ± 0.05 a10.02 ± 0.06 b
Available carbohydrates53.90 ± 0.19 b48.76 ± 0.15 b
Total dietary fiber1.27 ± 0.11 a6.48 ± 0.08 b
Insoluble dietary fiber0.51 ± 0.18 a3.25 ± 0.23 b
Soluble dietary fiber0.76 ± 0.14 a3.23 ± 0.17 b
Energy value (kJ/100 g)
(kcal/100 g)		1096.70
258.27		1057.76
249.70
Results represent mean values of repeated measurements ± SD (n = 3); values in rows marked with different letters are significantly different (p < 0.05).
Table 5. Mineral content of bread with 15% carob pulp powder and 10% dry sourdough compared to the control sample.
Table 5. Mineral content of bread with 15% carob pulp powder and 10% dry sourdough compared to the control sample.
MineralsNRV 1 (mg)15% NRV 2
(mg/100 g)		CP0DS0 (mg/100 g)CP15DS10 (mg/100 g)
K2000300130.63 ± 24.27 a247.44 ± 12.52 b
Ca80012026.54 ± 2.11 a62.17 ± 4.59 b
Mg37553.2517.04 ± 0.90 a32.83 ± 1.17 b
Fe142.101.43 ± 0.24 a1.95 ± 0.53 a
Zn101.501.67 ± 0.55 a1.39 ± 0.35 a
Mn20.300.33 ± 0.01 a0.76 ± 0.01 b
Cu10.150.18 ± 0.00 a0.21 ± 0.00 b
1 NRV—Nutrient Reference Values; 2 15% NRV—Significant Amount of minerals (EU Regulation 1169/2011). Results represent mean values of repeated measurements ± SD (n = 3); values in rows marked with different letters are significantly different (p < 0.05).
Table 6. Polyphenol content and antioxidant activity of bread with 15% carob pulp powder and 10% dry sourdough compared to the control sample.
Table 6. Polyphenol content and antioxidant activity of bread with 15% carob pulp powder and 10% dry sourdough compared to the control sample.
Sample 1TP
(mg GAE/100 g)		TF
(mg CE/100 g)		DPPH
(µM TE/100 g)		FRAP
(µM Fe2+/100 g)		ABTS
(µM TE/100 g)
CP0DS0C11.52 ± 0.45 a38.29 ± 1.76 a32.92 ± 4.32 a154.95 ± 6.69 a17.09 ± 9.08 a
CP0DS0W12.67 ± 0.88 a45.45 ± 1.09 b40.85 ± 1.14 b157.66 ± 4.71 a27.40 ± 9.10 a
CP15DS10C117.77 ± 2.71 b51.78 ± 0.99 c402.69 ± 16.24 c857.53 ± 35.37 b450.04 ± 33.73 b
CP15DS10W140.55 ± 8.41 c60.14 ± 2.93 d498.15 ± 10.06 d1260.40 ± 57.95 c727.35 ± 80.57 c
1 C—crumb; W–whole bread. Results represent mean values of repeated measurements ± SD (n = 3); values in columns marked with different letters are significantly different (p < 0.05).
Table 7. Microbiological profile of bread with 15% carob pulp powder and 10% dry sourdough and control sample during 14 days of storage (~23 °C and 40% RH).
Table 7. Microbiological profile of bread with 15% carob pulp powder and 10% dry sourdough and control sample during 14 days of storage (~23 °C and 40% RH).
DayTBC (log CFU/g)TSC (log CFU/g)TMC (log CFU/g)
CP0DS0CP15DS10CP0DS0CP15DS10CP0DS0CP15DS10
1<1.0 a<1.0 a<1.0 a<1.0 a1.00 ± 0.00 a<1.0 a
45.30 ± 0.09 b<1.0 a2.34 ± 0.00 b1.30 ± 0.00 a2.57 ± 0.11 b<1.0 a
75.21 ± 0.03 b2.45 ± 0.18 a<1.0 a<1.0 a<1.0 a<1.0 a
115.73 ± 0.09 b4.66 ± 0.04 a2.98 ± 0.02 a3.22 ± 0.53 b<1.0 a<1.0 a
145.80 ± 0.00 b5.50 ± 0.00 a2.59 ± 0.00 b<1.0 a<1.0 a<1.0 a
TBC—total bacterial count; TSC—total spore count; TMC—total mold count; CFU—colony forming unit. Results represent mean values of repeated measurements ± SD (n = 3); values in rows within individual parameters marked with different letters are significantly different (p < 0.05).
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Zahorec, J.; Šoronja-Simović, D.; Petrović, J.; Šereš, Z.; Pavlić, B.; Božović, D.; Perović, L.; Martić, N.; Bulut, S.; Kocić-Tanackov, S. Development of Functional Bread: Exploring the Nutritional, Bioactive and Microbial Potential of Carob (Ceratonia siliqua L.) Pulp Powder. Processes 2024, 12, 2882. https://doi.org/10.3390/pr12122882

AMA Style

Zahorec J, Šoronja-Simović D, Petrović J, Šereš Z, Pavlić B, Božović D, Perović L, Martić N, Bulut S, Kocić-Tanackov S. Development of Functional Bread: Exploring the Nutritional, Bioactive and Microbial Potential of Carob (Ceratonia siliqua L.) Pulp Powder. Processes. 2024; 12(12):2882. https://doi.org/10.3390/pr12122882

Chicago/Turabian Style

Zahorec, Jana, Dragana Šoronja-Simović, Jovana Petrović, Zita Šereš, Branimir Pavlić, Danica Božović, Lidija Perović, Nikola Martić, Sandra Bulut, and Sunčica Kocić-Tanackov. 2024. "Development of Functional Bread: Exploring the Nutritional, Bioactive and Microbial Potential of Carob (Ceratonia siliqua L.) Pulp Powder" Processes 12, no. 12: 2882. https://doi.org/10.3390/pr12122882

APA Style

Zahorec, J., Šoronja-Simović, D., Petrović, J., Šereš, Z., Pavlić, B., Božović, D., Perović, L., Martić, N., Bulut, S., & Kocić-Tanackov, S. (2024). Development of Functional Bread: Exploring the Nutritional, Bioactive and Microbial Potential of Carob (Ceratonia siliqua L.) Pulp Powder. Processes, 12(12), 2882. https://doi.org/10.3390/pr12122882

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