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Journal of Food Science and Technology logoLink to Journal of Food Science and Technology
. 2023 Dec 12;61(5):969–982. doi: 10.1007/s13197-023-05893-y

Exploring the biofunctionalities of lactic fermented cactus pear (Opuntia elatior Mill.) fruit beverage: an exotic superfood

Mital R Kathiriya 1, Yogesh Vekariya 1, Subrota Hati 1,✉
PMCID: PMC10933228  PMID: 38487287

Abstract

Cactus pear fruit is known with many health benefits in ethnomedicine of countries like Mexico, Portugal, Chine, India etc. The study was aimed to develop biofunctional lactic fermented cactus pear fruit beverage to add values to the medicinal fruit. The processing parameters such as quantity of freeze dried cactus pear fruit powder, sucrose and incubation time were optimised using response surface methodology. The optimized product was then subjected to proximate compositional, physicochemical, biofunctional and microbial analysis. The lactic fermented cactus pear fruit beverage was prepared by mixing 12% [w/v] freeze dried cactus pear fruit powder and 3% sucrose in water, then pasteurised and inoculated with 3% Lactobacillus fermentum MTCC 25515 and Lactobacillus rhamnosus M9, then incubated at 37 °C for 6 h. The moisture content of the beverage was 87.77% and major constituent was carbohydrate (9.58% per wet matter basis). The 100 mL beverage contains 89.84 mg GAE phenolic compounds, 5.86 mg QE flavonoids, 71.82 mg betacyanin, 28.08 mg betaxanthin, 10.59 mg ascorbic acid. The beverage also exhibited 58% ABTS antioxidant activity. The beverage was shelf stable for 20 days at 7 ± 1 °C. Such a biofunctional beverage loaded with antioxidant potential can be consumed as refreshing drink.

Graphical abstract

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Keywords: Cactus pear fruit beverage, Lactic fermentation, Biofunctional, Antioxidant activity, Lactobacillus, Ascorbic acid

Introduction

Cactus pear is also called as prickly pear, member of the family Cactaceae, genus Opuntia. It was used for centuries by ancient civilizations to cure diseases and heal wounds. The fruit is available in different color like purple-red, orange-yellow, green (Scarano et al. 2020). In India, mainly three species are found i.e. Opuntia dillenii Haw. in the south, Opuntia vulgaris Mill. in the north and Opuntia elatior Mill. in west part such as Kutch and Surashtra (Chauhan et al. 2013). Cactus pear fruits are known to contain nutrients, bioactive compounds, phenolic compounds, minerals and vitamins. The native tribal people of Mexico utilized cactus to cure diarrhea, dysentery, superficial wounds, inflammation and boost immunity (Shedbalkar et al. 2010). The cactus pear seeds were used as an astringent in traditional Portuguese medicine. (Kumar et al. 2018). In USA, cactus pear was used as traditional medicines for its hypoglycemic and hypolipidemic actions (Shedbalkar et al. 2010). While, in Indian ethnomedicine, the cactus pear fruit is used to treat the anaemia, skin infections, urinary tract infection, diseases related to gastro intestinal tract (Chauhan et al. 2015). As per FSSAI Notification dated 23rd December 2016, Opuntia vulgaris and Opuntia dillenii fruit varieties are considered as Health supplement and nutraceutical for medical purpose under Schedule IV. In modern medicine, cactus pear’s extracts were used cure gastric ulcer, aging (Kumar et al. 2018), rheumatism, lumbago, hemoptysis, gonorrhea, lower cholesterol levels and prevent diabetes (Shedbalkar et al. 2010).

Cactus pear fruit is usually consumed fresh but, to increase its commercial value, it can be fermented with lactic acid bacteria. Fermentation is a metabolic process by which glucose is converted into cellular energy and lactic acid in media. Lactic acid bacteria (LAB) are utilised as natural or selected starters in fruit juice fermentation. Starter cultures helps in the rapid inhibition of spoilage and harmful bacteria while producing processed fruit with improved sensory and nutritional quality (Panda et al. 2017). Certain unwanted organic compounds like 3, 5-dihydroxy-2 methyl; furfuryl alcohol; 2-propenenitrile, 2-(acetyloxy); 2, 2-diethyl-3 methyloxazolidine; acetaldehyde and furan present in unfermented cactus pear fruit juice were disintegrated after lactic fermentation (Panda et al. 2017). Di Cagno et al (2016) utilised autochthonous Leuconostoc mesenteroides with capacity to acidify cactus pear fruit juice and ability to synthesize exopolysaccharides to develop fermented cactus pear fruit puree. Verón et al (2019) prepared fermented cactus pear juice using autochthonous and potentially probiotic Lactobacillus plantarum S-811 strain. They found that strain rapidly reduced the pH from 5.5 to 3.7 and produced 0.51% (w/v) lactic acid. The fermented juice showed antioxidant mechanisms in yeast cells against H2O2.

Although the lactic fermentation process has been around for generations, plant-based food fermentations are still largely unknown in comparison to dairy-based food fermentations. In addition to that, fruit juice fermentation results in major changes that affect the end product's microbiological safety, organoleptic properties, storage stability and economic benefit. The present investigation is therefore taken up to develop biofunctional lactic fermented cactus pear fruit beverage using potent indigenous Lactobacillus cultures.

Materials and methods

Maintenance of starter culture

Two indigenous lactobacilli, Limosilactobacillus fermentum MTCC 25515 and Lacticaseibacillus rhamnosus M9 were isolated from Meghalayan traditional fermented rice beverage. Both the lactobacilli cultures were propagated in sterilized MRS broth. For preservation and maintenance, the cultures were grown at 37 °C for 18–24 h and their 1.0 mL was transferred to the 2.0 mL cryovials containing sterile 1.0 mL 30% glycerol, mixed well, kept in ice cold water for 15 min, preserved at −30 °C for further study.

Preparation of freeze dried cactus pear fruit powder

The reddish purple variety of cactus pear (Opuntia elatior Mill.) authenticated by the Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat (India) was used for the study as it is available in the local market. In addition to that, it is also known to possess excellent source of health beneficial pigments such as betacyanin, betaxanthin and other phytochemicals (Chauhan et al. 2013). The 50 kg of fresh cactus pear fruits were purchased from the local market of Surat, Gujarat during November month (Winter season). The fruits were sorted based on the visual inspection and only ripe fruits without blemishes or damage were selected for the freeze dried cactus pear fruit powder preparation. The average weight of a fruit was 23.82 ± 4.51 g. The screened fruits were washed under tap water. Then skins of fruits were removed and pulp was collected in the large vessel. The juice was obtained from the pulp by straining initially with fine sieve and then by using clean and dry muslin cloth with pore size 0.2 mm. The juice was then frozen at -30 °C for overnight and afterwards transferred to the freeze dryer to make cactus pear fruit powder (CPP). The CPP was prepared in bulk, packed in air tight bag and stored at −10 °C. The fruit powder was used whenever required during the study.

Preliminary experiment

Preliminary trials were conducted to determine the optimum level of starter culture addition, type of lactobacilli strain and temperature of incubation for the rest of the investigation. After preliminary experimental trials, 3% (w/v) starter inoculum, combination of two lactobacilli (Limosilactobacillus fermentum MTCC 25515 and Lacticaseibacillus rhamnosus M9) in equal proportion and 37 °C as incubation temperature were selected for the further study. In addition to that, preliminary trials were also conducted to decide the optimum level of cactus pear fruit powder, level of sucrose and incubation period for the optimization of process parameters in the study. After preliminary experimental trials, 10–15% (w/v) cactus pear fruit powder, 1–3% (w/v) sucrose and 4–8 h incubation period were chosen for the further study.

Optimization of process parameters

The process for the development of lactic fermented cactus pear beverage was optimized using response surface methodology by Design-Ease® Software. The central composite rotatable design (CCRD), one of the most important experimental designs used in process optimization investigation was applied in this study with the objective to develop an empirical model of the process and to obtain a more precise estimate of the optimum operating conditions for the factors involved. The selected range for the independent variables was 10–15% for CPP, 1–3% for sucrose and 4–8 h for incubation time. The complete design consisted of 20 experiments including 8 factorial experiments, 6 axial experiments and 6 replicates in the centre point (Table 1). Sensory evaluation of the beverage was carried out using 9-point hedonic scale. Ten panelists (5 males and 5 females), in the age group between 22 and 50 years, were selected from among the faculty and technical staffs through personal communication and willingness to undertake this test. Instructions were given to the panelists to rinse mouth with potable water after evaluation of every sample. A 9-point hedonic score system was used with the following individual scores: 1-Dislike extremely, 2-Dislike very much, 3-Dislike moderately, 4-Dislike slightly, 5-Neither like nor dislike, 6-Like slightly, 7-Like moderately, 8-Like very much and 9-Like extremely (Ranganna 1987). They were asked to carefully examine and give score to the attributes such as flavor, consistency, color & appearance and overall acceptability of samples as per their level of liking. Lactic fermented cactus pear fruit beverage was prepared as given in Fig. 1. The scores of flavour, consistency, colour & appearance and overall acceptability for all the 20 experiments suggested by the RSM were fed into the design expert software to obtain the optimised solution for the three variables. Selection of optimum condition was determined by setting the goal and importance of the variables as well as responses through multi-response optimization. For the validation of model, the product was prepared based on the predicted levels of cactus pear fruit powder, sucrose and incubation period by design expert. It was then subjected to sensory analysis using 9-point hedonic scale. The comparison between predicted and actual values of sensory responses for optimized product was carried out by paired t-test for non-significant difference.

Table 1.

Effect of three independent variables on sensory score (9-point hedonic scale) of lactic fermented cactus pear fruit beverage

Run Std Independent variables Responses
A: CPP (%) B: IT (h) C: Sucrose (%) Flavor Consistency C & A OA
1 7 10.00 8.00 3.00 6.25 6.70 6.30 6.45
2 13 12.50 6.00 0.00 6.00 6.00 6.20 6.03
3 4 15.00 8.00 1.00 7.00 7.10 7.25 7.12
4 6 15.00 4.00 3.00 8.50 8.20 8.30 8.33
5 19 12.50 6.00 2.00 8.90 8.80 8.70 8.83
6 10 17.50 6.00 2.00 6.30 6.00 6.20 6.17
7 18 12.50 6.00 2.00 9.00 8.95 8.90 8.91
8 14 12.50 6.00 4.00 7.20 7.10 7.20 7.15
9 20 12.50 6.00 2.00 8.99 9.00 8.76 8.92
10 12 12.50 10.00 2.00 7.40 7.85 7.90 7.55
11 16 12.50 6.00 2.00 8.80 8.95 9.00 8.88
12 5 10.00 4.00 3.00 6.50 6.25 6.35 6.37
13 17 12.50 6.00 2.00 9.00 8.85 9.00 8.95
14 3 10.00 8.00 1.00 6.55 6.50 6.45 6.48
15 15 12.50 6.00 2.00 8.90 9.00 8.80 8.86
16 1 10.00 4.00 1.00 6.10 6.10 6.00 6.05
17 8 15.00 8.00 3.00 7.70 7.90 7.65 7.85
18 9 7.50 6.00 2.00 6.00 6.50 6.00 6.26
19 11 12.50 2.00 2.00 7.60 7.60 7.50 7.57
20 2 15.00 4.00 1.00 7.00 7.10 6.90 7.00
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CPP  Cactus pear fruit powder, IT  Incubation time, C & A  Color and appearance, OA =Overall acceptability

Fig. 1.

Fig. 1

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Generalised flowchart for the preparation of biofunctional lactic fermented cactus pear fruit beverage

Compositional analysis

Moisture content of lactic fermented cactus pear fruit beverage was determined using the gravimetric method (FSSA 2015). Soxhlet method suggested by Karabagias et al. (2020) was used for fat analysis. The Kjeldahl method specified by Borchani et al. (2019) was used to determine the protein content of lactic fermented cactus pear fruit beverage. The obtained values of total nitrogen were multiplied by a factor of 6.25 to get the percentage of total protein. Total carbohydrate in beverage was measured by the phenol sulfuric method as suggested by Berrabah et al. (2019). The ash and crude fiber in the beverage were estimated using FSSA (2016) method. The minerals were estimated in instrument ICP-Optical Emission Spectrometer (Make: Perkin Elmer, Model: optima 7000 DV, made in India), Winlab 32 software version 5.1, Peristaltic pump: 1.5 mL/min.

Physicochemical analysis

Well mixed 10 mL sample was put into a beaker and pH was measured at 25 °C, using digital pH meter (OAKTON pH700, India). Viscosity of the beverage was determined using ‘Brookfield’ viscometer (LV, DVE Viscometer, USA) at temperature of 20 ± 2 °C. The S-61 spindle was used to measure the viscosity and torque was maintained between 30 and 70% (del Socorro Cruz-Cansino et al. 2015). The beverage color was evaluated by measuring the lightness (L*), chroma (C*) and hue values in Chroma meter, CR-400 (Konica Minolta, Inc., Japan). For titratable acidity, 10 mL homogenous lactic fermented cactus pear fruit beverage sample was taken in a clean and dry beaker. It was titrated against 0.1 [N] NaOH solution to an endpoint of pH 8.2. The mL of 0.1 [N] NaOH required to neutralize the acid in the sample was noted (AOAC 1995).

Biofunctional analysis

Betacyanins and betaxanthins quantification was performed by following Moßhammer et al (2006) method. The total phenolic compounds in the beverage was measured by Folin-Ciocalteu method (Chaalal et al. 2013). Total flavonoid in the beverage was checked by using colorimetric method adopted by Yeddes et al (2013). The Ascorbic acid content in the lactic ferments cactus pear beverage was estimated by following the HPLC method suggested by Pertuzatti et al (2015). ABTS activity of the beverage was determined by following the Çam et al. (2009) method.

Microbial analysis

Lactobacilli counts of lactic fermented cactus pear fruit beverage was determined by Randazzo et al (2016) method. Enterobacteriaceae counts in the beverage was by following the method described by Di Cagno et al (2016). Yeast and mold count were by the BIS (1999) method.

Shelf stability of optimized beverage at 7 °C

The lactic fermented cactus pear fruit beverage was packed in polyethylene terephthalate (PET) bottle, stored at 7 ± 1 °C and subjected to shelf life study. Uninoculated cactus pear beverage was taken as control. The samples were removed from the refrigerator after every four days for determining the alteration in sensory attributes, pH, mL 0.1 [N] NaOH per 10 mL, viscosity, color, betalain (betacyanin and betaxanthin), total phenolic compounds, flavonoid, ascorbic acid, antioxidant activity, lactobacilli count, Enterobacteriaceae count, Yeast and mold count.

Statistical analysis

The process parameter optimisation was carried out by response surface methodology using Design Expert software (Stat-Ease, Inc., Minneapolis, MN. The coefficients of determination (R2) and analysis of variance (ANOVA) were used to evaluate the goodness of fit of the regression model. Three-dimensional RSM analyses were used to estimate the optimal extraction conditions of the three independent variables and each dependent variable. The optimised model was tested by the paired t-test at 5% level of significance. The changes obtained due to sensory evaluation, physico-chemical characteristics, biofunctional properties and microbial counts were analyzed statistically by following analysis of variance (F-test) at 5% levels of significance using one/two factor experiment with completely randomized design with equal replications (Gomez and Gomez 1984). Significant F-test assures that the observed difference among the treatment means is real and not due to chance. All these calculations including standard deviation were done using Microsoft Excel 2019 (Microsoft Corp., USA).

Results and discussion

Optimization of process parameters

Effect of various attributes on sensory scores (9-point hedonic scale) of the beverage is given in Table 1. Scores of flavour, consistency, colour & appearance and overall acceptability were well fitted in quadratic model when data were computed in the design expert software using response surface methodology (central composite rotatable design) as statistical tool. The experimental data were analyzed by multiple regression analysis and the coefficients of model were used for the significance levels. Table 2 shows the linear, interaction and quadratic effects of each independent parameter. In case of flavour response two linear (A, C), all interaction and all quadratic variables were significant at p < 0.01, whereas one linear variable (B) was significant at p < 0.05. In case of consistency response two linear (A, C), two interaction (AB, AC) and all quadratic variables were significant at p < 0.01, one linear (B) was significant at p < 0.05 whereas, one interaction parameter (BC) was nonsignificant. In case of color & appearance response two linear (A, C), one interaction (AC) and all quadratic variables were significant at p < 0.01, one interaction parameter (BC) was significant at p < 0.05 whereas, one linear (B) and one interaction (BC) parameters were nonsignificant. In case of overall acceptability response two linear (A, C), all interaction and all quadratic variables were significant at p < 0.01 whereas, one linear parameter (B) was nonsignificant. The flavour, consistency, color & appearance and overall acceptability scores were quadratic with a good regression coefficient (R2) 0.9890, 0.9792, 0.9862 and 0.9847, respectively. The optimum levels suggested by the software Design Expert for level of CPP, incubation time and sucrose were 12%, 6 h and 3% respectively. By carrying out the experiments, the optimum conditions suggested through response surface analysis were confirmed. Both, predicted and actual values of sensory responses for optimized product were statistically at par (Table 3). In a similar study conducted by Di Cagno et al (2009), fermented tomato juice with 1% (w/v) sucrose was prepared. The tomato juice (TJ) was pasteurised and inoculated with Lactobacillus plantarum. The inoculated juice was incubated at 25 °C for 17 h. Fermented TJ exhibited better flavor profile, color, health-promoting properties and viscosity with respect to unfermented TJ.

Table 2.

Comparison of predicted and actual values of responses for optimized product

Response Flavour1 Consistency1 Color & Appearance1 Overall acceptability1
Predicted Value* 8.76 8.72 8.68 8.71
Actual Value# 8.72 8.74 8.62 8.70
P Value 0.79 0.73 0.67 0.87
Cal. t-Value@ 0.27 0.35 0.45 0.17
Significance NS NS NS NS
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1Sensory score on 9-point hedonic scale,

*Predicted values of design expert 12.0.11.0,

#Actual values are average of seven trials for optimized product,

@t-values found non-significant at 5% level of significance, Tabulated t-value = 2.447 (Cal. t-value less than tabulated t-value),

NS  Non-significant

Table 3.

Results of regression coefficients for four response variables

Factor Flavor score Consistency score Color & appearance score Overall acceptability score
Linear A 1.0875** 1.0906** 1.0875** 1.1323**
B −0.0625* 0.0594* 0.0563 −0.0085
C 0.7937** 0.7719** 0.8000** 0.7773**
Interaction AB −0.1250** −0.1313** −0.0875 −0.0787**
AC −0.2375** −0.2938** −0.3000** −0.2463**
BC −0.1875** −0.0188 −0.1875* −0.1371**
Quadratic A2 −1.2041** −1.2818** −1.2027** −1.2509**
B2 −0.3666** −0.3006** −0.3027** −0.3397**
C2 −0.8416** −0.8443** −0.8527** −0.8501**
R2 0.9890* 0.9792 0.9862 0.9847
Intercept 8.91 8.92 8.83 8.90
APV 111.46 132.39 58.03 220.49
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**Significant at 1 per cent level (p < 0.01)

*Significant at 5 per cent level (p < 0.05)

A Cactus Pear Powder (CPP), B Incubation Time (IT), C Sucrose, R2 Coefficient of determination, APV Adequate Precision Value

Proximate compositional, physicochemical, biofunctional and microbial analysis of the optimized product

The optimized product and control (unfermented beverage) were subjected to proximate compositional, physicochemical, biofunctional and microbial analysis (Table 4). The moisture content in our product was 87.77%. Chauhan et al (2013) found 86.08% moisture content in the fruit juice of Opuntia elatior Mill. Sabtain et al (2021) also reported 87.07%, moisture content in cactus pear juice (variety-Opuntia ficus-indica). The moisture content in our fermented cactus pear beverage is similar to the fresh cactus pear juice in previously reported studies (Chauhan et al. 2013; Sabtain et al. 2021). While, Randazzo et al (2016) reported 14.07% total soluble solids in fermented cactus pear beverage. The fat content in control and fermented beverage was 0.14 and 0.12%, respectively. Sabtain et al (2021) reported 0.40%, fat content in cactus pear (Opuntia ficus-indica). The reported fat content in cactus pear fruit juice, was found between 0.09 to 0.70% (Piga 2004; Aruwa et al. 2018). Fat content in our product was in the range of previously mentioned data, indicating no effect of lactic fermentation on fat. The protein determined in this study was 0.76% in unfermented and 0.54% in fermented beverage. Our result of protein in unfermented juice agrees with the 0.34% protein in Opuntia macrocentra Engelm. Fruit. On the contrary, 0.09 and 0.05% protein was found in unfermented and fermented cactus pear juice, respectively (Veron et al., 2019). The carbohydrate content of the current study was 11.52% in unfermented beverage. Which was found to reduce significantly (p < 0.05) after lactic fermentation to 9.58%. Similar reduction was observed by Verón et al. (2019). They reported that, carbohydrate content was 12.73% in unfermented and 10.70% in lactic fermented cactus pear (Opuntia ficus-indica) fruit juice. Whereas, Panda et al (2017) found 1.75% total carbohydrate in lactic fermented cactus pear (variety ‘skinners court’) fruit juice. As the compositional parameters of cactus pear fruit juice vary with the variety, cultivation practices, environment and geographic location. That’s why the results of fat and protein content of our product was comparatively different from other reported studies. Moreover, after lactic fermentation, protein content was reduced significantly (p < 0.05), while fat remained unaffected in the studied beverage. The difference in carbohydrate content with the different study could be due to variation found in initial carbohydrate content i.e. before fermentation, type of starter culture used, incubation time varied from one study to another, method of carbohydrate estimation, addition of sugar to the juice to improve sweetness etc. The crude fiber of studied beverages was 2.21% and 1.50% in unfermented and fermented one, respectively. Lactic fermentation significantly (p < 0.05) reduced the crude fiber in the fermented beverage. Similar findings were reported earlier, where fermentation induced loss of crude fibre in beet, carrot and cucumber were observed (Kiczorowski et al. 2022). Losses of nutrients such as protein, carbohydrate and crude fibre during the lactic fermentation were related to the nutritional needs of the growing lactic acid bacteria and meeting their metabolic requirements in the carrier medium (Dallal et al. 2017). The effect of fermentation on ash content of the tested beverage was non- significant. The ash content in unfermented and fermented beverage was 0.76 and 0.82%, respectively.

Table 4.

Proximate compositional, physicochemical, biofunctional and microbial analysis of control (unfermented beverage) and optimized lactic fermented cactus pear fruit beverage

Parameter Unfermented pasteurized cactus pear fruit beverage Fermented pasteurized cactus pear fruit beverage
Composition (Wet matter basis)
Moisture % 85.51 ± 0.20b 87.77 ± 0.13a
Fat, % 00.14 ± 0.19a 00.12 ± 0.24a
Protein, % 00.76 ± 0.29a 00.54 ± 0.34b
Carbohydrate, % 11.52 ± 0.15a 09.58 ± 0.11b
Crude fibre, % 02.21 ± 0.23a 01.50 ± 0.31b
Ash, % 00.76 ± 0.33a 00.82 ± 0.42a
Iron, mg/100 mL 01.56 ± 0.18a 01.40 ± 0.11a
Calcium, mg/100 mL 50.00 ± 3.11a 48.72 ± 2.56a
Magnesium, mg/100 mL 31.38 ± 5.48a 28.63 ± 4.56a
Phosphorus, mg/100 mL 26.60 ± 3.11a 24.13 ± 3.43a
Potassium, mg/100 mL 97.10 ± 3.18a 95.76 ± 3.25a
Sodium, mg/100 mL 00.93 ± 0.25a 01.11 ± 0.34a
Manganese, mg/100 mL 01.58 ± 0.22a 01.43 ± 0.17a
Zinc, mg/100 mL 00.17 ± 0.01a 00.14 ± 0.10a
Copper, mg/100 mL 00.10 ± 0.03a 00.02 ± 0.10a
Physicochemical parameters
pH (at 25 °C) 4.80 ± 0.13a 3.79 ± 0.12b
Titratable acidity, mL 0.1 [N] NaOH per 10 mL 1.23 ± 0.15b 2.70 ± 0.26a
Viscosity (at 20 °C), cP 8.91 ± 0.38a 9.55 ± 0.58a
Lightness (L*) 18.31 ± 0.86b 19.61 ± 0.42a
Chroma (C*) 16.53 ± 0.35a 13.84 ± 0.37b
Hue 346.15 ± 4.51a 345.25 ± 3.63a
Biofunctional properties
Betacyanin, mg/100 mL 67.81 ± 1.65a 71.82 ± 1.59a
Betaxanthins, mg/100 mL 25.82 ± 1.16a 28.08 ± 0.62a
Total phenolic compounds, mg GAE /100 mL 99.47 ± 1.66a 89.84 ± 1.58b
Total flavonoid content, mg QE/100 mL 6.05 ± 0.24a 5.86 ± 0.21a
Ascorbic acid, mg/100 mL 9.38 ± 0.37b 10.59 ± 0.12a
Antioxidant activity (ABTS), % Inhibition 82.00 ± 3.14a 58.02 ± 2.56a
Microbial counts
Lactobacilli counts, log CFU/mL – 8.57 ± 0.17
Enterobacteriaceae counts, CFU/mL Absent/ml Absent/ml
Yeast and mold counts, CFU/mL Absent/ml Absent/ml
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Values are mean ± SD, n = 3; Values with different superscripts in a row are significantly different (P < 0.05); GAE  gallic acid equivalent; QE  quercetin equivalent; CFU  colony forming unit

Cactus pear fruit juice is known to contain good amount of minerals. The iron, calcium, magnesium, phosphorus, potassium, sodium, manganese, zinc and copper content in our beverage as milligram per 100 mL were 1.40, 48.72, 28.63, 24.13, 95.76, 1.11, 1.14, 0.14 and 0.02, respectively. The results reported by previous researchers showed the iron, calcium, magnesium, phosphorus, potassium, sodium, manganese, zinc and copper content in various cactus pear fruit as milligram per 100 mL were 0.40–1.50, 12.80–59.00, 16.10–98.40, 2.82–32.80, 90–217, 0.60–1.10, 1.08, 0.78 and 1.53, respectively (El-Mostafa et al. 2014; Aruwa et al. 2018; Ramírez-Rodríguez et al. 2020; Barba et al. 2022). Our results on mineral content were found in these reported range indicating negligible effect of lactic fermentation on minerals. The content of mineral in our beverage was characterized by high amounts of potassium, followed by calcium, magnesium, phosphorus, manganese, iron, sodium, zinc and copper. The studied beverage had high potassium and low sodium content which could be beneficial for people with kidney problems and hypertension. In addition to that, lactic fermented fruit beverages were reported to improves the bioavailability of minerals in the host (Parvez et al. 2006).

The pH of lactic fermented cactus pear juice was found less than 4 as reported by several studies (Di Cagno et al. 2016; Verón et al. 2017, 2019). The pH of studied lactic fermented beverage also reduced from 4.80 to 3.79 after fermentation. As the pH reduced, the titratable acidity found to increase during fermentation. The titratable acidity of studied beverage was 2.71 mL of 0.1 [N] NaOH. Whereas, comparatively higher acidity i.e. 13.20 mL of 0.1 [N] NaOH per 10 mL lactic fermented cactus pear juice was reported (Di Cagno et al. 2016). The reason for huge difference in acidity was because pH of unfermented cactus pear of Di Cagno et al. was 6.01 and after fermentation at 25 °C for 12 h, pH dropped to less than 4. Hence the amount of 0.1 N NaOH required for the neutralisation would be more compared to studied beverage where pH was dropped from 4.80 to 3.79 after fermentation. The viscosity of our product (9.55 cP) was comparatively lower than the following reported value. The viscosity of reported fermented cactus pear juice was 14.5 cP (Song et al. 2022) and lactic fermented cactus pear puree was 8.11 (Di Cagno et al. 2016). In another study, viscosity of fresh cactus pear juice was 53.62 cP at 100 rpm (Chauhan et al. 2013). This may be due to difference in starter culture used, length of incubation time, initial viscosity of the cactus pear juice before fermentation etc.

Cactus pear fruit is known to possess nitrogen containing pigment compounds such as red-violet betacyanins and yellow-orange betaxanthin (Jimenez-Aguilar et al. 2015). The variation in this colored pigments in the cactus pear fruit influence the fruit color. Betacyanins and betaxanthins in 100 mL lactic fermented cactus pear (Opuntia ficus-indica) fruit puree was 14.93 mg and 8.43 mg, respectively (Di Cagno et al. 2016), while 100 mL lactic fermented cactus pear (Opuntia ficus-indica) fruit juice had 1.87 mg and 1.77 mg, respectively (Veron et al. 2019). In our beverage, betacyanins and betaxanthin was 71.82 mg and 28.08 mg per 100 mL, respectively. These observations were comparatively more than the Di Cagno et al (2016) and Veron et al. (2019) results. We used reddish purple variety (Opuntia elatior), which is known to possess comparatively higher betacyanin and betaxanthin than the other variety. Also, the fermentation process had non significant effect on the level of colored pigments in the studies beverage. Betacyanins and betaxanthin are not only essential for the Opuntia’s fruit coloring, but they are also important bioactive compounds with excellent antioxidant activity (Jiménez-Aguilar et al. 2015). We obtained 99.47 and 89.84 mg total phenolic compounds in 100 mL unfermented and fermented beverage, respectively. The level of phenolic compounds in the studied fermented beverage was affected significantly when compared with the unfermented beverage. Similar result was obtained by Verón et al. (2017) they found 80.80 mg total phenolic compounds in 100 mL lactic fermented cactus pear fruit juice. In contrast, Randazzo et al (2016) reported 37.41 mg total phenolic compounds in 100 mL cactus pear based kefir like beverage. The variation in phenolic compounds in various reported studies, may be due to type of starter culture used, length of fermentation period, variety of cactus pear fruit, demographic location. Moreover, it is reported that microorganism possesses phenolic acid decarboxylases, which reduced the concentration of polyphenols (Alamed et al. 2009) in the carrier medium.

The flavonoid content of unfermented and fermented beverage was 6.05 and 5.86 mg QE per 100 mL, respectively in the current study. Non significant effect of lactic fermentation was observed on the content of flavonoids. Kuti (2004) reported the ∼9.35 mg in 100 mL unfermented purple-skinned cactus pear fruits. The flavonoids are reported to possess various beneficial effects including antioxidant, antiviral and antibacterial (Silva et al. 2021). Our beverage had 9.38 mg ascorbic acid in 100 mL control sample which increased significantly (p < 0.05) to 10.59 mg after lactic fermentation. The ascorbic acid content as mg in 100 mL lactic fermented cactus pear fruit juice reported by many authors was in the range of 3.11 to 17.00 (Di Cagno et al. 2016; Panda et al. 2017; Verón et al. 2019). In the present study, the antioxidant (ABTS) activity of unfermented and lactic fermented beverage was 82.00% and 58.02%, respectively. Lactic fermentation had non significant effect on the antioxidant activity of the beverage. Veron et al. (2019) also observed the non significant effect of lactic fermentation on the antioxidant activity of the cactus pear juice. As per their report, the antioxidant activity (ABTS) of unfermented and lactic fermented juice was 2.4 µg GAE and 2.6 µg GAE per mL, respectively. Panda et al (2017) reported 105 µM Trolox equivalent per mL antioxidant activity by DPPH assay in lactic fermented cactus pear fruit juice. The lactobacilli count in optimised beverage was 8.57 log CFU/mL. Our result was in agreement with Di Cagno et al (2016), Randazzo et al (2016) and Veron et al. (2019). They reported lactobacilli count > 8 log CFU/mL in lactic fermented cactus pear fruit juice. While Enterobacteriaceae and yeast & mold were not detected in the freshly prepared beverage. Presence of lactobacilli in larger number may contribute to food functionality through their enzyme portfolio that promotes the synthesis of various metabolites such as organic acids aromatic compounds, exopolysaccharides, bacteriocins and release of bioactive compounds, which are mainly cryptic in the raw matrix (Gobbetti et al. 2010). They are also responsible for the increase in food safety and shelf life.

Shelf stability of lactic fermented cactus pear fruit beverage stored at 7 °C

Sensory analysis

Scores obtained for the different sensory attributes are given in Fig. 2. The optimized product was liked by the sensory panelists and significant (P < 0.05) difference was seen in all attributes, during storage. Optimized product received comparatively higher sensory scores as compared to control throughout the shelf life study. The optimized product with acidity 6.50 mL 0.1 N NaOH and pH 3.10 was accepted by the sensory panellist upto 20 days because of very high acidity perception on the tongue with overall acceptability score < 6. Similar observation was obtained by Di Cagno et al (2016), who prepared cactus pear fruit puree fermented with Leuconostoc mesenteroides. Fermented puree was preferred with superior odor, aroma pleasantness, smoothness with the highest acceptability level over unfermented one. Di Cagno et al (2016) also reported 21 days of shelf stability of the fermented cactus pear product with acidity 5.9 mL 0.1 N NaOH and pH 3.95 under refrigeration condition.

Fig. 2.

Fig. 2

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Changes in sensory scores of control and optimized product during storage. Each bar represents the average of three independent experiments. Error bars are standard deviations. (P < 0.05, mean ± SD, n = 3)

Physiochemical and biofunctional properties

The effect of storage on physiochemical and biofunctional properties of the optimized beverage and control is shown in Tables 5 and 6, respectively. The pH of the optimized product was significantly (P < 0.05) lower than the control as it was lactic fermented product. pH was found to reduce significantly (P < 0.05) throughout the storage in control and optimized product. Marked reduction in pH during refrigerated storage indicated the activity of starter culture at low temperature. Our result matches with the following studies. Di Cagno et al (2016) reported the 3.95 pH of freshly prepared fermented cactus pear puree, which decreased significantly to 3.73 on day 20 when stored at 4 °C, while pH of unfermented puree was 5.41. In another study, pH of the cactus pear juice was reduced from 5.4 to 3.1 after fermentation with Lactobacilli S-04 strain (Verón et al. 2017).

Table 5.

Changes in physicochemical parameters of the product during storage at 7 °C

Days pH Titratable acidity (mL 0.1 N NaOH) Viscosity (cP) Lightness Chroma Hue
Control 0 4.80 ± 0.13a 1.23 ± 0.15i 8.91 ± 0.38a 18.31 ± 0.86f 16.53 ± 0.35d 346.15 ± 4.51a
4 4.80 ± 0.13a 1.37 ± 0.15hi 9.00 ± 0.21a 18.03 ± 0.63f 16.93 ± 0.44c 346.66 ± 5.13a
8 4.79 ± 0.14a 1.43 ± 0.21ghi 9.17 ± 0.88a 18.55 ± 1.17ef 21.37 ± 0.36b 349.46 ± 9.16a
12 4.79 ± 0.14a 1.57 ± 0.15ghi 9.43 ± 0.69a 19.10 ± 0.30def 22.31 ± 0.47ab 352.23 ± 4.29a
16 4.79 ± 0.10a 1.63 ± 0.15gh 9.61 ± 0.56a 21.46 ± 0.86ab 22.33 ± 0.46ab 353.24 ± 7.11a
20 4.78 ± 0.10a 1.73 ± 0.15 g 9.71 ± 0.20a 21.98 ± 0.26a 23.04 ± 0.33a 354.24 ± 6.43a
Optimised product 0 3.79 ± 0.12b 2.70 ± 0.26f 9.55 ± 0.58a 19.61 ± 0.42cde 13.84 ± 0.37 g 345.25 ± 3.63a
4 3.50 ± 0.13c 4.00 ± 0.30e 9.74 ± 0.30a 19.84 ± 0.25 cd 14.31 ± 0.83 fg 345.30 ± 4.90a
8 3.28 ± 0.13d 5.23 ± 0.15d 9.88 ± 0.66a 20.16 ± 0.30 cd 14.60 ± 0.70 fg 346.74 ± 6.14a
12 3.18 ± 0.11e 5.60 ± 0.26c 9.96 ± 0.83a 20.61 ± 0.26bc 14.72 ± 0.65 fg 347.50 ± 5.73a
16 3.13 ± 0.11e 6.23 ± 0.21ab 10.04 ± 0.14a 20.70 ± 0.35bc 15.14 ± 0.57f 347.91 ± 4.96a
20 3.10 ± 0.13e 6.50 ± 0.26a 10.14 ± 0.16a 22.16 ± 0.13a 15.43 ± 0.60e 348.74 ± 5.53a
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Values are mean ± SD, n = 3, values with different superscripts in a column are significantly different (P < 0.05), cP  centipoise

Table 6.

Changes in biofunctional properties of the product during storage at 7 °C

Days Betacyanin (mg/100 mL) Betaxanthin (mg/100 mL) TPC (mg GAE/100 mL) Flavonoid content (mg QE/100 ml) Ascorbic acid (mg/100 mL) ABTS activity (% Inhibition)
Control 0 67.81 ± 1.65a 25.82 ± 1.16a 99.47 ± 1.66a 6.05 ± 0.24a 9.38 ± 0.37 cd 82.00 ± 3.14a
4 66.54 ± 1.98a 25.32 ± 0.87a 96.01 ± 1.49ab 5.95 ± 0.14ab 8.54 ± 0.57ef 77.80 ± 3.43a
8 65.15 ± 2.41a 24.04 ± 0.69a 93.39 ± 4.56abc 5.88 ± 0.14ab 8.14 ± 0.40f 67.94 ± 2.91a
12 63.91 ± 1.70a 22.86 ± 1.06a 90.13 ± 4.34bc 5.81 ± 0.17abc 7.39 ± 0.40 g 60.17 ± 2.89a
16 62.39 ± 1.63a 22.08 ± 0.65a 89.44 ± 4.16bc 5.71 ± 0.14abc 6.38 ± 0.23 h 52.78 ± 3.41a
20 61.45 ± 1.95a 21.55 ± 0.68a 87.88 ± 2.75bc 5.64 ± 0.20bc 5.80 ± 0.33 h 57.75 ± 3.01a
Optimised product 0 71.82 ± 1.59a 28.08 ± 0.62a 89.84 ± 1.58 cd 5.86 ± 0.21ab 10.59 ± 0.12a 58.02 ± 2.56a
4 70.62 ± 2.21a 27.69 ± 0.72a 81.20 ± 1.18de 5.49 ± 0.35 cd 10.26 ± 0.11ab 51.45 ± 2.54a
8 69.57 ± 2.43a 27.40 ± 0.46a 76.30 ± 2.60e 5.27 ± 0.28d 10.11 ± 0.15ab 49.25 ± 2.84a
12 69.07 ± 2.55a 26.84 ± 0.95a 75.19 ± 1.81e 5.13 ± 0.23de 9.86 ± 0.12bc 45.64 ± 3.15a
16 68.71 ± 1.57a 26.52 ± 0.84a 67.43 ± 4.47f 4.82 ± 0.21ef 9.75 ± 0.11bcd 41.70 ± 3.23a
20 68.10 ± 2.62a 26.35 ± 0.57a 62.68 ± 3.62f 4.67 ± 0.18f 9.15 ± 0.13de 32.14 ± 2.99a
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Values are mean ± SD, n = 3, values with different superscripts in a column are significantly different (P < 0.05); TPC Total phenolic compounds GAE Gallic acid equivalent; QE Quercetin equivalent

The mL 0.1 [N] NaOH required to neutralize acids per 10 mL sample indicates the amount of organic acids produced in the product due to microbial activity and it was measured as titratable acidity. The titratable acidity was comparatively (P < 0.05) more in optimized product than control. It increased significantly (P < 0.05) in control and optimized product from 1.23 and 2.70 mL, respectively on day 0 to 1.73 and 6.50 mL, respectively on day 20. The results of titratable acidity were positively affected by the pH of the optimized product during storage study. The titratable acidity of lactic fermented cactus pear fruit puree, increased significantly during fermentation and storage (Di Cagno et al. 2016). Randazzo et al (2016) also found highly significant difference in titratable acidity between fermented and unfermented cactus pear based kefir like beverage.

The viscosity of control and optimized product was 9.55 cP and 8.91 cP, respectively on day 0 and 10.14 cP and 9.71 cP, respectively on day 20. After fermentation and during the storage period, the viscosity of optimized product remained noticebly (P < 0.05) greater than control. Whereas, there was no significant effect on viscosity with the storage time in both control and optimized product. The higher viscosity of fermented product could be due excretion of exopolysaccharides like metabolites by the starter, which may increase the viscosity of the final product. In contrast to our result, fermented cactus pear fruit puree viscosity reduced markedly (P < 0.05) than unfermented. While, opposite trend observed in stored products (Di Cagno et al. 2016).

Color is an important attribute of juice quality because it can decide it’s acceptability by the consumer. In the present study, lactic fermented beverage received significantly (P < 0.05) higher L* value compared to unfermented one. Lightness value of fermented and control beverage increased significantly (P < 0.05) during storage. Randazzo et al (2016) also observed significant increase (P < 0.01) in L* of fermented cactus pear juice than unfermented. Di Cagno et al (2016) also found significant (P < 0.05) increase in L* values of lactic fermented cactus pear puree during storage at 4 °C for 21 days. Unfermented cactus pear puree also showed the significantly higher L* value after 21 days than fresh sample. In contrast, del Socorro Cruz-Cansino et al. (2015) observed non significant (P < 0.05) difference in L* value of pasteurized purple cactus pear juice during storage at 4 °C for 28 days. Chrome (C*) indicates the level of saturation, purity or visual intensity of color and is defined as the level of deviation from gray to a pure chromatic color, therefore, high values represent a high color saturation (del Socorro Cruz-Cansino et al. 2015). After fermentation, C* was non-significant with the control. Marked increase (P < 0.05) in C* was found for optimized product and control, during storage. Moreover, significant (p < 0.05) increase in C* noticed in control opposite to optimized product, indicating that color brilliance was greater with low betaxanthin. This finding is consistent with significant high chroma value in the Opuntia ficus-indica cv. Rossa (red–purple variety) compared to Opuntia ficus-indica cv. Gialla (yellow-orange) variety of cactus pear (Stintzing et al. 2003). Hue represents tonality and hue values were non significant in optimized product and control, after fermentation as well as during storage. The hue angle of kefir like beverage prepared using various fruits (cactus pear, pomegranate and quince), increased after fermentation (Randazzo et al. 2016). This study agrees with the results obtained for our product.

Biofunctional properties

Betacyanin and betaxanthin are the most widely found color pigments in Opuntia spp. Fruits. Betacynin gives red–purple and betaxanthins gives yellow-orange color to the fruit. These pigments must be preserved during processing, as they are responsible for the final attractive colour of the product and have antioxidant activity (Ramírez-Ramos et al. 2015). Betacyanin content of optimized product and control on day 0 was 71.82 and 67.81 mg per 100 mL respectively. Betaxanthin content of optimized product and control on day 0 was 28.08 and 25.82 mg per 100 mL respectively. Betacyanins and betaxanthins were found to reduce non significantly with the progression of storage period. In short, optimized beverage had comparatively more level than control, both in fresh as well as stored samples. The pigment compounds were not affected by the fermentation as well as storage. The pasteurization can partially break the betalamic acid bond which re-associate to precursory amino acids such as prolin, leading to a partial regeneration of this pigment in the juice (Castellar et al. 2003). Betanin content for the majority of samples were > 200 mg/L and remained above this value during storage probably due to acidic pH of juice. Besides, betalains can retain their color at pH ranges of 3–7 and remain stable for up to 392 days if stored at 4 °C (Castellar et al. 2003). Additionaly, organic acids in cactus pear fruit juice indicated preservative effect on color pigments (Moßhammer et al. 2006).

Plant phenolic compounds are known to possess antioxidative potential. The total phenolic compounds (TPC) of optimised product and control immediately after preparation was 89.84 mg GAE and 99.47 mg GAE in 100 mL, respectively. The TPC of both the samples reduced significantly (P < 0.05) with the increase in storage period. Moreover, control showed significantly higher (P < 0.05) TPC compared to optimised product, in both fresh and stored samples. Our results were in agreement with the following study. Kefir like beverages prepared from various fruit (apple, grape, kiwi, pomegranate, cactus pear, quince and watermelon) showed lower levels of TPC than corresponding unfermented fruit juices. Randazzo et al (2016) also observed significant (P < 0.05) reduction in TPC after fermentation from 54.6 mg to 37.4 mg in 100 mL cactus pear fruit juice. Phenolic acid decarboxylases enzyme secreted by lactobacilli were reported to reduce the TPC during fermentation as well as refrigerated storage in the carrier medium (Alamed et al. 2009; Lodha et al. 2021). In contrast, non significant difference in TPC, between fermented and unfermented cactus pear juice was reported by many authors (Panda et al. 2017; Allendez et al. 2020; Verón et al. 2019).

The predominant flavonoids in cactus pears fruits were quercetin, kaempferol and isorhamnetin (Kuti 2004). The total flavonoids consisted of 96.8% quercetin (9.05 mg/100 g fruit) in purple-skinned and 93.1% quercetin (5.10 mg/100 g fruit) in red-skinned cactus pear. Majority of the cactus pear fruit flavonoids was in the fruit pulp tissues (Kuti 2004). In the present study, flavonoids reduced significantly (P < 0.05) in control and optimized beverage during refrigerated storage. Some enzymes secreted by lactobacilli have been reported to exhibit a type of β-glucosidase activity and used to convert flavonol glycosides into aglycones (Quines-Lagmay et al. 2020), which may explain the significant (p < 0.05) decrease in flavonoid of optimized beverage during storage. Among the cactus pear fruit varieties examined, total flavonoid contents ranged from as low as ∼0.98 mg per 100 g fresh weight in yellow-skinned cactus pear to as high as ∼9.35 mg per 100 g fresh weight in purple-skinned cactus pear (Kuti 2004). We used reddish purple variety (Opuntia elatior) of cactus pear fruit and our results were nearer to the reported value.

The ascorbic acid content in freshly prepared 100 mL fermented samples and control was 10.59 mg and 9.38 mg, respectively and the values reduced significantly (P < 0.05) with the storage period. The ascorbic acid was significantly (P < 0.05) higher in the optimised sample after fermentation and storage, than control. Our results agree with following reported values. The total ascorbic acid content ranged from 1.00 to 11.10 mg per 100 g purple-skinned cactus pear fruit and 2.30 to 79.20 mg per 100 g red-skinned fruit (Kuti 2004). Di Cagno et al (2016) reported that the ascorbic acid content was significantly (P < 0.05) higher in 100 g lactic fermented cactus pear puree (17 mg) as compared to unfermented (10.8 mg) and non significant reduction was observed in fermented puree with the progression in storage period from day 0 to 21 at 4 °C (16 mg). However, significant (P < 0.05) reduction was observed in unfermented cactus pear puree during storage (5.1 mg). It is reported that certain starter cultures were found to synthesise ascorbic acid during fermentation (Adetuyi and Ibrahim 2014). In addition to that, ascorbic acid was preserved at low pH (in lactic fermented product during storage) as it prevents autooxidation of ascorbic acid after changes in oxidation reduction potential (Verón et al. 2019). In contrast to our results, the ascorbic acid content declined significantly from 10 to 6 mg in 100 mL juice during lactic fermentation at 28 °C for 48 h (Panda et al. 2017). As ascorbic acids are sensitive to certain chemicals and enzyme, could be the reason for the reduction in concentration after fermentation.

The antioxidant potential of cactus pear fruit is linked to greater concentrations of ascorbic acid, TPC, flavonoids and betalains. The ABTS activity of optimised product and control was 58.02% and 82.00%, respectively. The ABTS activity of optimised product was found to reduced significantly (P < 0.05) after fermentation as well during storage than control. Same trend was observed by Panda et al (2017). Lactic fermented cactus pear juice had 30% less antioxidant activity than unfermented one. Randazzo et al (2016) also observed the decrease in antioxidant activity of cactus pear based kefir like beverage. Marked reduction in TPC, ascorbic acid, flavonoids and betalain content of our optimised beverage during storage positively contributed to the antioxidant activity as these phytochemicals hugely affects the antioxidant capacity of the final product. The lactobacilli count of lactic fermented cactus pear fruit beverage decreased significantly (P < 0.05) from 8.57 log CFU/mL on day 0 to 8.07 log CFU/mL on day 20 at 7 °C. The viability of lactobacilli remained more than 8 log CFU/mL at the end of storage study. Di Cango et al. (2016), also observed the 1.5 log reduction in lactobacilli count in lactic fermented cactus pear fruit puree after 21 days of storage at 4 °C. The concomitant reduction in pH and rise in titratable acidity of the tested beverage could explain the reason for the decrease in lactobacilli count during storage study. The viable cell count reaches its maximum at its lowest tolerable pH, depending on the species and strain of the LAB (Hutkins et al. 1993). Also, continuous depletion of fermentable sugars will slow down the exponential growth of the bacteria. At plateau, almost all sugars are depleted, causing a rapid decrease in viability, and the cell enters the death phase due to accumulation of waste (Mojikon et al. 2022).

Conclusions

The developed biofunctional lactic fermented cactus pear fruit beverage with indigenous starter, possess desirable biofunctional potential. This product also has shelf stability of 20 days when packed in PET bottle and stored at 7 °C. The biofunctional properties of the product were well preserved with minor diminishes throughout the cold storage. Such a novel biofunctional fruit based beverage can be beneficial to the consumer owing to its antioxidant potential, presence of healthy bacteria and bioactive phytochemicals like polyphenols, flavonoids, color pigments and ascorbic acid. Further, specific characteristics such as anti-inflammation, antianemic, hypoglycemic and hypolipidemic actions needs to be tested in suitable animal models.

Acknowledgements

Authors are highly thankful to the Anand Agricultural University, Anand, Gujarat, India and Kamdhenu University, Anand, Gujarat, India for conducting the study.

Abbreviations

LAB

Lactic acid bacteria

MRS

De Man Rogosa and Sharpe

CPP

Cactus pear fruit powder

RSM

Response surface methodology

FSSA

Food safety standard authority

BIS

Bureau of Indian Standards

CFU

Colony forming unit

TPC

Total phenolic compounds

GAE

Gallic acid equivalent

QE

Quercetin equivalent

cP

Centipoise

Author’s contribution

Conceptualization: MRK, SH; Methodology: MRK, SH.; Formal analysis and investigation: MRK, YV; Writing: MRK, SH, YV; Original draft preparation: MRK, SH; Review and editing: MRK, SH, YV; Resources: MRK, SH; Supervision: SH. All the authors viewed and approved the final version of the manuscript. MRK, SH and YV contributed equally to this study.

Funding

Not Applicable.

Data availability

All data generated or analysed during this study are included in this published article (and its supplementary information files).

Declarations

Conflict of interest

All the authors declare that there is no conflict of interest.

Footnotes

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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