Next Article in Journal
The Effects of Pre-Fermentative Treatments on the Aroma of Krstač and Žižak Wines
Previous Article in Journal
Exploring Static Biological Aging as a Method for Producing Low-Alcohol ‘Fino’ Type White Wines
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Fermentation
Volume 11
Issue 10
10.3390/fermentation11100576
fermentation-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

High-Mountain Tuber Products Improve Selectively the Development and Detoxifying Capacity of Lactobacilli Strains as an Innovative Culture Strategy

by
Cecilia Hebe Orphèe
1,
María Inés Mercado
2,
Fernando Eloy Argañaraz Martínez
1,
Mario Eduardo Arena
1,2 and
Elena Cartagena
1,2,3,*
1
Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán (UNT), Ayacucho 471, Tucumán CP 4000, Argentina
2
Instituto de Morfología Vegetal, Área Botánica, Fundación Miguel Lillo, Miguel Lillo 251, Tucumán CP 4000, Argentina
3
Instituto de Biotecnología Farmacéutica y Alimentaria (INBIOFAL) Consejo Nacional de Investigaciones Científicas y Técnicas CONICET–UNT, Avenida Néstor Kirchner 1900, Tucumán CP 4000, Argentina
*
Author to whom correspondence should be addressed.
Fermentation 2025, 11(10), 576; https://doi.org/10.3390/fermentation11100576
Submission received: 2 August 2025 / Revised: 25 September 2025 / Accepted: 30 September 2025 / Published: 6 October 2025
Browse Figures
Review Reports Versions Notes

Abstract

The study provides valuable insights into the sustainable utilization of edible tuber peels from the high mountains of the Argentinian Puna, which constitutes promising reserves of bioactive phenolic compounds with the potential to enhance the biofunctional properties of lactic acid bacteria. Thirty-two extracts derived from peels of different varieties of tubers, such as Oxalis tuberosa Mol., Ullucus tuberosus Caldas, and Solanum tuberosum L. were incorporated into lactobacilli cultures and individually evaluated. These selectively enhance the development of the probiotic strain Lactiplantibacillus plantarum ATCC 10241 and of Lacticaseibacillus paracasei CO1-LVP105 from ovine origin, without promoting the growth of a pathogenic bacteria set (Escherichia coli O157:H12 and ATCC 35218, Salmonella enterica serovar Typhimurium ATCC 14028, and S. corvalis SF2 and S. cerro SF16), in small amounts. To determine the main phenolic group concentrated in the phytoextracts, a bio-guided study was conducted. The most significant results were obtained by O. tuberosa phytochemicals added to the culture medium at 50 µg/mL, yielding promising increases in biofilm formation (78% for Lp. plantarum and 43% for L. paracasei) and biosurfactant activity (112% for CO1-LVP105 strain). These adaptive strategies developed by bacteria possess key biotechnological significance. Furthermore, the bio-detoxification capacity of phenol and o-phenyl phenol, particularly of the novel strain CO1-LVP105, along with its mode of action and genetic identification, is described for the first time to our knowledge. In conclusion, lactobacilli strains have potential as fermentation starters and natural products, recovered from O. tuberosa peels, and added into culture media contribute to multiple bacterial biotechnological applications in both health and the environment.

1. Introduction

Lactic acid bacteria (LAB) have a long history of application in fermented foods due to their beneficial influence on nutritional, organoleptic, and shelf-life characteristics. Lactic acid fermentation is one of the most extensively investigated processes because of the significant application potential of LAB biomass in the food, chemical, and pharmaceutical industries. LAB have traditionally been the main probiotics used in food processing as starter cultures, pharmaceuticals, and biological control agents. More recently, the enormous relevance currently attributed to the beneficial intestinal microbiome for human health has led to increased interest in LAB. The nutraceutical industry is actively promoting the use of Lactobacillus in food as probiotics [1,2,3]. Nutraceuticals are substances that provide medical or health-promoting benefits, including the prevention and treatment of diseases, such as food supplements, herbal products, probiotics, and prebiotics, which are now of great importance to both the pharmaceutical and food industries [4,5].
Toxic ingredients in food (bacterial toxins, mycotoxins, pesticides or disinfectants, heavy metals, and natural antinutrients such as phytates, oxalates, and cyanide-generating glycosides) can lead to serious food-related diseases. LAB, as essential components of fermented foods and probiotics, are a main biological strategy against a great diversity of food-related toxins due to their detoxifying properties through adsorption or biodegradation processes [6].
Native Andean tubers, or “Andean treasures”, play a crucial role in the nutrition and economy of farmers in the Puna region of Argentina. Since ancient times, they have been used for nutritional and medicinal purposes. Andean tubers grow at high altitudes under challenging conditions of drought, freezing temperatures, and UV exposure, holding potential for exportation and further research in terms of adaptation and use in other regions of the world [7]. Many of them are categorized as neglected and underutilized species despite offering high vitamin, micronutrient, and starch content, in addition to dietary fibers, antioxidants, and antimicrobials associated with medicinal properties and health benefits [8]. Moreover, S. tuberosum peels exhibit healing properties for wounds and burns [9,10]. Andean potatoes possess potential for effective nutraceutical formulations, owing to the presence of antioxidant polyphenols, including chlorogenic acid, caffeoylquinic acid and its derivatives, flavonoids, flavonones, anthocyanins, and coumarins, among others. High-mountain tubers constitute natural sources of antioxidants, establishing their value as functional foods. Many of their constituents can neutralize free radicals, which are important in the fight against cancer, cardiovascular, and neurovascular diseases [11].
Recently, the scientific community has increasingly focused on the recovery of bioactive compounds from agro-industrial and processing wastes, such as leaves, peels, seeds, pulp, and shells, to reduce their environmental impact and align with the concept of a circular bio-economy. Potato peel compounds can be usefully employed as ingredients in functional foods or nutraceuticals [12].
The objectives of this study were as follows: (1) to test phytochemical-rich tuber peel extracts as selective growth/biofilm promoters of LAB, and (2) to evaluate the detoxification capacity of Lacticaseibacillus paracasei CO1-LVP105. For this purpose, a bioguided chemo-spectroscopic characterization of the phytochemicals obtained from peels of different varieties of edible tubers from the Argentinian Puna (such as Oxalis tuberosa Mol., Ullucus tuberosus Caldas, and Solanum tuberosum L.) was performed.
This research provides insights into the sustainable utilization of these peels as a natural source of selective promoters of a probiotic Lactiplantibacillus plantarum strain and of an environmental isolate of L. paracasei CO1-LVP105 from ovine origin, without stimulating the growth of a pathogenic bacteria set. In addition, the biodetoxifying power and genetic identification of the novel indigenous strain CO1-LVP105, as well as its surfactant capacity, were also determined.

2. Materials and Methods

2.1. Natural Product Source

Tubers were collected in the high-mountain plateau of the Puna desert (2500–4000 m) and obtained in the Feria Nacional de la Papa Andina (Alfarcito, Salta, Argentina). Species were identified and classified by Dr. María Inés Mercado (Foundation Miguel Lillo) in collaboration with Dr. Andrea Clausen and Dr. Ariana Digilio of the Banco Activo de Germoplasma EEA-INTA-Balcarce, Argentina.
The selected species were identified by their morphological characteristics as Oxalis tuberosa Mol. var. oca rosa and var. oca blanca, Ullucus tuberosus Caldas, and five varieties of Solanum tuberosum L. subsp. andigena var. miskila azul or Negra, var. miskila colorada, var. chila, var. cuarentona, and var. castilla blanca.

Natural Product Extraction

Thirty-two dried extracts were prepared from peels of O. tuberosa (var. oca rosa and oca blanca), U. tuberosus, and S. tuberosum L. subsp. andigena (var. miskila azul or negra, miskila colorada, cuarentona, chila, and castilla blanca).
Full ethanol extract was obtained by the maceration method using 96% ethanol (EE), aqueous extract was obtained by infusion at 50 °C during 30 min (AE), and their sub-extracts were obtained by partition of AE with ethyl acetate (EAS) and ethanol (ES), which were prepared from each sample. Then, the solvent was removed using a rotary evaporator, and the extraction yields were calculated and listed in Table 1.

2.2. Bacterial Strains

2.2.1. Probiotic and Environmental Bacteria

A food origin-probiotic bacterium, Lactiplantibacillus plantarum ATCC 10241 [13], and a wild isolate of Lacticaseibacillus paracasei CO1-LVP105 found in the soil and healthy sheep stool from a farm were employed for the bioassays. L. paracasei species is widely regarded as safe and has received GRAS/QPS status.
The culture media employed were PTYg (15 g/L peptone, 10 g/L triptone, 10 g/L yeast extract, and 5 g/L glucose, pH 6.0 ± 0.1) and PTYg, modified by the addition of natural products in concentrations of 25–100 µg/mL (pH 6.0 ± 0.2). Stock cultures were preserved in appropriate PTYg broth containing 20% glycerol (v/v) (−80 °C).

2.2.2. Pathogenic Bacteria

Gram-negative bacteria included Escherichia coli ATCC 35218, E. coli O157:H12, and Salmonella enterica serovar Typhimurium ATCC 14028, while indigenous pathogenic isolates of S. corvalis (SF2) and S. cerro (SF16) were also employed. All pathogenic strains were manipulated under Biosafety Level 2 (BSL-2) conditions at the Faculty of Biochemistry, Chemistry and Pharmacy (Universidad Nacional de Tucumán, Argentina) following institutional biosafety procedures. As only bacterial strains were used, no ethical approval was required under current regulations.

2.3. Genotypic Characterization of the Novel Isolate

The wild isolate’s entire genomic DNA was extracted using a PrestoTM Mini gDNA Bacteria Kit Quick Protocol from Geneaid in accordance with the instructions. Two primers were used to amplify the 16s rDNA; one of them was 27F (5′-GTGCTGCAGAGAGTTTGATCCTGGCTCAG-3′) and the other was 1492R (5′-CACGGATCCTACGGGTACCTTGTTACGACTT-3′) [14]. The thermocycler from My Cycler (Bio-Rad Laboratories, Hercules, CA, USA) was utilized to conduct the polymerase chain reaction. For the PCR mixture, 1.5 mmol/L MgCl 2.5 µL 10× reaction buffer, 100 µmol/L dNTPs, 0.5 µmol/L of each primer, 4 µL bacterial DNA, and 1.5 U Taq polymerase (Invitrogen, Carlsbad, CA, USA) were used. An initial denaturation at 94 °C for 5 min, followed by 30 cycles of denaturation at 94 °C for 1 min, hybridization at 52 °C for 2 min, and extension at 72 °C for 2 min with a final extension step at 72 °C for 7 min was performed. Amplification products were separated by electrophoresis (80 v) on a 0.8% (w/v) agarose gel stained with SYBR Gel DNA Safe Stain (Invitrogen, Carlsbad, CA, USA) in 1× TAE buffer (40 mmol/L Tris-acetate, 1 mmol/L EDTA, pH 8).
MACROGEN Inc. (Sejong, Republic of Korea) sequenced the PCR products after purifying them with the PCR Purification AccuPrep Kit (Bioneer, Hayward, CA, USA). 16S rRNA gene sequences were edited using Chromas Pro software (version 1.5, Technelysium Pty. Ltd., Brisbane, QLD, Australia, 2003–2009) and analyzed with DNAMAN software (version 2.6, Lynnon-Biosoft, San Ramon, CA, USA).
To determine sequence homologies, these were compared to the obtained sequences with those from the GenBank/EMBL/DDBJ database using BLAST (version 2.17.0 Technelysium Pty Ltd., Brisbane, QLD, Australia) (http://blast.ncbi.nlm.nih.gov/Blast.cgi (accessed on 5 June 2025)), and the closest relative was determined.
The 16S ribosomal DNA sequence of the isolated strain was deposited in GenBank, Accession number: PV763827.

2.4. Bacterial Growth

Extracts of plant species (EE, AE, EAS, ES) were sixfold assayed (n = 6 in two independent experiments) into a polystyrene microplate at 25, 50 and 100 μg/mL and dissolved in dimethyl sulfoxide (DMSO)-H2O (50:50). Control cultures contained the diluted culture (180 µL) and 20 µL of the solution of DMSO-H2O.
The bacterial cells (105 CFU/mL) were exposed to a maximum of 0.25% DMSO and cultured in liquid medium at 37 °C. Bacterial growth was measured as absorbance at 600 nm using a microplate reader (Power Wave XS2, BioTek Instruments, Winooski, VT, USA).

2.5. Biofilm Biomass Quantification

Crystal violet (CV)-based assay was used to quantify the development of biofilm on a polystyrene microplate, which was based on a protocol previously published [15]. The micromethod implemented allowed the use of small volumes (µL) and amounts (µg) of samples, and a 0.1% solution of CV was employed.
The removal of crystal violet binding to biofilm from each well was performed using 200 µL of ethanol, and a microtiter plate reader (Power Wave XS2, BioTek Instruments, Winooski, VT, USA) was used to determine the absorbance (560 nm) of ethanol solutions.
The results were calculated using the following formula:
Biofilm formation (%) = (As/A0) × 100,
where A0 is the absorbance of the control wells (biofilm biomass in untreated medium, measured at 560 nm) and As is the absorbance of each sample (biofilm biomass under test conditions, measured at 560 nm). The result is expressed as a percentage (%) of the control.

Specific Biofilm Index Determination

The specific biofilm index (SBI), which expresses the amount of biofilm that each bacterium forms, was determined as the relationship between the formed biofilm (measured at 560 nm) and the bacterial growth (measured at 600 nm) after 24 h, according to the following formula:
SBI = A560 nm − Ac560 nm/A600 nm,
where A: Absorbance measurement. Ac: Stained control wells containing only bacteria-free medium (to eliminate unspecific or abiotic OD values). The degree of biofilm production is classified in three categories according to this useful index: weak (SBI ≤ 0.5), moderate (0.5 > SBI ≤ 1), and strong (SBI > 1) [16].

2.6. Surfactant Activity

The oil dispersion test is a rapid and sensitive method for detecting surface-active substances. Thus, it is a useful tool for discovering the LAB surfactant activities. After 24 h of incubation, the bacterial cells were removed by centrifugation (3500 rpm, 15 min at 4 °C, 2193× g), and the supernatants were filtered through a 0.22 µm pore-size filter to obtain cell-free supernatants. For the assay, 20 µL of mineral oil was placed in a crystallizer of 250 mm in diameter, containing deionized water (100 mL), over millimeter paper according to the protocol developed by Cartagena et al. [17]. Then, 10 µL of each cell-free supernatant was placed in the center of the oil film. If a surfactant is present, the oil is displaced, and a clearing zone is formed. The diameter of this clearing zone on the surface of the oil correlates with the biosurfactant content and its activity [17,18,19]. The clear halos (mm) visualized under visible light were measured fivefold compared to the control. Polysorbate 80 (Tween 80, Merck, Darmstadt, Germany) was used as a reference standard. The PTYg and modified PTYg media (both without bacteria) showed no activity.

2.7. Liquid–Liquid Interfacial Activity

The emulsifying properties of the cell-free supernatants were evaluated using the emulsification index (E24) test. This test was conducted in quintuplicate (n = 5 in two independent experiments) using a micro-method previously described [20], which employs mineral oil as the oil phase and tween 80 as the reference standard. Briefly, a volume of the oil phase was added to an equal volume of cell-free supernatant, and the obtained mixture was vigorously vortexed for 2 min and left to stand for 24 h. Then, the height of the emulsion layer and the total height of both phases were measured. The E24 index was calculated as follows:
E24 (%) = height of the emulsion layer (mm)/total height (mm) × 100

2.8. Chemical and Spectrochemical Analysis

Qualitative reactions with ferric chloride solution (1%) and aluminum chloride solution (5%) were preliminarily implemented to characterize phenolic and flavonoid compounds [21] in the phytoextracts selected from a bioguided study. These reactions are often used in organic chemistry to identify the phenolic functional group. UV spectroscopy analysis was also carried out. This technique allows the detection of conjugated double bonds, aromatic compounds, and α,β-unsaturated carbonyls, according to the absorption maxima [22]. UV spectra were obtained using a Shimadzu UV-Vis 160 A spectrometer (Shimadzu Scientific Instruments Inc., Columbia, MD, USA). Extracts were dissolved in EtOH (spectroscopic grade, Sigma-Aldrich, Buenos Aires, Argentina) and placed in quartz cuvettes for UV measurements.

Quantification of Phenolic Compounds

For the bioguided phytochemical analysis, stock solutions of 30 mg/mL of each sub-extract of O. tuberosa var. oca rosa (EAS and ES) were used. Total extractable phenols and non-flavonoid compounds were determined colorimetrically using Folin–Ciocalteu’s reagent at 765 nm [23]. A standard curve was performed with gallic acid (2–20 µg/mL) as the standard (R2 = 0.9965, p < 0.05), and the results were expressed in mg of gallic acid equivalents (GAE) per g of sub-extract obtained from dry peels (mg GAE/g).
The total flavonoid content was determined using sodium nitrite (5%) and aluminum chloride (10%). The technique is based on the formation of specific colored complexes between the flavonoids and the reagents (NaNO2 and AlCl3), whose color intensity is measured spectrophotometrically at 510 nm [19]. For quantification, a quercetin standard curve (4–80 µg/mL) was used (R2 = 0.9939, p < 0.05), and the results were expressed as mg of quercetin equivalents (QE) per g of sub-extract (mg QE/g).
Orthodiphenols were assessed by UV-Visible spectrophotometry using the sodium molybdate method at a wavelength of 375 nm [24]. A standard curve was performed with caffeic acid (2–20 µg/mL) (R2 = 0.9981, p < 0.05), and the results were expressed as mg of caffeic acid equivalents (CAE) per g of extract (mg CAE/g).
Hydroxycinnamic derivatives were measured at 328 nm using caffeic acid (0.5–5 µg/mL) as the standard, and the results were expressed as mg of caffeic acid equivalents (CAE) per g of sub-extract (mg CAE/g) (R2 = 0.9972, p < 0.05) [25].
The evaluation of the total anthocyanin content was performed using the pH differential method, and the results were expressed as mg of cyanidin-3-glucoside equivalents per g of sub-extract (mg C3GLE/g) according to Carullo et al. [26].

2.9. Lactobacilli Fermentation into Modified Media

LAB were statically grown in 10 mL of PTYg supplemented with a solution of phenol at 100 μg/mL (pH 6.0 ± 0.2) for 7 days at 37 ± 1 °C in borosilicate glass tubes (10 mL) in duplicate. In the same way, o-phenyl phenol (OPP) was incorporated into the L. paracasei CO1-LVP105 culture medium (PTYg), modified by adding EAS of O. tuberosa var. oca rosa (50 µg/mL, pH 6.0 ± 0.1), which was selected and evaluated for its bioactivity. The inoculum was adjusted to 106 CFU/mL. Control experiments (without the addition of phenolic compounds or O. tuberosa EAS) were also performed.

2.10. Specific Enzyme Inhibition

The enzymatic activity of phenol oxidases in the L. paracasei CO1 culture was investigated with the oxidase test (Merck, Darmstadt, Germany) and corroborated by the addition of phenol solutions in concentrations ranged from 5 to 50 μg/mL, and combinations with dithiothreitol and ascorbic acid (as reducing agents) that were individually incorporated into the cultures in order to determine the enzyme inhibition [17,20].

2.11. Electron Microscopy

To determine the cell morphology of L. paracasei, Scanning Electron Microscopy (SEM) was employed. Phenol (PhOH) was chosen as a representative mutagenic substance [27] for the treatment. The samples (T and C) were fixed in Karnovsky’s solution (formaldehyde 2.66%, glutaraldehyde 1.66%, and phosphate buffer 0.1 M, pH 7.4) and incubated overnight at 4 °C for morphological and ultrastructural studies. The fixed samples were washed and dehydrated using the routine technique. The ZEISS Crossbeam 340 (Carl ZEISS NTS, Oberkochen, Germany) field emission scanning electron microscope was used to examine the samples previously coated with gold.

2.12. TLC and GC-MS Analysis

After seven days of incubation, the compounds (PhOH and OPP) remaining from biodegradation were extracted (from modified cultures with a solution of O. tuberosa EAS and unmodified) and analyzed by thin layer chromatography (TLC), using SiO2 plates (GF254, Merck, Darmstadt, Germany), CHCl3:CH3COOC2H5 (92.5:7.5%) as the mobile phase, and UV radiation to detect chromophores (such as phenols). Finally, an analysis by gas chromatography-mass spectrometry (GC-MS) was performed.
A Thermo Scientific TSQ 9000 triple quadrupole GC-MS/MS system (Thermo Scientific, Waltham, MA, USA) equipped with a DB-5 capillary column (5% phenyl-methylpolysiloxane, film thickness, 0.25 μm, inner diameter, 0.25 mm) was employed. The initial temperature of the column was 60 °C for 3 min. A temperature program was applied, ranging from 60 °C to 300 °C at a rate of 10 °C/min, followed by a final hold at 300 °C for 5 min. The carrier gas was helium (flow 1 mL/min). The main volatile constituents were determined by comparing their mass spectra with those of the WILEY/NIST mass spectral database (National Institute of Standards and Technology, Gaithersburg, MD, USA) and with authentic standards.

3. Results and Discussion

3.1. Identification of a Novel Lacticaseibacillus paracasei Strain

Based on the 16S rRNA gene sequence of the isolated strain, the comparative analysis using different databases (GenBank/EMBL/DDBJ) allowed the identification of a species with a high identity score. This sequence was deposited in GenBank, Accession number PV763827. The isolated strain of Lacticaseibacillus paracasei was lyophilized and incorporated into the LIVAPRA strain collection with the CO1-LVP105 code.

3.2. Effects of Tuber Peel Extracts Added into Culture Media on the Non-Pathogenic Bacteria Development

3.2.1. Food Origin-Probiotic Lactiplantibacillus plantarum ATCC 10241

In our study, all natural products derived from Andean tuber peels did not inhibit planktonic growth and biofilms of the food-derived probiotic Lp. plantarum ATCC 10241 when they were added to the culture medium, and, in many cases, increased these parameters. Interestingly, the oca rosa EAS (50 µg/mL) stimulated biofilm formation by 75.94% after 24 h of incubation (Figure 1), while S. tuberosum subsp. andigena var. chila extracts ES (100 µg/mL) and EE (50 µg/mL) stimulated it in 69.31% and 50.22%, respectively. In the case of ES from the variety blanca, the biofilm increment was 61.22% at the highest concentration of 100 µg/mL. Likewise, the best growth-promoting effects were determined with AE (100 µg/mL) and ES (25 µg/mL) of oca blanca, with increases of 59.94% and 54.16%, respectively (Figure S1A–G, Supplementary Materials).

3.2.2. Lacticaseibacillus paracasei CO1-LVP105

In line with the results obtained in our study of the Lp. plantarum ATCC 10241 strain, many extracts demonstrated potential as L. paracasei CO1-LVP105 growth and biofilm promoters (p < 0.05). The highest levels of biofilm stimulation were determined when the oca rosa natural products, EAS at 50 µg/mL (43%) and AE at 25 µg/mL (39%), were added to the culture media (Figure 2). The aqueous extract of U. tuberosus (50 µg/mL) also increased biofilm biomass, reaching 36.19%. At the maximum concentration of 100 µg/mL, EE and AE of miskila colorada stimulated 62.27% and 51.52%, respectively. Finally, increases of 40.52% and 38.59% with EE of cuarentona and ES of chila were observed at 100 µg/mL (Figure S2A–G, Supplementary Materials).
Regarding the growth promotion of this strain, AE, EE, EAS (50 µg/mL), and ES (25 µg/mL) from the oca blanca peels strongly increased bacterial growth (60.86%, 54.72%, 54.39%, and 54.97%, respectively), while the miskila negra EAS (25 µg/mL) produced a growth augmentation of 46.10% (Figure S2A–G).
It is important to note that the Andean tubers’ extracts did not exert inhibitory effects on the LAB. Only oca rosa and chila phytochemicals at the highest concentration (100 µg/mL) weakly reduced the L. paracasei CO1-LVP105 growth (up to 17%) with respect to the control culture (p < 0.05). In comparison, reductions in biofilm formation were only produced by chila ES (66.72%) and miskila colorada AE (50%) in concentrations of 25 µg/mL, as shown in Figure S2A–G (Supplementary Materials). However, these latter phytoextracts at 100 µg/mL increased biofilm biomass by 38.59% and 51.52% (respectively) due to a likely dose-dependent stress effect that triggered quorum detection mechanism (QS), as previously reported for other natural products [20].
Overall, phytochemicals concentrated in EAS, AE, and EE were the most potent at certain concentrations. According to Table 1 and Table 2, they represent a good source of flavonoids, ferulic acid, and coumarins, as is well known [11,28]. However, to our knowledge, this is the first study to investigate the main metabolites of Andean tuber peels that promote the development of beneficial bacteria.
Andean tubers or their bioactive constituents, such as phenolic compounds [29,30], may act as prebiotics, that is, “a substrate that is selectively utilized by host microorganisms conferring a health benefit” [31]. Thus, the combination of LAB with Andean tuber peel extracts could also serve as “synbiotic”, that is, a mixture of probiotics and prebiotics (dietary supplements), which selectively stimulates the growth and/or activates the metabolism of one or a limited number of health-promoting bacteria in the gastrointestinal tract, thus improving host welfare [32].
On the other hand, in many cases, the biofilm’s stimulating effects did not result in a significant increase in growth but rather in a probable adaptive behavior to chemical stress, mediated by a cell–cell communication mechanism (quorum sensing). In our study, similar patterns were observed: some Andean tuber peel extracts at specific concentrations enhanced biofilm formation of Lp. plantarum and L. paracasei CO1-LVP105 without concomitant increases in planktonic growth. This behavior is in line with the adaptive response to environmental stressors described by Verni et al. [20], indicating that selected polyphenolic compounds may trigger biofilm-related phenotypes without directly stimulating bacterial proliferation.
Capable of colonizing almost every environment, the microorganisms have evolved a wide range of biological responses to environmental stressors, and biofilms constitute a protective physical barrier that confers tolerance to antimicrobial agents (disinfectants and antibiotics) by reducing the diffusion of those toxic compounds. Moreover, they effectively reduce the grazing by protozoa. Biofilms are multicellular complex microbial communities held together by a self-produced extracellular matrix. They form highly diverse and complex structures that can attach to interfaces, grow, and aggregate in layers. Particularly, biofilm-growing probiotic bacteria can improve thermotolerance and freeze-drying resistance, and one of their important features is their capacity to replace pathogenic biofilms by annulling competitors [33].

3.3. Impact on Gram-Negative Bacteria

Phytoextracts selected for their bioactivity did not display stimulant effects on Gram-negative bacteria (Table S1). Some of these natural products significantly decreased biofilm formation (p < 0.05). As a result, Escherichia coli ATCC 35218 biofilm biomass was reduced by 41.64% for the U. tuberosus EAS (3EAS) at 25 µg/mL, and 42.64% by EE of S. tuberosum subsp. andigena var. cuarentona (7EE), while EAS of the miskila colorada variety diminished the biofilm of E. coli O157:H12 by 70.58% at 100 µg/mL. Coherently, these extracts decreased the E. coli O157:H12 specific biofilm index (SBI) from 2.48 (control) to 0.73 (5EAS), and for the ATCC strain, from 0.87 (control) to 0.53 and 0.51 by 3EAS and 7EE, respectively (Table S1, Supplementary Materials).
For Salmonella strains, significant effects were observed on the pathogenic strain S. cerro (SF-16) with reductions of 79.86% and 42.24% by 3EAS and the miskila colorada AE (5AE), respectively, at the highest concentration tested. Similarly, SBI decreased from 3.44 to 0.78 and 1.99 by modifying cultures with 3EAS and 5AE. In addition, 3EAS, 7EE, and EE of miskila colorada (5EE) at 100 µg/mL partially inhibited the biofilm biomass of the S. corvalis strain (approximately 40%). For the S. enterica ATCC 14028 biofilm, the reduction was only 22.75% by 5EE at 50 µg/mL, as shown in Table S1.
The polyphenolic-rich extracts from Andean tuber peels evaluated in this study significantly enhanced the growth and biofilm formation of Lp. plantarum ATCC 10241 and L. paracasei CO1-LVP105, while not stimulating and in some cases reducing biofilm formation of Gram-negative enteric pathogens. Given their composition (flavonoids, ferulic acid, and coumarins), these extracts could act as selective prebiotic substrates that favor colonization and persistence of beneficial lactobacilli while limiting pathogen adhesion. This selective stimulation, combined with the biosurfactant and detoxifying activities exhibited by the studied LAB strains, would help beneficial bacteria occupy intestinal niches, strengthen mucosal barriers, and competitively exclude pathogens. Such a mechanism provides a plausible and realistic way in which these extracts could support gut ecology and host health [34]. Previous studies have also reported selective antibacterial and antipathogenic effects of diverse plant-derived compounds, which support our findings [20,35].

3.4. Spectrochemical Data

Phenolic compounds absorb strongly in the ultraviolet region, and their UV spectra can be used in the detection. O. tuberosa and U. tuberosus extracts exhibited typical UV absorptions of flavonoid compounds at 300–390 nm (band I) and a band II at 250–280 nm (Table 1). These bands are ascribable to electronic transitions within the aromatic three-ring system of the ligand molecule. Specifically, band I is due to the absorption of the cinnamoyl system (ring B), while band II is due to that of the benzoyl moiety (ring A) [36]. In all extracts, UV absorption peaks of phenolic compounds, such as ferulic acid (287–312 nm) and coumarin (290–300 nm), were also recorded [37].
These results were consistent with the qualitative chemical test data, as shown in Table 1. Recently, Vescovo and co-authors (2025) claimed that potato peels represent a rich source of bioactive compounds, including phenolics among other constituents, as a byproduct of significant agro-industrial value [12].

Phenolic Compounds: Quantitative Bioguided Analysis

The quantitative analysis of O. tuberosa var. oca rosa, whose sub-extract was selected for its stimulant effects on lactobacilli strains, showed a rich polyphenolic composition (Table 2). In fact, total phenolics, non-flavonoid phenolics, flavonoids, hydroxycinnamic acids, and orthodiphenols were found in higher percent in the EAS than ES, resulting in the first sub-extract at 50 µg/mL being the most active (Figure 1).
Recently, Tapia and co-authors (2024) reported lower percentages of phenolic compounds in grape waste products (pomace) in general [23]. Therefore, the peels of the oca rosa tuber constitute a promising natural source of polyfunctional phenolic compounds.

3.5. Toxic Compounds Biodegradation

Phenol was chosen as a representative mutagenic disinfectant [27] for its treatment, as well as o-phenylphenol (OPP), which is usually applied in disinfections, veterinary hygiene, food, and animals, being very toxic to aquatic life with lasting effects, and causing serious eye damage and skin and respiratory irritations [38].
Phenol was degraded by L. paracasei CO1-LVP105 and Lp. plantarum ATCC 10241 (even at 100 μg/mL) after seven days of static incubation at 37 °C (pH 5.0 ± 0.2). Using two chromatographic methods, TLC-UV and GC-MS (sensibility limits ~10−9 and 10−12, respectively), no traces of phenol were detected in the obtained chromatographic profiles, indicating the ability of these LAB strains to remove the mutagenic substance.
In contrast, only 20% of the OPP solution (100 µg/mL) was degraded by the wild-type strain CO1-LVP105, while this same culture, previously stimulated with 1EAS at 50 µg/mL, removed the OPP solution up to 73% and improved the bacterial degrading activity by 53%, which was inferred from the area estimation of the OPP peaks obtained by GC-MS, as shown in Table 3.
This differential behavior is likely due to structural features and lipophilicity, with OPP being more stressful and stimulating to surface activity than phenol (Table 4). Biosurfactants facilitate the solubilization and metabolism of lipophilic substances. This process is more difficult for OPP (n-octanol-water partition coefficient = 3.06) than for phenol (n-octanol-water partition coefficient = 1.47).
Similarly, a biofilm-producing strain, L. paracasei CE75, degraded phenol (100 µg/mL) and 4-hydroxyacetophenone (25 µg/mL) by 99.89%, as previously reported [20].
It is noteworthy that the SEM analysis showed a strong increase in biofilm biomass in the presence of phenol (25 µg/mL), in comparison to control cells (Figure 3A), and a higher number of adhering cells, which are arranged in the form of solid aggregates in treated cells (Figure 3B–E). Microphotographs also revealed that treated cells present a rough surface with a considerable number of variable-sized protrusions (Figure 3D,E) as an adaptive response to chemical stress. These results are in agreement with previous studies [20,39].

3.6. Surfactant Activity

Phenolic compounds added to the cultures enhanced the oil-spreading activity of the supernatant, particularly when OPP or PhOH at 100 μg/mL were individually added to the L. paracasei CO1-LVP105 cultures, which had been previously incubated with the oca rosa EAS at 50 μg/mL. Halos were approximately four and two times larger than that of the control supernatant (80 ± 5 mm), and higher than tween 80 (50 ± 3 mm). Probiotic Lp. plantarum ATCC 10241 strains also increased their surfactant activity in the presence of PhOH (24%), as shown in Table 4.

3.7. Liquid–Liquid Interfacial Activity

The emulsification activity of LAB supernatants from cultures previously incubated with phenol (100 μg/mL), EAS of oca rosa (50 μg/mL) or the mixture was greater (66–71%) than tween 80 (50%); in particular, the cell-free supernatant from L. paracasei CO1-LVP105 culture, previously stimulated with phenolic compounds, exhibited an emulsification activity of 71% (Table 4) and better than the control culture supernatant (50%).
It is important to note that these substances and their solvent system, previously added to the culture media, did not exert any surface activity by themselves. Therefore, the substantial increase in supernatant surface activity would be due to an increase in surface-active substances resulting from the induction of their biosynthesis in the LAB cultures, as reported in our previous studies of L. paracasei CE75 [20]. The extracellular biosurfactants’ increase optimized the biodegradation process, as explained in Section 3.5.
A great number of bacteria are able to produce extracellular and/or cell-bound biosurfactants [40,41]. Indeed, Lara and co-authors (2025) identified a LAB strain, Lp. plantarum Tw226, as a cell-bound surfactant producer. This biosurfactant has a glycolipopeptide nature and is useful as an emulsifier of food ingredients such as corn oil. Studies on biosurfactants produced by LAB as essential oils’ emulsifiers are quite scarce [41]. Therefore, its approach in L. paracasei CO1-LVP105, as well as studies of functional and safety properties, would be essential in future studies.

3.8. Phenol Oxidase Activity

Phenol and OPP biodegradation was carried out by phenol oxidases, determined according to Lee et al. [42], in the Lacticaseibacillus paracasei CO1-LVP105 cultures by adding specific enzymatic inhibitors that increased the susceptibility of this bacterium to phenol, and by the oxidase test. The occurrence of (poly)phenol oxidases in LAB was previously reported by Matthews et al. [43].
These results showed congruence with our previous studies of the Flourensia fiebrigii phytochemicals that demonstrated stimulant effects on L. paracasei subsp. paracasei CE75 strain, as well as enzymes and glycoprotein surfactants involved in the detoxifying process [17,20].
The safety profile of Lacticaseibacillus paracasei CO1-LVP105 is supported by comparison with other strains of the same species. For instance, L. paracasei strain F-19e has been evaluated under GRAS Notice No. 810 by the U.S. Food & Drug Administration, including whole-genome analyses, absence of virulence and antibiotic resistance genes, and human feeding studies with infants, children, and adults, without adverse outcomes [44]. Likewise, L. paracasei subsp. paracasei Q-1, isolated from cow dung, showed no hemolytic, gelatinase, or mucin-degrading activity in vitro, and in vivo toxicity studies in mice (14-day acute and 28-day sub-chronic) revealed no effects on organ indices or histology [45].
These data contextualize the origin of CO1-LVP105 from healthy animals and its lack of observed pathogenic traits under our experimental conditions, although whole-genome sequencing of CO1-LVP105 remains a planned future work to confirm its safety more precisely.

4. Conclusions

The study provides valuable and comprehensive information on the Andean tuber peels as natural sources of phytochemicals, mainly phenolic compounds, and their selective stimulating effects on probiotic and environmental bacteria without promoting the development of pathogenic bacteria.
A novel indigenous strain was also identified as Lacticaseibacillus paracasei CO1-LVP105, and our results suggest a promising detoxifying activity of phenol, in high concentrations of 100 µg/mL, just like that of Lactiplantibacillus plantarum ATCC 10241. In addition, small amounts of EAS from O. tuberosa var. oca rosa added to L. paracasei CO1-LVP105 culture medium increased the bacterial surfactant activity (112%) and its OPP detoxification capacity (34.4%), mediated by phenol oxidases that were determined herein.
Given the nutraceutical and biotechnological potential of Andean tuber peels, it is essential to promote global strategies for their sustainable applications as an innovative technological strategy for the LAB fermentation.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/fermentation11100576/s1, Genotypic characterization of the novel isolate. Figure S1 (A–G). Effects of Andean tuber peels on the growth and biofilm formation of Lactiplantibacillus plantarum ATCC 10241. Figure S2 (A–G). Effects of Andean tuber peels on the growth and biofilm formation of Lacticaseibacillus paracasei CO1-LVP105. Table S1. Impact of the selected phytoextracts on Gram-negative bacteria.

Author Contributions

C.H.O.: Performed the experiments and analyzed the data. M.I.M.: Performed the experiments, analyzed the data, and wrote the paper. F.E.A.M.: Performed the experiments and analyzed the data. M.E.A.: Performed the experiments, analyzed the data, and wrote and revised the paper. E.C.: Conceived and designed the experiments, analyzed the data, and wrote and revised the paper. All authors have read and agreed to the published version of the manuscript.

Funding

The authors acknowledge the financial support from the Secretaria de Ciencia, Arte e Innovación Tecnológica-SCAIT-UNT (PIUNT 26D-715, 26D-708, PICT2021-A00439) and the Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data will be made available on request.

Acknowledgments

The authors gratefully acknowledge María Ángela Jure for providing the pathogenic bacteria, as well as Cintia Mariana Romero (CONICET-UNT) and Miguel Gilabert (CONICET-UNT) for their valuable help in the GC-MS measurements.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Neylon, E.; Nyhan, L.; Zannini, E.; Monin, T.; Münch, S.; Sahin, A.W.; Arendt, E.K. Food Ingredients for the future: In-depth analysis of the effects of lactic acid bacteria fermentation on spent barley rootlets. Fermentation 2023, 9, 78. [Google Scholar] [CrossRef]
  2. Abedin, M.M.; Chourasia, R.; Phukon, L.C.; Sarkar, P.; Ray, R.C.; Singh, S.P.; Rai, A.K. Lactic acid bacteria in the functional food industry: Biotechnological properties and potential applications. Crit. Rev. Food Sci. Nutr. 2024, 64, 10730–10748. [Google Scholar] [CrossRef] [PubMed]
  3. Nuñez, I.M.; Verni, M.C.; Argañaraz Martinez, F.E.; Babot, J.D.; Terán, V.; Danilovich, M.E.; Cartagena, E.; Alberto, M.R.; Arena, M.E. Novel lactic acid bacteria strains from regional peppers with health-promoting potential. Fermentation 2024, 10, 209. [Google Scholar] [CrossRef]
  4. Dable-Tupas, G.; Otero, M.C.B.; Bernolo, L. (Eds.) Functional Foods and Nutraceuticals: Bioactive Components, Formulations, and Innovations; Springer Nature: Berlin/Heidelberg, Germany, 2020. [Google Scholar]
  5. AlAli, M.; Alqubaisy, M.; Aljaafari, M.N.; AlAli, A.O.; Baqais, L.; Molouki, A.; Abushelaibi, A.; Lai, K.-S.; Lim, S.-H.E. Nutraceuticals: Transformation of Conventional Foods into Health Promoters/Disease Preventers and Safety Considerations. Molecules 2021, 26, 2540. [Google Scholar] [CrossRef]
  6. Petrova, P.; Arsov, A.; Tsvetanova, F.; Parvanova-Mancheva, T.; Vasileva, E.; Tsigoriyna, L.; Petrov, K. The complex role of lactic acid bacteria in food detoxification. Nutrients 2022, 14, 2038. [Google Scholar] [CrossRef]
  7. Chantre-López, A.R.; Solanilla-Duque, J.F.; Ascacio-Valdes, J.A.; Salazar-Sánchez, M.d.R.; Flores-Gallegos, A.C.; Nery-Flores, S.D.; Silva-Belmares, S.Y.; Rodríguez-Herrera, R. Andean tubers: Traditional medicine and other applications. Plant Sci. Today 2024, 11, 342–352. [Google Scholar] [CrossRef]
  8. Choquechambi, L.A.; Callisaya, I.R.; Ramos, A.; Bosque, H.; Mújica, A.; Jacobsen, S.-E.; Sørensen, M.; Leidi, E.O. Assessing the nutritional value of root and tuber crops from Bolivia and Peru. Foods 2019, 8, 526. [Google Scholar] [CrossRef]
  9. Camire, M.E.; Kubow, S.; Donnelly, D.J. Potatoes and human health. Crit. Rev. Food Sci. Nutr. 2009, 49, 823–840. [Google Scholar] [CrossRef]
  10. Rosas-Cruz, G.P.; Silva-Correa, C.R.; Calderón-Peña, A.A.; Villarreal-La Torre, V.E.; Aspajo-Villalaz, C.L.; CruzadoRazco, J.L.; Rosario-Chávarri, J.; Rodríguez-Soto, J.; Pretel-Sevillano, O.; Sagástegui-Guarniz, W.; et al. Wound healing activity of an ointment from Solanum tuberosum L. “Tumbay yellow potato” on Mus musculus Balb/c. Pharmacogn. J. 2020, 12, 1268–1275. [Google Scholar] [CrossRef]
  11. Campos, D.; Noratto, G.; Chirinos, R.; Arbizu, C.; Roca, W.; Cisneros-Zevallos, L. Antioxidant capacity and secondary metabolites in four species of Andean tuber crops: Native potato (Solanum sp.), mashua (Tropaeolum tuberosum Ruiz & Pavon), oca (Oxalis tuberosa Molina) and ulluco (Ullucus tuberosus Caldas). J. Sci. Food Agric. 2006, 86, 1481–1488. [Google Scholar] [CrossRef]
  12. Vescovo, D.; Manetti, C.; Ruggieri, R.; Spizzirri, U.G.; Aiello, F.; Martuscelli, M.; Restuccia, D. The Valorization of potato peels as a functional ingredient in the food industry: A comprehensive review. Foods 2025, 14, 1333. [Google Scholar] [CrossRef] [PubMed]
  13. Mis-Solval, K.E.; Jiang, N.; Yuan, M.; Joo, K.H.; Cavender, G.A. The Effect of the ultra-high-pressure homogenization of protein encapsulants on the survivability of probiotic cultures after spray drying. Foods 2019, 8, 689. [Google Scholar] [CrossRef] [PubMed]
  14. Lane, D.J. 16S/23S rRNA sequencing. In Nucleic Acid Techniques in Bacterial Systematics; Stackebrandt, E., Goodfellow, M., Eds.; Wiley: New York, NY, USA, 1991; pp. 115–175. [Google Scholar]
  15. O’Toole, G.A.; Kolter, R. Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: A genetic analysis. Mol. Microbiol. 1998, 28, 449–461. [Google Scholar] [CrossRef] [PubMed]
  16. Martinez-Medina, M.; Naves, P.; Blanco, J.; Aldeguer, X.; Blanco, J.E.; Blanco, M.; Ponte, C.; Soriano, F.; Darfeuille-Michaud, A.; Garcia-Gil, L.J. Biofilm formation as a novel phenotypic feature of adherent-invasive Escherichia coli (AIEC). BMC Microbiol. 2009, 9, 202. [Google Scholar] [CrossRef]
  17. Cartagena, E.; Orphèe, C.H.; Verni, M.C.; Arena, M.E.; González, S.N.; Argañaraz, M.I.; Bardón, A. Patent: Medio de Cultivo Promotor y Bacterias No Patógenas Detoxificantes de Compuestos Mutagénicos/Carcinogénicos. Instituto Nacional de la Propiedad Industrial-INPI Nº 20190102418, 25 March 2021. [Google Scholar]
  18. Walter, V.; Syldatk, C.; Hausmann, R. Screening Concepts for the Isolation of Biosurfactant Producing Microorganisms. In Biosurfactants. Advances in Experimental Medicine and Biology; Sen, R., Ed.; Springer: New York, NY, USA, 2010; pp. 1–13. [Google Scholar]
  19. Sambanthamoorthy, K.; Feng, X.; Patel, R.; Patel, S.; Paranavitana, C. Antimicrobial and antibiofilm potential of biosurfactants isolated from lactobacilli against multi-drug-resistant pathogens. BMC Microbiol. 2014, 14, 197. [Google Scholar] [CrossRef]
  20. Verni, M.C.; Orphèe, C.H.; González, S.N.; Bardón, A.; Arena, M.E.; Cartagena, E. Flourensia fiebrigii SF Blake in combination with Lactobacillus paracasei subsp. paracasei CE75. A novel antipathogenic and detoxifying strategy. LWT 2022, 156, 113023. [Google Scholar]
  21. Jia, Z.S.; Tang, M.C.; Wu, J.M. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem. 1999, 64, 555–559. [Google Scholar] [CrossRef]
  22. Chen, Z. Application of UV-vis spectroscopy in the detection and analysis of substances. Trans. Mater. Biotechnol. Life Sci. 2024, 3, 131–136. [Google Scholar] [CrossRef]
  23. Tapia, P.E.; Silva, A.M.; Delerue-Matos, C.; Moreira, M.; Rodrigues, F.; Torres Carro, R.; Santi, M.D.; Ortega, M.G.; Blázquez, M.A.; Arena, M.E.; et al. Exploring the phytochemical composition and the bioactive properties of malbec and torrontés wine pomaces from the Calchaquíes Valleys (Argentina) for their sustainable exploitation. Foods 2024, 13, 1795. [Google Scholar] [CrossRef]
  24. Singh, P.K.; Singh, J.; Medhi, T.; Kumar, A. Phytochemical screening, quantification, FT-IR analysis, and in silico characterization of potential bioactive compounds identified in hr-lc/ms analysis of the polyherbal formulation from Northeast India. ACS Omega 2022, 7, 33067–33078. [Google Scholar] [CrossRef]
  25. Ben Hmida, R.; Frikha, N.; Bouguerra Neji, S.; Kit, G.; Medina, F.; Bouaziz, M. Synthesis of high added value compounds through catalytic oxidation of 2-phenylethanol: A kinetic study. Int. J. Chem. Kinet. 2020, 52, 124–133. [Google Scholar] [CrossRef]
  26. Carullo, G.; Spizzirri, U.G.; Loizzo, M.R.; Leporini, M.; Sicari, V.; Aiello, F.; Restuccia, D. Valorization of red grape (Vitis vinifera cv. sangiovese) pomace as functional food ingredient. Ital. J. Food Sci. 2020, 32, 367–385. [Google Scholar]
  27. ECHA. European Chemicals Agency Documents. 2021. Available online: https://echa.europa.eu/documents/10162/de108693-1d4f-abfe-818e-9cea812ab4c2 (accessed on 21 April 2025).
  28. Lachman, J.; Hamouz, K. Red and purple coloured potatoes as a significant antioxidant source in human nutrition—A review. Plant Soil Environ. 2005, 51, 477–482. [Google Scholar] [CrossRef]
  29. Campos, D.; Betalleluz-Pallardel, I.; Chirinos, R.; Aguilar-Galvez, A.; Noratto, G.; Pedreschi, R. Prebiotic effects of yacon (Smallanthus sonchifolius Poepp. & Endl.), a source of fructooligosaccharides and phenolic compounds with antioxidant activity. Food Chem. 2012, 135, 1592–1599. [Google Scholar] [CrossRef]
  30. Verediano, T.A.; Viana, M.L.; Vaz-Tostes, M.D.G.; Costa, N. The potential prebiotic effects of yacon (Smallanthus sonchifolius) in colorectal cancer. Curr. Nutr. Food Sci. 2021, 17, 167–175. [Google Scholar] [CrossRef]
  31. Gibson, G.R.; Hutkins, R.; Sanders, M.E.; Prescott, S.L.; Reimer, R.A.; Salminen, S.J.; Scott, K.; Stanton, C.; Swanson, K.S.; Cani, P.D.; et al. Expert consensus document: The international scientific association for probiotics and prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics. Nat. Rev. Gastroenterol. Hepatol. 2017, 14, 491–502. [Google Scholar] [CrossRef] [PubMed]
  32. Palai, S.; Derecho, C.M.P.; Kesh, S.S.; Egbuna, C.; Onyeike, P.C. Prebiotics, probiotics, synbiotics and its importance in the management of diseases. In Functional Foods and Nutraceuticals: Bioactive Components, Formulations and Innovations; Springer International Publishing: Cham, Switzerland, 2020; pp. 173–196. [Google Scholar]
  33. Berlanga, M.; Guerrero, R. Living together in biofilms: The microbial cell factory and its biotechnological implications. Microb. Cell Fact. 2016, 15, 165–176. [Google Scholar] [CrossRef] [PubMed]
  34. Frassinetti, S.M.G.; Moccia, E.; Longo, V.; Di Gioia, D. Antimicrobial and antibiofilm activity of Cannabis sativa L. seeds extract against Staphylococcus aureus and growth effects on probiotic Lactobacillus spp. LWT 2020, 124, 109149. [Google Scholar]
  35. Cartagena, E.; Arena, M.E.; Verni, M.C.; Bardón, A. Natural terpenoids as a valuable resource of selective health-beneficial biofilm promoters. In Biofilms: Advances in Research and Applications; Rowland, S., Ed.; Nova Science Publishers: New York, NY, USA, 2021; pp. 159–186. [Google Scholar]
  36. Santonoceta, G.D.G.; Sgarlata, C. pH-Responsive cobalt(II)-coordinated assembly containing quercetin for antimicrobial applications. Molecules 2023, 28, 5581. [Google Scholar] [CrossRef]
  37. Holser, R. Principal component analysis of phenolic acid spectra. Int. Sch. Res. Not. 2012, 2012, 493203. [Google Scholar] [CrossRef]
  38. Scientific Committee on Consumer Safety. 2015. Available online: https://ec.europa.eu/health/scientific_committees/consumer_safety/docs/sccs_o_177.pdf (accessed on 2 September 2025).
  39. Parlindungan, E.; Dekiwadia, C.; Tran, K.T.; Jones, O.A.; May, B.K. Morphological and ultrastructural changes in Lactobacillus plantarum B21 as an indicator of nutrient stress. LWT 2018, 92, 556–563. [Google Scholar] [CrossRef]
  40. Lvova, K.; Martínez-Arcos, A.; López-Prieto, A.; Vecino, X.; Moldes, A.B.; Cruz, J.M. Optimization of the operational conditions to produce extracellular and cell-bound biosurfactants by Aneurinibacillus aneurinilyticus using corn steep liquor as a unique source of nutrients. Fermentation 2023, 9, 351. [Google Scholar] [CrossRef]
  41. Lara, V.M.; Gliemmo, M.F.; Vallejo, M.; García González, M.d.C.; Alfaro Rodríguez, M.d.C.; Campos, C.A. Use of the glycolipopeptid biosurfactant produced by Lactiplantibacillus plantarum Tw226 to formulate functional Cinnamon bark essential oil emulsions. Foods 2025, 14, 1540. [Google Scholar] [CrossRef]
  42. Lee, M.Y.; Lee, M.K.; Park, I. Inhibitory effect of onion extract on polyphenol oxidase and enzymatic browning of taro (Colocasia antiquorum var. esculenta). Food Chem. 2007, 105, 528–532. [Google Scholar] [CrossRef]
  43. Matthews, A.; Grimaldi, A.; Walker, M.; Bartowsky, E.; Grbin, P.; Jiranek, V. Lactic acid bacteria as a potential source of enzymes for use in vinification. Appl. Environ. Microbiol. 2004, 70, 5715–5731. [Google Scholar] [CrossRef]
  44. Arla Foods Ingredients Group P/S.; Heimbach, J.T. GRAS Notice No. 810: Lactobacillus paracasei subsp. paracasei Strain F-19e—Intended Use in Powdered Milk-Based Infant Formula; Scientific Procedures. U.S. Food & Drug Administration. 2019. Available online: https://www.fda.gov/media/129477/download (accessed on 21 April 2025).
  45. Wang, F.; Chen, L.; Li, Y.; Gao, G.; Wang, Y.; Shi, R.; Zhang, H.; Chen, Y.; Cao, R.; Shi, Q.; et al. Lacticaseibacillus paracasei subsp. paracasei Q-1 exhibits good safety and effectively prevents Escherichia coli K99-induced diarrhea in mice. Probiotics Antimicrob. Proteins 2025. Advance online publication. [Google Scholar] [CrossRef]
Fermentation 11 00576 g001
Figure 1. Oxalis tuberosa effects on growth and biofilm formation of Lactiplantibacillus plantarum ATCC 10241. EAS: Ethyl acetate sub-extract. AE: Aqueous extract. ES: Ethanol sub-extract. EE: Ethanol extract. Asterisks (* for growth bars, and ** for biofilm bars) indicate significant differences (n = 6 in two independent experiments), according to Tukey’s test (p < 0.05).
Figure 1. Oxalis tuberosa effects on growth and biofilm formation of Lactiplantibacillus plantarum ATCC 10241. EAS: Ethyl acetate sub-extract. AE: Aqueous extract. ES: Ethanol sub-extract. EE: Ethanol extract. Asterisks (* for growth bars, and ** for biofilm bars) indicate significant differences (n = 6 in two independent experiments), according to Tukey’s test (p < 0.05).
Fermentation 11 00576 g001
Fermentation 11 00576 g002
Figure 2. O. tuberosa effects on growth and biofilm formation of Lacticaseibacillus paracasei CO1-LVP105. EAS: Ethyl acetate sub-extract. AE: Aqueous extract. ES: Ethanol sub-extract. EE: Ethanol extract. Asterisks (* for growth bars, and ** for biofilm bars) indicate significant differences (n = 6 in two independent experiments), according to Tukey’s test (p < 0.05).
Figure 2. O. tuberosa effects on growth and biofilm formation of Lacticaseibacillus paracasei CO1-LVP105. EAS: Ethyl acetate sub-extract. AE: Aqueous extract. ES: Ethanol sub-extract. EE: Ethanol extract. Asterisks (* for growth bars, and ** for biofilm bars) indicate significant differences (n = 6 in two independent experiments), according to Tukey’s test (p < 0.05).
Fermentation 11 00576 g002
Fermentation 11 00576 g003
Figure 3. (A–E) Scanning electron microphotographs of Lacticaseibacillus paracasei CO1-LVP105 cultures grown in the presence (treated) and absence (control) of phenol at 25 µg/mL. (A): Control (2500×). (B–E): Treated (1000×, 2500×, 5000× and 10,000×, respectively).
Figure 3. (A–E) Scanning electron microphotographs of Lacticaseibacillus paracasei CO1-LVP105 cultures grown in the presence (treated) and absence (control) of phenol at 25 µg/mL. (A): Control (2500×). (B–E): Treated (1000×, 2500×, 5000× and 10,000×, respectively).
Fermentation 11 00576 g003
Table 1. Phytochemicals from Andean tuber peels’ extracts, yields, and comparative UV spectroscopy data of the selected bioactive extracts.
Table 1. Phytochemicals from Andean tuber peels’ extracts, yields, and comparative UV spectroscopy data of the selected bioactive extracts.
Andean Plant Tubers
(Names)		Peel Extracts
(Codes)		Extract Yields
(mg/g of Dry Peels)		Phenolic Compounds
(FeCl3 and AlCl3 Reagents)		UV SpectroscopyAssignments
λ (nm)Abs
Oxalis tuberosa var.
oca rosa		1AE11.9Positive 348.51.003Flavonoids (cinnamoyl group)
325.51.120Ferulic acid and coumarins
303.51.225
1EAS5.0Positive323.00.774Flavonoids (cinnamoyl group)
282.41.243Flavonoids (benzoyl group), Ferulic acid, and coumarins
Ullucus tuberosus3AE72.9Positive348.50.631Flavonoids (cinnamoyl)
267.51.147Flavonoids (benzoyl group)
3EAS1.1Positive351.20.704Flavonoids (cinnamoyl)
277.21.191Flavonoids (benzoyl group), Ferulic acid and, coumarins
Solanum tuberosum subsp. andigena
var. miskila colorada		5AE20.0Positive343.00.791Flavonoids (cinnamoyl)
300.50.461Ferulic acid and coumarins
5EAS1.6Positive341.50.617Flavonoids (cinnamoyl group)
321.50.964Ferulic acid and coumarins
300.50.447
EE24.7Positive322.01.110Flavonoids (cinnamoyl group)
305.50.965Ferulic acid and coumarins
294.50.956Flavonoids (benzoyl group)
Solanum tuberosum subsp. andigena var.
cuarentona		7EE31.5Positive322.01.843Flavonoids (cinnamoyl group)
305.51.619Ferulic acid and coumarins
AE: Aqueous extract. EAS: Ethyl acetate sub-extract. EE: Ethanol extract. All extractions were performed in single batch.
Table 2. Quantification of Phenolic Compounds of O. tuberosa var. oca rosa.
Table 2. Quantification of Phenolic Compounds of O. tuberosa var. oca rosa.
Phytochemical GroupEAS
mg/g Dry Peel Sub-Extract		ES
mg/g Dry Peel Sub-Extract
Total phenolics (GAE)226.39 ± 18.25 a29.45 ± 0.28 f
Non-flavonoid phenolics (GAE)123.85 ± 10.25 b17.66 ± 0.39 e
Total flavonoids (QE)398.04 ± 18.02 c111.27 ± 2.08 g
Hydroxycinnamic acids (CAE)21.40 ± 0.95 d5.75 ± 0.21 h
Orthodiphenols (CAE)18.07 ± 0.65 e2.34 ± 0.51 i
Orthodiphenols (GAE)29.42 ± 1.06 f3.80 ± 0.83 j
Anthocyannins (C3GE)<LOQ<LOQ
GAE: Gallic acid equivalents. QE: Quercetin equivalents. CAE: Caffeic acid equivalents. C3GE: Cyanidin-3-glucoside equivalents. LOQ: Limit of quantification. Results are expressed as means ± standard deviations. Different letters indicate significant differences between samples (p < 0.05).
Table 3. GC-MS of o-phenyl phenol (OPP) data under different conditions (retention times, area, comparative peaks, and mass spectrum).
Table 3. GC-MS of o-phenyl phenol (OPP) data under different conditions (retention times, area, comparative peaks, and mass spectrum).
Culture Media Under Different ConditionsGC-MS
OPP Retention Time		GC-MS
Peak Area		OPP Comparative Peaks (Magnified) and Mass Spectrum
Solution of 100 µg/mL OPP standard (abiotic control)12.76 min
(peak in blue)		16.969.230.572.786Fermentation 11 00576 i001
Solution of 100 µg/mL OPP standard plus L. paracasei CO1-LVP105 12.74 min
(peak in magenta)		13.504.203.129.604
Solution of 100 µg/mL OPP standard plus L. paracasei CO1-LVP105 promoted with 1EAS 12.71 min
(peak in gray)		4.652.244.706.499
Table 4. LAB surfactant and emulsifying activities.
Table 4. LAB surfactant and emulsifying activities.
SupernatantsOil Spreading Halos (mm)E24 Index (%)
Lacticaseibacillus paracasei CO1-LVP10580 ± 5 a 54 ± 5 h
L. paracasei CO1-LVP105 + PhOH88 ± 1b67 ± 6 i
L. paracasei CO1-LVP105- EAS + PhOH 169 ± 2 c71 ± 0 j
L. paracasei CO1-LVP105 + OPP160 ± 0 d ND
L. paracasei CO1-LVP105-EAS + OPP 320 ± 0 e ND
Lactiplantibacillus plantarum ATCC 10241105 ± 2 f53 ± 3 h
Lp. plantarum ATCC 10241 + PhOH130 ± 1 g66 ± 5 i
Lp. plantarum ATCC 10241 -EAS+ PhOH129 ± 5 g 67 ± 4 i
Tween 8050 ± 3 h50 ± 0 h
OPP: O-phenyl phenol at 100 µg/mL. EAS: Ethyl acetate extract of O. tuberosa var. oca rosa at 50 µg/mL. PhOH: Phenol solution at 100 µg/mL. ND: Not determined. Results are expressed as means ± standard deviations. Different letters indicate significant differences between samples (p < 0.05).
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Orphèe, C.H.; Mercado, M.I.; Argañaraz Martínez, F.E.; Arena, M.E.; Cartagena, E. High-Mountain Tuber Products Improve Selectively the Development and Detoxifying Capacity of Lactobacilli Strains as an Innovative Culture Strategy. Fermentation 2025, 11, 576. https://doi.org/10.3390/fermentation11100576

AMA Style

Orphèe CH, Mercado MI, Argañaraz Martínez FE, Arena ME, Cartagena E. High-Mountain Tuber Products Improve Selectively the Development and Detoxifying Capacity of Lactobacilli Strains as an Innovative Culture Strategy. Fermentation. 2025; 11(10):576. https://doi.org/10.3390/fermentation11100576

Chicago/Turabian Style

Orphèe, Cecilia Hebe, María Inés Mercado, Fernando Eloy Argañaraz Martínez, Mario Eduardo Arena, and Elena Cartagena. 2025. "High-Mountain Tuber Products Improve Selectively the Development and Detoxifying Capacity of Lactobacilli Strains as an Innovative Culture Strategy" Fermentation 11, no. 10: 576. https://doi.org/10.3390/fermentation11100576

APA Style

Orphèe, C. H., Mercado, M. I., Argañaraz Martínez, F. E., Arena, M. E., & Cartagena, E. (2025). High-Mountain Tuber Products Improve Selectively the Development and Detoxifying Capacity of Lactobacilli Strains as an Innovative Culture Strategy. Fermentation, 11(10), 576. https://doi.org/10.3390/fermentation11100576

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop