Characterization of Exopolysaccharides Isolated from Donkey Milk and Its Biological Safety for Skincare Applications

Kefiran is a heteropolysaccharide that is considered a postbiotic and is obtained by kefir grains fermented in cow’s milk, while little is known about the donkey milk (DM) variety. Postbiotics are recognised as having important human health benefits that are very similar to probiotics but without th...

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Main Authors: Chiara La Torre, Pierluigi Plastina, Diana Marisol Abrego-Guandique, Paolino Caputo, Cesare Oliviero Rossi, Giorgia Francesca Saraceno, Maria Cristina Caroleo, Erika Cione, Alessia Fazio
Format: Article
Language:English
Published: MDPI AG 2024-09-01
Series:Polysaccharides
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Online Access:https://www.mdpi.com/2673-4176/5/3/31
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author Chiara La Torre
Pierluigi Plastina
Diana Marisol Abrego-Guandique
Paolino Caputo
Cesare Oliviero Rossi
Giorgia Francesca Saraceno
Maria Cristina Caroleo
Erika Cione
Alessia Fazio
author_facet Chiara La Torre
Pierluigi Plastina
Diana Marisol Abrego-Guandique
Paolino Caputo
Cesare Oliviero Rossi
Giorgia Francesca Saraceno
Maria Cristina Caroleo
Erika Cione
Alessia Fazio
author_sort Chiara La Torre
collection DOAJ
description Kefiran is a heteropolysaccharide that is considered a postbiotic and is obtained by kefir grains fermented in cow’s milk, while little is known about the donkey milk (DM) variety. Postbiotics are recognised as having important human health benefits that are very similar to probiotics but without the negative effects associated with their ingestion. Donkey is a monogastric animal, as are humans, and when used as an alternative food for infants who suffer from cow milk protein allergies, DM could therefore display more biocompatibility. In this study, the DM kefiran was extracted by ultrasound from kefir grains cultured in donkey milk and fully characterized for its structural and physicochemical properties by Fourier-transform infrared spectroscopy (FT-IR), High-Performance Liquid Chromatography- Refractive Index (HPLC-RI), Scanning electron microscope (SEM), Differential Scanning Calorimeters (DSC) and rheological analyses. In addition, tests were conducted on keratinocytes cell lines and human red blood cells to assess the nontoxicity and haemolysis degree of the polymer. The extraction yield of the DM kefiran was 6.5 ± 0.15%. The FT-IR analysis confirmed the structure of the polysaccharide by showing that the stretching of the C-O-C and C-O bonds in the ring, which formed two bands at 1157 and 1071 cm<sup>−1</sup>, respectively, and the anomeric band at 896 cm<sup>−1</sup> indicates the β configuration and vibrational modes of glucose and galactose. Results were confirmed by HPLC-RI analysis indicating that the ratio glucose/galactose was 1:0.87. Furthermore, the SEM analysis showed a porous and homogeneous structure. The rheological analysis confirmed the pseudoplastic nature of the polymer, while the DSC analysis highlighted excellent thermal resistance (324 °C). Finally, DM kefiran was revealed to have biologically acceptable toxicity, showing a haemolytic activity of less than 2% when using fresh human red blood cells and showing no cytotoxicity on human keratinocytes. Therefore, kefiran obtained by DM shows an excellent biocompatibility, establishing it as a promising polymer for bioengineering human tissue for regenerative applications.
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spelling doaj-art-97e7e5e486a64dcea43b25c8a6e4e95a2025-08-20T01:55:48ZengMDPI AGPolysaccharides2673-41762024-09-015349350310.3390/polysaccharides5030031Characterization of Exopolysaccharides Isolated from Donkey Milk and Its Biological Safety for Skincare ApplicationsChiara La Torre0Pierluigi Plastina1Diana Marisol Abrego-Guandique2Paolino Caputo3Cesare Oliviero Rossi4Giorgia Francesca Saraceno5Maria Cristina Caroleo6Erika Cione7Alessia Fazio8Department of Pharmacy, Health and Nutrition Sciences, University of Calabria, 87036 Arcavacata di Rende, CS, ItalyDepartment of Pharmacy, Health and Nutrition Sciences, University of Calabria, 87036 Arcavacata di Rende, CS, ItalyDepartment of Health Sciences, University of Magna Graecia Catanzaro, 88100 Catanzaro, CZ, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, 87036 Arcavacata di Rende, CS, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, 87036 Arcavacata di Rende, CS, ItalyDepartment of Pharmacy, Health and Nutrition Sciences, University of Calabria, 87036 Arcavacata di Rende, CS, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, 87036 Arcavacata di Rende, CS, ItalyDepartment of Pharmacy, Health and Nutrition Sciences, University of Calabria, 87036 Arcavacata di Rende, CS, ItalyDepartment of Pharmacy, Health and Nutrition Sciences, University of Calabria, 87036 Arcavacata di Rende, CS, ItalyKefiran is a heteropolysaccharide that is considered a postbiotic and is obtained by kefir grains fermented in cow’s milk, while little is known about the donkey milk (DM) variety. Postbiotics are recognised as having important human health benefits that are very similar to probiotics but without the negative effects associated with their ingestion. Donkey is a monogastric animal, as are humans, and when used as an alternative food for infants who suffer from cow milk protein allergies, DM could therefore display more biocompatibility. In this study, the DM kefiran was extracted by ultrasound from kefir grains cultured in donkey milk and fully characterized for its structural and physicochemical properties by Fourier-transform infrared spectroscopy (FT-IR), High-Performance Liquid Chromatography- Refractive Index (HPLC-RI), Scanning electron microscope (SEM), Differential Scanning Calorimeters (DSC) and rheological analyses. In addition, tests were conducted on keratinocytes cell lines and human red blood cells to assess the nontoxicity and haemolysis degree of the polymer. The extraction yield of the DM kefiran was 6.5 ± 0.15%. The FT-IR analysis confirmed the structure of the polysaccharide by showing that the stretching of the C-O-C and C-O bonds in the ring, which formed two bands at 1157 and 1071 cm<sup>−1</sup>, respectively, and the anomeric band at 896 cm<sup>−1</sup> indicates the β configuration and vibrational modes of glucose and galactose. Results were confirmed by HPLC-RI analysis indicating that the ratio glucose/galactose was 1:0.87. Furthermore, the SEM analysis showed a porous and homogeneous structure. The rheological analysis confirmed the pseudoplastic nature of the polymer, while the DSC analysis highlighted excellent thermal resistance (324 °C). Finally, DM kefiran was revealed to have biologically acceptable toxicity, showing a haemolytic activity of less than 2% when using fresh human red blood cells and showing no cytotoxicity on human keratinocytes. Therefore, kefiran obtained by DM shows an excellent biocompatibility, establishing it as a promising polymer for bioengineering human tissue for regenerative applications.https://www.mdpi.com/2673-4176/5/3/31exopolysaccharidekefirandonkeyphysicochemical propertiesmonosaccharideshaemolysis
spellingShingle Chiara La Torre
Pierluigi Plastina
Diana Marisol Abrego-Guandique
Paolino Caputo
Cesare Oliviero Rossi
Giorgia Francesca Saraceno
Maria Cristina Caroleo
Erika Cione
Alessia Fazio
Characterization of Exopolysaccharides Isolated from Donkey Milk and Its Biological Safety for Skincare Applications
Polysaccharides
exopolysaccharide
kefiran
donkey
physicochemical properties
monosaccharides
haemolysis
title Characterization of Exopolysaccharides Isolated from Donkey Milk and Its Biological Safety for Skincare Applications
title_full Characterization of Exopolysaccharides Isolated from Donkey Milk and Its Biological Safety for Skincare Applications
title_fullStr Characterization of Exopolysaccharides Isolated from Donkey Milk and Its Biological Safety for Skincare Applications
title_full_unstemmed Characterization of Exopolysaccharides Isolated from Donkey Milk and Its Biological Safety for Skincare Applications
title_short Characterization of Exopolysaccharides Isolated from Donkey Milk and Its Biological Safety for Skincare Applications
title_sort characterization of exopolysaccharides isolated from donkey milk and its biological safety for skincare applications
topic exopolysaccharide
kefiran
donkey
physicochemical properties
monosaccharides
haemolysis
url https://www.mdpi.com/2673-4176/5/3/31
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