Acemannan and Fructans from Aloe Vera as promising prebiotics

Transcription

1 Acemannan and Fructans from Aloe Vera as promising prebiotics María Paz Quezada 1,3, Gabriela González H.,2,4,5, Carlos Salinas 2, Martin Gotteland, Miguel Allende 3 & Liliana Cardemil 1 1 LBMV, Facultad de Ciencias, Universidad de Chile. 2 Departamento de Nutrición, Facultad de Medicina, Universidad de Chile 3 CRG, Facultad de Ciencias, Universidad de Chile. 4 Departamento de Biotecnología, Universidad de La Frontera 5 Facultad de Ciencias Biológicas, Universidad Andrés Bello. lcardemi@gmail.com

2 Aloe barbadensis Miller, is also known as Aloe vera. It belongs to the order of Asparagales in the family of the Asphodelaceae. It was previously classified in the Liliaceae family. It is a monocot succulent plant, (xerophyte) and originally from África. Aloe vera is a CAM plant (crassulacean acid metabolism), and of course, adapted to the arid and semi-arid environments. Aloe barbadensis Miller is the species with higher commercial value among more 300 species of Aloe due to the medicinal and cosmetic applications. Aloe vera cultivadas en el campo Experimental de Las Cardas de la Universidad de Chile (Región de Coquimbo).

3 Arid Environments Low water availability Great temperature fluctuations between day and night (5 45 C) Hihg levels of solar radiation And poor soil conditions. Water stress Oxidative stress Heat stress Response mechanisms Osmotic adjustment (Carbohydrate), Expression of Heat Shock Genes and accumulation of Heat Shock Proteins (HSP), good activity of Superoxide Dismutase (SOD).

4 Aloe barbadensis Miller Fructans Acemannan

5 Structural Modification of Aloe vera polysaccharides Normal irrigation conditions Cortex Fructans Glucomannan LINEAR POLYSACCHARIDES Severe water deficit Cortex Fructans as Neofructans Glucomannan of higher DP MORE BRANCHED AND LONGER POLYSACCHARIDES

6 Figure 1. Polysaccharide structures present in Aloe vera plants. I, acemannan, II, fructans present in water stressed plants of Aloe vera.

7 Methodology Acemannan Cortex Fructans Fructans Lactobacillus Controled L. Fertirrigation plantarum L. casei L. fermentum L. plantarumn22-33 Bifidobacterium B. animalis BB-12 B. catenulatum N173-2 B. longum N180-3 B. bifidum N364-3 glucomannan Gel

8 Lactobacillus Kinetics of bacterial growth for Lactobacillus species. The growth was measured as the increase in optical density (OD) from the beginning (0 hours) and during 18 hours of culture at 37 C.

9 Lactobacillus plantarum N (18 Hr). 1 : Paired t test (P < 0,05). 2 : One-way ANOVA, TUKEY (P < 0,05). Sample Concentration (g/l) Variation between 0 hr and 18 hr 1 (%) Variation Variation at 18 hr 2 Glucose 10 *** 633 a Fructans 10 *** 271 a Acemannan 3 * 91 bc Fructans + Acemannan * 154 b Commercial FOS 10 * 79 c

10 Lactobacillus casei L (18 Hr). 1 : Paired t test (P < 0,05). 2 : One-way ANOVA, TUKEY (P < 0,05). Sample Concentration (g/l) Variation between 0 hr and 18 hr 1 % Variation Variation at 18 hr 2 Glucose 10 * 634 a Fructans 10 *** 304 b Acemannan 3 ** 57 c Fructans + Acemannan Commercial FOS * 88 d 10 ** 77 cd

11 Bifidobacterium Kinetic of bacterial growth in Bifidobacterium species. The rate of growth of the strains was measured by the increase in the optical density (OD) from the beginning (0 hours) and during 72 hours of culture at 37 C.

12 Bifidobacterium longum N180-3 (72 Hrs). 1 : Paired t test (P < 0,05). 2 : One-way ANOVA, TUKEY(P < 0,05). Sample Concentration (g/l) Variation between 0 hr and 72 hr 1 (%) Variation Significance at 72 hr 2 Glucose 10 ns -11 a Fructans 10 ** 498 b Acemannan 3 ** 238 c Fructans + Acemannan Commercial FOS *** 373 b 10 *** 257 c

13 Bifidobacterium bifidum N364-3 (72 Hrs). 1 : Paired t test (P < 0,05). 2 : One-way ANOVA, TUKEY(P < 0,05). Sample Concentration (g/l) Variation between 0 hr and 72 hr 1 Percent variation (%) Significance at 72 hr Glucose 10 ns -8 a Fructans 10 *** 318 b Acemannan 3 * 167 c Fructan + Acemannan Commercial FOS *** 298 b 10 *** 275 c

14 1) Fresh stools from three healthy volunteers (23 26 years old). 2) Body mass index between 18 and 24.9 kg/m 2. 3) Without previous intake of antibiotics, prebiotics or probiotics.

15 Glucose 10 g/l Fructans 10 g/l Acemannan 3 g/l Comm FOS 10 g/l - C Metodología Methodology Subject 1 Subject 2 Subject 3 5g 5g 5g Feces with carbohydrates were fermented in a bioreactor model B-Braun MU-200 for 48 hrs under Anaerobiotic medium FM. Measuring the DNA 16S from Bacterias by qpcr Short chain Fatty acids quantification (SCFA) by GC- FID

16 Total bacterial population Lactobacillus sp Bifidobacteria sp. Quantification of the microbiota from human stool after 48h of fermentation. Quantification was performed by qpcr using selected primers at 0 and 48h of fermentation in the presence of glucose, fructans and acemannan.

17 Concentration of SCFA and BCFA produced by fermentation of human feces in the presence of different carbon sources. The human feces were fermented in bioreactors and the SCFA and BCFA produced were quantified by gas chromatography, using different carbon sources.

18 Percentage composition of the SCFA and of BCFA produced by fermentation of human feces in a bioreactor. (A) Percent of each SCFA and BCFA in the feces before fermentation (0 h). (B) Percent of each SCFA and BCFA after 48 h of fermentation.

19 Extension to other organs Limph node Blood Vessel Muscular layer Submucose Mucose Polyps

20 CONCLUSIONS 1. The two polysaccharides obtained from Aloe vera from the fertigation treatment (water stressed plants) have a great prebiotic potential, better than Commercial FOS which is the most used commercial prebiotic. 2. The polysaccharide of water stressed plants increases the growth of selected intestinal strains of Lactobacillus sp. The fructans and acemannan of Aloe vera subjected to water restrictions, increase the growth of Bifidobacterium sp being more effective the combination of both in some strains. 3. The glucomannan of water stressed plants induce the synthesis of butyric acid which is the most potent inhibitor of colon polyp formation. Butyric acid increases several times the basal level. 4. Both polysaccharides complement each other in their prebiotic properties. While fructans increase the growth of Lactobacillus strains, both of them increase de growth of Bifidobacterium sp. On the other hand glucomannan increases the amount of butyric acid.

21 Colaborators Martin Gotteland Miguel Allende