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Acemannan and Fructans from Aloe Vera as promising prebiotics
María Paz Quezada1,3, Gabriela González H.,2,4,5, Carlos Salinas2,
Martin Gotteland, Miguel Allende3 & Liliana Cardemil1
1LBMV, Facultad de Ciencias, Universidad de Chile.2Departamento de Nutrición, Facultad de Medicina, Universidad de Chile
3CRG, Facultad de Ciencias, Universidad de Chile.4Departamento de Biotecnología, Universidad de La Frontera5Facultad de Ciencias Biológicas, Universidad Andrés Bello.
e-mail: lcardemi@gmail.com
Aloe barbadensis Miller, is also knownas Aloe vera. It belongs to the orderof Asparagales in the family of theAsphodelaceae. It was previouslyclassified in the Liliaceae family.
It is a monocot succulent plant,(xerophyte) and originally fromÁfrica. Aloe vera is a CAM plant(crassulacean acid metabolism), andof course, adapted to the arid andsemi-arid environments.
Aloe barbadensis Miller is the specieswith higher commercial valueamong more 300 species of Aloe dueto the medicinal and cosmeticapplications. Aloe vera cultivadas en el campo
Experimental de Las Cardas de laUniversidad de Chile (Región deCoquimbo).
AridEnvironments
• Low water availability•Great temperature fluctuations between day and night (5° ↔ 45 °C)
•Hihg levels of solar radiation•And poor soil conditions.
Waterstress
Heatstress
Oxidativestress
Response mechanisms
Osmotic adjustment (Carbohydrate), Expression of Heat Shock Genes and accumulation of Heat Shock Proteins
(HSP), good activity of Superoxide Dismutase (SOD).
Normal irrigation conditions
Severe water deficit
Fructans
Glucomannan
Fructans as Neofructans
Glucomannan of higherDP
LINEAR POLYSACCHARIDES
MORE BRANCHED AND LONGER
POLYSACCHARIDES
Structural Modification of Aloe vera polysaccharides
Cortex
Cortex
Figure 1. Polysaccharide structures present in Aloe vera plants. I, acemannan, II, fructans
present in water stressed plants of Aloe vera.
Fructans
glucomannan
Cortex
Gel
ControledFertirrigation
LactobacillusL. plantarum 46-1-12
L. casei 54-2-33L. fermentum 55-233-3
L. plantarumN22-33
BifidobacteriumB. animalis BB-12
B. catenulatum N173-2B. longum N180-3B. bifidum N364-3
Methodology
Acemannan Fructans
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.
Lactobacillus plantarum N 223-3 (18 Hr). 1: Paired t test (P < 0,05).2: One-way ANOVA, TUKEY (P < 0,05).
SampleConcentration
(g/L)
Variationbetween
0 hr and 18 hr1
(%)Variation
Variationat 18 hr2
Glucose 10 *** 633 a
Fructans 10 *** 271 a
Acemannan 3 * 91 bc
Fructans + Acemannan 10 + 3 * 154 b
Commercial
FOS10 * 79 c
SampleConcentration
(g/L)
Variationbetween 0 hr
and 18 hr1
% VariationVariationat 18 hr2
Glucose 10 * 634 a
Fructans 10 *** 304 b
Acemannan 3 ** 57 c
Fructans +
Acemannan10 + 3 * 88 d
Commercial
FOS10 ** 77 cd
Lactobacillus casei L54-2-33 (18 Hr). 1: Paired t test (P < 0,05).2: One-way ANOVA, TUKEY (P < 0,05).
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.
SampleConcentration
(g/L)
Variationbetween 0 hr
and 72 hr1
(%)Variation
Significanceat 72 hr2
Glucose 10 ns -11 a
Fructans 10 ** 498 b
Acemannan 3 ** 238 c
Fructans +
Acemannan10 + 3 *** 373 b
Commercial
FOS10 *** 257 c
Bifidobacterium longum N180-3 (72 Hrs). 1: Paired t test (P < 0,05). 2: One-way ANOVA, TUKEY(P < 0,05).
SampleConcentration
(g/L)
Variationbetween 0 hr
and 72 hr1
Percentvariation
(%)
Significanceat 72 hr
Glucose 10 ns -8 a
Fructans 10 *** 318 b
Acemannan 3 * 167 c
Fructan +
Acemannan10 + 3 *** 298 b
Commercial
FOS10 *** 275 c
Bifidobacterium bifidum N364-3 (72 Hrs). 1: Paired t test (P < 0,05). 2: One-way ANOVA, TUKEY(P < 0,05).
1) Fresh stools from three healthy
volunteers (23−26 years old).
2) Body mass index between 18 and
24.9 kg/m2.
3) Without previous intake of antibiotics,
prebiotics or probiotics.
Subject 1
Subject 2
Subject 3
Glu
co
se
10
g/
L
Fru
cta
ns
10
g/
L
Acem
an
nan
3 g
/L
Co
mm
FO
S1
0 g
/L
Feces with carbohydrates were fermented in a bioreactor model B-Braun MU-200 for 48 hrs
under Anaerobiotic medium FM.
5g
Measuringthe DNA 16S
from Bacterias by qPCR
Short chainFatty acids
quantification(SCFA) by GC-
FID
5g
5g
Metodología
-C
Methodology
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.
Total bacterial population
Lactobacillus sp Bifidobacteria sp.
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.
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.
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.
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