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).

Fructans

Acemannan

Aloe barbadensis Miller

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.

Extension to other organs

Limph node

Blood Vessel

Muscular layer

SubmucoseMucose

Polyps

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.

Martin Gotteland

Miguel Allende

Colaborators