Fructooligosaccharides: Applications and Health benefits- a
review
SHUBHANGI S. KHANVILKAR, SHALINI S. ARYA*
*Corresponding author
Institute of Chemical Technology, Food Engineering and Technology Department
NM Parikh Marg, Matunga, Mumbai, 400 019, India
KEYWORDS: Non- digestible, Enzymes, Functional ingredient, Product formulation, Industrial production, Health benefits
ABSTRACT:
Fructooligosaccharides (FOSs) are non-digestible carbohydrates with functional and physiological attributes like low sweetness,
non-carcinogenicity, low caloric value, prebiotic, hypolipidemic and hypocholestrolemic properties. Although
fructooligosaccharides are present in trace amounts in natural foods like onions, asparagus, wheat, banana, tomato and honey,
commercial production is accomplished using microbial transferase such as fructosyl transferase or β-fructofuranosidase. The
industrial production of fructooligosaccharides (FOS) is expanding rapidly due to the food and pharmaceutical importance of
these compounds. Due to its health benefits and functional properties, FOS will continue to exist in market as functional food
ingredient for Product formulations.
ABBREVATIONS:
FOS- Fructooligosaccharides, DP- Degree of polymerization, DPav- Average Degree of polymerization, FTase- Fructosyl
Transferase, FFase- β-fructofuranosidase, SmF- Submerged Fermentation, SSF- Solid State Fermentation, SCFA- Short Chain
fatty Acids, NDO- Non Digestible Carbohydrate
INTRODUCTION
Fructooligosaccharides (FOS) are composed of a mixture of Kestose (GF2), Nystose (GF3) and Fructosyl nystose (GF4),
obtained from sucrose by enzymatic addition of fructosyl moieties, and having an average degree of polymerization (DPav) of 3.6,
and are named as short-chain fructooligosaccharides. They are different from natural fructans having degree of polymerization
(DP) (only 10 % of native chicory Inulins have a DP between 2 and 5) [1], and from oligofructoses formed by inulin hydrolysis (DP
from 2 to 7, DPav 4) by the systematic presence of a glucose moiety [2].
FOS is found in trace amounts as natural components in fruits, vegetables, and honey. FOS is present in more than 36,000 plant
sources as reserve carbohydrates [3]. It is present in various plant sources such as asparagus, jerusalem artichoke, chicory,
sugar garlic, onion, wheat, honey, banana, barley, tomato, and rye [3]. Its concentration varies from 0.15% to 0.75% in natural
sources. The highest concentration is present in jerusalem artichoke (up to 20%) and chicory (between 5 and 10%). These
fructose oligomers have beneficial functional properties, low caloric value, non-cariogencity and can be used in diabetic
formulation. In addition, it exhibits many health benefits such as decrease levels of phospholipids, cholesterol and triglycerides,
helps in gut absorption of minerals such as calcium and magnesium, and stimulate the growth of bifidobacteria in the human
colon [4-5-6]. Therefore, utilization of fructooligosaccharides in food and pharmaceutical formulations is of great interest. Thus ,
developing economically feasible process of production of fructooligosaccharides to get high content FOS has become major
research area.
CHEMISTRY
FOS can be obtained in two different ways, which result in slightly different end products. First method involves transfructosylation
of sucrose by fructosyl transferase to produce short chain FOS, while the second method involves enzymatic hydrolysis of the
polysaccharide inulin (inulin oligofructose).
In the first method, The FOS formed in this process are the fructose oligomers that consist of a chain of two to four fructosyl
moieties linked by a β (2→1) glycosidic bond and a single glucose unit at the non-reducing end, linked by an α(1→2) glycosidic
bond [7]. These are named l-Kestose (Glu-Fru,), l- nystose (Glu-Fru,) and l F-fructofuranosyl nystose (Glu-Fru,). These types of
FOS are considered as short chain FOS. The transfructosylation of sucrose takes place via the cleavage of the β-(2→1)
glycosidic bond and the transfer of the fructosyl moiety onto any acceptor other than water, such as sucrose or a FOS. This
synthesis is a complex process in which several reactions occur simultaneously, both in parallel and in series, because Sc-FOS
are also potential substrates of fructosyltransferases (Figure 1). Glucose formed as by-products in the reaction acts as inhibitor for
the further production. Therefore, by-products and un-reacted sucrose formed during reaction is removed using chromatographic
procedures to get high purity product. These short chains FOS are quickly fermented in the colon to give prebiotic benefits [8].
In the second method, FOS production takes place by the controlled enzymatic hydrolysis of the polysaccharide inulin (inulin
oligofructose). The FOS mixture obtained by this process is different from the mixture produced by the transfructosylation
process. The oligosaccharide mixture obtained is longer than that produced by transfructosylation of sucrose. Additionally, not all
of the β(1→2)-linked fructosyl chains end with a terminal glucose in the oligofructoses prepared by inulin hydrolysis. The inulin
extracted from chicory roots contains some fructooligosaccharides in addition to polysaccharides. These longer-chain
oligosaccharide products have applications as fat replacers [8].
ENZYME SOURCES OF FOS SYNTHESIS
The enzyme source of FOS synthesis can be classified into two classes; one is plants such as asparagus, jerusalem artichoke
sugar beet, onion, etc.[10] and other consists of bacterial and fungal origins such as Aspergillus sp., Aureobasidium sp.,
Artrobacter sp., Fusarium sp. etc. [10]. The production yield of FOS using plant enzymes is low and dependent on seasonal
conditions; therefore, industrial production depends chiefly on fungal enzymes from either Aureobasidium sp. or A. niger.
Fructosyl Transferase (FTase) or β-fructofuranosidase (FFase) is responsible for the microbial production of FOS. FTase
produces FOS (GFn) from sucrose (GF) in a disproportionate mode, thereby forming 1-kestose (GF2) initially, then1-nystose
(GF3), followed by 1-fructofuranosyl nystose (GF4). Microbial FTases are derived from bacterial and fungal sources.
PRODUCTION OF FOS
FERMENTATIVE METHODS OF MICROBIAL PRODUCTION OF FRUCTOOLIGOSACCHARIDES
There are two methods of FTase production by fermentation- Submerged Fermentation (SmF) and Solid State Fermentation
(SSF). Production of enzymes by SSF has several advantages over SmF with respect to simplicity in operation, high productivity
during fermentation, less contamination and concentrated product formation. SSF requires less space capital and operating costs,
simpler equipment and easier downstream processing compared to SmF. In addition, it utilizes agro-industrial residues as
substrates, which are converted in to bulk chemicals and fine products of high commercial value. [11]
CONTINUOUS PRODUCTION OF FOS
A continuous process using an immobilized enzyme or an immobilized microorganism is preferred due to high production rate
compared to batch process. The immobilization of biocatalyst is more advantageous due to easy separation of enzyme and
product, the enhancement of volumetric productivity of the reactor.
Immobilization of FTases by chitosan and alginate for efficient production of FOS was studied by Ganaie et al (2014). In this study
the authors had carried out enzyme immobilization with sodium alginate and chitosan to form gel beads. The alginate beads were
more stable by maintaining its spherical conformation in presence of 0.3% (w/v) sodium alginate and 0.1% (w/v) of CaCl 2 solution
to obtain a product with highest transfructosylating activity. Further, the study described the effect of sodium alginate & CaCl2
concentration on entrapment of FTase for transfructosylating activity and effect of temperature and pH on free and immobilized
FTase. The yield obtained was as high as 67.75% (w/w) FTase entrapped alginate beads and 42.79% (w/w) by chitosan beads in
36 h of enzyme substrate reaction [12].
Production of FOS using β-fructofuranosidase immobilized onto chitosan-coated magnetic nanoparticles was reported by Chen et
al., 2013[13]. The study described immobilization of β-Fructofuranosidase (EC 3.2.1.26) enzyme onto chitosan-coated magnetic
nano-particles due to its various advantages such as a higher specific surface area which permit binding of a larger amount of
enzymes, mass transfer resistance is relatively low, and the immobilized enzyme can be selectively separated from a reaction
mixture by application of a magnetic field [14]. The immobilized enzyme retained 55% of its initial activity after 10 batches of FOS
production and it achieved 59.5% yield on a dry weight basis from 50% (w/v) sucrose.
FOS and β–fructofuranosidase production by Aspergillus japonicas immobilized on lignocellulosic materials studied by Mussatto
et al, 2009[15]. Authors have reported immobilization of FFase on various lignocellulosic materials including brewer’s spent grain,
wheat straw, corn cobs, coffee husks, cork oak and loofa sponge. It was found that corn cobs was the best support material gave
highest results of immobilization with highest productivity and yield than those obtained by free cell system
Authors have reported Immobilization conditions of Pectinex Ultra SP-L to produce FOS [16]. Pectinex Ultra SP-L is a commercial
enzyme preparation containing FTase activity and ability to produce short chain FOS. It was immobilized onto anionic ion
exchange resin. In the experiments, the optimal immobilization conditions were determined. Moreover, an integrated reactor
system was constructed for simultaneous FOS production synthesis and glucose elimination to enhance the product yield. The
system achieved ~74% Product yield.
PROPERTIES OF FRUCTOOLIGOSACCHARIDES
FOS delivers nutritional advantages as well as technological properties which makes it important food ingredient.
Sweetness:
FOS exhibits sweetness about one third of sucrose, deliver small amount of energy, estimated to be 1.5kcal/g [17]. It can be used
in various kinds of foods instead of sucrose to avoid excessive sweetness.
Non – cariogenicity:
FOS is not utilized by oral microflora unlike starch and simple sugars to form acids, which serve as a matrix for plaque formation
and ultimately results in dental caries [18]. Hence, FOS is presently utilized as non – cariogenic sugar substitutes in chewing
gums, confectionery, yoghurts and drinks,
Solubility: It is highly water soluble which makes it useful in variety of food applications for enhancing nutritional value of product
[10].
Viscosity of FOS solution: The viscosity FOS solution is higher than that of sucrose at same concentration. The higher
molecular weight of oligosaccharides provides increased viscosity compared with mono and disaccharides, results in improved
body and mouthfeel [11].
Thermal/ freezing stability: It is highly stable than sucrose and control the amount of browning due to maillard reactions in heat -
processed foods. It can be used to alter the freezing temperature of frozen foods. It is highly stable at refrigerated temperatures
over one year. [10]
pH range: It is highly stable in the normal pH range for food (4.0 – 7.0) [11].
Water activity: FOSs is highly hygroscopic. It provides high moisture retaining capacity, preventing excessive drying, and a low
water activity, which is convenient in controlling microbial contamination [19].
BULKING AGENT: Bulking agents can be any reduced-calorie or non-calorie carbohydrate. This can be achieved by replacing
the usual sweetening carbohydrate with an intense sweetener such as aspartame, phenylalanine or sucralose, thereby masking
the aftertaste produced by some of these intense sweeteners [19] plus a bulking agent
APPLICATIONS OF FOS IN FOOD FORMULATIONS
FOS provides improved organoleptic, nutritional, functional properties which make it important functional food ingredient.
Fructooligosaccharides synthesized by transfructosylation from sucrose are used as prebiotic ingredients while the longer chain
oligosaccharides derived from controlled enzymatic hydrolysis of inulin are used as fat replacers. An example of the use of FOS in
food formulations includes jam products, Ice cream, confectionery applications.[5]
Padma Ishwarya et al., (2013) studied the effect of addition of FOS in syrup and powder form in biscuits to provide nutritional and
technological benefits. FOS in powder form had better stability and comparatively less adverse effects on the biscuit
properties[20].
Renuka et al., (2009) studied the effect of fortification of FOS in selected fruit juice beverages. The physicochemical and sensory
properties of fruit juices were studied during 6 months storage at ambient and refrigerated conditions. The FOS retained in 6
month storage at refrigerated temperature was significant than the ambient temperature. FOS fortified fruit beverages was
acceptable up to 4 and 6 months of storage at ambient or refrigerated storage[21].
HEALTH BENEFITS
LOW CALORIFIC VALUE
The presence of β-glycosidic linkage makes FOS nondigestible by human digestive enzymes hence it is not utilized in the body as
an energy source. The caloric contribution of FOS is due to colonic fermentation producing short-chain fatty acids (SCFA). It has
energy contribution to food of about 1.5 kcal/ g.
The addition FOS leads to caloric reduction which can be achieved by replacing the usual sweetening carbohydrate with an
intense sweetener and FOS. Therefore, it can be utilized in low calorie diet ,sweets, diabetic food. In sweet foods, it can be used
along with intense artificial sweeteners to mask its bitter after taste [19].
DIETARY FIBER EFFECT
FOS produces a soluble fiber-like effect and fits well within the concept of dietary fibre [22]. It shows various physiological effects
being soluble-fermentable fiber which includes resistance to hydrolysis by digestive enzymes, fermentation by the intestinal micro
flora, reduced gastrointestinal transit time, increased fecal weight, lowered fecal pH, predictable reduction in caloric value,
reduced plasma cholesterol and delay or decrease in glucose absorption. In addition, it may strengthen mucosal protection and
reduce the risk of gastrointestinal diseases.
PREBIOTIC EFFECT
FOS is not hydrolyzed by the human digestive enzymes; it undergoes fermentation in the colon. It encourages the growth of
favorable bacteria in the colon; this in turn discourages the growth of potentially putrefactive microorganisms in the colon resulting
in a healthy gut environment.
In human clinical study on pre biotic effect of FOS, the effect of ingestion of low dose of FOS (5 g/day) was investigated [23].
There was a selective growth of certain groups of bacteria such as bifidobacteria was obsereved, thus confirming the prebiotic
properties of oligofructose.
EFFECT ON MINERAL ABSORPTION
Colonic fermentation of FOS leads to decrease in pH in the colon and this facilitates the absorption of mineral ions from the
intestine, mainly calcium and magnesium [5].
In vivo study to assess the effect of mineral absorption; Lobo et al., 2006 studied the effect of FOS on mineral absorption on
bone parameters in healthy growing rats. In this study, the rats were fed with 5% FOS for 23 days and the mineral absorption for
3 periods of 5 day each was examined. It was observed that FOS intake led to a significant increase in Ca and Mg intestinal
apparent absorption in healthy growing rats. This effect resulted in a increased bone mineralization, which, in turn, increased the
femur resistance to fracture. These results show an important role of FOS in the maintenance of healthy bones [24].
It was assumed that the effect may be due to enhancement of passive and active mineral transport across the intestinal
epithelium, facilitated by an increase in certain metabolites of the intestinal flora and a reduction in pH [26].
ROLE OF FOS IN DEFENSE FUNCTIONS
FOS is known to prevent growth of pathogenic microorganisms by encouraging the growth of beneficial bacteria. This effect is
attributed to the low pH environment created during fermentation of FOS in the colon and due to the secretion of antibiotic like
substances by the beneficial bacteria.
In vivo Study
Buddington, Kelly-Quagliana, Buddington, & Kimura et al. (2002) studied the effect of FOS diet on colon health of mice infected
with virulent strains of Listeria monocytogenes and Salmonella typhimurium. It was observed from the study that supplementing
the diet of mice with inulin and oligofructose reduced the densities of Candida in the small intestine of mice, 7 days after infection.
Mice infected systemically with virulent strains of Listeria monocytogenes and Salmonella typhimurium after being fed a diet with
inulin and oligfructose (at 100 g/kg) had lower mortality due to enhanced T-lymphocyte functions than mice fed a diet with
cellulose as the source of fiber. It also lowered the incidence of the infection of Listeria and Salmonella and growth of tumors after
exposure to carcinogens and transplanted tumor cells[26].
LIPID METABOLISM
FOS reduces the level of serum lipids, serum cholesterol, hypotriglyceridemic effect; thus reducing the risk of diabetes and
obesity [27]. There are mechanisms have been proposed to explain lipid metabolism by FOS.The first involves modification of the
glucose or insulin concentrations. FOS reduces the glucose peaks in blood that occur after eating and, as a result, there is a large
reduction in the production of glucose and insulin induced lipidic enzymes [28]. The second involves the production of SCFAs in
the colon. Propionate inhibits the cholesterogenesis and lipogenesis pathways, while acetate stimulates the same. It seems, then,
that the pattern of FOS fermentation and, specially, the quotient between acetate and propionate that reaches the liver may act as
one of the potential lipid reducing characteristics of FOS [28].The third mechanism proposed is that serum cholesterol is reduced
due to the precipitation and excretion of bile acids in the intestine, which implies that the liver uses cholesterol to synthesize bile
acids. On the other hand, changes in the concentration of serum cholesterol have been related to changes in intestinal microflora.
Some strains of Lactobacillus acidophilus assimilate the cholesterol present in the medium, while others inhibit cholesterol
absorption through the intestinal wall [29]. There has been a interrelation between the level of lipids in serum and the incidence of
cardiovascular diseases, which makes FOS a possible tool for their prevention [28].
ANTI-DIABETES EFFECT
Alles et al.,(1999) studied Consumption of FOS on serum lipids, serum acetate, blood glucose in 20 patients with type 2
diabetes. In a single-blind, crossover, randomized study design, patients consumed either glucose as a placebo (4 g/d) or FOS
(15 g/d) for 20 d each. It was concluded that 20 d of dietary supplementation with FOS had no major effect on serum lipids, or
serum acetate, blood glucose in patients with type 2 diabetes [30].
ANTICANCEROUS EFFECT
Studies with inulin and FOS have shown reduction of chemically induced aberrant crypts and prevention of colon cancer.
According to Pool-Zobel, van Loo, Rowland, and Roberfroid, (2002), in rats, a prebiotic effect resulting in the proliferation of
bifidobacteria (with the major metabolites lactate or acetate) as well as of other bacteria could be responsible for the observed
anticancer effects.
CONCLUSION
FOS has emerged as functional food ingredient in market due to growing demand for healthy food. Research work has been done
on production of FOS using different microbial FTase for production yield. Development of continuous methods for FOS
production using immobilization has led to increased industrial production. Nutritional and Functional properties has resulted in
incorporation in variety of products such as Bakery products, Beverages, Confectionery, Fruit based and dairy products. The
health benefits associated with the consumption of these FOS has led to its increased popularity as food ingredients and they are
also being promoted as functional food ingredient for product formulations.
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