FIELD OF INVENTION
[001] The present disclosure broadly relates to the field of fortified food products and nutritional supplements and particularly refers to a prebiotic composition that improves the overall nutrient absorption profile.
BACKGROUND OF INVENTION
[002] The human gastrointestinal (GI) tract harbors a wide variety of microorganisms. The microbial communities present inside the GI tract provides metabolic, immunologic, and protective functions that play a crucial role in human health (Jumpertz R, et al, Am J Clin Nutr. 2011 Jul; 94(l):58-65). [003] The type of bacteria that are commonly present in intestine are obligate anaerobes. These obligates anaerobes may be categorized as either harmful or beneficial bacteria. Gut bacteria comprising Clostridium perfringens, C. difficile, Salmonella, etc. can be potentially harmful when the ecosystem undergoes abnormal changes. Dysbiosis of the gut bacteria communities in patients or animal models may cause many diseases such as diarrhea, systemic infections, liver damage and carcinoma etc. (Yu-Jie Zhang, et al, Int J Mol Sci. 2015 Apr 2;16(4):7493-519.) In contrast, the beneficial bacteria promote the host's health by stimulating the immune system, synthesizing vitamins, inhibiting the growth of harmful bacteria and improving digestion and absorption of essential nutrients. (Gibson GR, and RoberfroidMB (1995), J Nutr 125: 1401-1412)
[004] The antibiotics may have a modest effect in decreasing disease activity or for treating bacterial overgrowth. However, these antibiotics may also kill populations of beneficial intestinal bacteria that are not harmful and that aid in nutrient absorption.
[005] Various approaches have been deployed to stimulate the growth of beneficial bacteria in GI tract that helps in ameliorating the quality of life. For instance, US7115297B2 discloses a nutritionally fortified liquid composition for consumption by humans and animals comprising safe water; encapsulated additives and water-soluble indigestible fibre.

[006] US20100239559A1 relates to a nutritional product composition comprising an enzyme selected from the group consisting of a lipase, an amylase, a protease, and any combination thereof, wherein said enzyme formulated for sustained stability in an aqueous medium; and a nutritional supplement.
[007] Although several attempts have been made to improve the nutrient absorption in body, there remains a need in the art to develop a composition that results in increasing the bio-availability of all nutrients available for absorption and its utilization in human body.
SUMMARY OF THE INVENTION
[008] In an aspect of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75.
[009] In another aspect of the present disclosure, there is provided a process for preparing a composition comprising: (a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i) obtaining galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition.
[0010] In one another aspect of the present disclosure, there is provided a fermented product obtained by fermentation of the composition comprising:(a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75.
[0011] In yet another aspect of the present disclosure, there is provided a process to obtain a fermented product obtained by fermentation of the composition comprising:(a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i) obtaining

galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition; and (iv) incubating the composition in presence of at least one microorganism for a time-period in a range of 10-18 hours, at a temperature in a range of 25-37°C, to obtain the fermented product.
[0012] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.
BRIEF DESCRIPTION OF ACCOMPANYING DRAWINGS
[0013] The following drawings form a part of the present specification and are
included to further illustrate aspects of the present disclosure. The disclosure may be
better understood by reference to the drawings in combination with the detailed
description of the specific embodiments presented herein.
[0014] Figure 1 depicts the PCR cycling conditions used for real-time PCR
quantification of the genes, in accordance with an embodiment of the present
disclosure.
[0015] Figure 2 depicts the expression changes in SI OOF gene involved in nutrient
absorption, in accordance with an embodiment of the present disclosure.
[0016] Figure 3 depicts the expression changes in CDX-2 gene involved in nutrient
absorption, in accordance with an embodiment of the present disclosure.
[0017] Figure 4 depicts the expression changes in FCNF4A gene involved in nutrient
absorption, in accordance with an embodiment of the present disclosure.
[0018] Figure 5 depicts the expression changes in SLC6A19 gene involved in
nutrient absorption, in accordance with an embodiment of the present disclosure.
DETAILED DESCRIPTION OF THE INVENTION

[0019] Those skilled in the art will be aware that the present disclosure is subject to
variations and modifications other than those specifically described. It is to be
understood that the present disclosure includes all such variations and modifications.
The disclosure also includes all such steps, features, compositions, and compounds
referred to or indicated in this specification, individually or collectively, and any and
all combinations of any or more of such steps or features.
Definitions
[0020] For convenience, before further description of the present disclosure, certain
terms employed in the specification, and examples are delineated here. These
definitions should be read in the light of the remainder of the disclosure and
understood as by a person of skill in the art. The terms used herein have the meanings
recognized and known to those of skill in the art, however, for convenience and
completeness, particular terms and their meanings are set forth below.
[0021] The articles "a", "an" and "the" are used to refer to one or to more than one
(i.e., to at least one) of the grammatical object of the article.
[0022] The terms "comprise" and "comprising" are used in the inclusive, open sense,
meaning that additional elements may be included. It is not intended to be construed
as "consists of only".
[0023] Throughout this specification, unless the context requires otherwise the word
"comprise", and variations such as "comprises" and "comprising", will be
understood to imply the inclusion of a stated element or step or group of element or
steps but not the exclusion of any other element or step or group of element or steps.
[0024] The term "including" is used to mean "including but not limited to".
"Including" and "including but not limited to" are used interchangeably.
[0025] For the purposes of the present document, the term "prebiotic composition"
used herein refers to non-digestible food ingredients that beneficially affect the host
by selectively stimulating the growth and/or activity of one or a limited number of
bacteria in the gastrointestinal tract and thereby exerting a health-promoting effect.
[0026] As used herein, galacto-oligosaccharide (GOS) refers to oligosaccharides
composed of different galactosyl residues with degree of polymerisation in a range
of2to6.

[0027] Fermentable fibre refers to fibres that are readily fermented by bacteria colonizing the gastrointestinal tract.
[0028] For the purpose of the present disclosure, inulin refers to naturally occurring polysaccharide consisting of chain-terminating glucosyl moieties and a repetitive fructosyl moiety, which are linked by (3(2,1) bonds. The degree of polymerization of inulin ranges from 2 to 60; lactulose refers to a synthetic disaccharide synthesized from fructose and galactose that is used for the purpose of treating constipation and some liver diseases; xylooligosaccharide refers to mixtures of oligosaccharides containing [3-1,4-linked xylose residues; fructooligosaccharide refers to linear chains of fructose units, wherein the fructose units are linked by (3(2-1) bonds. The number of fructose units ranges from 2 to 60 and often terminate in a glucose unit; isomaltooligosaccharides are a mixture a-(l —> 6) and a-(l —> 4)-linked glucose oligomers and serves as alternative sweeteners; glucomannan refers to water soluble polysaccharide comprising [3-(l—>4)-linked D-mannose and D-glucose units; resistant starch refers to starch that resists digestion in the small intestine; pectin refers to structural heteropolysaccharide comprising galacturonic acid monomer units that are linked via a-(l—>4)-glycosidic bond forming a backbone. [0029] As used herein, diluent, carrier, preservatives to the generally known components used in the art.
[0030] For the purpose of the present disclosure, the genes such as HNF4A (hepatocyte nuclear factor) positively impacts expression of other genes like CDX-2 and SLC6A19. [Turner et al, 2013, JBC, 288(47), 33813-33823]; SLC40A1 (iron transporter); S100G (calcium binding protein); SLC6A19 (neutral amino acid transporter); CDX2 (regulator of vitamin D receptor gene expression [Yamamoto et al, 1999 J. Bone Miner. Res., 14(2): 240-7]; CUBN (vitamin B12 receptor), AMN (protein involved in vitamin B12 absorption) [Kiela et al, Best Pract Res Clin Gastroenterol. 2016 Apr; 30(2): 145-159.] used for evaluating the effect of the composition of the present disclosure in improving the nutrient absorption profile. [0031] Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only

the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited.
[0032] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference.
[0033] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally-equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein. [0034] In recent years, various efforts have been made to increase the nutritive quality of food products, in order to meet the demand of consumers. The nutritive quality of these food products not only depends on its nutritive content, but also depends on the amount of nutrient available for absorption and its utilization in human body to treat various diseases. Some of the strategies that have been conventionally used to improve mineral absorption including the use of fortification techniques in processed food, bio-fortification, supplementation and reduction in anti-nutritional factors. However, the afore-mentioned strategies fail to provide the overall increase in absorption of nutrients (including both macronutrients and micronutrients) due to poor bio-availability of these nutrients from diet rich in legumes, cereals, etc. The poor bio-availability of these nutrients leads to a deficiency of nutrients in a subject.
[0035] To overcome the afore-mentioned problems, the purpose of the present disclosure is to provide a combination of prebiotics. The present disclosure discloses a composition having a combination of galacto-oligosaccharide and at least one fermentable fibre. Galacto-oligosaccharide and the at least one fermentable fibre present in a weight ratio range of 1:0.75-1:1.75, when fermented with Bacillus subtilis, synergistically improves the overall nutrient absorption of nutrients,

including but not limited to calcium, zinc, neutral amino acids, vitamins, iron, and combinations thereof.
[0036] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75. In another embodiment of the present disclosure, galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.9-1:1.4. In one another embodiment of the present disclosure, galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio of 1:1. [0037] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.8-1:1.25.
[0038] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, and wherein galacto-oligosaccharide has a weight percentage in a range of 0.35-0.5% with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.35-0.5% with respect to the composition. In another embodiment of the present disclosure, galacto-oligosaccharide has a weight percentage in a range of 0.38-0.5%) with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.40-4.8% with respect to the composition. In one another embodiment of the present disclosure, galacto-oligosaccharide has a weight percentage in a range of 0.40-4.8%) with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.38-0.5%) with respect to the composition. [0039] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre selected from a group consisting of inulin, lactulose, xylo-oligosaccharide, fructo-oligosaccharide, isomalto-oligosaccharides, glucomannan, resistant starch, pectin, and combinations thereof, wherein galacto-oligosaccharide to the at least one

fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75. In another embodiment of the present disclosure, the at least one fermentable fibre is inulin. [0040] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre selected from a group consisting of inulin, lactulose, xylooligosaccharide, fructooligosaccharide, isomaltooligosaccharides, glucomannan, resistant starch, pectin, and combinations thereof, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, and wherein galacto-oligosaccharide has a weight percentage in a range of 0.35-0.5% with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.35-0.5% with respect to the composition.
[0041] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide; (b) at least one fermentable fibre; and (c) at least one excipient selected from a group consisting of diluent, carrier, preservatives and combinations thereof, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-l: 1.75. [0042] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide; (b) at least one fermentable fibre; and (c) at least one excipient selected from a group consisting of diluent, carrier, preservatives and combinations thereof, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, and wherein galacto-oligosaccharide has a weight percentage in a range of 0.35-0.5%) with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.35-0.5%) with respect to the composition.
[0043] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide (GOS); (b) at least one fermentable fibre selected from a group consisting of inulin, lactulose, xylooligosaccharide, fructooligosaccharide, isomaltooligosaccharides, glucomannan, resistant starch, pectin, and combinations thereof; and (c) at least one excipient selected from a group consisting of diluent, carrier, preservatives and combinations thereof, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a

range of 1:0.75-l: 1.75, and wherein galacto-oligosaccharide has a weight percentage
in a range of 0.35-0.5% with respect to the composition, and the at least one
fermentable fibre has a weight percentage in a range of 0.35-0.5% with respect to the
composition.
[0044] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) galacto-oligosaccharide (GOS); and (b) at least one fermentable
fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a
weight ratio in a range of 1:0.-1:0.8-l: 1.25, and wherein galacto-oligosaccharide has
a weight percentage in a range of 0.35-0.5%) with respect to the composition, and the
at least one fermentable fibre has a weight percentage in a range of 0.35-0.5%) with
respect to the composition.
[0045] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) galacto-oligosaccharide (GOS); and (b) at least one fermentable fibre
selected from a group consisting of inulin, lactulose, xylooligosaccharide,
fructooligosaccharide, isomaltooligosaccharides, glucomannan, resistant starch,
pectin, and combinations thereof, wherein galacto-oligosaccharide to the at least one
fermentable fibre has a weight ratio in a range of 1:0.8-1:1.25.
[0046] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) galacto-oligosaccharide (GOS); (b) at least one fermentable fibre;
and (c) at least one excipient selected from a group consisting of diluent, carrier,
preservatives and combinations thereof, wherein galacto-oligosaccharide to the at
least one fermentable fibre has a weight ratio in a range of 1:0.8-l: 1.25.
[0047] In an embodiment of the present disclosure, there is provided a process for
preparing a composition: (a) galacto-oligosaccharide; and (b) at least one
fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable
fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i)
obtaining galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; and
(iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to
obtain the composition.
[0048] In an embodiment of the present disclosure, there is provided a process for
preparing a composition: (a) galacto-oligosaccharide; and (b) at least one

fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.8-1:1.25, said process comprising: (i) obtaining galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; and (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition.
[0049] In an embodiment of the present disclosure, there is provided a process for preparing a composition: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i) obtaining galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; and (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition, and wherein galacto-oligosaccharide has a weight percentage in a range of 0.35-0.5% with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.35-0.5% with respect to the composition.
[0050] In an embodiment of the present disclosure, there is provided a process for
preparing a composition: (a) galacto-oligosaccharide; and (b) at least one
fermentable fibre, selected from a group consisting of inulin, lactulose,
xylooligosaccharide, fructooligosaccharide, isomaltooligosaccharides,
glucomannan, resistant starch, pectin, and combinations thereof, wherein galacto-
oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of
1:0.75-1:1.75, said process comprising: (i) obtaining galacto-oligosaccharide; (ii)
obtaining at least one fermentable fibre; and (iii) contacting galacto-oligosaccharide
and the at least one fermentable fibre to obtain the composition.
[0051] In an embodiment of the present disclosure, there is provided a process for
preparing a composition: (a) galacto-oligosaccharide; and (b) at least one
fermentable fibre, selected from a group consisting of inulin, lactulose,
xylooligosaccharide, fructooligosaccharide, isomaltooligosaccharides,
glucomannan, resistant starch, pectin, and combinations thereof, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i) obtaining galacto-oligosaccharide; (ii)

obtaining at least one fermentable fibre; and (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition, and wherein galacto-oligosaccharide has a weight percentage in a range of 0.35-0.5% with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.35-0.5% with respect to the composition.
[0052] In an embodiment of the present disclosure, there is provided a fermented product obtained by fermentation of the composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75.
[0053] In an embodiment of the present disclosure, there is provided a fermented product obtained by fermentation of the composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.8-1:1.25.
[0054] In an embodiment of the present disclosure, there is provided a fermented product obtained by fermentation of the composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, and wherein galacto-oligosaccharide has a weight percentage in a range of 0.35-0.5%) with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.35-0.5%) with respect to the composition.
[0055] In an embodiment of the present disclosure, there is provided a fermented product obtained by fermentation of the composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre selected from a group consisting of inulin, lactulose, xylooligosaccharide, fructooligosaccharide, isomaltooligosaccharides, glucomannan, resistant starch, pectin, and combinations thereof, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75.

[0056] In an embodiment of the present disclosure, there is provided a fermented product obtained by fermentation of the composition comprising: (a) galacto-oligosaccharide; (b) at least one fermentable fibre; and (c) least one excipient selected from a group consisting of diluent, carrier, preservatives and combinations thereof, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75.
[0057] In an embodiment of the present disclosure, there is provided a process for preparing a fermented product obtained by fermentation of the composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i) obtaining galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition; (iv) incubating the composition in presence of at least one microorganism for a time-period in a range of 10-18 hours, at a temperature in a range of 25-37°C, to obtain the fermented product. In another embodiment of the present disclosure, the composition is incubated in presence of at least one microorganism for a time-period in a range of 12-17 hours, at a temperature in a range of 27-37°C, to obtain the fermented product.
[0058] In an embodiment of the present disclosure, there is provided a process for preparing a fermented product obtained by fermentation of the composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i) obtaining galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition; (iv) incubating the composition in presence of at least one microorganism selected from a group consisting of Bacillus subtilis, Lactobacillus, Lactococcus, Saccharomyces, Bifidobacterium, and combinations thereof for a time-period in a range of 10-18 hours, at a temperature in a range of 25-37°C, to obtain the fermented product. In

another embodiment of the present disclosure, the at least one microorganism is Bacillus subtilis, wherein the strain of Bacillus subtilis is CNCM 1-2745. [0059] In an embodiment of the present disclosure, there is provided a process for preparing a fermented product obtained by fermentation of a composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i) obtaining galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition; (iv) incubating the composition in presence of at least one microorganism for a time-period in a range of 10-18 hours, at a temperature in a range of 25-37°C, to obtain the fermented product, and wherein incubating the composition in presence of at least one microorganism is done in a growth medium. In another embodiment of the present disclosure, incubating the composition in presence of at least one microorganism is done in de Man, Rogosa and Sharpe (MRS) medium. [0060] In an embodiment of the present disclosure, there is provided a process for preparing a fermented product obtained by fermentation of a composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, said process comprising: (i) obtaining galacto-oligosaccharide; (ii) obtaining at least one fermentable fibre; (iii) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition; (iv) incubating the composition in presence of at least one microorganism selected from a group consisting of Bacillus subtilis, Lactobacillus, Lactococcus, Saccharomyces, Bifidobacterium, and combinations thereof for a time-period in a range of 10-18 hours, at a temperature in a range of 25-37°C, to obtain the fermented product, and wherein incubating the composition in presence of at least one microorganism is done in a growth medium.
[0061] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight

ratio in a range of 1:0.75-1:1.75, and wherein the composition increases the overall nutrient absorption status, and wherein nutrients includes, but is not limited to calcium, zinc, neutral amino acids, iron, vitamins B6, vitamin B12, and combinations thereof.
[0062] In an embodiment of the present disclosure, there is provided a fermented obtained by fermentation of a composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-l: 1.75, and wherein the fermented product increases the overall nutrient absorption status, and wherein nutrients includes, but is not limited to calcium, zinc, neutral amino acids, iron, vitamins and combinations thereof.
[0063] In an embodiment of the present disclosure, there is provided a composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, wherein the composition is used in at least one substance selected from a group consisting of dairy products, beverages, bakery products, nutraceuticals and combinations thereof.
[0064] In an embodiment of the present disclosure, there is provided a fermented product obtained by fermentation of a composition comprising: (a) galacto-oligosaccharide; and (b) at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75, wherein the composition is used in at least one substance selected from a group consisting of dairy products, beverages, bakery products, nutraceuticals and combinations thereof.
[0065] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.
EXAMPLES
[0066] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively

to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply. The working and non-working examples as depicted in the forthcoming sections highlight the criticality of the working percentages of different components in achieving the composition of the present disclosure. It is further specified that the presence of galacto-oligosaccharide (GOS) and at least one fermentable fibre, in a weight ratio range of 1:0.75-l: 1.75 is critical so as to achieve the composition in providing a synergistic effect in improving the overall nutrient absorption profile. The presence of any of the components specified above outside the disclosed weight ratio range(s) substantially affects the desired result.
Materials and Methods
[0067] The components- galacto-oligosaccharide (GOS) and inulin (fermentable fibre) were obtained from FrieslandCampina and Sigma, respectively; SYBR green master mix was procured from Thermo Fisher Scientific, Cat. No. 4309155.
Example 1
Process of preparing the composition of the present disclosure
Galacto-oligosaccharide and inulin were obtained. 1% solution (w/v) of galacto-oligosaccharide and inulin were prepared in de Man, Rogosa and Sharpe (MRS) medium (in the absence of glucose) and were subsequently contacted in appropriate volumes for different ratios to obtain the composition of the present disclosure.
Example 2
Process for preparing the fermented product of the present disclosure

The composition as obtained in Example 1 was contacted and incubated with 5% 0.8 OD Bacillus subtilis CNCM 1-2745 for a time period in a range of 12-16 hours at a temperature of 37°C, to obtain the fermented product of the present disclosure.
Example 3
Incubation of Caco-2 cell lines with the composition and mRNA expression
study
[0068] To study the effect of the fermented product on Caco-2 cells, the fermented
product as obtained in example 2 were centrifuged at 4500 rpm for around 10
minutes and supernatant was collected. Supernatant was filter sterilized using 0.2um
filter under sterile conditions. The filtered supernatants were treated at v/v
concentration of 10% on fully confluent Caco-2 cell line for a time period in a range
of 12-16 hours.
[0069] To study the effect of the fermented product on the expression of genes
involved in nutrient absorption, real time PCR was performed. For this purpose, the
treated Caco-2 cells were lysed, and mRNA was extracted using Qiagen RNeasy
Mini Kit (Cat. No. 74104). mRNA that was isolated was quantified using nanodrop
and 250 ng of isolated mRNA was converted into cDNA using Applied Biosystems
High Capacity cDNA reverse transcription kit (SKU No. 4368814). The cycling
conditions that were maintained for the purpose of cDNA synthesis were: a) initial
incubation of cDNA was carried out at a temperature of 25°C for about 10 minutes;
b) secondary incubation of cDNA was carried out at temperature of 37°C for 120
minutes; c) terminal incubation of cDNA was carried out at a temperature of 85°C
for 5 minutes.
[0070] Real-time PCR was performed on Applied Biosystems 7500 instrument using
primers specific for various nutrient receptors/transporters. GAPDH levels were also
evaluated for background normalization. Table 1 shows the list of primers that can
be used for the purpose of the present disclosure.
Table 1

[0071] Real-time analysis was performed in triplicates, using SYBR green chemistry. It can be depicted from Figure 1, that the cycling conditions maintained in PCR for the purpose of quantitative analysis of the transcripts consisted of a holding stage, cycling stage and melt curve stage, wherein CT (cycle threshold was adjusted to 0.2. Fold regulation of the genes was calculated by deriving -AACT values.
[0072] The genes that were evaluated using real time PCR were: FINF4A (hepatocyte nuclear factor), S100G (calcium binding protein), CDX2 (regulator of vitamin D receptor gene expression), and SLC6A19 (neutral amino acid transporter). Changes in the expression levels of the afore-mentioned genes were further used as representatives of the overall nutrient absorption profile.

Example 4
Effect of GOS and inulin on S100G and CDX2
[0073] Different ratios of galacto-oligosaccharide (GOS) and inulin were used to evaluate their effect on the expression level of the genes used as representatives of the overall nutrient absorption profile. The change in the expression level of the genes such as S100G (calcium binding protein) and CDX2 (regulator of vitamin D receptor gene expression) involved in nutrient absorption is determined by the fold change in the genes.
[0074] Table 2 shows the fold change in S100G and CDX2 involved in nutrient absorption at different ratios of GOS and inulin. Table 2
[0075] It can be observed from Table 2, Figure 2 and 3 that when components such
as GOS, inulin, lactulose, FOS, XOS fermented individually with Bacillus subtilis,
none of these individual components caused upregulation of genes such as S100G
and CDX2.
[0076] Further it was observed that when the GOS and inulin present at a weight
ratio of 1:1 in combination I (0.5g/100ml of GOS and 0.5g/100ml of inulin), was
fermented with Bacillus subtilis, showed a synergistic enhancement in expression of
genes i.e. S100G (Figure 2) and CDX2 (Figure 3) with a fold change of 1.72 and 1.9
respectively.
[0077] When GOS and inulin present at various weight ratios of 1:2, 2:1, 1:3 and
3:1 in combination(s) II (0.33g/100ml of GOS and 0.66g/100ml of inulin ), III

(0.66g/100ml of GOS and 0.33g/100ml of inulin), IV (0.25g/100ml of GOS and 0.75g/100ml of inulin), and V (0.75g/100ml of GOS and 0.25g/100ml of inulin) respectively, were fermented by Bacillus subtilis, it was observed that none of these combinations (II, III, IV, and V) exhibited any substantial upregulation of genes i.e. S100GandCDX2.
[0078] It can be inferred from Table 2, Figure 2 and 3, that the GOS and inulin present in combination I (1:1), caused maximum increase in expression levels of S100G and CDX2 as compared to GOS and inulin present in combination II (1:2), combination III (2:1), combination IV (1:3) and combination V (3:1). Therefore, GOS and inulin present in combination I, was considered as the working example of the present disclosure, whereas, GOS and inulin present in combination II, III, IV, and V, were considered as the non-working examples of the present disclosure.
Example 5
Effects of combination of GOS and inulin on HNF4A and SLC6A19
[0079] The combination of GOS and inulin were used at various ratio(s) for the
purpose of evaluating their effect in the expression levels of HNF4A, and SLC6A19.
[0080] Table 3 shows the fold change in HNF4A and SLC6A19 involved in
nutrient absorption at different ratios of GOS and inulin.

[0081] It can be observed from Table 3, Figure 4 and Figure 5, that when components such as GOS, inulin, lactulose, FOS, XOS were fermented individually with Bacillus subtilis, none of these individual components caused the upregulation of genes such as HNF4A (hepatocyte nuclear factor) and SLC6A19 (neutral amino acid transporter).
[0082] Further it was observed that when GOS and inulin present at a weight ratio of 1:1 in combination VI (0.5g/100ml of GOS and 0.5g/100ml of inulin) was fermented with Bacillus subtilis, there was a synergistic enhancement in expression of genes i.e. HNF4A and SLC6A19 with fold change of 1.93 and 1.69 respectively. [0083] When a combination of GOS and inulin were present at weight ratios of 1:2, 2:1, 1:3 and 3:1 in combinations VII (0.33g/100ml of GOS and 0.66g/100ml of inulin), VIII (0.66g/100ml of GOS and 0.33g/100ml of inulin), IX (0.25g/100ml of GOS and 0.75g/100ml of inulin), and X (0.75g/100ml of GOS and 0.25g/100ml of inulin) respectively, and were fermented by Bacillus subtilis, it was observed that none of these combination at said weight ratio(s) exhibited any substantial upregulation of genes i.e. HNF4A and SLC6A19.
[0084] It can be inferred from Table 3, Figure 4 and 5, that GOS and inulin present in combination VI (1:1) caused maximum increase in expression levels of HNF4A (Figure 4) and SLC6A19 (Figure 5) as compared to GOS and inulin present in combination VII (1:2), combination VIII (2:1), combination IX (1:3), and combination X (3:1). Therefore, GOS and inulin present in combination VI was considered as the working example of the present disclosure, whereas, GOS and inulin present in combinations VII, VIII, IX, and X, were considered as the non-working examples of the present disclosure.
[0085] Overall, the combination of galacto-oligosaccharide (GOS) and inulin (fermentable fibre) in the disclosed weight ratio range of 1:0.75-1:1.75 is essential for synergistically improving the overall nutrient absorption profile. The combination of GOS and inulin (fermentable fibre) cause maximum increase in expression levels of genes, including but not limited to S100G, CDX2, HNF4A and SLC6A19. As exemplified by the afore-mentioned examples, the presence of GOS

and a fermentable fibre outside said weight ratio range does not exhibit the desired result.
Advantages of the present disclosure
[0086] The present disclosure discloses a composition comprising galacto-oligosaccharide (GOS) and at least one fermentable fibre, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75. The composition was fermented with Bacillus subtilis to provide a fermented product. The fermented product provides synergistic enhancement of overall nutrient absorption, wherein nutrients including, but not limited to calcium, zinc, neutral amino acids, iron, vitamin D, vitamin B12, and combinations thereof. Apart from improving the overall nutrient profile, the composition also improves an overall health of an individual consuming the afore-mentioned products having the composition of the present disclosure. Further, the composition can be incorporated into various products that includes dairy products, beverages, bakery products, nutraceuticals, and combinations thereof. The process for preparing the composition and the fermented product is also quite simple and rapid.

I/We Claim:
1. A composition comprising:
a) galacto-oligosaccharide (GOS); and
b) at least one fermentable fibre,
wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.75-1:1.75.
2. The composition as claimed in claim 1, wherein galacto-oligosaccharide to the at least one fermentable fibre has a weight ratio in a range of 1:0.8-l: 1.25.
3. The composition as claimed in claim 1, wherein galacto-oligosaccharide has a weight percentage in a range of 0.35-0.5% with respect to the composition, and the at least one fermentable fibre has a weight percentage in a range of 0.35-0.5% with respect to the composition.
4. The composition as claimed in claim 1, wherein the at least one fermentable
fibre is selected from a group consisting of inulin, lactulose, xylooligosaccharide,
fructooligosaccharide, isomaltooligosaccharides, glucomannan, resistant starch,
pectin, and combinations thereof.
5. The composition as claimed in claim 1, wherein the composition further
comprises at least one excipient selected from a group consisting of diluent,
carrier, preservatives and combinations thereof.
6. A process for preparing the composition as claimed in claim 1, said process
comprising:
a) obtaining galacto-oligosaccharide;
b) obtaining at least one fermentable fibre; and
c) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition.

7. A fermented product obtained by fermentation of the composition as claimed in claim 1.
8. A process to obtain the fermented product as claimed in claim 7, said process comprising:

a) obtaining galacto-oligosaccharide;
b) obtaining at least one fermentable fibre;

c) contacting galacto-oligosaccharide and the at least one fermentable fibre to obtain the composition;
d) incubating the composition in presence of at least one microorganism for a time-period in a range of 10-18 hours, at a temperature in a range of 25-37°C, to obtain the fermented product.

9. The process as claimed in claim 8, wherein the at least one microorganism is selected from a group consisting of Bacillus subtilis, Lactobacillus, Lactococcus, Saccharomyces, Bifidobacterium, and combinations thereof
10. The process as claimed in claim 8, wherein incubating the composition in presence of at least one microorganism is done in a growth medium.