Description:FIELD OF INVENTION

The present disclosure, in general, relates to compositions for fermentation media and processes for growing microbial biomass. Particularly, it relates to fermentation media and processes for growing biomass of fructooligosaccharides producing microorganisms.

BACKGROUND

Fructooligosaccharides (FOS) also known as oligofructose or oligofructan are naturally occurring carbohydrates with numerous desirable characteristics. They are naturally indigestible and promote growth of good microbes in the gastrointestinal tract of humans and thus are referred to as prebiotics. They are subtly sweet and low-calorie and used as alternative sweetener. They are also used in the food industry as adjuvants in due to their organoleptic and technological properties.

FOS are found in several kinds of plants and vegetables such as wheat, banana, onion, and asparagus roots. They are industrially produced by two different processes. The first process involves inulin hydrolysis. The second process involves sucrose biotransformation by the enzymes fructosyltransferase (FTase) or ß-fructofuranosidase from bacterial and fungal sources.

The industrial production of FOS allows their mass production to meet global demand. Due the increased demand, there is a need to develop more productive, efficient, effective, and economical processes for production of FOS.

SUMMARY

The present disclosure relates to a fermentation medium for growing biomass of a fructooligosaccharides (FOS) producing microorganism, the medium comprising 2-20% ¬(v/v) of an FOS solution as a carbon source, wherein the FOS solution comprises 6-9% (w/v) of fructose, 50-60% (w/v) of glucose, 10-20% (w/v) of sucrose, and 18-20% (w/v) of FOS.

The present disclosure also relates to a process for growing biomass of an FOS producing microorganism, the process comprising culturing the microorganism in a fermentation medium comprising 2-30% (v/v) of an FOS solution as a carbon source, wherein the FOS solution comprises 6-9% (w/v) of fructose, 50-60% (w/v) of glucose, 10-20% (w/v) of sucrose, and 18-20% (w/v) of FOS.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 depicts the impact of a fermentation medium in accordance with an embodiment of the present disclosure on biomass yield.

FIG. 2 depicts the impact of a fermentation medium in accordance with an embodiment of the present disclosure on enzyme activity.

DETAILED DESCRIPTION

For the purpose of promoting an understanding of the principles of the disclosure, reference will now be made to embodiments and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the disclosure is thereby intended, such alterations and further modifications in the disclosed composition and method, and such further applications of the principles of the disclosure therein being contemplated as would normally occur to one skilled in the art to which the disclosure relates.

It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the disclosure and are not intended to be restrictive thereof.

Reference throughout this specification to “one embodiment” “an embodiment” or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, appearances of the phrase “in one embodiment”, “in an embodiment” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

The present disclosure relates to compositions for fermentation media. The disclosed fermentation media contain an FOS solution as a carbon source. The fermentation media are suitable for growing biomass of an FOS producing microorganism.

The fermentation medium in accordance with the present disclosure comprises 2-20% ¬(v/v) of an FOS solution as a carbon source, wherein the FOS solution comprises 6-9% (w/v) of fructose, 50-60% (w/v) of glucose, 10-20% (w/v) of sucrose, and 18-20% (w/v) of FOS.

The medium contains one or more additional components necessary for growth of the microorganism. It may contain one or more additional carbon source, one or more nitrogen source, one or more phosphorous nutrient, one or more of inorganic nutrient, and/or salt.

In accordance with an embodiment, the additional carbon source is in the range of 5-15% (w/v). Any suitable additional carbon source may be used. A combination of two or more additional carbon sources may be used. In accordance with various embodiments, the additional carbon source is selected from glucose, sucrose, maltose, fructose, glycerol, starch, lactose, galactose, sugarcane juice, jaggery, molasses, sugar beet, wheat, jowar and/or sweet sorghum, and a combination thereof.

The nitrogen source may be present in the range of 0.5-5% (w/v). Any suitable nitrogen source may be used. A combination of two or more nitrogen sources may also be used. In accordance with various embodiments, the nitrogen source is selected from peptone, yeast extract, casein, malt extract, powder of peanut, powder of soybean, corn steep solid, yeast powder, ammonium nitrate, and/or ammonium chloride and/or soya. In various embodiments, the nitrogen source is in the range of 0.5 to 3% (w/v). In accordance with some embodiments, the nitrogen source is the yeast extract. In a specific embodiment, the yeast extract is in the range of 0.5 to 3% (w/v).

The phosphorous source may be present in the range of 0.02-1.5% (w/v). Any suitable phosphorous source may be used. A combination of two or more phosphorus sources may also be used. In accordance with an embodiment, the phosphorous source is selected from di-potassium hydrogen phosphate, potassium di-hydrogen phosphate, di-sodium hydrogen phosphate, glycerophosphate, and a combination thereof. In specific embodiments, the phosphorous source comprises di-potassium hydrogen phosphate and/or potassium di-hydrogen phosphate. In an embodiment, the phosphorous source comprises 0.1-1% (w/v) of di-potassium hydrogen phosphate and 0.02-0.5% (w/v) of potassium di-hydrogen phosphate.

The inorganic nutrient may be present in the range of 0.05-0.5% (w/v). Any suitable inorganic nutrient may be added in the medium as per the specific needs and requirements. A combination of two or more inorganic nutrients may also be added. In accordance with various embodiments, the inorganic nutrient is selected from magnesium sulphate, ferrous sulphate, ferric chloride, boric acid, potassium iodide, zinc sulphate, manganese sulphate, and a combination thereof. In accordance with a specific embodiment, the inorganic nutrient is magnesium sulphate.

In accordance with an embodiment, the medium comprises 0.02-0.5% (w/v) of sodium chloride.

In some embodiments, the medium comprises 0.005-0.3% (w/v) of an antifoam agent. Any suitable antifoam agent may be used. A combination of two or more antifoam agents may also be used. In accordance with various embodiments, the antifoam agent is selected from polypropylene glycol, silicone-based oil, a silicone-based emulsion, and a combination thereof.

The medium may further contain other organic and/or inorganic materials so as to support or accelerate growth of the microorganism and increase the biomass yield. Examples of such materials include but are not limited to vitamins such ascorbic acid; inducers, such as methyl jasmonic acid, arachidonic acid, amino citrate, ceric ammonium nitrate, potassium permanganate, pyruvic acid, p-coumaric acid, vanadic sulfate, hippuric acid, c-naphthylacetic acid, 6-benzylamine purine, silver nitrate, and/or cinnamic acid; and precursors, such as-phenylalanine, benzamide, sodium benzoate, sodium acetate, propionamide, benzoic acid, and acetamide.

In accordance with some embodiments, the FOS solution is a reconstituted FOS solution. In accordance with some other embodiments, the FOS solution is a crude FOS solution generated as a byproduct during production of FOS. In accordance with an embodiment, the byproduct is generated after a chromatographic purification of the raw product obtained during the FOS production. For example, a typical process of FOS production broadly includes biomass cultivation, biotransformation, color removal, Sequential Simulated Moving Bed (SSMB) chromatographic purification, ionic impurity removal, evaporation and spray drying. The biotransformation usually results in a 45 to 65 brix low pure FOS solution having 50-65% purity as a raw product. This low pure FOS solution is subjected to the SSMB chromatographic purification by passing it through a resin such as styrene divinylbenzene copolymer resin to obtain pure FOS solution of 95-96% purity. The byproduct of the SSMB chromatographic purification may be used as the crude FOS solution for the purposes of the disclosed medium.

In accordance with an embodiment, the medium has pH in the range of 6.0-7.0. The pH may be adjusted using 1N NaOH and/or KOH or 1N HCL and/or H2SO4 solution.
The FOS producing microorganism can be any suitable microorganism. In accordance with an embodiment, the microorganism is a strain of Aspergillus spp. and/or Auerobasidium spp. In accordance with some embodiments, the microorganism is Aspergillus niger and/or Auerobasidium pullulans.

The present disclosure also relates to a process for growing biomass of an FOS producing microorganism. The process comprises culturing the microorganism in a fermentation medium comprising 2-30% (v/v) of an FOS solution as a carbon source, wherein the FOS solution comprises 6-9% (w/v) of fructose, 50-60% (w/v) of glucose, 10-20% (w/v) of sucrose, and 18-20% (w/v) of FOS.

In accordance with an embodiment, the microorganism is combined with the medium at a concentration of 3-5% (w/v).

In accordance with an embodiment, the microorganism is at least one of Aspergillus spp. and Aureobasidium spp. In accordance with some embodiments, Aspergillus spp. is Aspergillus niger and Aureobasidium spp. is Aureobasidium pullulans.

The medium used in the process contains one or more additional components necessary for growth of the microorganism. It may contain one or more additional carbon source, one or more nitrogen source, one or more phosphorous nutrient, one or more of inorganic nutrient, and/or salt.

In accordance with an embodiment, the medium used in the process comprises the additional carbon source in the range of 5- 15% (w/v). Any suitable additional carbon source may be used. A combination of two or more additional carbon sources may also be used. In accordance with various embodiments, the additional carbon source is selected from glucose, sucrose, maltose, fructose, glycerol, starch, lactose, galactose, sugarcane juice, jaggery, molasses, sugar beet, wheat, jowar and/or sweet sorghum, and a combination thereof.

The nitrogen source may be present in the range of 0.5-5% (w/v) in the medium. In an embodiment, the nitrogen source is in the range of 0.5 to 3% (w/v). Any suitable nitrogen source may be added to the medium. A combination of two or more nitrogen sources may also be used. In accordance with various embodiments, the nitrogen source is selected from peptone, yeast extract, casein, malt extract, powder of peanut, powder of soybean, corn steep solid, yeast powder, ammonium nitrate, and/or ammonium chloride and/or soya. In accordance with an embodiment, the nitrogen source is the yeast extract. In some embodiments, the yeast extract is in the range of 0.5 to 3% (w/v).

The phosphorous source may be present in the range of .02-1.5% (w/v) in the medium. Any suitable phosphorous source may be added the medium. A combination of two or more phosphorous sources may also be used. In accordance with an embodiment, the phosphorous source is selected from di-potassium hydrogen phosphate, potassium di-hydrogen phosphate, di-sodium hydrogen phosphate, glycerophosphate, and a combination thereof. In specific embodiments, the phosphorous source comprises di-potassium hydrogen phosphate and/or potassium di-hydrogen phosphate. In some embodiments, the phosphorous source comprises 0.1-1% (w/v) of di-potassium hydrogen phosphate and 0.02-0.5% (w/v) of potassium di-hydrogen phosphate.

The medium used in the disclosed process may include any suitable inorganic nutrient. A combination of two or more inorganic nutrients may also be used. The inorganic nutrient may be present in the range of 0.05-0.5% (w/v) in the medium. In accordance with various embodiments, the inorganic nutrient is selected from magnesium sulphate, ferrous sulphate, ferric chloride, boric acid, potassium iodide, zinc sulphate, manganese sulphate, and a combination thereof. In accordance with some embodiments, the inorganic nutrient is magnesium sulphate.

In accordance with an embodiment, the medium used in the process comprises 0.02-0.5% (w/v) of sodium chloride.
In another embodiment, the medium used in the process comprises 0.005-0.3% (w/v) of an antifoam agent. Any suitable antifoam agent may be used. A combination of two or more antifoam agents may be used. In accordance with various embodiments, the antifoam agent is selected from polypropylene glycol, silicone-based oil, a silicone-based emulsion, and a combination thereof.

The medium used in the process may comprise other organic and/or inorganic materials so as to support or accelerate growth of the microorganism and increase the biomass yield. Examples of such materials include but are not limited to vitamins such ascorbic acid; inducers, such as methyl jasmonic acid, arachidonic acid, aminocitrate, ceric ammonium nitrate, potassium permanganate, pyruvic acid, p-coumaric acid, vanadic sulfate, hippuric acid, c-naphthylacetic acid, 6-benzylamine purine, silver nitrate, and/or cinnamic acid; and precursors, such as-phenylalanine, benzamide, sodium benzoate, sodium acetate, propionamide, benzoic acid, and acetamide.

In accordance with an embodiment, the FOS solution is a reconstituted FOS solution. In accordance with another embodiment, the FOS solution is a crude FOS solution generated as a byproduct during production of FOS. In accordance with an embodiment, the byproduct is generated after a chromatographic purification of the raw product obtained during the FOS production as described above.

In accordance with an embodiment, the medium has pH in the range of 6.0-7.0. The pH may be adjusted using 1N NaOH and/or KOH or 1N HCL and/or H2SO4 solution.

The culturing may be carried out under conventional and/or known growth conditions in the art. In accordance with various embodiments, the culturing may be carried out under aerobic conditions at a temperature of around 25°C. In some embodiments, the temperature is in the range of 25-30°C. In accordance with an embodiment, the culturing is carried out for 20-28 hours. In accordance with an embodiment, the culturing is carried out at a stirring rate of 100-400 rpm.
The culturing may be carried out using any known apparatus, for example, a bottle, Erlenmeyer flask or conventional fermentation tanks, such as 7L, 35L, and 50L tanks.

The disclosed invention will be illustrated by means of the following examples. The examples are intended to illustrate various embodiments of the invention and are not meant to limit the scope of the claimed invention. Those skilled in the art will recognize that variations and modifications may be made to the described embodiments without departing from the broader scope of the invention as set forth in the claims.

EXAMPLES

Materials used in the Examples: The following materials were used in the Examples provided below.

1. Crude FOS Solution: The crude FOS solution was a byproduct of FOS production process carried out in the Tata Chemicals Ltd - Nutraceuticals plant. The FOS production was carried out using a strain of Aureobasidum pullulans. The process broadly included biomass cultivation, biotransformation, color removal, SSMB chromatographic purification, ionic impurity removal, evaporation and spray drying. The biotransformation resulted in a 35 brix low pure FOS solution having 55-60% purity as a raw product. Said 35 brix of low pure FOS solution was subjected to SSMB chromatographic purification by passing it through styrene divinylbenzene copolymer resin. resin to obtain pure FOS solution of 95-96% purity. Prior to the purification, the resin was regenerated with 5% KOH solution. After the purification, the crude FOS solution produced as a byproduct was collected.

2. Reconstituted FOS Solution: 20% reconstituted FOS solution was prepared by adding 0.2% Fructose, 3.0% Glucose, 0.9% Sucrose, and 0.9% FOS powder in 100 ml water.

3. Glucose-Yeast-Peptone (GYP) Agar Medium: GYP agar medium was prepared by adding 1 g of glucose, 0.6 g of yeast extract, 1 g of peptone, and 20 g of agar in 100 ml water.

4. Seed Inoculation Medium: Seed inoculum medium was prepared by adding 1 g of glucose, 0.6 g of yeast extract, and 1 g of peptone in 100 ml water. The pH of this medium was adjusted to 6.5 using 1N NaOH or 1N HCL solution.

5. Base Medium: Base medium was prepared by adding 1 g of peptone, 0.5 g of di-potassium hydrogen phosphate, 0.2 g potassium di-hydrogen phosphate, 0.2 g of magnesium sulphate, 0.25 g of sodium chloride, and 0.001 g of PPG in 100 ml water. The pH of the medium was adjusted to 6.5 using 1N NaOH or 1N HCL solution.

6. Microbial Strains: Cultures of Auerobasidium pullulans strain MCC 0127 (hereinafter referred to as the “AP strain”) and Aspergillus niger strain MCC 0252 (hereinafter referred to as the “AN strain”).

Example 1: Impact of crude FOS solutions having different FOS concentrations on biomass yield

AP strain cultures were maintained in the GYP agar medium. The cultures were then inoculated in the seed inoculation medium.

Crude FOS solutions having different concentrations were obtained after the SSMB chromatographic purification. Said solutions were subjected to High Pressure Chromatography (“HPLC”) to check the distribution of various sugars in each solution. The results are provided in Table 1 below.

Table 1: The Crude FOS Solutions having Different FOS Concentrations

FOS% in the Crude FOS Solution Glucose Fructose Sucrose FOS
2% 77 18 3 2
5% 76 17 5 5
10% 73 14 8 10
15% 62 8 19 15
20% 49 16 20 20
25% 52 9 19 25
30% 48 8 18 30
35% 47 4 15 35

Said solutions were used to investigate the impact on the biomass yield. The constant solid concentration of 5% (w/v) of different concentration of FOS was taken individually along with the base media and dissolved in the water to obtain 8 different fermentation media, A1-A8. The pH of A1-A8 was adjusted to 6.5 using 1N NaOH or 1N HCL solution. 10 ml of grown inoculum was transferred to different flasks containing 90 ml of A1-A8. The flasks were incubated at 27oC and 100 rpm for 24-30 hours. After the incubation period, the flasks were harvested, and biomass was collected. The collected biomass was used to check the biomass yield, which is depicted in Table 2 below.
Table 2: Impact of the crude FOS solutions having different FOS concentrations on Biomass Yield

Fermentation Medium Biomass Yield (g/L)
A1
(Base medium + 2% Crude FOS Solution + water) 20.65
A2
(Base medium + 5% Crude FOS Solution + water) 24.65
A3
(Base medium + 10% Crude FOS Solution + water) 30.35
A4
(Base medium + 15% Crude FOS Solution + water) 37.98
A5
(Base medium + 20% Crude FOS + water) 39.5
A6
(Base medium + 25% Crude FOS Solution + water) 38.67
A7
(Base medium + 30% Crude FOS Solution + water) 37.67
A8
(Base medium+ 25% Crude FOS Solution + water) 35.7

Example 2: Impact of different types of sugars and FOS on biomass yield

AP strain cultures was maintained in the GYP agar medium. The cultures were then inoculated in the seed inoculation media. Nine different media, F1-F9 were prepared by adding different sugars and FOS in the base medium as shown in Table 3.
Table 3: Media F1-F9

Medium Fructose (% w/v) Glucose (% w/v) Sucrose (% w/v) 20% Crude FOS Solution (% v/v) 20% Reconstituted FOS Solution (% v/v)
F1 5 - - - -
F2 4.1 - - - 0.9
F3 - 5 - -
F4 - 4.1 - - 0.9
F5 - - 5 - -
F6 - - 4.1 - 0.9
F7 - - - 5 -
F8 - - - - 5
F9 0.2 3 0.9 - 0.9

10 ml of the grown seed inoculum was transferred each of the 90 ml media F1-F9 in flasks. The flasks were incubated at 27°C for 24 hours at 80-150 rpm. After 24 hours, biomass in each flask was harvested and the biomass quantity was measured. The results are provided in Table 4.

Table 4: Impact of Different Types of Sugars and FOS on the Biomass Yield
Medium Biomass Yield (g/l)

F1 19.8
F2 25.4
F3 21.4
F4 26.5
F5 23.67
F6 25.6
F7 39.5
F8 23.4
F9 38.4

Example 3: Impact of the different media on different microbial cultures
10 ml grown inoculum of the AN strain and the AP strain were transferred to 90 ml each of F5, F7 and F9. The flasks were incubated at 27°C and 100 rpm for 24-30 hours. After the incubation period biomass was collected from each flask and biomass quantity was checked. The results are provided in Table 5.

Table 5: Biomass Yield
Microorganism Medium Biomass Yield (g/l)

AP Strain F5 23.67
AN Strain 40
AP Strain F7 39.5
AN Strain 62
AP Strain F8 38.4
AN Strain 58

Example 4: Evaluation of biomass yield with crude FOS solution
The 20% crude FOS solution was used to test biomass yield at 35 L working capacity. Accordingly, fermentation media containing 20% crude FOS solution (FM-1) and sucrose (FM-2) respectively were prepared. The details of the media composition are provided in Table 6. The media were sterilized at 121°C for 20 minutes and then cooled down to 27°C.

Table 6: Fermentation Media Compositions
S. No. Components FM-1 FM-2
1 Sucrose (g) 1750 -
2 Yeast extract (g) 350 350
3 K2HPO4 (g) 175 175
4 KH2PO4 (g) 70 70
5 NaCl (g) 87.5 87.5
6 MgSO4.7H2O (g) 70 70
7 PPG (g) 3.5 3.5
8 Water (l) 31.5 -
9 20% Crude FOS Solution (5% v/v) - 31.5
10 Final volume (Media and Inoculum) (l) 35 35
11 pH 6.5 6.5

Overnight grown AP strain inoculum was transferred to 31.5 l sterile FM-1 and FM-2 media. The temperature and the stirring rate were maintained at 27°C and 100 respectively. Samples were taken at different intervals to check biomass growth in each media. The results are provided in Table 7 below and depicted in FIG. 1.
Table 7: Biomass Yield
Duration Biomass Yield (g/L)

FM-1 FM-2
0 h 0.7 0.204
6 h 1.2 1.5
12 h 6.7 8.9
18 h 12.6 25.7
24 h 25.3 33.5
28 h 31 49

Example 5: Evaluation of FOS synthesis with crude FOS solution and sugar solution

The harvested biomass obtained in Example 5 was used to evaluate FOS synthesis. 0.3-0.8% of wet biomass was mixed with 50-65% of sugar solution and the solution was made up to 35 L and transferred to 50 L fermenter. The temperature and stirring rate were maintained at 50-60°C at 100-125 rpm respectively. Samples were taken at different intervals to analyse FOS and other content using HPLC. The results are provided in Table 8 below.

Table 8: FOS Synthesis
Duration (Hours) FM-1 FM-2
Fructose Glucose Sucrose FOS Fructose Glucose Sucrose FOS
4 0.46 13.94 49.43 36.17 0.25 11.39 57.51 30.85
8 0.57 17.95 36.2 45.28 0.78 17.83 34.86 46.53
12 0.61 21.22 26.88 51.29 0.96 22.48 20.82 55.74
16 0.7 23.29 21.42 54.59 1.11 24.72 15.17 59
24 0.84 24.78 17.53 56.85 0.99 27.3 10.96 60.75
28 0.8 25 15.69 58.51 1.25 28.45 10.03 60.27
36 1.05 28.09 9.54 62.37 1.31 28.4 10.14 60.15

Example 6: Effect of media on enzyme activity

The enzyme activity (IU) was estimated for one unit of Fructosyltransferase (FTase). The enzyme activity indicates that the quantity of enzyme capable of releasing 1µmol of reducing sugars (Glucose and Fructose) from sucrose per minute under standard assay conditions.

The enzyme activity was assessed using wet biomass obtained from FM1 and FM2. Initially, 100 mg of wet biomass was dissolved in 10 ml of water. Subsequently, 50 ml of a 50 brix sucrose solution was transferred to a clean, dried 250 ml flask. A known dilution of biomass from FM1 and FM2 was added to the flask, and the mixture was incubated at 55°C for 30 minutes, maintaining a shaker rpm of 150 during the incubation. After incubation, 1 ml of the reaction mixture was transferred to a clean, dried 50 ml test tube. The reactions were halted by adding 1 ml of DNS solution, and the test tubes were placed in a boiling water bath for 5 minutes.

Following incubation, the absorbance of the reaction mixture solution was determined at 595 nm using UV-spectrophotometry. The absorbance values obtained were utilized to calculate the concentration of reducing sugars, with glucose serving as the standard. The results are provided in Table 9 below and depicted in FIG.2.

Table 9. Effect of Media on Enzyme Activity
Duration Enzyme Activity (IU/g)
FM-1 FM-2
0 h 0 0
6 h 654 1400
12 h 675 1541
18 h 788 2028
24 h 850 2231
28 h 1100 2367

INDUSTRIAL APPLICABILITY

The present disclosure provides cost effective, sustainable, and economical fermentation media and processes for growing biomass of FOS producing microorganisms. The disclosed fermentation media and processes also allow the use of crude FOS solution which is a byproduct obtained during of industrial production of FOS. The disclosed fermentation media and processes provides enhanced biomass yield.
, Claims:¬¬We Claim:

1. A fermentation medium for growing biomass of a fructooligosaccharides producing microorganism, the medium comprising 2-20% ¬(v/v) of an fructooligosaccharides solution as a carbon source, wherein the fructooligosaccharides solution comprises 6-9% (w/v) of fructose, 50-60% (w/v) of glucose, 10-20% (w/v) of sucrose, and 18-20% (w/v) of fructooligosaccharides.

2. The medium as claimed in claim 1, comprising an additional carbon source selected from the group consisting of glucose, sucrose, maltose, fructose, glycerol, starch, lactose, galactose, sugarcane juice, jaggery, molasses, sugar beet, wheat, jowar and/or sweet sorghum, and a combination thereof.

3. The medium as claimed in claim 1, comprising 0.5-5% (w/v) of a nitrogen source selected from the group consisting of yeast extract, yeast powder, peptone, casein, malt extract, powder of peanut, powder of soybean, corn steep solid, ammonium nitrate, ammonium chloride, soya, soya protein and a combination thereof.

4. The medium as claimed in claim 3, comprising 0.5-3% (w/v) of the yeast extract.

5. The medium as claimed in claim 1, comprising 0.02-1.5% (w/v) of a phosphorous source selected from the group consisting of dipotassium hydrogen phosphate, potassium di hydrogen phosphate, di-sodium hydrogen phosphate, glycerophosphate, and a combination thereof.

6. The medium as claimed in claim 1, comprising 0.05-0.5% (w/v) of an inorganic nutrient selected from the group consisting of magnesium sulphate, ferrous sulphate, ferric chloride, boric acid, potassium iodide, Zinc sulphate, manganese sulphate, and a combination thereof.
7. The medium as claimed in claim 1, comprising 0.02-0.5% (w/v) of sodium chloride.

8. The medium as claimed in claim 1, comprising 0.005-0.3% (w/v) of an antifoam agent, the antifoam agent selected from the group consisting of polypropylene glycol, silicone-based oil, a silicone-based emulsion, and a combination thereof.

9. The medium as claimed in claim 1, wherein the fructooligosaccharides solution is a crude fructooligosaccharides solution generated as a byproduct during production of fructooligosaccharides.

10. The medium as claimed in claim 9, wherein the byproduct is generated after a chromatographic purification of a raw product generated during the production of fructooligosaccharides.

11. The medium as claimed in claim 1, wherein the fructooligosaccharides solution is a reconstituted fructooligosaccharides solution.

12. The medium as claimed in claim 1 having pH in the range of 6.0-7.0.

13. The medium as claimed in claim 1, wherein the microorganism is at least one of Aspergillus spp. and Aureobasidium spp.

14. A process for growing biomass of a fructooligosaccharides producing microorganism, the process comprising culturing the microorganism in a fermentation medium comprising 2-30% (v/v) of a fructooligosaccharides solution as a carbon source, wherein the fructooligosaccharides solution comprises 6-9% (w/v) of fructose, 50-60% (w/v) of glucose, 10-20% (w/v) of sucrose, and 18-20% (w/v) of fructooligosaccharides.

15. The process as claimed in claim 14, wherein the microorganism is combined with the medium at a concentration of 3-5% (w/v).
16. The process as claimed in claim 15, wherein the microorganism is at least one of Aspergillus spp. and Aureobasidium spp.

17. The process as claimed in claim 16, wherein the microorganism is at least one of Aspergillus niger and Aureobasidium pullulans.

18. The process as claimed in claim 14, wherein the medium comprises an additional carbon source selected from the group consisting of glucose, sucrose, maltose, fructose, glycerol, starch, lactose, galactose, sugarcane juice, jaggery, molasses, sugar beet, wheat, jowar and/or sweet sorghum, and a combination thereof.

19. The process as claimed in claim 14, wherein the medium comprises 0.5-5% (w/v) of a nitrogen source selected from the group consisting of yeast extract, yeast powder, peptone, casein, malt extract, powder of peanut, powder of soybean, corn steep solid, ammonium nitrate, ammonium chloride, soya, soya protein and a combination thereof.

20. The process as claimed in claim 19, wherein the medium comprises 0.5-3% (w/v) of the yeast extract.

21. The process as claimed in claim 14, wherein the medium comprises 0.02-1.5% (w/v) of a phosphorous source selected from the group consisting of di-potassium hydrogen phosphate, potassium di-hydrogen phosphate, di-sodium hydrogen phosphate, glycerophosphate, and a combination thereof.

22. The process as claimed in claim 14, wherein the medium comprises 0.05-0.5% (w/v) inorganic nutrient selected from the group consisting of magnesium sulphate, ferrous sulphate, ferric chloride, boric acid, potassium iodide, zinc sulphate, manganese sulphate, and a combination thereof.

23. The process as claimed in claim 14, wherein the medium comprises 0.02-0.5% (w/v) of sodium chloride.

24. The process as claimed in claim 14, wherein the culturing is carried out for 20-28 hours.

25. The process as claimed in claim 14, wherein the culturing is carried out at a stirring rate in the range of 100- 400 rpm.

26. The process as claimed in claim 14, wherein the culturing is carried out at a temperature in the range of 25-30°C.

27. The process as claimed in claim 14, wherein pH of the medium is in the range of 6-7.

28. The process as claimed in claim 14, wherein the fructooligosaccharides solution is a reconstituted fructooligosaccharides solution.

29. The process as claimed in claim 14, wherein the fructooligosaccharides solution is a crude fructooligosaccharides solution generated as a byproduct during production of fructooligosaccharides.

30. The process as claimed in claim 29, wherein the byproduct is generated after a chromatographic purification of a raw product generated during the production of fructooligosaccharides.

Dated this 13th day of December 2023