FIELD OF INVENTION
[001] The present disclosure relates to a methods of waste treatment, and in particular relates to microbial treatment process for digesting potato starch-containing substrates. 5
BACKGROUND OF INVENTION
[002] Potato is one of the most widely cultivated crop in India, and is also consumed and processed for various value addition products. Potatoes are processed in the industries for a variety of products including chips, mashed potatoes, fries, and
10 dehydrated products such as granules and flakes. The above-mentioned products generate wastes in the form of peel, pulp, and reject (FAO.2008., Proceedings of workshop to commemorate the international year of potato. RAP publication, Bangkok). The amount of waste generated by potato processing industries constitutes 12-20% of raw material processed (National Dairy Development Board
15 {NDDB}. 2014 Dairy knowledge portal.
http://www.dairyknowledge.in/article/potato-waste.)
[003] Cereal starch granules are reported to be easily digested by amylases of plant, microbe and human origin. Potato starch granules on the other hand are very resistant to hydrolysis except when gelatinized. Industries like potato chips processing plants
20 generate a huge amount of waste (vis-à-vis starch granules) which at times creates havoc for conventional effluent treatment plants. The conventional treatment of potato wastes including landfill, incineration, and animal feed are no longer preferred due to the secondary pollution caused by them. Also, dumping of the waste leads to undesirable microbial growth, thus creating an undesirable odor and an unhygienic
25 environment. Previous studies (Jacob S. et al., International journal of environmental science and technology. 2016 Oct 1;13(10):2363-74.) have attempted to study the anaerobic degradation of industrial potato wastes using consortia of microorganisms. [004] It is pertinent to note that gelatinisation of the potato starch further involves expenditure of energy and resources and considering the huge amounts of wastes

generated by the industry, an alternate solution to the problem needs to be developed, that can effectively degrade the non-gelatinized starch.
SUMMARY OF THE INVENTION
[005] In an aspect of the present invention, there is provided a process for digesting 5 potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate.
10 [006] 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
15 of the claimed subject matter.
BRIEF DESCRIPTION OF ACCOMPANYING DRAWINGS
[007] 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.
20 [008] Figure 1 illustrates Gram stained optical micrographs of microbial consortium in hydrolyzed potato starch extract after A) 18 h and B) 10 h. Inset in each image shows the complete consumption of starch with absence of blue colour, in accordance with an embodiment of the present disclosure. [009] Figure 2 illustrates iodine test of the samples, iodine was added to the culture
25 at 4 h and 5h and the mixture was serially diluted 1:1 to visually estimate the amount of starch remaining, in accordance with an embodiment of the present disclosure.

[0010] Figure 3 illustrates genus level profile of consortium based on 16S rRNA based metagenomics analysis, in accordance with an embodiment of the present disclosure.
[0011] Figure 4 illustrates Gram stained optical micrographs of microbial 5 consortium in hydrolyzed potato starch extract after A) 5 h and B) 10 h, in accordance with an embodiment of the present disclosure.
[0012] Figure 5 illustrates optical micrographs for A) Non-gelatinized and B) Gelatinized starch granules stained with iodine, in accordance with an embodiment of the present disclosure.
10 [0013] Figure 6 illustrates optical micrographs from Gram staining in non-gelatinized potato starch extract inoculated with bacillus at A) 24 h and B) 48 h post inoculation at pH 7.4 and C) 24 h and B) 48 h post inoculation at pH 6.0. Images from iodine test for presence of starch at 48 h for E) pH 7.4 and F) pH 6.0, Iodine was added to the culture at 24 h and 48 h and the mixture was serially diluted 1:1 to
15 visually estimate the amount of starch remaining, in accordance with an embodiment of the present disclosure.
[0014] Figure 7 illustrates a schematic for microbial digestion of potato extract, in accordance with an embodiment of the present disclosure.
20 DETAILED DESCRIPTION OF THE INVENTION
[0015] 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
25 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
[0016] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are delineated here. These
30 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. [0017] 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. 5 [0018] 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”.
[0019] Throughout this specification, unless the context requires otherwise the word “comprise”, and variations such as “comprises” and “comprising”, will be
10 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. [0020] The term “including” is used to mean “including but not limited to”. “Including” and “including but not limited to” are used interchangeably. [0021] Ratios, concentrations, amounts, and other numerical data may be presented
15 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. For example, a
20 temperature ranges of about 25-80 ℃ should be interpreted to include not only the explicitly recited limits of about 25 ℃ to about 80 ℃, but also to include sub-ranges, such as 25-75 ℃, 30-80 ℃, and so forth, as well as individual amounts, including fractional amounts, within the specified ranges, such as 25.2 ℃, and 79.5 ℃, for example.
25 [0022] To address the problems encountered in digesting the potato starch-containing substrates, the present disclosure discloses a process where an effluent comprising potato starch can be treated separately using microbial digestion with a microbe having the ability to digest potato starch. Since the use of enzymes alone is an expensive proposition, the use of a self-propagating microbial culture, having the
30 ability to produce extracellular enzymes to digest starch is an economically viable and powerful solution.

[0023] The present disclosure discloses a process whereby a gelatinized potato starch or non-gelatinized potato starch is treated with a strain of Bacillus cereus primarily contributes to the starch digestion and eventually the primary inoculum allows microbial consortia to build which ultimately digests the potato starch in a 5 time period in a range of 10-48 hours.
[0024] 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
10 disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference.
[0025] 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
15 are clearly within the scope of the disclosure, as described herein.
[0026] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second
20 mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch-containing substrate. In another embodiment of the present disclosure, incubating the second mixture is done for a time period in a range of 12-36 hours, for digesting potato starch-containing substrate. [0027] In an embodiment of the present disclosure, there is provided a process for
25 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is
30 obtained at a temperature in a range of 25-80℃. In another embodiment of the present disclosure, the first mixture comprising gelatinized potato starch is obtained

at a temperature in a range of 60-80℃. In yet another embodiment of the present disclosure, the first mixture is obtained at a temperature in a range of 70-80℃. In an alternate embodiment of the present disclosure, the first mixture comprising non-gelatinized potato starch is obtained at a temperature in a range of 25-35℃. 5 [0028] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48
10 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15 minutes. In another embodiment of the present disclosure, the first mixture is stirred at a speed in a range of 550-650 rpm for a time period in a range of 11-14 minutes. [0029] In an embodiment of the present disclosure, there is provided a process for
15 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the strain of Bacillus
20 cereus is ATCC 14579.
[0030] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second
25 mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein incubating the second mixture is done at a pH in a range of 7.0 to 7.4. In another embodiment of the present disclosure, incubating the second mixture is done at a pH in a range of 7.1 to 7.3.
30 [0031] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting

a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture 5 comprises gelatinized potato starch, and the starch in the substrate is completely digested in a time period in a range of 10-12 hours. In another embodiment of the present disclosure, the starch in the substrate is completely digested in a time period of 10 hours. [0032] In an embodiment of the present disclosure, there is provided a process for
10 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture
15 comprises non-gelatinized potato starch, and the starch is completely digested in a time period in a range of 36-48 hours. In another embodiment of the present disclosure, the starch is completely digested in a time period in a range of 38-46 hours. [0033] In an embodiment of the present disclosure, there is provided a process for
20 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein incubating the
25 second mixture is done at a temperature in a range of 25-30℃. In another embodiment of the present disclosure, incubating the second mixture is done at a temperature in a range of 26-29℃.
[0034] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting
30 a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second

mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the substrate is an industry effluent.
[0035] In an embodiment of the present disclosure, there is provided a process for 5 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein incubating the
10 second mixture is done at a stirring speed in a range of 300-500 rpm. In another embodiment of the present disclosure, incubating the second mixture is done at a stirring speed in a range of 350-450 rpm.
[0036] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting
15 a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the
20 second mixture has a volume in a range of 500-1000 mL. In another embodiment of the present disclosure, the second mixture is aerated using an air pump. In yet another embodiment of the present disclosure, the second mixture is aerated at a bubbling rate in a range of 7-10 litre per minute, and wherein the second mixture has a volume in a range of 600-900 mL.
25 [0037] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus ATCC 14579 to obtain a second mixture; and (c) incubating the second mixture for a time period in a range
30 of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range

of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃.
[0038] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting 5 a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15
10 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃.
[0039] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus ATCC 14579 to obtain a
15 second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃, and the first mixture comprises gelatinized potato starch, and the starch in the
20 first mixture is completely digested in a time period in a range of 10-12 hours.
[0040] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second
25 mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃, and the first mixture comprises gelatinized potato starch, and the starch in the first
30 mixture is completely digested in a time period in a range of 10-12 hours. It can be

contemplated that once the substrate is contacted with water at a temperature in a range of 60-80℃, the obtained first mixture comprises gelatinized potato starch. [0041] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting 5 a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus ATCC 14579 to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range
10 of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃, and the first mixture comprises non-gelatinized potato starch, and the starch is completely digested in a time period in a range of 36-48 hours. It can be contemplated that once the substrate is contacted with water at a temperature in a range of 25-35℃, the obtained first mixture comprises non-gelatinized potato starch.
15 [0042] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48
20 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃, and the first mixture comprises non-gelatinized potato starch, and the starch is completely digested in a time period in a range of 36-48 hours.
25 [0043] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus ATCC 14579 to obtain a second mixture; and (c) incubating the second mixture for a time period in a range
30 of 10-48 hours, for digesting potato starch containing substrate, wherein incubating the second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range

of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL. [0044] In an embodiment of the present disclosure, there is provided a process for 5 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein incubating the
10 second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL. [0045] In an embodiment of the present disclosure, there is provided a process for
15 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus ATCC 14579 to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein incubating
20 the second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL, and the first mixture comprises gelatinized potato starch, and the starch in the first mixture is completely
25 digested in a time period in a range of 10-12 hours.
[0046] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second
30 mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein incubating the

second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL, and the first mixture 5 comprises gelatinized potato starch, and the starch in the first mixture is completely digested in a time period in a range of 10-12 hours.
[0047] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b)
10 contacting the first mixture with a strain of Bacillus cereus ATCC 14579 to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein incubating the second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second
15 mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL, and the first mixture comprises non-gelatinized potato starch, and the starch is completely digested in a time period in a range of 36-48 hours. [0048] In an embodiment of the present disclosure, there is provided a process for
20 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein incubating the
25 second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL, and the first mixture comprises non-gelatinized potato starch, and the starch is completely digested in a
30 time period in a range of 36-48 hours.

[0049] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second 5 mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃, and the first mixture comprises non-gelatinized potato starch, and the starch is
10 completely digested in a time period in a range of 36-48 hours, and incubating the second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL.
15 [0050] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48
20 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃, and the first mixture comprises gelatinized potato starch, and the starch in the substrate is completely digested in a time period in a range of 10-12 hours, and incubating the
25 second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL. [0051] In an embodiment of the present disclosure, there is provided a process for
30 digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b)

contacting the first mixture with a strain of Bacillus cereus ATCC 14579 to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range 5 of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-80℃, and the first mixture comprises non-gelatinized potato starch, and the starch is completely digested in a time period in a range of 36-48 hours, and incubating the second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second 10 mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, and wherein the second mixture has a volume in a range of 500-1000 mL. [0052] In an embodiment of the present disclosure, there is provided a process for digesting potato starch-containing substrate, said process comprising: (a) contacting a potato starch containing substrate with water to obtain a first mixture; (b) 15 contacting the first mixture with a strain of Bacillus cereus ATCC 14579 to obtain a second mixture; and (c) incubating the second mixture for a time period in a range of 10-48 hours, for digesting potato starch containing substrate, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15 minutes, and the first mixture is obtained at a temperature in a range of 25-20 80℃, and the first mixture comprises gelatinized potato starch, and the starch in the substrate is completely digested in a time period in a range of 10-12 hours, and incubating the second mixture is done at a pH in a range of 7.0 to 7.4, at a temperature in a range of 25-30℃, and at a stirring speed in a range of 300-500 rpm, and wherein the second mixture is aerated at a bubbling rate in a range of 5-12 litre per minute, 25 and wherein the second mixture has a volume in a range of 500-1000 mL.
[0053] In an embodiment of the present disclosure, there is provided a process as described herein, wherein the substrate is an industry effluent. [0054] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations 30 are possible.

[0055] 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 5 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 10 described, as such methods and conditions may apply.
[0056] The examples as presented herein describe the best working process of the present disclosure. The examples have been described to explain the process for digesting gelatinized as well as non-gelatinized potato starch.
15 EXAMPLE 1
Studies with gelatinized potato starch
[0057] The pure Bacillus in the present process is a strain of Bacillus cereus. One particular strain used in the present process is Bacillus cereus ATCC 14579. The pure strain was inoculated in a freshly prepared potato starch extract to obtain a
20 mixture, the mixture was stirred and aerated by bubbling air continuously under ambient conditions. Complete starch hydrolysis was observed within 18 h as indicated by iodine test (Figure 1). Closer investigation revealed that starch was completely consumed by 10 h wherein approximately half of the total starch was consumed by 5 h (Figure 1 and 2). The 16S rRNA based metagenomics profiling of
25 the consortia growing in the hydrolyzed extract had the genus level profile as shown in Figure 3.
[0058] The microbial consortium during the digestion of gelatinized potato starch extract at 5 h and 10 h using Gram staining was studied (Figure 4). It was found that at 5 h, the inoculated Gram-positive Bacillus was identified as the major bacterium,
30 which was primarily responsible for digesting the starch. At 10 h, apart from the inoculated Bacillus, other Gram-negative bacteria were also visualised. Hence, from

the above paragraphs it is clear that pure inoculum of the Bacillus into the potato starch extract seems to primarily digest the starch in the sample. As the digestion progresses, a microbial consortium develops which continues digestion of other components in the digested potato starch extract. 5 [0059] The digestion of a fresh batch of potato extract was tested by adding hydrolysed potato starch extract into the fresh batch. In such a scenario, microbial consortia already developed while digesting the potato starch initiates the digestion of a fresh batch of potato extract. In the present study, it was observed that in such a scenario, complete hydrolysis of the starch was observed only after 18 h.
10 [0060] It can be appreciated that if the digestion of the potato starch is initiated with Bacillus cereus ATCC 14579 strain, starch was completely digested by 10 hours, whereas if the digestion of the potato starch is initiated by microbial consortia, complete hydrolysis is observed only after 18 hours. Therefore, the process using the strain as disclosed in the present disclosure results in faster and efficient starch
15 hydrolysis, thereby providing an assertive solution to address the problem of industrial potato starch effluent.
EXAMPLE 2
Studies with non-gelatinized starch
[0061] The next step was to study the hydrolysation of non-gelatinized potato starch
20 extract by the present process involving the bacterium. The fundamental difference between non-gelatinized and gelatinized starch is that gelatinization causes swelling of the starch granules (Figure 5) which makes the starch granules much more susceptible to microbial digestion. [0062] The non-gelatinized potato starch extract was either adjusted to pH 6.0 or
25 used at pH 7.4. The Bacillus strain (ATCC 14579) was inoculated in the extracts and the mixture was stirred and aerated by bubbling air continuously under ambient condition. Complete starch hydrolysis was observed at the end of 48 h for digestion initiated at pH 7.4 as indicated by iodine test. In case of digestion initiated at pH 6.0, no starch consumption was observed (Figure 6).
30 [0063] Overall, the present disclosure discloses a process of treating potato starch-containing substrates using a primary inoculum of Bacillus cereus strain. Although

the present disclosure is intended to cover the process involving all the strains of Bacillus cereus, a particular strain ATCC 14579 was used. The process involves treating of the substrate with water followed by incubation with a pure culture of Bacillus cereus ATCC 14579. The pure culture facilitates the growth of microbial consortia thus efficiently digesting the potato starch-containing substrate. The overall process that can be employed in this regard is mentioned as a schematic in Figure 7.
[0064] It can also be appreciated that the present process as disclosed herein works the best in presence of Bacillus cereus in general, and Bacillus cereus strain ATCC 14579 in particular. In case of Escherichia coli, the bacteria alone is not able to digest gelatinized or soluble starch like the Bacillus cereus strain disclosed in the present disclosure. Whereas, only if E. coli is grown in a medium (Luria-Bertani) supplemented with soluble starch, it is able to digest starch. Thus, it should be understood that using E. coli will not solve the purpose of digesting potato starch-containing substrates. Therefore, mere use of any microorganism in digesting potato starch does not work well and cannot be used in treating potato starch-containing substrate.
Advantages of the present disclosure
[0065] The present disclosure discloses provides a simple process for treating difficult-to-digest potato starch containing-substrates. The process leads to complete starch digestion in 36-48 hours in case of non-gelatinized starch, and in 10-12 hours in case of gelatinized starch. The process does not involve complex machinery. The present disclosure provides a solution to treatment of wastes generated by potato processing industry. The process as disclosed provides economic benefits in terms of using a self-propagating microbial culture instead of pure enzymes which are very costly.

1. A process for digesting potato starch-containing substrate, said process
comprising:
a) contacting a potato starch containing substrate with water to obtain a first mixture;
b) contacting the first mixture with a strain of Bacillus cereus to obtain a second mixture; and
c) incubating the second mixture for a time period in a range of 10-48
hours, for digesting potato starch containing substrate.
2. The process as claimed in claim 1, wherein the first mixture is obtained at a temperature in a range of 25-80℃.
3. The process as claimed in claim 1, wherein the first mixture is stirred at a speed in a range of 500-700 rpm for a time period in a range of 10-15 minutes.
4. The process as claimed in claim 1, wherein the strain of Bacillus cereus is ATCC 14579.
5. The process as claimed in claim 1, wherein incubating the second mixture is done at a pH in a range of 7.0 to 7.4.
6. The process as claimed in claim 1, wherein the first mixture comprises gelatinized potato starch, and the starch in the substrate is completely digested in a time period in a range of 10-12 hours.
7. The process as claimed in claim 1, wherein the first mixture comprises non-gelatinized potato starch, and the starch is completely digested in a time period in a range of 36-48 hours.
8. The process as claimed in claim 1, wherein incubating the second mixture is done at a temperature in a range of 25-30℃.
9. The process as claimed in claim 1, wherein the substrate is an industry effluent.

10. The process as claimed in claim 1, wherein incubating the second mixture is done at a stirring speed in a range of 300-500 rpm.