DESC:FORM-2
THE PATENTS ACT, 1970
(39 of 1970)
&
THE PATENTS RULES, 2003
COMPLETE SPECIFICATION
(See section 10 and rule 13)
Title: A PROTEASE COMPOSITION FOR DIGESTION OF WHEY PROTEIN AND
METHOD THEREOF
APPLICANT DETAILS:
(a) NAME: Zywie Ventures Pvt Ltd.
(b) NATIONALITY: IN
(c) ADDRESS: 5, Dashrath Singh Estate, Caves Road, Near Jogeshwari Caves,
Jogeshwari East, Mumbai 400060 India.
PREAMBLE TO THE DESCRIPTION:
The following specification (particularly) describes the nature of the invention (and the manner
in which it is to be performed):
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A PROTEASE COMPOSITION FOR DIGESTION OF WHEY PROTEIN AND
METHOD THEREOF
FIELD OF INVENTION
The present invention provides a protease composition. The Invention in particular provides a
Protease composition for digestibility of the whey protein and method thereof.
BACKGROUND OF THE INVENTION
The following background discussion includes information that may be useful in understanding
the present invention. It is not an admission that any of the information provided herein is prior
art or relevant to the presently claimed invention, or that any publication specifically or
implicitly referenced is prior art.
Milk is made of two proteins, casein and whey. Whey protein can be separated from the casein
in milk or formed as a by-product of cheese making.
There are different types of whey protein like whey protein concentrate (WPC), whey protein
isolate (WPI), and whey protein hydrolysate (WPH).Whey protein concentrate contains low
levels of fat and low levels of carbohydrates. Whey protein isolate are obtained after further
processing to remove all the fat and lactose. Whey protein hydrolysate is obtained after partial
hydrolysis – a process necessary for the body to absorb protein. It is important that consumed
protein is broken down by the body during digestion so that it can be effectively absorbed and
used to support body functions. Whey supplements provide high-quality dietary protein with
little or no fat. However, in addition to their benefits, they carry drawbacks, including digestive
problems.
As food trends come and go, the sustainability and nutritional value of whey protein remains
stable and continues to be the protein of choice among food manufacturers that have come to
rely on its functionality, price stability and use in a wide range of applications.
Protein breakdown by endogenous enzymes in the gastrointestinal (GI) tract results in
generation of peptides and amino acids that act as building blocks in essential biological
functions. Exogenous proteases possess immense potential as digestive enzyme supplements
that can assist protein digestion in the GI system.
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Hence, there is a need of a plant based protease composition that can help in the digestion of
whey protein efficiently.
OBJECTIVE OF THE PRESENT INVENTION:
An object of the present invention is to provide a protease composition.
Another object of the present invention is to provide a protease composition for digestibility of
the whey protein and method thereof.
Another object of the present invention is to provide an effective supplement composition for
protein digestion.
SUMMARY OF THE INVENTION:
In an aspect of the present invention there is provided a protease composition for digestibility
of the whey protein comprising:
a) plurality of plant based enzymes, wherein the composition comprises at least 2
plant based enzymes.
In an embodiment, there is provided a protease composition for digestibility of the whey protein
comprising fungal Acid Protease wherein the protease comprises 500 TU/mg, 25000 HUT/g
acid proteases.
In an embodiment, protease composition shows high efficacy of digestion which gives good
results in degree of hydrolysis.
In an embodiment, the protease composition displayed low molecular weight fragmented
peptides.
In an embodiment, whey protein isolate (WPI), whey protein concentrates (WPC) shows better
digestion with protease composition.
In an embodiment, the protease composition showed maximum Degree of hydrolysis (DH) up
to 15.77 ± 1.18 % after both Gastric digestion and 57.75 ± 3.55% after gastrointestinal
digestion.
In an embodiment, upon gastric and gastro-intestinal digestion, the protein samples with
protease enzyme were found to have fragmented peptides with lower molecular weight.
This together with the other aspects of the present invention along with the various features of
novelty that characterized the present disclosure is pointed out with particularity in claims
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annexed hereto and forms a part of the present invention. For better understanding of the
present disclosure, its operating advantages, and the specified objective attained by its uses,
reference should be made to the accompanying descriptive matter in which there are illustrated
exemplary embodiments of the present invention.
Description of the Drawings:
Figure 1: illustrates degree of hydrolysis (%) of protein samples (WPC, WPI and Plant Protein)
after gastric digestion with MDZenPro
Figure 2: illustrates degree of hydrolysis (%) of protein samples (WPC, WPI and Plant Protein)
after gastrointestinal digestion with MDZenPro
Figure 3: illustrates SDS-PAGE profile of undigested protein samples (WPC, WPI and plant
protein). Lane 1 - WPC, Lane 2 - WPI, Lane 3 - Plant Protein, Lane 4 – Ladder
Figure 4: illustrates SDS-PAGE profile of protein samples subjected to simulated gastric and
GI digestion (in absence and presence of exogenous enzymes viz. Commercial protease and
MDZenPro) Lane 1 - WPC, Lane 2 - WPC + MDZenPro, Lane 3 - WPC + Commercial
protease, Lane 4 - WPI, Lane 5 - WPI +MDZenPro, Lane 6 - Ladder, Lane 7 - Plant Protein,
Lane 8 - Plant Protein + MDZenPro
DETAIL DESCRIPTION OF THE INVENTION:
For the purpose of promoting an understanding of the principles of the invention, reference will
now be made to the embodiment illustrated in the drawings and specific language will be used
to describe the same. It will nevertheless be understood that no limitation of the scope of the
invention is thereby intended, such alterations and further modifications in the illustrated
system, and such further applications of the principles of the invention as illustrated therein
being contemplated as would normally occur to one skilled in the art to which the invention
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 invention and are not
intended to be restrictive thereof. Throughout the patent specification, a convention employed
is that in the appended drawings, like numerals denote like components.
Reference throughout this specification to “an embodiment, “another embodiment, “an
implementation, “another implementation” or similar language means that a particular feature,
structure, or characteristic described in connection with the embodiment is included in at least
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one embodiment of the present invention. Thus, appearances of the phrase “in an embodiment,
“in another embodiment, “in one implementation, “in another implementation, and similar
language throughout this specification may, but do not necessarily, all refer to the same
embodiment.
The terms "comprises", "comprising", or any other variations thereof, are intended to cover a
non-exclusive inclusion, such that a process or method that comprises a list of steps does not
include only those steps but may include other steps not expressly listed or inherent to such
process or method. Similarly, one or more devices or sub-systems or elements or structures
proceeded by "comprises... a" does not, without more constraints, preclude the existence of
other devices or other sub-systems or other elements or other structures or additional devices
or additional sub-systems or additional elements or additional structures.
The present invention provides a protease composition. The Invention in particular provides a
Protease composition for digestibility of the whey protein and method thereof.
In an aspect of the present invention there is provided a protease composition for digestibility
of the whey protein comprising:
a) plurality of plant based enzymes, wherein the composition comprises at least 2
plant based enzymes.
In an embodiment, at least 3 plant based enzymes present in a specific amount.
In an embodiment, the protease composition for digestibility of the whey protein comprises
fungal Acid Protease wherein the protease comprises 500 TU/mg, 25000 HUT/g acid proteases.
In an embodiment, protease composition shows high efficacy of digestion which gives good
results in degree of hydrolysis.
In an embodiment, the protease composition displayed low molecular weight fragmented
peptides.
In an embodiment, whey protein isolate (WPI), whey protein concentrates (WPC) show better
digestion with protease composition.
In an embodiment, the protease composition showed maximum Degree of hydrolysis (DH) up
to 15.77 ± 1.18 % after both Gastric digestion and 57.75 ± 3.55% after gastrointestinal
digestion.
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In an embodiment, upon gastric and gastro-intestinal digestion, the protein samples with
protease enzyme were found to have fragmented peptides with lower molecular weight.
The protease composition enhances protein digestibility for the population consuming whey
protein. Better Protein digestibility supports fast muscle growth by enhancing the degree of
hydrolysis and free amine group release. The protease composition enhances digestibility of
whey protein. The molecular weight is lower in case of samples with protease composition.
In an embodiment, the protease composition comprises plurality of plant based enzymes.
In an embodiment, the protease composition comprises at least 2 plant based enzymes.
In an embodiment, the protease composition comprises at least 3 plant based enzymes present
in a specific amount.
The protease composition shows high efficacy of digestion which gives good results in degree
of hydrolysis as compared to whey protein with other digestive enzymes.
The protease composition (being traded by Brand name MDZenPro) digests raw whey protein,
whey protein isolate and plant protein under INFOGEST simulated GI conditions. The gastric
and GI digested protein products were analyzed for determining the degree of hydrolysis. The
protein profiles were evaluated using SDS-PAGE. The results of the degree of hydrolysis
experiment reveals that MDZenPro protease blend facilitated gastric and GI digestion of
proteins. This increase in degree of hydrolysis was noted to be higher than that observed in
proteins that were not treated with the enzyme supplement. The SDS-PAGE profile further
supported these findings wherein, the MDZenPro treated protein samples displayed low
molecular weight fragmented peptides in contrast to the profile of undigested proteins.
The protease composition was further analysed for the degree of hydrolysis by OPA Method.
It was found that Whey protein isolate (WPI), Whey protein concentrates (WPC) and plant
proteins with MDZenPro showed better protein digestion as compared to samples without
protease. The results are mentioned below:
Sample WPI + protease composition of the present invention showed maximum Degree of
hydrolysis (DH) up to 15.77 ± 1.18 % after both Gastric digestion and 57.75 ± 3.55% after
gastro intestinal digestion.
The SDS-PAGE analysis shows that the SDS profile pattern confirms that upon gastric and
gastro-intestinal digestion, the protein samples with protease enzyme were found to have
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fragmented peptides with lower molecular weight when compared with undigested protein
profile. The fragmentation in the protein samples substantiates the digestion of protein samples.
In an embodiment, the method of preparing protease composition for digestibility of the whey
protein as described above, comprises the steps of:
a) fungal Acid Protease wherein the protease comprises 500 TU/mg, 25000 HUT/g acid
proteases, wherein fungal acid protease is produced by controlled fermentation of selected
strain of Aspergillus oryzae;
b) reducing the particle size to 30 mm;
c) mixing with whey protein;
d) maintaining pH 4 – 9 and Temperature of 40°C
The Invention is further described with the help of non-limiting examples:
The below examples show results of the experiment/tests to evaluate the degree of hydrolysis
and electrophoretic analysis of the present composition / MDZenPro, provided by the present
invention. The results indicates that MDZenPro has a positive effect on protein digestion under
simulated GI conditions which is based on plant protease.
The results shows that MDZenPro is a digestive aid for augmenting the breakdown of proteins.
Example 1:
Analysis of Degree of Hydrolysis by OPA Method
MDZenPro was studied to determine its ability to augment the in-vitro gastric and GI digestion
of whey protein and plant protein samples. Degree of hydrolysis for WPC, WPI and PP under
simulated GI conditions were found to be 8.3%, 14.4% and 6.6% respectively. It was observed
that the degree of hydrolysis of WPC, WPI and plant proteins that were treated with the
MDZenPro was higher as compared to the protein samples that were not treated with the
enzymes (Figure 1 and Figure 2). The untreated set showed a protein digestion of less than
10%. Hence, when compared to an untreated set, the set with MDZenPro showed
approximately 59% faster digestibility. Protein sample WPI treated with MDZenPro showed
maximum degree of hydrolysis after both gastric digestion (15.77%) and GI digestion
(57.75%). Whey protein is one of the most popular protein supplements available in the market
owing to its amino acid profile. In the present study, the degree of hydrolysis of WPC samples
after treatment with a commercially available protease and MDZenPro under simulated GI
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conditions were found to be comparable, thereby validating the high efficiency of MDZenPro
(Table 1).
Example 2:
Analysis of Protein Hydrolysis using SDS-PAGE
The profile of undigested and digested proteins (WPC, WPI and plant protein) were studied
using SDSPAGE. The SDS-PAGE profile pattern confirmed that upon simulated gastric and
GI digestion, the protein samples treated with MDZenPro were hydrolysed into fragmented
peptides (Figure 3). The WPC protein profiles obtained after hydrolysis with MDZenPro and
commercial protease were comparable (Figure 4). These peptides possessed lower molecular
weight as compared to the undigested protein samples (Figure 3 and Figure 4). The
fragmentation of the protein samples after treatment with exogenous MDZenPro product
further substantiates the digestion of protein samples. ,CLAIMS:We Claim:
1. A protease composition for digestibility of the whey protein comprising fungal Acid
Protease wherein the protease comprises 500 TU/mg, 25000 HUT/g acid proteases.
2. The protease composition for digestibility of the whey protein as claimed in claim 1,
wherein protease composition shows high efficacy of digestion which gives good
results in degree of hydrolysis.
3. The protease composition for digestibility of the whey protein as claimed in claim 1,
wherein the protease composition displayed low molecular weight fragmented peptides.
4. The protease composition for digestibility of the whey protein as claimed in claim 1,
wherein whey protein isolate (WPI), whey protein concentrates (WPC) show better
digestion with protease composition.
5. The protease composition for digestibility of the whey protein as claimed in claim 1,
wherein the protease composition showed maximum Degree of hydrolysis (DH) up to
15.77 ± 1.18 % after both Gastric digestion and 57.75 ± 3.55% after gastrointestinal
digestion.
6. The protease composition for digestibility of the whey protein as claimed in claim 1,
wherein upon gastric and gastro-intestinal digestion, the protein samples with protease
enzyme were found to have fragmented peptides with lower molecular weight.
7. A method of preparing protease composition for digestibility of the whey protein as
claimed in claim 1, comprising steps of:
a) fungal Acid Protease wherein the protease comprises 500 TU/mg, 25000 HUT/g
acid proteases, wherein fungal acid protease is produced by controlled fermentation of
selected strain of Aspergillus oryzae;
b) reducing the particle size to 30 mm;
c) mixing with whey protein;
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d) maintaining pH 4 – 9 and Temperature of 40°C