Field of the invention
The present invention relates to a protein composition derived from Amla {Emhelica officinalis) seed, which has immense practical utility as a material for foodstuffs. More particularly the invention relates to a process for extraction of protein from Amla seed. The isolated Amla protein fits safe application as material for preparation of cosmetic products, pharmaceuticals and foodstuffs.
Background of the invention
Amla is known for being a good source of nutrients and having high therapeutic value. It has been known for long that the fruit from Amla is edible and it serves as an excellent source of nutrition. Amla fruit is vastly used in varied treatment methods including Ayurvedic procedure of medicament. Amla as a whole is known for its therapeutic properties and finds immense usage in the area of nutraceutical and cosmetic preparations as it is considered to be a good source of antioxidants and hence acts as an immune-booster. Various parts of Amla plant are used in the above said preparations, however, little is known of the properties of Amla seed, which is majorily a by-product of the Amla based industries. There have not yet been any researches performed on Amla seed. The amla seed and its product are not disclosed in any literature. However, there does exist some information on certain legume and oil seed. For example US Patent 5,993,795 discloses a protein composition derived from sesame seed.
US 4, 366,097 discloses a novel protein isolation procedure from certain legume and oil seeds typically rapeseed by extracting protein from the source material with water and then diluting the resulting protein solution with water. This dilution forms a dispersion of protein aggregates, which are settled from the dispersion.
Therefore, there exists a need to research in the area of Amla seed and its usages. The subject imposed on the present invention therefore consists in providing a protein

composition of amla seed, its method of isolation and usage thereof in food, pharmaceutical and cosmetic preparations.
Summary of the invention
According to an embodiment of the invention, there is provided a protein composition derived from Amla (Embelica officinalis) seed, which has immense practical utility as a material for foodstuffs. The isolated Amla protein fits safe application as material for preparation of cosmetic products, pharmaceuticals and foodstuffs.
According an embodiment of the present invention there is provided a process for isolation of protein from Amla seeds. The isolated proteins from amla seed are useful for preparing protein liquid hydrolysate and protein hydrolysate in powder form.
According to another embodiment of the present invention the process for isolation of protein from amla seeds comprises the step of first breaking the seed and then extracting the proteins from the same. The broken seed contains large percentile of water and small percentile of solids and these solids contain appreciable amounts of proteins. The step of extracting or digesting the protein from the broken seed material is carried by subjecting the inside material of seed to digestion with a suitable enzyme, which method comprises the sequence provided here below:
- .Raising the moisture of the seed substance to a level of 25-85%, preferably between 45-65%, forming the slurry with a moisture laden seed materials with demineralised water
- Maintaining the temperature of the slurry between 45 to 65° C
- Maintaining the pH of the system from 7.5 to 11

- The solubilisation of proteins in the alkaline environment, by enzymatic activity of enzymes like alkaline proteases, Papain pancreatin etc, either alone or in combination thereof and in varying proportions thereof said proportions ranging from 0.1 to 99 %
- The alpha amino proteins levels are arrived from 10 to 65%, of the total proteins.

- Preferably one or more preservatives are added, such as sodium salt of methyl, ethyl and or propyl parabens, either alone or in combination thereof in varying proportions ranging from 0.5 to 99 %.
- Sodium salt of benzoate is also added from 0.5 to 2.5 % of the total system
According to yet another embodiment the protein solution may be concentrated by a suitable evaporation technique to a desired strength, and filtered by a suitable filtering technique described below:
Filtration of the hydrolysed system is done after allowing the system to rest overnight. Filtration if done in two steps; first the crude filtration to remove the coarser materials and finally through sparkler filters or similar filtration assemblies like Nutche filter, plate and Frame filters, leaf Filters, Continuous ejection type high- speed Centrifuges etc either alone or in combination thereof, to result in a sparkling Clear filtrate (Brix 5-18%). Concentration through a suitable concentration method is done to result in a clear solution of 20-30%.
According to another embodiment the Clear filtrate of 5-18% brix is concentrated to desired level of solids, through a suitable evaporation method such as either Falling film evaporator, or a triple effect evaporator or lypholiser, or Vacuum evaporator or by direct heating at low temperature etc, either alone or in various combinations thereof to result to a concentration of 30-45 %.
According to another embodiment of the present invention, the protein hydrolysate of amla seed protein thus obtained contains total solids 20-30 %, total protein 4.8 to 7.8 %. Alpha amino protein 10 - 50 % of the total proteins, specific gravity 1.07 to 1.12 and pH is 5-6.
According to yet another embodiment of the invention the filtered and concentrated protein mass obtained in the above step is subjected to drying. The drying may be carried out using a suitable drying method such as Spray drying, Foam mat drying, Drum Drying,

Vacuum Drying, tray drying etc, either alone or in combination of the above methods in varying proportions.
According to another embodiment of the invention the amla seed protein powder obtained after the drying contains 1.62 to 2.32 % nitrogen, 10.13 to 13.95 % protein. Whereas bulk density of the powder is 0.65 to 0.90.
In accordance with yet another embodiment of the invention the amla seed proteins can be utilized in preparation of compositions such as that of food, cosmetics, pharmaceuticals, nutraceutical and beverages.
According to yet another embodiment of the invention there is provided an economically feasible method of extraction of highly digested proteins in the aqueous phase from the seeds of amla.
According to yet another embodiment of the invention there is provided a process for extraction of amla seed proteins using enzymatic hydrolysis thereby allowing complete control of the minimum development of adverse taste and colour. The process can be controlled such that it can be stopped at a desirable level of hydrolysis.
Detailed description of the invention:
The present invention provides for an economically feasible method of extraction of highly digested proteins in the aqueous phase from the seeds of amla.
Although the process is described with respect to the extraction of proteins from Amla seed, from cultivated and wild varieties of Amla. depending upon the climatic, locational, seasonal, water quality, air quality etc, and a various combination of them ranging from 0.5 to 99% of their combinations, this is equally viable for seeds of other herbs as well.

This process is based on enzymatic hydrolysis of Amla seed proteins allowing complete control of the minimum development of adverse taste and colour. The process can be controlled such that it can be stopped at a desirable level of hydrolysis.
The steps involved in the process are described as under:
Enzymatic hydrolysis:
During Hydrolysis the incorporation of water molecules to the substance reduces molecular size of the substance.
Hydrolysis can take place through alkaline, acidic, and /or enzymatic route.
While in hydrolyzing the protein substance with acidic or alkaline route, it is difficult to control the hydrolysis, which results in development of unpleasant odour, dark colour and degradation /denaturation of proteins, it is easy to control hydrolysis avoiding un-necessary complications mentioned above, with Enzymatic Hydrolysis.
While Enzymatic hydrolysis results in high quality of Hydrolysate (Hydrolysed product), it is a very sensitive method and must be handled skillfully:
• Selection of appropriate enzyme /enzyme - mix.
• Selection of proper pH.
• Selection of proper temperature range.
• Working for particular time duration, depending upon the progress of the hydrolysis.
• The degree of hydrolysis depends upon the end use, it is therefore very important to understand the selection of enzymes and other working conditions according to end use.

» Different enzymes work at different pH range, it is therefore very important to select working conditions of enzymes before selecting enzyme/enzymes -mix.
• Other parameters like temperature, time, concentration of enzymes, and water quantity in the system; water quality (hard water may impact the working of enzymes) are very important factors
• If selection of enzymes and other working parameters are understood correctly, the result will deliver a quality product with desirable result (devoid of un-necessary by-product/s).
• The term Amla is used in specification means a particular matter, slurry, paste of one or more variety of Amla and seeds of other herbal substances.
• According to this invention there is provided a process for extraction of proteins from seed of Amla to make aqueous base proteins, comprising of following steps:
Liquid hydrolysate:
1. Raising the moisture of the seed substance to a level 25-85%, preferably between 45-65%, forming the slurry with a moisture laden seed materials with Demineralised water.
2. Maintaining the temperature of the slurry between 45 to 65 °C.
3. Maintaining the pH of the system from 7.5 to 11.
4. The solubilisation of proteins in the alkaline environment, by enzymes like alkaline proteases (AP), Papain (PAP) & Pancreatin (PAN) etc, either alone or in combination of them in varying proportion ranging from 0.1 to 99 %.
5. The alpha amino proteins levels are arrives from 10 to 65%, of the total proteins.

6. Termination of enzymatic activity is done either by heating the system to 70 -85 °C. to a particular time period ranging from 0.5 to 2.5 hrs, in the mild acidic conditions. A chelating agent like EDTA is added to sediment the denatured enzyme.
7. Preferably one or more preservatives, such as sodium salt of methyl, ethyl and or propyl parabens, either alone or in combination of them in varying proportions ranging from 0.5 to 99 %.
Sodium salt of benzoate is also added from 0.5 to 2.5 % of the total system.
8. Filtration of the hydrolysed system is done, after allowing the system to rest overnight.
Filtration is done in two steps; first the crude filtration to remove the coarser materials and
finally through sparkler filters or similar filtration assemblies like Nutche filter, plate and
Frame filters, leaf Filters, Continuous ejection type high- speed Centrifuges etc either
alone or in combination of them, to result a sparkling Clear filtrate (Brix 5-18%).
Concentration through a suitable concentration method is done to result the clear solution
of 20-30%.
Protein hydrolysate of Amla seed protein thus obtained has following specifications:
Colour : Brownish - green clear liquid
Clarity : sparkling clear
pH : 5-6
Solids : 20-30%
Total nitrogen : 0.76-1.25%
Total Protein : 4.8-7.8%
Alpha amino protein : 10-50%) of the total proteins.
Specific Gravity : 1.07-1.12
Microbial count : within limits (absence of pathogens)

Powder hydrolysate:
1.Clear filtrate of 5-18% brix is concentrated to desired level of solids, through either Falling film evaporator, or a triple effect evaporator or lypholiser, or Vacuum evaporator or by direct heating at low temperature etc, either alone or in various combination of them to result to a concentration of 30-45 %.
2. Concentrate obtained in the above step is subjected to preferably spray drying . The drying can also be done by Foam mat drying, Drum Drying, Vacuum Drying, tray drying etc, either alone or in combination of the above methods in varying proportions.
The powder thus obtained by spray drying has following specifications:
Colour : Off White to light Greenish -brown
Free flowing characteristics: Free flowing powder
Bulk Density : 0.65 -0.90
Solubility, in water : Freely soluble in water
Nitrogen Content : 1.62-2.32%
Protein content : 10.13-13.95%
Microbial content : Within limits, absence of pathogens.
The following examples shall only illustrate some of the embodiments of the invention
Example 1:
12 kgs of Cultivated Amla seed powder was soaked in 100 litre of Demineralised water, having conductivity of 140. 15-50 g of sodium metabisulphite (SMB) was added at moderate temperature Of 40 to 65°C.
The system was allowed to stay over at room temperature.

Next morning 200-500 g of sodium hydroxide (SHO) solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 10.5.
Suitable enzyme Alkaline Proteases (AP). Papain (PAP) and Pancreatin (PAN) were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-500 rpm for 1-4 hrs.
Temperature of the system was reduced to 45-65 °C and pH of the system was lowered to 8.5 with help of mild acid like citric acid or mineral acid like hydrochloric acid or sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to 99%.
Enzymes like Papain, pancreatin or Neutrase were added either alone or in combination of them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate (STS) was added from 20 -150 g, along with the enzymes
Temperature of the slurry was lowered to 45-65°C, the system was continued with above conditions for 12-60 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-85°C or by adding the metal chelating agent, EDTA or a combination of them.

Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-600 mesh.
20-60 g each of sodium methyl paraben (SMP), sodium propyl paraben (SPP) and sodium benzoate (SB) are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate.
Clear filtrate of about 7% brix obtained was concentrated to about 15-30% brix, through a suitable concentrate device, has following specification:
Colour : Brownish - green clear liquid
Clarity : sparkling clear
pH : 5-6
Solids : 15-30%
Total nitrogen : 0.58 -1.25%
Total Protein : 3.63-7.8%
Alpha amino protein : 10-50% of the total proteins.
Specific Gravity : 1.06-1.12
Microbial count : within limits (absence of pathogens)
Example 2:
8 kg. Wild Amla seed powder and 5 kg. of Cultivated Amla seed powder were soaked in 110 liters of Demineralised water, having conductivity of 140. 20-60 g. of sodium metabisulphite was added at moderate temperature Of 40 to 68°C.
The system was allowed to stay over at room temperature.

Next morning 200-600 g of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 11.0.
Suitable enzyme Alkaline Proteases, Papain were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-500 rpm for 1-4 hrs.
Temperature of the system was reduced to 45-65 °C and Ph of the system was lowered to 8.5 with help of mild acid like citric acid or mineral acid like hydrochloric acid or sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to 99%.
Enzymes like Papain, pancreatin or Neutrase were added either alone or in combination of them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate was added from 20 -160 g, along with the enzymes
Temperature of the slurry was lowered to 45-68°C, the system was continued with above conditions for 12-55 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-85°C or by adding the metal chelating agent, EDTA or a combination of them.

Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300 mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-600 meshes.
20-60 g each of sodium methyl paraben, sodium propyl paraben and sodium benzoate are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate.
Clear filtrate of about 7% brix obtained, was concentrated to about 15-30% brix, through a suitable concentrate device, has following specification:
• Colour : Brownish - green clear liquid
• Clarity : sparkling clear
• pH : 5-6
• Solids : 17-30%
• Total nitrogen : 0.60-1.25%
• Total Protein : 3.75-7.8%
• Alpha amino protein : 10-50% of the total proteins.
• Specific Gravity : 1.065-1.12
• Microbial count : within limits (absence of pathogens)
Example 3:
6 kgs Wild Amla seed powder and 6 kgs of Cultivated Amla seed powder were soaked in 100 liters of Demineralised water, having conductivity of 140. 18-60 g of sodium metabisulphite was added at moderate temperature Of 40 to 70 °C.
The system was allowed to stay over at room temperature.

Next morning 180-600 g of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 10.8.
Suitable enzyme Alkaline Proteases, Pancreatin were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-500 rpm for 1-4 hrs.
Temperature of the system was reduced to 45-65 °C and pH of the system was lowered to
8.0 with help of mild acid like citric acid or mineral acid like hydrochloric acid or
sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to
99%.
Enzymes like Papain, pancreatin or Neutrase were added either alone or in combination of
them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate was added from 20-165 g, along with the enzymes
Temperature of the slurry was lowered to 45-68°C, the system was continued with above conditions for 12-50 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-90 °C or by adding the metal chelating agent, EDTA or a combination of them.
Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300 mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-500 meshes.

20-60 g each of sodium methyl paraben. sodium propyl paraben and sodium benzoate are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate.
Clear filtrate of about 8 % brix obtained, was concentrated to about 20-30% brix, through a suitable concentrate device, has following specification:
• Colour : Brownish - green clear liquid
• Clarity : Sparkling clear
• pH : 5-6
• Solids : 20-30%
• Total nitrogen : 0.65-1.25%
• Total Protein : 4.06-7.8%
• Alpha amino protein : 10-50% of the total proteins.
• Specific Gravity : 1.065-1.12
• Microbial count : Within limits (absence of pathogens)
Example 4:
4 kgs Wild Amla seed powder and 8kgs of Cultivated Amla seed powder were soaked in 110 litre of demineralised water, having conductivity of 140. 20-60 g of sodium metabisulphite was added at moderate temperature Of 40 to 68°C. The system was allowed to stay over at room temperature.
Next morning 200-600 g of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 11.0.

Suitable enzyme Alkaline Proteases, Papain were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-500 rpm for 1 -4 hrs.
Temperature of the system was reduced to 45-65 °C and Ph of the system was lowered to 8.5 with help of mild acid like citric acid or mineral acid like hydrochloric acid or sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to 99%.
Enzymes like Papain, pancreatin or Neutrase were added either alone or in combination of them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate was added from 20 -160 g, along with the enzymes
Temperature of the slurry was lowered to 45-68°C, the system was continued with above conditions for 12-55 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-85°C or by adding the metal chelating agent, EDTA or a combination thereof.
Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300 mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-600 mesh.
20-60 g each of sodium methyl paraben, sodium propyl paraben and sodium benzoate are added in the solution form.

The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate.
Clear filtrate of about 7% brix obtained, was concentrated to about 15-30% brix, through a suitable concentrate device, has following specification:
Colour : Brownish - green clear liquid
Clarity : sparkling clear
pH : 5-6
Solids : 17-30%
Total nitrogen : 0.62 -1.25%
Total Protein : 3.87-7.8%
Alpha amino protein : 10-50% of the total proteins.
Specific Gravity : 1.06-1.12
Microbial count : within limits (absence of pathogens)
Example 5:
12 kgs Wild Amla seed powder and 2 kgs of Cultivated Amla seed powder were soaked in 110 liters of Demineralised water, having conductivity of 140. 18-70 g of sodium metabisulphite was added at moderate temperature Of 40 to 75 °C.
The system was allowed to stay over at room temperature.
Next morning 180-600 g. of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 11.2.
Suitable enzyme Alkaline Proteases, Pancreatin were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.

System was allowed to stirrer at 100-500 rpm for 1-4 hrs.
Temperature of the system was reduced to 45-68 °C and pH of the system was lowered to 8.5 with help of mild acid like citric acid or mineral acid like hydrochloric acid or sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to 99%.
Enzymes like papain, pancreatin or Neutrase were added either alone or in combination of them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate was added from 20 -175 g, along with the enzymes
Temperature of the slurry was lowered to 45-68°C, the system was continued with above conditions for 12-50 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-85 °C or by adding the metal chelating agent, EDTA or a combination thereof
Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300 mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-600 mesh.
20-60 g each of sodium methyl paraben, sodium propyl paraben and sodium benzoate are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate.

Clear filtrate of about 8 % brix obtained, was concentrated to about 20-30% brix, through a suitable concentrate device, has following specification:
• Colour : Brownish - green clear liquid
• Clarity : Sparkling clear
• pH : 5-6
• Solids : 20-30%
• Total nitrogen : 0.68-1.25%
• Total Protein : 4.25 -7.8%
• Alpha amino protein : 10-50% of the total proteins.
• Specific Gravity : 1.07-1.12
• Microbial count : Within limits (absence of pathogens)
Example 6:
2 Kgs of Wild Amla seed powder and 11 kgs of Cultivated Amla seed powder were soaked in 110 liters of Demineralised water, having conductivity of 140. 20-60 g of sodium metabisulphite was added at moderate temperature Of 40 to 68°C.
The system was allowed to stay over at room temperature.
Next morning 200-600 g of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80 C, the pH of the slurry was in between 7.5 to 10.5.
Suitable enzyme Alkaline Proteases, Pancreatin were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-500 rpm for 1-4 hrs.

Temperature of the system was reduced to 45-65°C and pH of the system was lowered to 8.5 with help of mild acid like citric acid or mineral acid like hydrochloric acid or sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to 99%.
Enzymes like papain, pancreatin or Neutrase were added either alone or in combination of them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate was added from 20 -160 g, along with the enzymes
Temperature of the slurry was lowered to 45-68°C, the system was continued with above conditions for 12-55 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-80 °C or by adding the metal chelating agent, EDTA or a combination of them.
Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-600 meshes.
20-60 g each of sodium methyl paraben, sodium propyl paraben and sodium benzoate are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate.
Clear filtrate of about 7% brix obtained, was concentrated to about 18-30% brix, through a suitable concentrate device has following specification:

Colour : Brownish green clear liquid
Clarity : sparkling clear
pH : 5-6
Solids : 19-30%
Total nitrogen : 0.65 -1.25%
Total Protein : 4.06-7.8%
Alpha amino protein : 10-50% of the total proteins.
Specific Gravity : 1.07-1.12
Microbial count : within limits (absence of pathogens)
Example 7:
12 kgs of Cultivated Amla seed powder and 1 kg of Wild Amla seed powder were soaked in 100 litre of Demineralised water, having conductivity of 140. 15-50 g of sodium metabisulphite was added at moderate temperature Of 40 to 65°C.
The system was allowed to stay over at room temperature.
Next morning 200-500 g of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 10.5.
Suitable enzyme Alkaline Proteases, papain and Pancreatin were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-600 rpm for 1-4 hrs.
Temperature of the system was reduced to 45-65 °C and pH of the system was lowered to 8.7 with help of mild acid like citric acid or mineral acid like hydrochloric acid or sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to 99%.

Enzymes like papain, pancreatin or Neutrase were added either alone or in combination of them in varying proportions of them in varying proportions from 0.5 to 99 %. Sodium thio sulphate was added from 20 -150 g, along with the enzymes.
Temperature of the slurry was lowered to 45-65°C, the system was continued with above conditions for 12-60 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-85°C or by adding the metal chelating agent, EDTA or a combination of them.
Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300 mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-600 meshes.
20-60 g each of sodium methyl paraben, sodium propyl paraben and sodium benzoate are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate. Clear filtrate of about 7 % brix obtained, was concentrated to about 15-30 % brix, through a suitable concentrate device has following specification:
• Colour : Brownish - green clear liquid
• Clarity : sparkling clear
• pH : 5-6
• Solids : 15-30%
• Total nitrogen : 0.60-1.25%

• Total Protein : 3.75-7.8%
• Alpha amino protein : 10-50% of the total proteins.
• Specific Gravity : 1.06-1.12
• Microbial count : within limits (absence of pathogens)
Example 8:
4 kgs Wild Amla seed powder and 8 kgs of Cultivated Amla seed powder were soaked in 100 liters of Demineralised water, having conductivity of 140. 18-60 g of sodium metabisulphite was added at moderate temperature Of 40 to 75 °C.
The system was allowed to stay over at room temperature.
Next morning 180-600 g of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 10.8.
Suitable enzyme Alkaline Proteases, Papain were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-500 rpm for 1-4 firs.
Temperature of the system was reduced to 45-65 °C and pH of the system was lowered to 8.0 with help of mild acid like citric acid or mineral acid like hydrochloric acid or sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to 99%.
Enzymes like papain & pancreatin were added either alone or in combination of them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate was added from 20-160 g, along with the enzymes

Temperature of the slurry was lowered to 45-68°C, the system was continued with above conditions for 12-50 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-90 °C or by adding the metal chelating agent, EDTA or a combination of them.
Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300 mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-600 mesh.
20-55 g each of sodium methyl paraben, sodium propyl paraben and sodium benzoate are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate.
Clear filtrate of about 8 % brix obtained, was concentrated to about 20-30% brix through a suitable concentrate device has following specification:
• Colour : Brownish - green clear liquid
• Clarity : Sparkling clear
• pH : 5-6 Solids : 20-30%
• Total nitrogen : 0.63 -1.25%
• Total Protein : 3.94-7.8%
• Alpha amino protein : 10-50% of the total proteins.
• Specific Gravity : 1.065-1.12

• Microbial count : Within limits (absence of pathogens)
•
Example 9:
10 kgs of Wild Amla seed powder and 3 kgs of Cultivated Amla seed powder were soaked
in 110 liters of Demineralised water, having conductivity of 140. 20-70 g of sodium
metabisulphite was added at moderate temperature Of 40 to 70 °C.
The system was allowed to stay over at room temperature.
Next morning 200-600 g of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 11.0.
Suitable enzyme Alkaline Proteases, Pancreatin were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-600 rpm for 1-4 hrs.
Temperature of the system was reduced to 45-65 °C and Ph of the system was lowered to
8.5 with help of mild acid like citric acid or mineral acid like hydrochloric acid or
sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to
99%.
Enzymes like papain, pancreatin or Neutrase were added either alone or in combination of
them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate was added from 20 -160g, along with the enzymes
Temperature of the slurry was lowered to 45-68°C, the system was continued with above conditions for 12-55 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.

When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-80 °C or by adding the metal chelating agent, EDTA or a combination of them.
Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300 mesh) then through high rpm (5,000-12,000) centrifuge with a cloth bag of about 300-600 meshes.
20-60 g each of sodium methyl paraben, sodium propyl paraben and sodium benzoate are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate. Clear filtrate of about 7% brix obtained, was concentrated to about 18-30% brix, through a suitable concentrate device, has following specification:
Colour : Brownish - green clear liquid
Clarity : sparkling clear
pH : 5-6
Solids : 19-30%
Total nitrogen : 0.64 -1.25%
Total Protein : 4.0-7.8%
Alpha amino protein : 10-50% of the total proteins.
Specific Gravity : 1.07-1.12
Microbial count : within limits (absence of pathogens)
Example 10:
2 kgs Wild Amla seed powder and 9 kgs of Cultivated Amla seed powder were soaked in 110 litre of Demineralised water, having conductivity of 140. 20-60 g of sodium metabisulphite was added at moderate temperature Of 40 to 68°C.

The system was allowed to stay over at room temperature.
Next morning 200-600 g of sodium hydroxide solution (30-50%) was added to the system with agitation, the temperature of the system was raised to 65-80°C, the pH of the slurry was in between 7.5 to 11.0.
Suitable enzyme Alkaline Proteases, papain were added either alone or in combination of them in varying proportion ranging from 0.5 to 99 %.
System was allowed to stirrer at 100-500 rpm for 1 -4 hrs.
Temperature of the system was reduced to 45-65 °C and pH of the system was lowered to 8.5 with help of mild acid like citric acid or mineral acid like hydrochloric acid or sulphuric acid alone or a combination of them in varying proportions ranging from 0.5 to 99%.
Enzymes like papain, pancreatin or Neutrase were added either alone or in combination of them in varying proportions of them in varying proportions from 0.5 to 99%.
Sodium thio- sulphate was added from 20 -160 g, along with the enzymes.
Temperature of the slurry was lowered to 45-68°C, the system was continued with above conditions for 12-55 hrs.
In the meantime periodic checks were conducted to measure the degree of hydrolysis, at different time intervals.
When the system has reached the desired degree of hydrolysis, the hydrolysis is terminated, either by heating the system to 70-80°C or by adding the metal chelating agent. EDTA or a combination of them.

Slurry is then passed through the series of filters including prefilters (electrically operated shifters having 200-300 mesh) then through high rpm (5,000-14,000) centrifuge with a cloth bag of about 300-600 meshes.
20-60 g each of sodium methyl paraben. sodium propyl paraben and sodium benzoate are added in the solution form.
The system is then finally filtered through filter pads of various mesh sizes, to result to sparkling clear filtrate.
Clear filtrate of about 7% brix obtained, was concentrated to about 20-35% brix, through a suitable concentrate device has following specification:
• Colour : Brownish - green clear liquid
• Clarity : sparkling clear
• pH : 5-6
• Solids : 20-30%
• Total nitrogen : 0.66-1.30%
• Total Protein : 4.13-8.13%
• Alpha amino protein : 10-50% of the total proteins.
• Specific Gravity : 1.06-1.12
• Microbial count : within limits (absence of pathogens)
The abbreviations used in the specification:
1. ASP: Amla Seed Proteins
2. SIB: Sodium metabisulphite
3. DMW: Demineralised water
4. SHO: Sodium Hydroxide
5. STS: Sodium thio Sulphate

6. PAP: Papain
7. PAN: Pancreatin
8. AP: Alkaline Proteases
9. SMB: Sodium Methyl Paraben
10. SPP: Sodium Propypl Paraben
11. Sodium Benzoate

We claim:
1. A process for isolation of proteins from amla (Embelica officinalis) seeds
comprising the steps of;
raising the moisture level of the seed substance to form a slurry, wherein the
moisture level of the slurry ranges from 25 to 85 % ;
maintaining the pH of slurry in the range of 7.5 to 11;
solubulizing proteins in alkaline environment by a combination of alkaline
proteases in varying proportion ranging from 0.1 to 99% to obtain liquid protein
hydrolysate
said hydrolysate is then filtered, concentrated and dried to obtain isolated amla
seed protein in powder form.
2. A process as claimed in claim 1 wherein the temperature of the slurry is maintained between 45 to 65°C.
3. A process as claimed in claim 1 to 2 wherein the alkaline proteases are selected from a group of papain, pancreatin, neutrase or a combination thereof.
4. A process as claimed in claim 1 to 3 wherein the enzymatic activity of alkaline proteases is deactivated by heating the slurry to a temperature ranging from 70-85°C, for a particular time ranging from 05 to 2.5 hours in mild acidic condition.
5. A process as claimed in claim 1 to 4 wherein a chelating agent like EDTA is added to sediment the denatured enzyme thereby obtaining a liquid protein hydrolysate.
6. A process as claimed in claims 1 to 5 wherein one or more preservatives such as sodium salt or methyl, ethyl and propyl parabens either alone or in combination thereof in varying proportions ranging from 0.5 to 99%.

7. A process as claimed in claim 1 to 6 wherein sodium salt is optionally added in the range of 0.5 to 2.5 of the liquid protein hydrolysate.
8. A process as claimed in claim 1 to 7 wherein the liquid protein hydrolysate obtained is allowed to rest overnight.
9. A process as claimed in claim 8 wherein a step-wise filtration of liquid protein hydrolysate is carried out, said steps comprising removal of coarse material first and later passing the filtrate through sparkler filters or similar filtration assemblies to achieve a filtrate having a Total Soluble Solid (TSS) content of 5-18% which is further concentrated to a TSS content of 20-30%.
10. The process as claimed in claim 9 wherein the filtration assembly used may be a nutche filter, plate and frame filter, leaf filter, continuous ejection type high speed centrifuge wherein the filtration assemblies are used either alone or in combination thereof.
11. A process as claimed in claim 9 and 10 wherein the filtered protein hydrolysate is further concentrated to get a TSS content ranging between 30-45 %.
12. The process as claimed in claim I 1 wherein the concentration is carried out either in a falling film evaporator or triple effect evaporator or lypholizer or direct vacuum evaporator or by direct heating at low tempaerature either alone or in combination thereof
13. A process as claimed in claim 11 and 12 wherein the concentrated protein hydrolysate is dried to obtain a powder.
14. The process as claimed in claim 13 wherein the drying is either spray drying or foam mat drying or drum drying or vacuum drying or tray drying either alone or in combination thereof.

15. The product obtained from claims 1 to 10 has total nitrogen 0.76 to 1.25%, total proteins 4.8 - 7.8 %, alpha amino acid 10-50%, pH 5-6, total solids 20-30 % and having a specific gravity of 1.07-1.12.
16. The powder obtained from claim 13 and 14 has a bulk density of 0.65 - 0.90, nitrogen content is 1.62-2.32 %, protein content-10.13 to 13.95 %.
17. The product obtained from claims 1 to 16 wherein said product is utilized in the preparation of compositions such as food, cosmetics, pharmaceuticals, nutraceutical, and beverages.