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Extraction of proteins by a two solvent method

a technology of protein extraction and solvent, applied in the field of extracting and fractionating algal products, can solve the problems of increasing the cost of extraction of petroleum oil from the earth, increasing the cost of extraction, and reducing the efficiency of extraction process, so as to reduce reduce the cost of extraction process. the effect of removing the solvent and reducing the amount of hydrophobic solven

Inactive Publication Date: 2012-10-25
HELIAE DEVMENT
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  • Abstract
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AI Technical Summary

Benefits of technology

[0013]In some embodiments of the invention, a single solvent and water are used to extract and fractionate components present in an oleaginous material. In other embodiments, these components include, but are not limited to, proteins, polar lipids, and neutral lipids. In still other embodiments, more than one solvent is used. In still other embodiments, a mixture of solvents is used. The disclosed methods allow for separation and fractionation of the components of the wet algal biomass. Another aspect of the systems and methods described herein is the ability to accomplish preliminary refining, because the hydrophobic solvent in the system selectively separates the products further into lipids and non-lipid components. The amount of amphipathic solvent used will depend on the amount of water in the system. The amount of hydrophobic solvent is minimized in order to reduce later processing to remove the solvent.
[0014]In some embodiments, the methods and systems described herein are useful for extracting coproducts of lipids from oleaginous material. Examples of such coproducts include, without limitation, proteinaceous material, chlorophyll, and carotenoids. Embodiments of the present invention allow for the simultaneous extraction and fractionation of algal products from algal biomass in a manner that allows for the production of both fuels and nutritional products.

Problems solved by technology

Extracting petroleum oil from the earth is expensive, dangerous, and often at the expense of the environment.
Furthermore, world wide reservoirs of oil are dwindling rapidly.
Conventional physical extraction processes, such as extrusion, do not work well with algae given the thickness of the cell wall and the small size (about 2 to about 20 nm) of algal cells.
Furthermore, the large amounts of polar lipids in algal oil, as compared to the typical oil recovered from seeds, lead to refining issues.
Due to the amount of energy required to heat the algal mass to dry it sufficiently, the algal feed to biofuel process is rendered uneconomical.
These steps may not work with the existing equipment due to the single cell micrometric nature of algae.
Furthermore, algal oil is very unstable due to the presence of double bonded long chain fatty acids.
The high temperatures used in conventional extraction methods cause degradation of the oil, thereby increasing the costs of such methods.
This process is energy intensive.
The use of heat to dry and hexane to extract produces product of lower quality as this type of processing causes lipid and protein degradation.
Most disruptive methods result in emulsions, requiring an expensive cleanup process.
Non-disruptive methods provide low yields.
While sometimes used to extract algal products, milking may not work with some species of algae due to solvent toxicity and cell wall disruption.
This complication makes the development of a generic process difficult.

Method used

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  • Extraction of proteins by a two solvent method
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  • Extraction of proteins by a two solvent method

Examples

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example 1

[0227]Green microalgae Scendesmus dimorphus (SD) were cultured in outdoor panel photobioreactors. SD samples of varying lipid contents were harvested. After removal of bulk water by centrifugation, the algal samples were stored as 3-5 cm algae cakes at −80° C. until use. A pre-calculated amount of wet algal biomass (15 grams dry algae weight equivalent) and 90 mL of ethanol solvent was added into a three-neck flask equipped with condenser, mechanical stirring and a thermocouple. In one experiment, the mixture was refluxed for 10 min under microwave irradiance. In a second, the mixture was refluxed for 1 h with electronic heating. Afterwards, the mixture was cooled to room temperature and separated into a diffusate and retentate by filtration.

[0228]The total lipids of algal samples were analyzed using a chloroform-methanol-water system according to Bligh and Dyer's lipid extraction method. This total lipid value was used as reference for the lipid recovery calculation. Total lipids w...

example 2

Protein Extraction from Algal Biomass

[0230](1) Acid Leaching: Algal biomass was soaked in water at pH 4.5 for 1 hour. The samples were then centrifuged at 3000 rpm for three minutes, and the supernatant removed. The remaining solids were washed 3 times with dilute acid (pH 4.5) and freeze dried.

[0231](2) Alkaline extraction: Algal biomass was soaked in water at pH 11 for 1 hour. following the addition of pH-adjusted water. The samples were then centrifuged at 3000 rpm for three minutes, and the supernatant removed. The supernatant was neutralized with acid (pH 4.5) following the centrifugation. The remaining solids were washed 3 times with dilute acid (pH 4.5) and freeze dried.

[0232]The results of acid leaching and alkaline extraction are shown below in Table 4.

TABLE 4Protein PurityProtein Yield(% weight ofProcess(% weight)protein yield)Alkaline Extraction1645Acid Leaching7032.5

[0233]Protein yield was calculated on a weight basis, comparing the weight of the freeze dried solids to t...

example 3

Extraction of Proteins from Saltwater Algal Biomass

[0234]The saltwater algal culture initially made up of about 1-10% w / w solids in saltwater was heated to 50° C. and maintained at this temperature for 1 hr. The resulting slurry was centrifuged to separate the liquid phase from the solid phase. The liquid extract was determined to be rich in globulin proteins (about 10% of the total proteins present in the original algal biomass).

[0235]The solids were then suspended in fresh water and heated to about 50° C. and maintained for about 1 hour. The resulting slurry was centrifuged again to separate the liquid from the solid phase. The liquid phase was determined to be rich in albumin proteins (about 10% of the total proteins present in the original algal biomass).

[0236]The solids were then suspended in ethanol to achieve a 70% w / w mixture. This mixture was heated to about 75° C. and maintained at that temperature for about 1 hour. The resulting slurry was centrifuged to separate the liqu...

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Abstract

A method for separating proteins from plant material, in particular, intact algal cells, using an amphipathic solvent set and a hydrophobic solvent set. Some embodiments include dewatering intact algal cells and then extracting proteins from the algal cells. The methods provide for single and multistep extraction processes which allow for efficient separation of algal proteins from a wet algal biomass. These proteins are high value products which can be used as renewable sources of food and food additives. Neutral lipids remaining in the algal biomass after extraction of proteins can be used to generate renewable fuels.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of U.S. patent application Ser. No. 13 / 286,904, filed Nov. 1, 2011, which is a continuation-in-part of U.S. patent application Ser. No. 13 / 081,197, filed Apr. 6, 2011, entitled Extraction with Fractionation of Oil and Proteinaceous Material from Oleaginous Material, now U.S. Pat. No. 8,142,659, which claims the benefit of U.S. Provisional Application No. 61 / 321,290, filed Apr. 6, 2010, entitled Extraction with Fractionation of Oil and Proteinaceous Material from Oleaginous Material, and U.S. Provisional Application No. 61 / 321,286, filed Apr. 6, 2010, entitled Extraction With Fractionation of Oil and Co-Products from Oleaginous Material, the entire contents of which are hereby incorporated by reference herein.FIELD OF THE INVENTION[0002]The invention is concerned with extracting and fractionating algal products, including, but not limited to, oils and proteins. In particular, the methods and systems discl...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K14/405
CPCC11B1/10C12N1/06C11C3/003C11B1/108
Inventor KALE, ANIKET
Owner HELIAE DEVMENT
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