Method of isolating proteins

Food or edible material: processes – compositions – and products – Fermentation processes – Of isolated seed – bean or nut – or material derived therefrom

Reexamination Certificate

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C426S044000, C426S052000, C426S656000

Reexamination Certificate

active

06284292

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to a method of isolating proteins from a proteinaceous vegetable material. More specifically the invention provides a method for isolating proteins from a proteinaceous vegetable material, which method comprises the steps of subjecting the proteinaceous vegetable material to the action of one or more carbohydrate degrading enzyme(s), thereby obtaining a mixture comprising proteins and hydrolyzed carbohydrates; and subjecting the mixture of step (i) to a separation process in order to separate proteins from the hydrolyzed carbohydrates.
BACKGROUND ART
Protein isolates are products of native, unhydrolyzed proteins, obtained by isolating proteins from a proteinaceous source, usually a proteinaceous vegetable source. Protein isolates are also referred to as protein concentrates or purified protein products. Protein isolates find various industrial utility, primarily in the food industry, e.g., for human and animal nutrition, especially products for human infants and young animals.
Methods of producing protein isolates by use of various hydrocarbon specific enzymes have been described. Thus, U.S. Pat. No. 4,478,856 describes a method for producing purified vegetable proteins, and U.S. Pat. No. 3,958,015 describes a method for concentrating soy proteins.
Protein isolates may also be produced by combining aqueous extraction and membrane isolation techniques. Such methods are described by e.g. Lawhon et al. [cf. e.g. Lawhon J T, Rhee K C & Lusas E W;
The Journal of the American Oil chemists Society
1981 58 (3) 377-384; and Lawhon J T, Manak L J, Rhee K C, Rhee K S & Lusas E W;
Journal of Food Science
1981 46 (3) 912-916+919]. Also U.S. Pat. No. 4,420,425 and U.S. Pat. No. 5,086,166 describe methods of processing oilseeds comprising solubilizing the proteins and separating the protein fraction by use of an ultrafiltration membrane.
By use of membrane isolation techniques, proteins are recovered from accompanying byproducts, in particular polysaccharides.
Methods of producing protein isolates by the combined action of carbohydrate degrading enzymes and separation techniques have never been described.
SUMMARY OF THE INVENTION
According to the invention it has now been found that the process of isolating vegetable proteins by separation techniques proceeds more efficiently and leads to products of improved quality if the vegetable proteinaceous material is subjected to the action of one or more carbohydrate degrading enzymes.
Accordingly the invention provides a method of isolating proteins from a proteinaceous vegetable material, which method comprises the steps of:
(i) subjecting the proteinaceous vegetable material to the action of one or more carbohydrate degrading enzyme(s), thereby obtaining a mixture comprising proteins and hydrolyzed carbohydrates; and
(ii) subjecting the mixture of step (i) to a separation process in order to separate the proteins from the hydrolyzed carbohydrates.
DETAILED DISCLOSURE OF THE INVENTION
The present invention provides a method of isolating proteins from a proteinaceous vegetable material, which method comprises the steps of subjecting the proteinaceous vegetable material to the action of one or more carbohydrate degrading enzyme(s), thereby obtaining a mixture comprising proteins and hydrolyzed carbohydrates, and subjecting the mixture to a separation process in order to separate the proteins from the hydrolyzed carbohydrates.
By the addition of carbohydrate degrading enzymes, the accompanying polysaccharides, which constitute a majority of the byproducts, are hydrolyzed into smaller fragments, thereby increasing differences in size between the main product (the proteins) and the byproducts. The enzyme treatment step according to the invention therefore increases the efficiency of the separation step. During the separation step, solutions having a higher dry matter content can be processed, and the quality of the products becomes improved, in particular with respect to purity and organoleptic properties, i.e. lack of undesirable flavor, odor, and color.
Protein Isolates
The product of the process of this invention is usually referred to as a protein isolate, a protein concentrate or a purified protein product. The proteins essentially are native proteins, that have not become hydrolyzed during the process, and that are not enzymatically modified proteins.
The proteins constitute more than 80% by weight of the dry matter content of the protein isolate obtained by the process of the invention, preferably more than 90% by weight.
The proteins isolated by the method of the invention are particularly useful for incorporation into food products.
Proteinaceous Vegetable Materials
The proteinaceous vegetable material subjected to the method of the invention may be any protein containing material of vegetable sources, and materials obtained therefrom. Preferably the vegetable proteinaceous material is a cereal, maize, rice, sorghum, wheat, soybean, faba bean, cowpeas, cassava, sesame, peanuts, peas, cofton, oilseed, and yams. The vegetable proteinaceous material may be derived from a vegetable source or vegetable material, e.g., by milling, crushing or grounding, such as flour, de-fatted soybean or soybean flakes.
Preferably the proteinaceous vegetable material is essentially free of fibers.
Carbohydrate Degrading Enzymes
The process of the invention comprises subjecting the proteinaceous vegetable material to the action of one or more carbohydrate degrading enzyme(s).
In a preferred embodiment one or more of the enzymes employed in the process is a glycosidase enzyme (EC 3.2).
In a more preferred embodiment one or more of the enzymes employed in the process is an amylase, in particular an &agr;-amylase or a &bgr;-amylase, an arabinanase, an arabinofuranosidase, a galactanase, an &agr;-galactosidase, a &bgr;galactosidase, a polygalacturonase, a pectin methyl esterase, a rhamnogalacturonase, a rhamnogalacturon acetyl esterase, a pectin lyase, a xylanase, a cellulase, a &bgr;-glucosidase, a cellobiohydrolase, a xylosidase, a mannanase, and/or a glucuronisidase.
In order to obtain an isolate of native proteins, the enzyme preparation should be substantially free of proteolytic enzymes, as these will degrade the protein in question, thereby turning this into a modified protein.
Microbial Sources
The glycosidase enzyme of the invention may be obtained from any known source. Preferably the glycosidase enzyme may be obtained from microbial sources, in particular from a filamentous fungus or a yeast, or from a bacteria.
In particular the amylase may be derived from a strain of Acremonium, a strain of Alcaligenes, in particular
Alcaligenes latus,
a strain of Aspergillus, in particular
Aspergillus kawachii
and
Aspergillus oryzae,
a strain of Bacillus, in particular
Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus polymyxa, Bacillus subtilis
and
Bacillus stearothermophilus,
a strain of Desulfurococcus, in particular
Desulfurococcus mucosus,
a strain of Fervidobacterium, a strain of Lactobacillus, a strain of Micrococcus, a strain of Pseudomonas, in particular
Pseudomonas amyloderamosa,
a strain of Pyrococcus, in particular
Pyrococcus furiosus
and
Pyrococcus woesei,
a strain of Pyrodictium, a strain of Sulfolobus, a strain of Staphylothermus, or a strain of Thermococcus.
The arabinanase may be derived from a strain of
Aspergillus aculeatus.
The galactanase may be derived from a strain of Aspergillus in particular
Aspergillus aculeatus,
a strain of Humicola, in particular
Humicola insolens,
a strain of Myceliophthora, in particular
Myceliophthora thermophila,
or a strain of Meripilus, in particular
Meripilus giganteus.
The galactosidase enzyme (&agr;-galactosidase or &bgr;-galactosidase) may be of bacterial origin and derived from a strain of
Escherichia coli,
or a strain of Bacillus, in particular
Bacillus stearothermophilus
and
Bacillus subtilis,
or it may be of fungal origin and derived from a strain of Aspergillus, in particular
Aspergillus aculeatu

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