CN102174071A - Method for extracting and preparing protein powder and enzymolysis product thereof from almond meal - Google Patents
Method for extracting and preparing protein powder and enzymolysis product thereof from almond meal Download PDFInfo
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- CN102174071A CN102174071A CN2011100051283A CN201110005128A CN102174071A CN 102174071 A CN102174071 A CN 102174071A CN 2011100051283 A CN2011100051283 A CN 2011100051283A CN 201110005128 A CN201110005128 A CN 201110005128A CN 102174071 A CN102174071 A CN 102174071A
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Abstract
The invention discloses a method for continuously extracting and preparing concentrated protein, separated protein and proteolysis products from almond meal. The method comprises the following steps of: degreasing and debitterizing residual oil meal obtained after almond oil is mechanically and coldly pressed, and then extracting with a low-concentration NaCl phosphate buffer solution to obtain the concentrated protein; preparing almond separated protein from the concentrated protein solution by applying an ultrafiltration membrane technology; preparing a proteolysis-almond polypeptide mixed solution from the separated protein solution by applying an enzymolysis technology; and ultrafiltering the polypeptide mixed solution by applying a membrane technology to obtain multifunctional active almond peptide with different class level molecular weights in sequence. The method has the advantages of simple process, strong operability and easiness in control of extracting conditions and can finish the extraction and preparation of the almond concentrated protein, the separated protein and the proteolysis products in sequence; and the obtained products have high purity and wide application and can be directly used as food additives, nutrition reinforcers, health care products, medicaments, and the like needed by the protein foods and different processing foods.
Description
Technical field
The present invention relates to the almond deep process technology, be specifically related to a kind of method of from the almond dregs of rice, extracting and prepare protein concentrate, protein isolate and proteolysis product continuously.
Background technology
Protein content reaches about 50% in the almond dregs of rice after the almond oil expression.Wherein contain 18 seed amino acids and 8 kinds of essential amino acids of needed by human body, the amino acid composition is reasonable, approaches international reference model, so almond protein is the high vegetable-protein of a kind of edibleness.Plant protein products mainly is divided into apolipoprotein (purity＜60%), protein concentrate (purity＞70%) and protein isolate (purity＞90%) in the world at present, has different functional performances respectively, adds in the food as functional ingredient.Different with functional requirement according to different foods to adding protein content, can be respectively applied in the processing of cereal product, meat product, milk-product.Every functional performance of proteolysis product further improves, and has different physiologically actives, can be developed as medicines and health protection, cosmetics etc.
Composition as food, the application of vegetable-protein relies on its functional performance, but proteinic functional, nutritive value and other biological are learned to tire and all are subjected to the influence of production technique, the protein concentrate, the protein isolate denaturation degrees that use conventional methods (as the alkali extraction and acid precipitation method) production are big, functional bad, limited in Application in Food.
Summary of the invention
With not enough, the purpose of this invention is to provide a kind of method of from the almond dregs of rice, extracting and prepare protein concentrate, protein isolate and proteolysis product continuously at above-mentioned problems of the prior art.This method has guaranteed the functional and biological value of protein concentrate and protein isolate, has improved the activity recovery of protein concentrate, protein isolate, provides guarantee for follow-up enzymolysis protein prepares the multifunction activity peptide.
The technical scheme that realizes the foregoing invention purpose is a kind of method of extracting concentrated protein powder from the almond dregs of rice, specifically comprises the following steps:
1) the cold press almond dregs of rice are pulverized after, with 75 ℃ of following lixiviates of sherwood oil of 3～5 times of amounts 2 times, each 30 minutes, carry out skimming treatment, vacuum-drying obtains degreasing dregs of rice powder;
2) with 95% ethanolic soln of degreasing dregs of rice powder with 3～5 times of amounts, lixiviate 2 times, each 110 minutes or doubly measure 50～60 ℃ warm water soaking 5～7 days with 3-5, change water every day 1～2 time, to carry out debitterize and handle, vacuum-drying obtains degreasing debitterize dregs of rice powder;
3) degreasing debitterize dregs of rice powder is added the NaCl phosphate buffer soln of 0.05～0.2mol/L of 25～45 times of amounts, 30～55 ℃ of following lixiviates 3 times, each 30～60min merges No. 3 extracting solutions and obtains almond protein concentrate solution, and the lyophilize of almond protein concentrate solution is obtained the almond concentrated protein powder.
A further object of the invention provide a kind of from the almond dregs of rice method of extraction separation protein powder, it is characterized in that, comprise the following steps:
1) the cold press almond dregs of rice are pulverized after, with 75 ℃ of following lixiviates of sherwood oil of 3～5 times of amounts 2 times, each 30 minutes, carry out skimming treatment, vacuum-drying obtains degreasing dregs of rice powder;
2) with 95% ethanolic soln of degreasing dregs of rice powder with 3～5 times of amounts, lixiviate 2 times, each 110 minutes or doubly measure 50～60 ℃ warm water soaking 5～7 days with 3-5, change water every day 1～2 time, to carry out debitterize and handle, vacuum-drying obtains degreasing debitterize dregs of rice powder;
3) degreasing debitterize dregs of rice powder is added the NaCl phosphate buffer soln of 0.05～0.2mol/L of 25～45 times of amounts, 30～55 ℃ of following lixiviates 3 times, each 30～60min merges No. 3 extracting solutions and obtains almond protein concentrate solution, and the lyophilize of almond protein concentrate solution is obtained the almond concentrated protein powder;
4) the almond concentrated protein powder being assigned to concentration with deionized water is 4～10%, at 0.01～0.25MPa, 30～55 ℃, ultrafiltration under pH6～8 conditions, the polysulfone membrane of 50kD～80kD is held back in selection, obtain almond protein isolate solution after the ultrafiltration, the lyophilize of almond protein isolate solution is obtained the almond separated protein powder.
A further object of the invention provides a kind of preparation method who extracts protein powder enzymolysis product from the almond dregs of rice, comprises the following steps:
1) the cold press almond dregs of rice are pulverized after, with 75 ℃ of following lixiviates of sherwood oil of 3～5 times of amounts 2 times, each 30 minutes, carry out skimming treatment, vacuum-drying obtains degreasing dregs of rice powder;
2) with 95% ethanolic soln of degreasing dregs of rice powder with 3～5 times of amounts, lixiviate 2 times, each 110 minutes or doubly measure 50～60 ℃ warm water soaking 5～7 days with 3-5, change water every day 1～2 time, to carry out debitterize and handle, vacuum-drying obtains degreasing debitterize dregs of rice powder;
3) degreasing debitterize dregs of rice powder is added the NaCl phosphate buffer soln of 0.05～0.2mol/L of 25～45 times of amounts, 30～55 ℃ of following lixiviates 3 times, each 30～60min, merge No. 3 extracting solutions and obtain almond protein concentrate solution, the lyophilize of almond protein concentrate solution is obtained the almond concentrated protein powder, and wherein protein content is＞70%;
4) the almond concentrated protein powder being assigned to concentration with deionized water is 4-10%, at 0.01～0.25MPa, 30～55 ℃, ultrafiltration under pH6～8 conditions, the polysulfone membrane of 50kD～80kD is held back in selection, obtain almond protein isolate solution after the ultrafiltration, the lyophilize of almond protein isolate solution is obtained the almond separated protein powder;
5) the almond separated protein powder being assigned to concentration with deionized water is 4～10%, at 40～60 ℃ of temperature, pH 6～9, stirs the proteolytic enzyme that adds 20～60mg/g almond separated protein powder down, stirs enzymolysis 120～420min down, obtains the almond protein enzymolysis solution;
6) pH with the almond protein enzymolysis solution transfers to 6～7, and is warming up to 90～100 ℃ rapidly, and the enzyme 10～15min that goes out is cooled to 20～30 ℃ of after-filtration;
7) with filtrate at 0.01～0.25MPa, 30～55 ℃, ultrafiltration under pH 6～7 conditions, the polysulfone membrane of 1kD～10kD is held back in selection, after the ultrafiltration successively collection＜1kD, 1KD～2KD, 2KD～3KD, 3KD～4KD, 4KD～5KD, 5KD～6KD, 6KD～7KD, 7KD～8KD, 8KD～9KD, 9KD～10KD,＞the almond peptide concentrated solution of 10KD;
8) lyophilize of various almond peptide concentrated solution is obtained the almond peptide powder of different purposes.
Compared with prior art, the method for extracting from the almond dregs of rice and preparing protein powder and enzymolysis product thereof of the present invention has the following advantages:
1) mild conditions such as the extraction that the inventive method adopted, separation, having guaranteed the activity of protein concentrate, protein isolate provides guarantee for follow-up enzymolysis protein prepares the multifunction activity peptide.
2) protein concentrate, protein isolate and the proteolysis product that adopts the inventive method to prepare can satisfy different foods to adding the different of protein content and functional requirement, can be used as the protein product of food, medicine, the needed different purposes of cosmetic industry, for the comprehensive utilization of the almond dregs of rice provides a new approach, and can improve the added value of the almond dregs of rice.
Embodiment
The present invention is described in further detail below in conjunction with specific examples:
Embodiment 1
The NaCl phosphate buffer soln (pH=7) that almond dregs of rice powder 1000g after the degreasing detoxification is added 30kg 0.08mol/L, 50 ℃ of following lixiviates 3 times, each 40min merges No. 3 extracting solutions and obtains almond protein concentrate solution, continues lyophilize and can obtain the almond concentrated protein powder; It is 5% that the almond concentrated protein powder is assigned to concentration with deionized water, selects to hold back the polysulfone membrane of 60kD, and at 0.012MPa, 40 ℃, almond protein isolate solution is collected in ultrafiltration under the pH7 condition, and lyophilize can obtain the almond separated protein powder; It is 5% that the almond separated protein powder is assigned to concentration with deionized water, 55 ℃ of temperature, pH 8, stir the Alcalase proteolytic enzyme that adds 50mg/g almond separated protein powder down, stir enzymolysis 300min down, obtain the almond protein enzymolysis solution, pH transfers to 7 with enzymolysis solution, and be warmed up to 100 ℃ rapidly, enzyme 15min goes out, be cooled to 25 ℃ of after-filtration, with filtrate at 0.012MPa, 40 ℃, ultrafiltration under the pH7 condition, the polysulfone membrane of 1kD～10kD is held back in selection, collection＜1kD successively after the ultrafiltration, 1KD～2KD, 2KD～3KD, 3KD～4KD, 4KD～5KD, 5KD～6KD, 6KD～7KD, 7KD～8KD, 8KD～9KD, 9KD～10KD, the almond peptide concentrated solution of＞10KD, lyophilize obtain the almond peptide powder of different purposes.
Embodiment 2
The NaCl phosphate buffer soln (pH=7) that almond dregs of rice powder 1000g after the degreasing detoxification is added the 0.05mol/L of 25kg, 45 ℃ of following lixiviates 3 times, each 30min, merge No. 3 extracting solutions and obtain almond protein concentrate solution, continue lyophilize and can obtain the almond concentrated protein powder, wherein protein content is＞70%;
It is 4% that the almond concentrated protein powder is assigned to concentration with deionized water, selects to hold back the polysulfone membrane of 50kD～80kD, at 0.01MPa, 30 ℃, almond protein isolate solution is collected in ultrafiltration under the pH6 condition, lyophilize can obtain the almond separated protein powder, and wherein protein content is＞90%; It is 4% that the almond separated protein powder is assigned to concentration with deionized water, 40 ℃ of temperature, pH 6.5, stir the Protamex proteolytic enzyme that adds 30mg/g almond separated protein powder down, stir enzymolysis 120min down, obtain the almond protein enzymolysis solution, pH transfers to 6 with enzymolysis solution, and be warmed up to 90 ℃ rapidly, enzyme 10min goes out, be cooled to 20 ℃ of after-filtration, with filtrate at 0.01MPa, 30 ℃, ultrafiltration under pH 6 conditions, the polysulfone membrane of 1kD～10kD is held back in selection, collection＜1kD successively after the ultrafiltration, 1KD～2KD, 2KD～3KD, 3KD～4KD, 4KD～5KD, 5KD～6KD, 6KD～7KD, 7KD～8KD, 8KD～9KD, 9KD～10KD, the almond peptide concentrated solution of＞10KD, lyophilize obtain the almond peptide powder of different purposes.
Embodiment 3
The NaCl phosphate buffer soln (pH=7) that almond dregs of rice powder 1000g after the degreasing detoxification is added 35kg 0.1mol/L, 55 ℃ of following lixiviates 3 times, each 50min merges No. 3 extracting solutions and obtains almond protein concentrate solution, continues lyophilize and can obtain the almond concentrated protein powder; It is 6% that the almond concentrated protein powder is assigned to concentration with deionized water, selects to hold back the polysulfone membrane of 70kD, and at 0.015MPa, 40 ℃, almond protein isolate solution is collected in ultrafiltration under the pH7 condition, and lyophilize can obtain the almond separated protein powder; It is 6% that the almond separated protein powder is assigned to concentration with deionized water, 50 ℃ of temperature, pH 6.5, stir the papoid that adds 20mg/g almond separated protein powder down, stir enzymolysis 200min down, obtain the almond protein enzymolysis solution, pH transfers to 7 with enzymolysis solution, and be warmed up to 100 ℃ rapidly, enzyme 12min goes out, be cooled to 25 ℃ of after-filtration, with filtrate at 0.015MPa, 30 ℃, ultrafiltration under the pH7 condition, the polysulfone membrane of 1kD～10kD is held back in selection, collection＜1kD successively after the ultrafiltration, 1KD～2KD, 2KD～3KD, 3KD～4KD, 4KD～5KD, 5KD～6KD, 6KD～7KD, 7KD～8KD, 8KD～9KD, 9KD～10KD, the almond peptide concentrated solution of＞10KD, lyophilize obtain the almond peptide powder of different purposes.
Embodiment 4:
The NaCl phosphate buffer soln (pH=7) that almond dregs of rice powder 1000g after the degreasing detoxification is added 40kg 0.15mol/L, 50 ℃ of following lixiviates 3 times, each 60min merges No. 3 extracting solutions and obtains almond protein concentrate solution, continues vacuum-drying and can obtain the almond concentrated protein powder; It is 8% that the almond concentrated protein powder is assigned to concentration with deionized water, selects to hold back the polysulfone membrane of 80kD, and at 0.02MPa, 30 ℃, almond protein isolate solution is collected in ultrafiltration under the pH7 condition, and vacuum-drying can obtain the almond separated protein powder; It is 5% that the almond separated protein powder is assigned to concentration with deionized water, 50 ℃ of temperature, pH 7, stir the compound protease that adds enzyme 30mg/g almond separated protein powder down, stir enzymolysis 360min down, obtain the almond protein enzymolysis solution, pH transfers to 7 with enzymolysis solution, and be warmed up to 90 ℃ rapidly, enzyme 15min goes out, be cooled to 25 ℃ of after-filtration, with filtrate at 0.02MPa, 30 ℃, ultrafiltration under the pH7 condition, the polysulfone membrane of 1kD～10kD is held back in selection, collection＜1kD successively after the ultrafiltration, 1KD～2KD, 2KD～3KD, 3KD～4KD, 4KD～5KD, 5KD～6KD, 6KD～7KD, 7KD～8KD, 8KD～9KD, 9KD～10KD, the almond peptide concentrated solution of＞10KD, lyophilize obtain the almond peptide powder of different purposes.
Embodiment 5
The NaCl phosphate buffer soln (pH=7) that almond dregs of rice powder 1000g after the degreasing detoxification is added 45kg 0.2mol/L, 55 ℃ of following lixiviates 3 times, each 50min merges No. 3 extracting solutions and obtains almond protein concentrate solution, continues vacuum-drying and can obtain the almond concentrated protein powder; It is 10% that the almond concentrated protein powder is assigned to concentration with deionized water, selects to hold back the polysulfone membrane of 80kD, and at 0.025MPa, 30 ℃, almond protein isolate solution is collected in ultrafiltration under the pH7 condition, and vacuum-drying can obtain the almond separated protein powder; It is 3% that the almond separated protein powder is assigned to concentration with deionized water, 45 ℃ of temperature, pH 7, stir the neutral protease that adds 60mg/g almond separated protein powder down, stir enzymolysis 180min down, obtain the almond protein enzymolysis solution, pH transfers to 7 with enzymolysis solution, and be warmed up to 100 ℃ rapidly, enzyme 10min goes out, be cooled to 25 ℃ of after-filtration, with filtrate at 0.025MPa, 30 ℃, ultrafiltration under the pH7 condition, the polysulfone membrane of 1kD～10kD is held back in selection, collection＜1kD successively after the ultrafiltration, 1KD～2KD, 2KD～3KD, 3KD～4KD, 4KD～5KD, 5KD～6KD, 6KD～7KD, 7KD～8KD, 8KD～9KD, 9KD～10KD, the almond peptide concentrated solution of＞10KD, lyophilize obtain the almond peptide powder of different purposes.
Claims (3)
1. a method of extracting concentrated protein powder from the almond dregs of rice is characterized in that, comprises the following steps:
1) the cold press almond dregs of rice are pulverized after, with 75 ℃ of following lixiviates of sherwood oil of 3～5 times of amounts 2 times, each 30 minutes, carry out skimming treatment, vacuum-drying obtains degreasing dregs of rice powder;
2) with 95% ethanolic soln of degreasing dregs of rice powder with 3～5 times of amounts, lixiviate 2 times, each 110 minutes or doubly measure 50～60 ℃ warm water soaking 5～7 days with 3-5, change water every day 1～2 time, to carry out debitterize and handle, vacuum-drying obtains degreasing debitterize dregs of rice powder;
3) degreasing debitterize dregs of rice powder is added the NaCl phosphate buffer soln of 0.05～0.2mol/L of 15～35 times of amounts, 30～55 ℃ of following lixiviates 3 times, each 30～60min merges No. 3 extracting solutions and obtains almond protein concentrate solution, and the lyophilize of almond protein concentrate solution is obtained the almond concentrated protein powder.
2. the method for an extraction separation protein powder from the almond dregs of rice is characterized in that, comprises the following steps:
1) the cold press almond dregs of rice are pulverized after, with 75 ℃ of following lixiviates of sherwood oil of 3～5 times of amounts 2 times, each 30 minutes, carry out skimming treatment, vacuum-drying obtains degreasing dregs of rice powder;
2) with 95% ethanolic soln of degreasing dregs of rice powder with 3～5 times of amounts, lixiviate 2 times, each 110 minutes or doubly measure 50～60 ℃ warm water soaking 5～7 days with 3-5, change water every day 1～2 time, to carry out debitterize and handle, vacuum-drying obtains degreasing debitterize dregs of rice powder;
3) degreasing debitterize dregs of rice powder is added the NaCl phosphate buffer soln of 0.05～0.2mol/L of 15～35 times of amounts, 30～55 ℃ of following lixiviates 3 times, each 30～60min, merge No. 3 extracting solutions and obtain almond protein concentrate solution, the lyophilize of almond protein concentrate solution is obtained almond concentrated protein powder %;
4) the almond concentrated protein powder being assigned to concentration with deionized water is 4～10%, at 0.01～0.25MPa, 30～55 ℃, ultrafiltration under pH6～8 conditions, the polysulfone membrane of 50kD～80kD is held back in selection, obtain almond protein isolate solution after the ultrafiltration, the lyophilize of almond protein isolate solution is obtained the almond separated protein powder.
3. a preparation method who extracts protein powder enzymolysis product from the almond dregs of rice is characterized in that, comprises the following steps:
1) the cold press almond dregs of rice are pulverized after, with 75 ℃ of following lixiviates of sherwood oil of 3～5 times of amounts 2 times, each 30 minutes, carry out skimming treatment, vacuum-drying obtains degreasing dregs of rice powder;
2) with 95% ethanolic soln of degreasing dregs of rice powder with 3～5 times of amounts, lixiviate 2 times, each 110 minutes or doubly measure 50～60 ℃ warm water soaking 5～7 days with 3-5, change water every day 1～2 time, to carry out debitterize and handle, vacuum-drying obtains degreasing debitterize dregs of rice powder;
3) degreasing debitterize dregs of rice powder is added the NaCl phosphate buffer soln of 0.05～0.2mol/L of 15～35 times of amounts, 30～55 ℃ of following lixiviates 3 times, each 30～60min, merge No. 3 extracting solutions and obtain almond protein concentrate solution, the lyophilize of almond protein concentrate solution is obtained the almond concentrated protein powder, and wherein protein content is＞70%;
4) the almond concentrated protein powder being assigned to concentration with deionized water is 4-10%, at 0.01～0.25MPa, 30～55 ℃, ultrafiltration under pH6～8 conditions, the polysulfone membrane of 50kD～80kD is held back in selection, obtain almond protein isolate solution after the ultrafiltration, the lyophilize of almond protein isolate solution is obtained the almond separated protein powder;
5) the almond separated protein powder being assigned to concentration with deionized water is 4～10%, at 40～60 ℃ of temperature, pH 6～9, stirs the proteolytic enzyme that adds 20～60mg/g almond separated protein powder down, stirs enzymolysis 120～360min down, obtains the almond protein enzymolysis solution;
6) pH with the almond protein enzymolysis solution transfers to 6～7, and is warming up to 90～100 ℃ rapidly, and the enzyme 10～15min that goes out is cooled to 20～30 ℃ of after-filtration;
7) with filtrate at 0.01～0.25MPa, 30～55 ℃, ultrafiltration under pH 6～7 conditions, the polysulfone membrane of 1kD～10kD is held back in selection, after the ultrafiltration successively collection＜1kD, 1KD～2KD, 2KD～3KD, 3KD～4KD, 4KD～5KD, 5KD～6KD, 6KD～7KD, 7KD～8KD, 8KD～9KD, 9KD～10KD,＞the almond peptide concentrated solution of 10KD;
8) lyophilize of various almond peptide concentrated solution is obtained the almond peptide powder of different purposes.
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