CN104195204A - Method for preparing alfalfa polypeptide - Google Patents
Method for preparing alfalfa polypeptide Download PDFInfo
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- CN104195204A CN104195204A CN201410374398.5A CN201410374398A CN104195204A CN 104195204 A CN104195204 A CN 104195204A CN 201410374398 A CN201410374398 A CN 201410374398A CN 104195204 A CN104195204 A CN 104195204A
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Abstract
The invention provides a method for preparing an alfalfa polypeptide. The method comprises the following steps of (1) extracting proteins from alfalfa leaves; (2) decolorizing and deodorizing; (3) carrying out enzymolysis; and (4) carrying out spray drying to obtain the alfalfa polypeptide, wherein in the step (1), the alfalfa crude proteins are obtained by a heating method, in the step (2), the leaching, decolorizing and deodorizing are carried out with ethanol, the enzyme preparations adopted in the step (3) are Protamex enzyme and Flavourzyme1000L enzyme and the content of the alfalfa polypeptide accounts for about 75%. Polypeptide powder has a faint grass fragrance, after polypeptide powder is dissolved, the polypeptide solution has fresh smell and faint grass fragrance, is free of bitter feeling and can be directly taken.
Description
Technical field
The present invention relates to the preparation method of field of health care food one peptide species, particularly a kind of preparation method of alfalfa polypeptide.
Background technology
Green plants leaf is a kind of renewable resource maximum on the earth, its rich in proteins and various VITAMIN, and rationally the protein in utilization and developing green plant and other nutritive ingredient are important channels that solves human protein's resource scarcity.Alfalfa is as a kind of perennial leguminous plants, and in China, cultivated area is wider.Since ancient times, clover both can be used as food and has entered dish and allay one's hunger, and can be used as again pharmacological agent illness.It is more complete that alfalfa is rich in rich in protein, VITAMIN, mineral substance, amino acid ratio of components, and foreign study shows that the biological value of alfalfa leaf protein is up to 80%, can compare favourably with cow's milk and meat.Since the eighties in 20th century, the U.S., Britain, Australia and the country such as Russian just attach great importance to technical study and the product development of extracting the active substances such as Mierocrystalline cellulose, protein, cauline leaf from various tame forage grasses, clover etc. is processed at many levels and fully utilized, the products such as clover Mierocrystalline cellulose, leaf protein are produced in batch production, and as base-material, for feed industry, grocery trade and pharmaceutical sector, obtained higher economic benefit.
The extracting method of alfalfa leaf protein is mostly heating flocculence, acidic process, alkaline process processing, enzyme process, fermentation method, organic solvent method etc. at present.In patent CN102351944A, adopt extracting alfalfa leaf protein by ultrasonic-assisted enzyme process, optimized the extraction cost of alfalfa leaf protein, further refining to its quality, be applicable to suitability for industrialized production.Zhang Zesheng etc. are studied the decolouring of alfalfa leaf protein and fishy smell elimination process, adopt the optimum process condition of dehydrated alcohol lixiviate decolouring, de-raw meat to be: liquid-solid volume mass was than 6: 1; 85 ℃ of extraction temperatures; Extraction time 4 hours; 3 grades of lixiviate progression.The careless fishy smell of gained leaf protein is very light, and protein mass mark is 57.01% (food and biotechnology journal, 2006,25 (4): 16).
Prior art is all alfalfa leaf proteins, there is not yet the preparation method of clover polypeptide.
Summary of the invention
The object of the invention is to make full use of the advantage that alfalfa is rich in high protein, a kind of preparation method who is rich in nutritive effect composition, has bioactive alfalfa polypeptide is provided.The preparation method of a kind of alfalfa polypeptide proposed by the invention comprises the following steps:
(1) extraction of alfalfa leaf protein; (2) decoloration deodorization; (3) enzymolysis; (4) spraying is dry, obtains.
The extracting method of the alfalfa leaf protein described in step (1) is:
Alfalfa cauline leaf is cut into 2cm~3cm segment after draining with tap water cleaning, alfalfa young stem and leaf and water are put into tissue mashing machine's making beating and are processed, after taking-up, will in whole juice and alfalfa residue immigration colloidal mill, process, to the water that drops into 1~2 times of alfalfa weight in colloidal mill, filter to obtain alfalfa juice during this time;
By the heating of alfalfa juice, Heating temperature is 75 ℃~85 ℃, and preferably 80 ℃, insulation is processed 10min and obtained deep green floss precipitation, removes supernatant liquor after standing separation, floss is precipitated to centrifugal treating and obtain centrifugal sediment, is dried to obtain alfalfa leaf protein.
Decoloration deodorization method described in step (2) is:
Alfalfa leaf protein and dehydrated alcohol mass ratio be controlled at 1: 5~and between 1: 10, extraction temperature is below 85 ℃, and extraction time 6h~10h, discards vat liquor, obtains throw out.
Mode of action described in step (3) is to add Protamex enzyme and Flavourzyme1000L enzyme, the consumption of Protamex enzyme is 0.5 ‰ of alfalfa weight~1.5 ‰, be preferably 1.0 ‰, the consumption of Flavourzyme1000L enzyme is 1.0 ‰ of alfalfa protein weight~2.5 ‰, is preferably 2.0 ‰.
The spraying drying conditions adopting in step (4) is that inlet temperature is 180 ℃~190 ℃, and temperature of outgoing air is 65 ℃~85 ℃.
The present invention has the following advantages and beneficial effect:
(1) content of peptides reaches 70%~75% left and right, and products obtained therefrom is pale powder, is easy to dissolve.
(2) gained alfalfa polypeptide powder smell has light green grass fragrance, after dissolving smell pure and fresh, have light green grass fragrance, without bitterness sense, can directly take.
(3) operating procedure of the present invention is simple, and starting material are easy to draw materials and are cheap, and in preparation process, reaction conditions is easy to control, and production cost is relatively cheap.Two kinds of zymins selecting in experiment are easy to obtain, and are applicable to suitability for industrialized production.
Embodiment
Describe embodiments of the invention below in detail, it should be noted that, the present embodiment is narrative, is not determinate, can not limit protection scope of the present invention with this.
Embodiment 1
The extraction of alfalfa leaf protein: get initial bloom stage alfalfa (second batch~the 3rd batch) top 8cm~10cm young stem and leaf partly for processing raw material, pick removal flavescence, wilt, rot, damaged by vermin leaf and other impurity, with tap water, clean up after (2 times~3 times) drain and be cut into 2cm~3cm segment, alfalfa young stem and leaf in mass ratio: water=1: 2 put into tissue mashing machine's making beating processes 5min, after taking-up, whole juice and alfalfa residue are moved in colloidal mill, gear setting is degree of card aftertreatment 10min the most carefully, during this time to the water that drops into one times of alfalfa weight in colloidal mill, and adopt 200 order nylon filtering cloths to filter to obtain alfalfa juice, by alfalfa juice 10000ml direct heating, in heat-processed, constantly carry out mechanical stirring, when temperature rises to 75 ℃, insulation is processed 10min and is obtained deep green floss precipitation, after standing separation, remove supernatant liquor, floss is deposited in whizzer and obtains centrifugal sediment with 4000r/min centrifugal treating 10min, under normal temperature condition, seasoning obtains alfalfa leaf protein, and its protein mass mark is 65.22%.
Alfalfa protein decoloration deodorization is processed: adopt 120 mesh sieves to pulverize gained alfalfa protein, at alfalfa leaf protein and dehydrated alcohol mass ratio, it is 1: 5, at 85 ℃ of extraction temperatures, 8h is processed in lixiviate, the alfalfa leaf protein canescence of light color obtaining, protein mass mark is 64.15, and products obtained therefrom still has lighter careless fishy smell.
Enzymolysis and dry: after decoloration deodorization is processed, alfalfa protein is put into enzymatic vessel, in enzymatic vessel, the mass ratio of alfalfa protein and water is 1: 1, treat 55 ℃ of tank Nei Wenduda, under nature pH value condition, add the Protamex enzyme (Chinese Novi's letter bio tech ltd) that is equivalent to alfalfa leaf protein weight 0.5 ‰, add the Flavourzyme1000L enzyme (Chinese Novi's letter bio tech ltd) that is equivalent to alfalfa leaf protein weight 1.0 ‰ simultaneously, in temperature, it is 55 ℃, enzymolysis processing 3h under nature pH value condition, enzyme 60min goes out under 100 ℃ of high temperature, filter and collect enzymolysis solution.Use dual-effect concentrator that the enzymolysis solution of collection is concentrated into concentrated solution relative density for 1.05g/ml, adopting inlet temperature is 180 ℃, it is dry that temperature of outgoing air is that the spraying drying conditions of 75 ℃ is sprayed, gained polypeptide dry powder water ratio < 4.0%, carries out abrasive dust by gained dry powder and crosses 120 mesh sieves and put into specified containers and store.
The alfalfa polypeptide powder making by above technique, after sampling, analytical test, content of peptides is 70.05%, and products obtained therefrom is pale powder, be easy to dissolve, after dissolving smell pure and fresh, do not there is careless fishy smell, but light green grass fragrance, almost without bitterness sense.
Embodiment 2
The preparation of alfalfa leaf protein: get initial bloom stage alfalfa (second batch~the 3rd batch) top 8cm~10cm young stem and leaf partly for processing raw material, pick removal flavescence, wilt, rot, damaged by vermin leaf and other impurity, with tap water, clean up after (2 times~3 times) drain and be cut into 2cm~3cm segment, alfalfa young stem and leaf in mass ratio: water=1: 2 put into tissue mashing machine's making beating processes 5min, after taking-up, whole juice and alfalfa residue are moved in colloidal mill, gear setting is degree of card aftertreatment 10min the most carefully, during this time to the water that drops into one times of alfalfa weight in colloidal mill, and adopt 200 order nylon filtering cloths to filter to obtain alfalfa juice, by alfalfa juice 10000ml direct heating, in heat-processed, constantly carry out mechanical stirring, when temperature rises to 80 ℃, insulation is processed 10min and is obtained deep green floss precipitation, after standing separation, remove supernatant liquor, floss is deposited in whizzer and obtains centrifugal sediment with 4000r/min centrifugal treating 10min, under normal temperature condition, seasoning obtains alfalfa leaf protein, and its protein mass mark is 68.16%.
Alfalfa protein decoloration deodorization is processed: adopt 120 mesh sieves to pulverize gained alfalfa protein, at alfalfa leaf protein and dehydrated alcohol mass ratio, it is 1: 8, at 85 ℃ of extraction temperatures, 10h is processed in lixiviate, the alfalfa leaf protein canescence of light color obtaining, protein mass mark is 67.13%, and products obtained therefrom still has lighter careless fishy smell.
Enzymolysis and dry: after decoloration deodorization is processed, alfalfa protein is put into enzymatic vessel, in enzymatic vessel, the mass ratio of alfalfa protein and water is 1: 1, treat 55 ℃ of tank Nei Wenduda, under nature pH value condition, add the Protamex enzyme (Chinese Novi's letter bio tech ltd) that is equivalent to alfalfa leaf protein weight 2.0 ‰, add the Flavourzyme1000L enzyme (Chinese Novi's letter bio tech ltd) that is equivalent to alfalfa leaf protein weight 1.0 ‰ simultaneously, in temperature, it is 55 ℃, enzymolysis processing 3h under nature pH value condition, enzyme 60min goes out under 100 ℃ of high temperature, filter and collect enzymolysis solution.Use dual-effect concentrator that the enzymolysis solution of collection is concentrated into concentrated solution relative density for 1.10g/ml, adopting inlet temperature is 185 ℃, it is dry that temperature of outgoing air is that the spraying drying conditions of 80 ℃ is sprayed, gained polypeptide dry powder water ratio < 4.0%, carries out abrasive dust by gained dry powder and crosses 120 mesh sieves and put into specified containers and store.
The alfalfa polypeptide powder making by above technique, after sampling, analytical test, content of peptides is 74.65%, and products obtained therefrom is pale powder, be easy to dissolve, after dissolving smell pure and fresh, do not there is careless fishy smell, but light green grass fragrance, almost without bitterness sense.
Embodiment 3
The preparation of alfalfa leaf protein: get initial bloom stage alfalfa (second batch~the 3rd batch) top 8cm~10cm young stem and leaf partly for processing raw material, pick removal flavescence, wilt, rot, damaged by vermin leaf and other impurity, with tap water, clean up after (2 times~3 times) drain and be cut into 2cm~3cm segment, alfalfa young stem and leaf in mass ratio: water=1: 2 put into tissue mashing machine's making beating processes 5min, after taking-up, whole juice and alfalfa residue are moved in colloidal mill, gear setting is degree of card aftertreatment 10min the most carefully, during this time to the water that drops into one times of alfalfa weight in colloidal mill, and adopt 200 order nylon filtering cloths to filter to obtain alfalfa juice, by alfalfa juice 10000ml direct heating, in heat-processed, constantly carry out mechanical stirring, when temperature rises to 85 ℃, insulation is processed 8min and is obtained deep green floss precipitation, after standing separation, remove supernatant liquor, floss is deposited in whizzer and obtains centrifugal sediment with 4000r/min centrifugal treating 10min, under normal temperature condition, seasoning obtains alfalfa leaf protein, and its protein mass mark is 66.24%, and products obtained therefrom still has lighter careless fishy smell.
Alfalfa protein decoloration deodorization is processed: adopt 120 mesh sieves to pulverize gained alfalfa protein, at alfalfa leaf protein and dehydrated alcohol mass ratio, be 1: 10, at 85 ℃ of extraction temperatures, 6h is processed in lixiviate, the alfalfa leaf protein canescence of light color obtaining, protein mass mark is 64.68%.
Enzymolysis and dry: after decoloration deodorization is processed, alfalfa protein is put into enzymatic vessel, in enzymatic vessel, the mass ratio of alfalfa protein and water is 1: 1, treat 55 ℃ of tank Nei Wenduda, under nature pH value condition, add the Protamex enzyme (Chinese Novi's letter bio tech ltd) that is equivalent to alfalfa leaf protein weight 1.5 ‰, add the Flavourzyme1000L enzyme (Chinese Novi's letter bio tech ltd) that is equivalent to alfalfa leaf protein weight 2.5 ‰ simultaneously, in temperature, it is 55 ℃, enzymolysis processing 3h under nature pH value condition, enzyme 60min goes out under 100 ℃ of high temperature, filter and collect enzymolysis solution.Use dual-effect concentrator that the enzymolysis solution of collection is concentrated into concentrated solution relative density for 1.15g/ml, adopting inlet temperature is 190 ℃, it is dry that temperature of outgoing air is that the spraying drying conditions of 85 ℃ is sprayed, gained polypeptide dry powder water ratio < 4.0%, carries out abrasive dust by gained dry powder and crosses 120 mesh sieves and put into specified containers and store.
The alfalfa polypeptide powder making by above technique, after sampling, analytical test, content of peptides is 73.26%, and products obtained therefrom is pale powder, be easy to dissolve, after dissolving smell pure and fresh, do not there is careless fishy smell, but light green grass fragrance, almost without bitterness sense.
Embodiment 4
The preparation of alfalfa leaf protein: get initial bloom stage alfalfa (second batch~the 3rd batch) top 8cm~10cm young stem and leaf partly for processing raw material, pick removal flavescence, wilt, rot, damaged by vermin leaf and other impurity, with tap water, clean up after (2 times~3 times) drain and be cut into 2cm~3cm segment, alfalfa young stem and leaf in mass ratio: water=1: 2 put into tissue mashing machine's making beating processes 5min, after taking-up, whole juice and alfalfa residue are moved in colloidal mill, gear setting is degree of card aftertreatment 10min the most carefully, during this time to the water that drops into one times of alfalfa weight in colloidal mill, and adopt 200 order nylon filtering cloths to filter to obtain alfalfa juice, by alfalfa juice 10000ml direct heating, in heat-processed, constantly carry out mechanical stirring, when temperature rises to 80 ℃, insulation is processed 10min and is obtained deep green floss precipitation, after standing separation, remove supernatant liquor, floss is deposited in whizzer and obtains centrifugal sediment with 4000r/min centrifugal treating 10min, under normal temperature condition, seasoning obtains alfalfa leaf protein, and its extraction yield is 68.05%.
Alfalfa protein decoloration deodorization is processed: adopt 120 mesh sieves to pulverize gained alfalfa protein, at alfalfa leaf protein and dehydrated alcohol mass ratio, it is 1: 6, at 85 ℃ of extraction temperatures, 8h is processed in lixiviate, the alfalfa leaf protein canescence of light color obtaining, protein mass mark is 66.96%, and products obtained therefrom still has lighter careless fishy smell.
Enzymolysis and dry: after decoloration deodorization is processed, alfalfa protein is put into enzymatic vessel, in enzymatic vessel, the mass ratio of alfalfa protein and water is 1: 1, treat 55 ℃ of tank Nei Wenduda, under nature pH value condition, add the Protamex enzyme (Chinese Novi's letter bio tech ltd) that is equivalent to alfalfa leaf protein weight 1.0 ‰, add the Flavourzyme1000L enzyme (Chinese Novi's letter bio tech ltd) that is equivalent to alfalfa leaf protein weight 2.0 ‰ simultaneously, in temperature, it is 55 ℃, enzymolysis processing 3h under nature pH value condition, enzyme 60min goes out under 100 ℃ of high temperature, filter and collect enzymolysis solution.Use dual-effect concentrator that the enzymolysis solution of collection is concentrated into concentrated solution relative density for 1.10g/ml, adopting inlet temperature is 180 ℃, it is dry that temperature of outgoing air is that the spraying drying conditions of 65 ℃ is sprayed, gained polypeptide dry powder water ratio < 4.0%, carries out abrasive dust by gained dry powder and crosses 120 mesh sieves and put into specified containers and store.
The alfalfa polypeptide powder making by above technique, after sampling, analytical test, content of peptides is 74.06%, and products obtained therefrom is pale powder, be easy to dissolve, after dissolving smell pure and fresh, do not there is careless fishy smell, but light green grass fragrance, almost without bitterness sense.
Embodiment 5
Embodiment 1-4 product is carried out to inspection for food hygiene according to the method for stipulating in GB 5009.3, GB 5009.4, GB/T 5009.11, GB 5009.12, GB/T 5009.17, GB4789.2-15, result shows: moisture content < 4.0%, ash content≤1.5%, inorganic arsenic (in AS, mg/kg) 0.22, plumbous (in Pb, mg/kg) 0.62, mercury (in Hg, mg/kg)≤0.05, total number of bacterial colony (cfu/g) 1.0*10
3, coliform (MPN/100g)≤35, mould (cfu/g)≤10, yeast (cfu/g)≤10, pathogenic bacterium do not detect.Meet the general mark of GB16740-1997 health (functional) food.
Claims (10)
1. a preparation method for alfalfa polypeptide, comprises the following steps: the extraction of (1) alfalfa leaf protein; (2) decoloration deodorization; (3) enzymolysis; (4) spraying is dry, obtains.
2. the preparation method of alfalfa polypeptide according to claim 1, is characterized in that, in step (3), enzyme used is Protamex enzyme and Flavourzyme1000L enzyme.
3. the preparation method of alfalfa polypeptide according to claim 2, is characterized in that, the consumption of described Protamex enzyme is 0.5 ‰ of alfalfa weight~1.5 ‰.
4. the preparation method of alfalfa polypeptide according to claim 3, is characterized in that, the consumption of described Protamex enzyme is 1.0 ‰ of alfalfa weight.
5. the preparation method of alfalfa polypeptide according to claim 2, is characterized in that, the consumption of described Flavourzyme1000L enzyme is 1.0 ‰ of alfalfa weight~2.5 ‰.
6. the preparation method of alfalfa polypeptide according to claim 5, is characterized in that, the consumption of described Flavourzyme1000L enzyme is 2.0 ‰ of alfalfa weight.
7. the preparation method of alfalfa polypeptide according to claim 1, is characterized in that, described step (1) adopts heating method, 75 ℃~85 ℃ of Heating temperatures, preferably 80 ℃.
8. the preparation method of alfalfa polypeptide according to claim 1, is characterized in that, described step (2) decoloration deodorization method is, alfalfa leaf protein and dehydrated alcohol mass ratio be controlled at 1: 5~and between 1: 10, preferably 1: 8.
9. the preparation method of alfalfa polypeptide according to claim 9, is characterized in that, extraction temperature is 85 ℃, and extraction time is 6h~10h, preferably 8h.
10. the preparation method of alfalfa polypeptide according to claim 1, is characterized in that, in described step (4), spraying drying conditions is that inlet temperature is 180 ℃~190 ℃, and temperature of outgoing air is 65 ℃~85 ℃.
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Cited By (1)
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CN105624248A (en) * | 2016-03-08 | 2016-06-01 | 陈忆凤 | Preparing method for alfalfa biological active peptide |
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CN102351944A (en) * | 2011-11-03 | 2012-02-15 | 齐齐哈尔大学 | Method for extracting alfalfa leaf protein by ultrasonic-assisted enzyme process |
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CN101077118A (en) * | 2007-06-12 | 2007-11-28 | 江南大学 | Method for preparing soluble heart trefoil protein |
CN102351944A (en) * | 2011-11-03 | 2012-02-15 | 齐齐哈尔大学 | Method for extracting alfalfa leaf protein by ultrasonic-assisted enzyme process |
Non-Patent Citations (2)
Title |
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PREVOT-D’ALVISE N.等: ""Development of a pilot process for the production of alfalfa peptide isolate"", 《J CHEM TECHNOL BIOTECHNOL》 * |
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105624248A (en) * | 2016-03-08 | 2016-06-01 | 陈忆凤 | Preparing method for alfalfa biological active peptide |
CN105624248B (en) * | 2016-03-08 | 2020-02-11 | 江苏天福莱集团有限公司 | Preparation method of alfalfa bioactive peptide |
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