CN105002247A - Micromolecule walnut peptide and preparation method thereof - Google Patents
Micromolecule walnut peptide and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a micromolecule walnut peptide and a preparation method thereof. The preparation method sequentially comprises the following steps of protein extraction, twice enzymolysis, filtering, purification, concentration and drying. In the twice enzymolysis process, alkaline protease with the mass being 1-3% that of the walnut protein is added into a walnut protein extracting solution for once enzymolysis; enzyme deactivation is performed on a once enzymolysis solution after the once enzymolysis is finished; one or more kinds of proteinase including the flavored proteinase, the neutral proteinase and the papain with the mass being 1-3% that of the walnut protein are added after the once enzymolysis solution obtained after enzyme deactivation is performed is cooled for secondary enzymolysis. According to the preparation method, the twice enzymolysis method is adopted, due to the specificity of the structure of the walnut protein structure, sufficient enzymolysis cannot be performed on the walnut protein through enzymolysis of one proteinase, and the twice enzymolysis method is adopted for ensuring that the enzymolysis is brought into play on the most suitable condition of each proteinase, and therefore the obtained walnut peptide content is high.
Description
Technical field
The present invention relates to biologically active substance and extract field, be specifically related to a kind of small molecules walnut peptide and preparation method thereof.
Background technology
Walnut is again English walnut, Qiang peach, long live etc., is Juglandaceae Juglans plants.In Semen Caryae Cathayensis, oleaginousness can reach 60%-70%, and wherein unsaturated fatty acid content is more than 90%, and Semen Caryae Cathayensis protein content is 14%-17%, digestibility can reach 87.2%.But Semen Caryae Cathayensis protective foods desirable is at present commercially actually rare, except being made into can, mooncake filling, walnut powder, Walnut Milk, walnut oil, the exploitation of other purposes still belongs to blank, and particularly deep processed product is less.The application of walnut in daily life and industrial production is also more and more extensive, but problems faced is also more and more obvious.Now, walnut oil manufacturer of China has reached tens of family, and a large amount of degreasing walnut slags is by as feed and fertilizer or abandon, waste discharge contaminate environment, and its added value is lower.
Degreasing walnut slag is the by product after walnut oil expression, about containing the protein of 30%-50%.Walnut protein primarily of albumin, sphaeroprotein, prolamine and gluten four proteinoid form, account for 6.81% of walnut protein total amount, 17.57%, 5.33% and 70.11% respectively.Scientific research in recent years finds, human consumption's protein is after digestive ferment effect, and non-principal absorbs with amino acid whose form, but absorb with the form of peptide, some peptides can not only provide growth in humans, grow needed for nutritive substance, there is different physiological roles simultaneously, as improve mineral substance transport and absorb, antibacterial and viral, improve immunizing power, anti-oxidant, decreasing cholesterol, antitumous effect, the anti-ageing effect of waiting for a long time of scavenging free radicals.
Both at home and abroad large quantity research shows, walnut protein obtains walnut peptide through biological enzymolysis to be had and improve immunizing power, anti-oxidant activity and the effect such as anticancer.
Summary of the invention
Technical problem to be solved by this invention is for problems of the prior art, provides a kind of preparation method of small molecules walnut peptide.
The technical scheme that the present invention solves the problems of the technologies described above is as follows: a kind of preparation method of small molecules walnut peptide, comprises the following steps successively: protein extraction, twice enzymolysis, filtration, purified concentration and drying;
Described twice enzymolysis process be specifically: in the walnut protein extracting solution of protein extraction procedure extraction, add the Sumizyme MP being equivalent to walnut protein quality 1%-3% carry out primary enzymolysis, primary enzymolysis temperature is 30 DEG C-60 DEG C, primary enzymolysis pH value in reaction is 8-9, and the primary enzymolysis time is 3h-6h;
After primary enzymolysis terminates, primary enzymolysis liquid is gone out ferment treatment;
By the primary enzymolysis liquid cooling through the ferment treatment that goes out, and the proteolytic enzyme that is a kind of or any several mixing added in flavor protease, neutral protease or the papoid being equivalent to walnut protein quality 1%-3% carries out secondary enzymolysis, secondary enzymolysis temperature is 40 DEG C-60 DEG C, secondary enzymolysis pH value in reaction is 6-7, and the secondary enzymolysis time is 3h-6h;
After secondary enzymolysis terminates, secondary enzymolysis liquid is gone out ferment treatment.
The invention has the beneficial effects as follows: the present invention adopts twice enzymolysis process, due to the singularity of walnut protein structure, fully enzymolysis can not be carried out to walnut protein with a kind of enzyme enzymolysis, therefore adopt twice enzymolysis process, to guarantee that each enzyme can play enzymolysis under oneself optimal condition, the walnut peptide egg content obtained like this is higher.
On the basis of technique scheme, the present invention can also do following improvement.
Further, the process of described protein extraction is: walnut is carried out skimming treatment, obtains degreasing walnut slag, after degreasing walnut ground-slag being broken to 40 order-60 orders, add the deionized water being equivalent to walnut slag amount 5 times-8 times, add alkali and regulate pH to 8.5-9, after soaking 1h-3h, centrifugal segregation lower sediment; Acid adding regulates the pH to 4-4.5 of centrifugate, and centrifugation after standing 1h-3h, gets lower sediment and be washed to neutrality, obtaining walnut protein extracting solution.
The beneficial effect of above-mentioned further scheme is adopted to be: the inventive method makes full use of the byproduct-degreasing walnut slag after peach-pit oil expression to prepare small-molecular peptides, and with low cost, adopt alkali extraction and acid precipitation method to extract walnut protein, extraction yield is greater than 85%.
Further, the alkali that described adjustment pH adds is sodium hydrogen carbonate solution or sodium hydroxide solution, and the acid that described adjustment pH adds is citric acid solution or glacial acetic acid solution.
Further, described filtration procedure is: in the enzymolysis solution through twice enzymolysis, add activated carbon, and is warming up to 100 DEG C, keeps 5min-10min, leaves standstill after being then cooled to 70 DEG C-80 DEG C, to remove color in enzymolysis solution and peculiar smell.
Further, the add-on of described activated carbon is the 2%-4% of walnut protein quality, and time of repose is 30min-2h.
Further, described purified concentration process is: by the walnut peptide solution centrifugal removing solid residue through filtering, be the macromole impurity that the daltonian ultrafiltration membrance filter of 1000 dalton-5000 removes in clear liquid by the clear liquid molecular weight obtained, be that 200 daltonian nanofiltration membrane remove inorganic salt and small molecular weight impurities with molecular weight again, it is 20%-30% that clear liquid after filtration is concentrated into solid content, namely obtains small molecules walnut peptide solution.
Adopt the beneficial effect of above-mentioned further scheme to be: adopt centrifugal collaborative Using Multistage Membranes separation system purifying walnut peptide, removal of impurities, desalination and concentrated effect are good, and low temperature nanofiltration concentrates the activity that ensure that product, product saltiness is low simultaneously, is applicable to large-scale industrial and produces.
Further, the mould of described ultra-filtration membrane is 0.5MPa-1MPa, and ultrafiltrate temperature is 25 DEG C-40 DEG C; The mould of described nanofiltration membrane is 1MPa-2MPa, and nanofiltration temperature is 25 DEG C-40 DEG C.
Further, described drying process is: the small molecules walnut peptide solution after purified concentration is carried out spraying dry, and spray-dired air outlet temperature is 70 DEG C-90 DEG C, and inlet temperature is 140 DEG C-180 DEG C, namely obtains small molecules walnut peptide powder after drying.
Further, ferment treatment process of going out described in is: primary enzymolysis liquid or secondary enzymolysis liquid are warming up to 80 DEG C-85 DEG C, and constant temperature 10min-20min carries out going out ferment treatment.
Adopt aforesaid method namely to obtain a kind of small molecules walnut peptide powder of the present invention, described small molecules walnut peptide is that molecular-weight average is less than 1000 daltonian oligopeptides, and color is white or faint yellow.
Embodiment
Be described principle of the present invention and feature with the following Examples, example, only for explaining the present invention, is not intended to limit scope of the present invention.
Embodiment 1
The preparation method of a kind of small molecules walnut peptide of the present embodiment, comprises the following steps:
1) protein extraction: the process of protein extraction is for carry out skimming treatment by walnut, obtain degreasing walnut slag, after degreasing walnut ground-slag is broken to 40 orders, add the deionized water of walnut slag amount 5 times, add sodium hydrogen carbonate solution or sodium hydroxide solution adjustment pH to 9, after soaking 2h, remove lower sediment with whizzer; Add the pH to 4.2 of citric acid solution or glacial acetic acid solution adjustment centrifugate, centrifugation after standing 3h, get lower sediment and be washed to neutrality, obtaining walnut protein extracting solution, adopting B or Kjeldahl determination to detect the walnut protein quality in walnut protein extracting solution.The present embodiment make full use of after peach-pit oil expression byproduct---degreasing walnut slag is to prepare small-molecular peptides, with low cost, adopt alkali extraction and acid precipitation method to extract walnut protein, extraction yield is greater than 85%; The walnut raw material of the present embodiment preferably adopts Semen Caryae Cathayensis.
2) twice enzymolysis: twice enzymolysis process is add the Sumizyme MP being equivalent to walnut protein quality 1% to carry out primary enzymolysis in walnut protein extracting solution, and primary enzymolysis temperature is 50 DEG C, and primary enzymolysis pH value is 8.5, and the primary enzymolysis time is 3h;
After primary enzymolysis terminates, primary enzymolysis liquid is warming up to 85 DEG C, constant temperature 10mi n carries out going out ferment treatment;
Be cooled to 20 DEG C by through the primary enzymolysis liquid of ferment treatment of going out, and add the neutral protease being equivalent to walnut protein quality 1% and carry out secondary enzymolysis, secondary enzymolysis temperature is 45 DEG C, and secondary enzymolysis pH value in reaction is 7, and the secondary enzymolysis time is 3h;
After secondary enzymolysis terminates, secondary enzymolysis liquid is warming up to 85 DEG C, constant temperature 20mi n carries out going out ferment treatment.
The present embodiment adopts twice enzyme process, due to the singularity of walnut protein structure, fully enzymolysis can not be carried out to walnut protein by a kind of enzyme or primary enzymolysis, therefore adopt multiple prozyme and twice enzymolysis process, to guarantee that each enzyme can play enzymolysis under oneself optimal condition, the walnut peptide egg content obtained like this is higher.
3) filter: filtration procedure is add activated carbon in the enzymolysis solution through twice enzymolysis, the add-on of activated carbon is 2% of walnut protein quality, and is warming up to 100 DEG C, keeps 5min, then 2h is left standstill after being cooled to 80 DEG C, to remove color in enzymolysis solution and peculiar smell.
4) purified concentration: by the walnut peptide solution centrifugal removing solid residue through filtering, be the macromole impurity that 3500 daltonian ultrafiltration membrance filters remove in clear liquid by the clear liquid molecular weight obtained, the mould of ultra-filtration membrane is 0.6MPa, and ultrafiltrate temperature is 30 DEG C; Be that 200 daltonian nanofiltration membrane remove inorganic salt and small molecular weight impurities with molecular weight again, the mould of nanofiltration membrane is 1.5MPa, and nanofiltration temperature is 30 DEG C.It is 30% that clear liquid after filtration is concentrated into solid content, namely obtains small molecules walnut peptide solution.
Adopt centrifugal collaborative Using Multistage Membranes separation system purifying walnut peptide, removal of impurities, desalination and concentrated effect are good, and low temperature nanofiltration concentrates the activity that ensure that product, and product saltiness is low simultaneously, are applicable to large-scale industrial and produce.
5) dry: the small molecules walnut peptide solution after purified concentration is carried out spraying dry, and spray-dired air outlet temperature is 80 DEG C, and inlet temperature is 160 DEG C, namely obtains small molecules walnut peptide powder after drying.
Embodiment 2
The preparation method of a kind of small molecules walnut peptide of the present embodiment, comprises the following steps:
1) protein extraction: the process of protein extraction is for carry out skimming treatment by walnut, obtain degreasing walnut slag, after degreasing walnut ground-slag is broken to 40 orders, add the deionized water of walnut slag amount 8 times, add sodium hydrogen carbonate solution or sodium hydroxide solution adjustment pH to 8.5, after soaking 1h, remove lower sediment with whizzer; Add the pH to 4 of citric acid solution or glacial acetic acid solution adjustment centrifugate, centrifugation after standing 1h, get lower sediment and be washed to neutrality, obtaining walnut protein extracting solution, adopting B or Kjeldahl determination to detect the walnut protein quality in walnut protein extracting solution.The present embodiment make full use of after peach-pit oil expression byproduct---degreasing walnut slag is to prepare small-molecular peptides, with low cost, adopt alkali extraction and acid precipitation method to extract walnut protein, extraction yield is greater than 85%; The walnut raw material of the present embodiment preferably adopts Semen Caryae Cathayensis.
2) twice enzymolysis: twice enzymolysis process is add the Sumizyme MP being equivalent to walnut protein quality 3% to carry out primary enzymolysis in walnut protein extracting solution, and primary enzymolysis temperature is 30 DEG C, and primary enzymolysis pH value is 8, and the primary enzymolysis time is 3h;
After primary enzymolysis terminates, primary enzymolysis liquid is warming up to 80 DEG C, constant temperature 10min carries out going out ferment treatment;
Be cooled to 20 DEG C by through the primary enzymolysis liquid of ferment treatment of going out, and add the flavor protease being equivalent to walnut protein quality 1% and carry out secondary enzymolysis, secondary enzymolysis temperature is 40 DEG C, and secondary enzymolysis pH value in reaction is 6, and the secondary enzymolysis time is 3h;
After secondary enzymolysis terminates, secondary enzymolysis liquid is warming up to 80 DEG C, constant temperature 10min carries out going out ferment treatment.
The present embodiment adopts twice enzyme process, due to the singularity of walnut protein structure, fully enzymolysis can not be carried out to walnut protein by a kind of enzyme or primary enzymolysis, therefore adopt multiple prozyme and twice enzymolysis process, to guarantee that each enzyme can play enzymolysis under oneself optimal condition, the walnut peptide egg content obtained like this is higher.
3) filter: filtration procedure is add activated carbon in the enzymolysis solution through twice enzymolysis, the add-on of activated carbon is 2% of walnut protein quality, and is warming up to 100 DEG C, keeps 5min, then 30min is left standstill after being cooled to 80 DEG C, to remove color in enzymolysis solution and peculiar smell.
4) purified concentration: by the walnut peptide solution centrifugal removing solid residue through filtering, be the macromole impurity that 1000 daltonian ultrafiltration membrance filters remove in clear liquid by the clear liquid molecular weight obtained, the mould of ultra-filtration membrane is 0.5MPa, and ultrafiltrate temperature is 25 DEG C; Be that 200 daltonian nanofiltration membrane remove inorganic salt and small molecular weight impurities with molecular weight again, the mould of nanofiltration membrane is 1MPa, and nanofiltration temperature is 25 DEG C.It is 20% that clear liquid after filtration is concentrated into solid content, namely obtains small molecules walnut peptide solution.
Adopt centrifugal collaborative Using Multistage Membranes separation system purifying walnut peptide, removal of impurities, desalination and concentrated effect are good, and low temperature nanofiltration concentrates the activity that ensure that product, and product saltiness is low simultaneously, are applicable to large-scale industrial and produce.
5) dry: the small molecules walnut peptide solution after purified concentration is carried out spraying dry, and spray-dired air outlet temperature is 70 DEG C, and inlet temperature is 180 DEG C, namely obtains small molecules walnut peptide powder after drying.
Embodiment 3
The preparation method of a kind of small molecules walnut peptide of the present embodiment, comprises the following steps:
1) protein extraction: the process of protein extraction is for carry out skimming treatment by walnut, obtain degreasing walnut slag, after degreasing walnut ground-slag is broken to 60 orders, add the deionized water of walnut slag amount 8 times, add sodium hydrogen carbonate solution or sodium hydroxide solution adjustment pH to 9, after soaking 1h, remove lower sediment with whizzer; Add the pH to 4.5 of citric acid solution or glacial acetic acid solution adjustment centrifugate, centrifugation after standing 3h, get lower sediment and be washed to neutrality, obtaining walnut protein extracting solution, adopting B or Kjeldahl determination to detect the walnut protein quality in walnut protein extracting solution.The present embodiment make full use of after peach-pit oil expression byproduct---degreasing walnut slag is to prepare small-molecular peptides, with low cost, adopt alkali extraction and acid precipitation method to extract walnut protein, extraction yield is greater than 85%; The walnut raw material of the present embodiment preferably adopts Semen Caryae Cathayensis.
2) twice enzymolysis: twice enzymolysis process is add the Sumizyme MP being equivalent to walnut protein quality 3% to carry out primary enzymolysis in walnut protein extracting solution, and primary enzymolysis temperature is 60 DEG C, and primary enzymolysis pH value is 9, and the primary enzymolysis time is 6h;
After primary enzymolysis terminates, primary enzymolysis liquid is warming up to 85 DEG C, constant temperature 20min carries out going out ferment treatment;
Be cooled to 25 DEG C by through the primary enzymolysis liquid of ferment treatment of going out, and add the papoid being equivalent to walnut protein quality 3% and carry out secondary enzymolysis, secondary enzymolysis temperature is 60 DEG C, and secondary enzymolysis pH value in reaction is 7, and the secondary enzymolysis time is 6h;
After secondary enzymolysis terminates, secondary enzymolysis liquid is warming up to 85 DEG C, constant temperature 20mi n carries out going out ferment treatment; The proteolytic enzyme of twice enzymolysis employing is at least one of Sumizyme MP, flavor protease, neutral protease and papoid.
The present embodiment adopts twice enzyme process, due to the singularity of walnut protein structure, fully enzymolysis can not be carried out to walnut protein by a kind of enzyme or primary enzymolysis, therefore adopt multiple prozyme and twice enzymolysis process, to guarantee that each enzyme can play enzymolysis under oneself optimal condition, the walnut peptide egg content obtained like this is higher.
3) filter: filtration procedure is add activated carbon in the enzymolysis solution through twice enzymolysis, the add-on of activated carbon is 4% of walnut protein quality, and is warming up to 100 DEG C, keeps 10min, then 2h is left standstill after being cooled to 80 DEG C, to remove color in enzymolysis solution and peculiar smell.
4) purified concentration: by the walnut peptide solution centrifugal removing solid residue through filtering, be the macromole impurity that 5000 daltonian ultrafiltration membrance filters remove in clear liquid by the clear liquid molecular weight obtained, the mould of ultra-filtration membrane is 1MPa, and ultrafiltrate temperature is 40 DEG C; Be that 200 daltonian nanofiltration membrane remove inorganic salt and small molecular weight impurities with molecular weight again, the mould of nanofiltration membrane is 2MPa, and nanofiltration temperature is 40 DEG C.It is 30% that clear liquid after filtration is concentrated into solid content, namely obtains small molecules walnut peptide solution.
Adopt centrifugal collaborative Using Multistage Membranes separation system purifying walnut peptide, removal of impurities, desalination and concentrated effect are good, and low temperature nanofiltration concentrates the activity that ensure that product, and product saltiness is low simultaneously, are applicable to large-scale industrial and produce.
5) dry: the small molecules walnut peptide solution after purified concentration is carried out spraying dry, and spray-dired air outlet temperature is 90 DEG C, and inlet temperature is 180 DEG C, namely obtains small molecules walnut peptide powder after drying.
Embodiment 4
The preparation method of a kind of small molecules walnut peptide of the present embodiment, comprises the following steps:
1) protein extraction: the process of protein extraction is for carry out skimming treatment by walnut, obtain degreasing walnut slag, after degreasing walnut ground-slag is broken to 50 orders, add the deionized water of walnut slag amount 7 times, add sodium hydrogen carbonate solution or sodium hydroxide solution adjustment pH to 8.9, after soaking 2h, remove lower sediment with whizzer; Add the pH to 4.2 of citric acid solution or glacial acetic acid solution adjustment centrifugate, centrifugation after standing 2h, get lower sediment and be washed to neutrality, obtaining walnut protein extracting solution, adopting B or Kjeldahl determination to detect the walnut protein quality in walnut protein extracting solution.The present embodiment make full use of after peach-pit oil expression byproduct---degreasing walnut slag is to prepare small-molecular peptides, with low cost, adopt alkali extraction and acid precipitation method to extract walnut protein, extraction yield is greater than 85%; The walnut raw material of the present embodiment preferably adopts Semen Caryae Cathayensis.
2) twice enzymolysis: twice enzymolysis process is add the Sumizyme MP being equivalent to walnut protein quality 2% to carry out primary enzymolysis in walnut protein extracting solution, and primary enzymolysis temperature is 40 DEG C, and primary enzymolysis pH value is 9, and the primary enzymolysis time is 4h;
After primary enzymolysis terminates, primary enzymolysis liquid is warming up to 83 DEG C, constant temperature 15min carries out going out ferment treatment;
Be cooled to 23 DEG C by through the primary enzymolysis liquid of ferment treatment of going out, and add the papoid being equivalent to walnut protein quality 2% and carry out secondary enzymolysis, secondary enzymolysis temperature is 50 DEG C, and secondary enzymolysis pH value in reaction is 6.5, and the secondary enzymolysis time is 4h;
After secondary enzymolysis terminates, secondary enzymolysis liquid is warming up to 82 DEG C, constant temperature 15mi n carries out going out ferment treatment.
The present embodiment adopts twice enzyme process, due to the singularity of walnut protein structure, fully enzymolysis can not be carried out to walnut protein by a kind of enzyme or primary enzymolysis, therefore adopt multiple prozyme and twice enzymolysis process, to guarantee that each enzyme can play enzymolysis under oneself optimal condition, the walnut peptide egg content obtained like this is higher.
3) filter: filtration procedure is add activated carbon in the enzymolysis solution through twice enzymolysis, the add-on of activated carbon is 4% of walnut protein quality, and is warming up to 100 DEG C, keeps 8min, then 30min-2h is left standstill after being cooled to 80 DEG C, to remove color in enzymolysis solution and peculiar smell.
4) purified concentration: by the walnut peptide solution centrifugal removing solid residue through filtering, be the macromole impurity that 2500 daltonian ultrafiltration membrance filters remove in clear liquid by the clear liquid molecular weight obtained, the mould of ultra-filtration membrane is 0.8MPa, and ultrafiltrate temperature is 30 DEG C; Be that 200 daltonian nanofiltration membrane remove inorganic salt and small molecular weight impurities with molecular weight again, the mould of nanofiltration membrane is 1.5MPa, and nanofiltration temperature is 30 DEG C.It is 25% that clear liquid after filtration is concentrated into solid content, namely obtains small molecules walnut peptide solution.
Adopt centrifugal collaborative Using Multistage Membranes separation system purifying walnut peptide, removal of impurities, desalination and concentrated effect are good, and low temperature nanofiltration concentrates the activity that ensure that product, and product saltiness is low simultaneously, are applicable to large-scale industrial and produce.
5) dry: the small molecules walnut peptide solution after purified concentration is carried out spraying dry, and spray-dired air outlet temperature is 80 DEG C, and inlet temperature is 160 DEG C, namely obtains small molecules walnut peptide powder after drying.
Embodiment 5
The preparation method of a kind of small molecules walnut peptide of the present embodiment, comprises the following steps:
1) protein extraction: the process of protein extraction is for carry out skimming treatment by walnut, obtain degreasing walnut slag, after degreasing walnut ground-slag is broken to 45 orders, add the deionized water of walnut slag amount 7 times, add sodium hydrogen carbonate solution or sodium hydroxide solution adjustment pH to 8.5, after soaking 1h, remove lower sediment with whizzer; Add the pH to 4.5 of citric acid solution or glacial acetic acid solution adjustment centrifugate, centrifugation after standing 2.5h, get lower sediment and be washed to neutrality, obtaining walnut protein extracting solution, adopting B or Kjeldahl determination to detect the walnut protein quality in walnut protein extracting solution.The present embodiment make full use of after peach-pit oil expression byproduct---degreasing walnut slag is to prepare small-molecular peptides, with low cost, adopt alkali extraction and acid precipitation method to extract walnut protein, extraction yield is greater than 85%; The walnut raw material of the present embodiment preferably adopts Semen Caryae Cathayensis.
2) twice enzymolysis: twice enzymolysis process is add the Sumizyme MP being equivalent to walnut protein quality 2.5% to carry out primary enzymolysis in walnut protein extracting solution, and primary enzymolysis temperature is 48 DEG C, and primary enzymolysis pH value is 8.5, and the primary enzymolysis time is 4h;
After primary enzymolysis terminates, primary enzymolysis liquid is warming up to 80 DEG C, constant temperature 15min carries out going out ferment treatment;
Be cooled to 20 DEG C by through the primary enzymolysis liquid of ferment treatment of going out, and add the flavor protease being equivalent to walnut protein quality 1% and carry out secondary enzymolysis, secondary enzymolysis temperature is 40 DEG C, and secondary enzymolysis pH value in reaction is 6, and the secondary enzymolysis time is 5h;
After secondary enzymolysis terminates, secondary enzymolysis liquid is warming up to 82 DEG C, constant temperature 18mi n carries out going out ferment treatment.
The present embodiment adopts twice enzyme process, due to the singularity of walnut protein structure, fully enzymolysis can not be carried out to walnut protein by a kind of enzyme or primary enzymolysis, therefore adopt multiple prozyme and twice enzymolysis process, to guarantee that each enzyme can play enzymolysis under oneself optimal condition, the walnut peptide egg content obtained like this is higher.
3) filter: filtration procedure is add activated carbon in the enzymolysis solution through twice enzymolysis, the add-on of activated carbon is 3% of walnut protein quality, and is warming up to 100 DEG C, keeps 10min, then 2h is left standstill after being cooled to 80 DEG C, to remove color in enzymolysis solution and peculiar smell.
4) purified concentration: by the walnut peptide solution centrifugal removing solid residue through filtering, be the macromole impurity that 1500 daltonian ultrafiltration membrance filters remove in clear liquid by the clear liquid molecular weight obtained, the mould of ultra-filtration membrane is 1MPa, and ultrafiltrate temperature is 40 DEG C; Be that 200 daltonian nanofiltration membrane remove inorganic salt and small molecular weight impurities with molecular weight again, the mould of nanofiltration membrane is 2MPa, and nanofiltration temperature is 40 DEG C.It is 30% that clear liquid after filtration is concentrated into solid content, namely obtains small molecules walnut peptide solution.
Adopt centrifugal collaborative Using Multistage Membranes separation system purifying walnut peptide, removal of impurities, desalination and concentrated effect are good, and low temperature nanofiltration concentrates the activity that ensure that product, and product saltiness is low simultaneously, are applicable to large-scale industrial and produce.
5) dry: the small molecules walnut peptide solution after purified concentration is carried out spraying dry, and spray-dired air outlet temperature is 80 DEG C, and inlet temperature is 150 DEG C, namely obtains small molecules walnut peptide powder after drying.
Adopt aforesaid method namely to obtain a kind of small molecules walnut peptide powder, the molecular-weight average of the small molecules walnut peptide that above-described embodiment method obtains is less than 1000 dalton, and color is white or faint yellow, free from extraneous odour.Any organic solvent is not added, technique simple, high energy-conservation, low cost, high hydrolysis efficiency in the production process of small molecules walnut peptide, and the Stability Analysis of Structures of spraying dry products obtained therefrom, functional.
Walnut peptide powder embodiment 3 method obtained adopts GB/B 22729-2008 to carry out molecular weight distribution detection, and detected result is as shown in table 1.
The molecular weight distribution table of table 1 walnut peptide powder
| Molecular weight ranges | Peak area percent (%, λ 220nm) | Number-average molecular weight | Weight-average molecular weight |
| >5000 | 0.34 | 7708 | 8701 |
| 5000～3000 | 0.43 | 3622 | 3701 |
| 3000～2000 | 1.43 | 2327 | 2356 |
| 2000～1000 | 9.97 | 1263 | 1311 |
| 1000～500 | 31.82 | 755 | 770 |
| 500～180 | 51.57 | 295 | 321 |
| ＜180 | 4.44 | / | / |
As shown in Table 1, adopt the molecular weight distribution of the small molecules walnut peptide powder that method obtains described in embodiment 3 between 180 dalton-1000 dalton, the small molecules walnut peptide good water solubility between this molecular weight area, is more conducive to absorption and the utilization of walnut peptide activeconstituents.
GB 5009.5-2010 is adopted to detect the peptide content of the small molecules walnut peptide powder that method described in embodiment 3 obtains, acid-soluble protein content, total nitrogen content and weight loss on drying.Detected result is as shown in table 2.
The peptide content of table 2 small molecules of the present invention Gly-His-Lys, acid-soluble protein content, total nitrogen content and weight loss on drying amount
| Peptide content (g/100g) | 91.05 |
| Acid-soluble protein content (g/100g) | 88.10 |
| Total nitrogen content (g/100g) | 16.05 |
| Weight loss on drying (%) | 4.28 |
As shown in table 2, the yield adopting method described in the present embodiment 3 to produce small molecules walnut peptide is more than 90%, wherein acid-soluble protein content is in the great majority, and is conducive to digesting and assimilating of human stomach, has antitumor, raising body immunity, anti-oxidant, anti-ageing different physiological roles of waiting for a long time.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. a preparation method for small molecules walnut peptide, is characterized in that, comprises the following steps successively: protein extraction, twice enzymolysis, filtration, purified concentration and drying;
Described twice enzymolysis process be specifically: in the walnut protein extracting solution of protein extraction procedure extraction, add the Sumizyme MP being equivalent to walnut protein quality 1%-3% carry out primary enzymolysis, primary enzymolysis temperature is 30 DEG C-60 DEG C, primary enzymolysis pH value in reaction is 8-9, and the primary enzymolysis time is 3h-6h;
After primary enzymolysis terminates, primary enzymolysis liquid is gone out ferment treatment;
By the primary enzymolysis liquid cooling through the ferment treatment that goes out, and the proteolytic enzyme that is a kind of or any several mixing added in flavor protease, neutral protease or the papoid being equivalent to walnut protein quality 1%-3% carries out secondary enzymolysis, secondary enzymolysis temperature is 40 DEG C-60 DEG C, secondary enzymolysis pH value in reaction is 6-7, and the secondary enzymolysis time is 3h-6h;
After secondary enzymolysis terminates, secondary enzymolysis liquid is gone out ferment treatment.
2. the preparation method of a kind of small molecules walnut peptide according to claim 1, it is characterized in that, the process of described protein extraction is: walnut is carried out skimming treatment, obtain degreasing walnut slag, after degreasing walnut ground-slag being broken to 40 order-60 orders, add the deionized water being equivalent to walnut slag amount 5 times-8 times, add alkali and regulate pH to 8.5-9, after soaking 1h-3h, centrifugal segregation lower sediment; Acid adding regulates the pH to 4-4.5 of centrifugate, and centrifugation after standing 1h-3h, gets lower sediment and be washed to neutrality, obtaining walnut protein extracting solution.
3. a kind of preparation method of small molecules walnut peptide according to claim 1 or 2, is characterized in that, the alkali that described adjustment pH adds is sodium hydrogen carbonate solution or sodium hydroxide solution, and the acid that described adjustment pH adds is citric acid solution or glacial acetic acid solution.
4. the preparation method of a kind of small molecules walnut peptide according to claim 1, it is characterized in that, described filtration procedure is: in the enzymolysis solution through twice enzymolysis, add activated carbon, and be warming up to 100 DEG C, keep 5min-10min, then leave standstill after being cooled to 70 DEG C-80 DEG C, to remove color in enzymolysis solution and peculiar smell.
5. the preparation method of a kind of small molecules walnut peptide according to claim 4, it is characterized in that, the add-on of described activated carbon is the 2%-4% of walnut protein quality, and time of repose is 30min-2h.
6. the preparation method of a kind of small molecules walnut peptide according to claim 1, it is characterized in that, described purified concentration process is: by the walnut peptide solution centrifugal removing solid residue through filtering, be the macromole impurity that the daltonian ultrafiltration membrance filter of 1000 dalton-5000 removes in clear liquid by the clear liquid molecular weight obtained, be that 200 daltonian nanofiltration membrane remove inorganic salt and small molecular weight impurities with molecular weight again, it is 20%-30% that clear liquid after filtration is concentrated into solid content, namely obtains small molecules walnut peptide solution.
7. the preparation method of a kind of small molecules walnut peptide according to claim 6, it is characterized in that, the mould of described ultra-filtration membrane is 0.5MPa-1MPa, and ultrafiltrate temperature is 25 DEG C-40 DEG C; The mould of described nanofiltration membrane is 1MPa-2MPa, and nanofiltration temperature is 25 DEG C-40 DEG C.
8. the preparation method of a kind of small molecules walnut peptide according to claim 1, it is characterized in that, described drying process is: the small molecules walnut peptide solution after purified concentration is carried out spraying dry, spray-dired air outlet temperature is 70 DEG C-90 DEG C, inlet temperature is 140 DEG C-180 DEG C, namely obtains small molecules walnut peptide powder after drying.
9. the preparation method of a kind of small molecules walnut peptide according to claim 1, is characterized in that, described in ferment treatment process of going out be: primary enzymolysis liquid or secondary enzymolysis liquid are warming up to 80 DEG C-85 DEG C, and constant temperature 10min-20min carries out going out ferment treatment.
10. a small molecules walnut peptide, it is characterized in that, adopt method i.e. obtained small molecules walnut peptide of the present invention as described in any one of claim 1-9, described small molecules walnut peptide is that molecular-weight average is less than 1000 daltonian oligopeptides, and color is white or faint yellow.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105111282A (en) * | 2015-09-11 | 2015-12-02 | 哈尔滨工业大学 | Walnut peptide having ACE inhibitory activity and preparation method thereof |
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| CN105567774A (en) * | 2016-01-22 | 2016-05-11 | 杏辉天力（杭州）药业有限公司 | Walnut oligopeptide powder and preparation method and application thereof |
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| CN110037163A (en) * | 2019-04-24 | 2019-07-23 | 王书敏 | A kind of preparation method of compound plant protein peptide |
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| CN111073941A (en) * | 2019-09-12 | 2020-04-28 | 天津芸熙生物技术有限公司 | Preparation process of sandalwood polypeptide |
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| CN115181775A (en) * | 2022-07-19 | 2022-10-14 | 张建明 | Extraction method of small molecular liver peptide |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102406050A (en) * | 2011-11-17 | 2012-04-11 | 中国农业大学 | Walnut low molecular weight polypeptide and preparation method thereof |
| CN104489239A (en) * | 2015-01-06 | 2015-04-08 | 石河子大学 | Method of preparing antihypertensive peptide with walnut dregs as raw material |
-
2015
- 2015-07-30 CN CN201510461228.5A patent/CN105002247A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102406050A (en) * | 2011-11-17 | 2012-04-11 | 中国农业大学 | Walnut low molecular weight polypeptide and preparation method thereof |
| CN104489239A (en) * | 2015-01-06 | 2015-04-08 | 石河子大学 | Method of preparing antihypertensive peptide with walnut dregs as raw material |
Non-Patent Citations (2)
| Title |
|---|
| 孔令明 等: "酶解条件对核桃多肽抗氧化活性的影响", 《食品研究与开发》 * |
| 杜蕾蕾 等: "核桃蛋白的分离纯化及功能性质的研究", 《中国油脂》 * |
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