CN109762862A - A kind of preparation method of high-purity lotus seed protein oligopeptide - Google Patents
A kind of preparation method of high-purity lotus seed protein oligopeptide Download PDFInfo
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Abstract
The present invention provides a kind of lotus seed protein oligopeptides and preparation method thereof, and the method includes (1) to impregnate, defibrination；(2) salt extracts: adding NaCl to 0.2~0.5mol/L by solid-liquid ratio 1:10~20, adjusts solution pH value to 8~10, filter and be centrifuged to obtain supernatant；(3) acid is heavy: the HCl that 0.25~0.5mol/L of concentration is added adjusts supernatant pH value to 4.3~4.7, staticly settles, is centrifuged, obtains lotus seed protein sediment；(4) digest: being added with water to concentration of substrate is 8%~16%, and temperature is set in 35 DEG C~45 DEG C, and PH is adjusted to 3~8, and protease is added, and the protease is the neutral proteinase and papain that mass ratio is 1:1.5~2.5, digests 2~4h；Enzyme deactivation；Dialysis；Film filtering；Drying.The oligomeric peptide yield of lotus seed protein produced by the present invention is up to 78.6%, and appearance luster and mouthfeel quality are all good.
Description
Technical field
The present invention relates to article processing techniques fields more particularly to a kind of lotus seed protein oligopeptide and preparation method thereof.
Background technique
Lotus seeds are the seeds of nymphaeaceae plant lotus, are produced in the ground such as Hunan, Hubei, Jiangxi, Fujian, resourceful.Lotus seeds
Protein content is higher, containing amino acid in 18, the content of albumen essential amino acid, ratio close to FAO/WHO recommendation pattern,
Amino acid content ratio compares balance, can reduce cholesterol and saturated fatty acid content in diet, keep the equilibrium of nutrition.It compares
Peas protein lacks methionine, and lotus seed protein has reached WHO required standard.Lotus seed protein is rich in lysine, and lysine exists
The case where being the first limiting amino acids in common food, shortage crowd's insufficiency of intake can be improved.Lotus seed protein oligopeptide is protected
The rational proportion of amino acid is stayed, and small-molecular peptides absorb faster, also have significant immunoloregulation function, and to chemical source
Caused hepatic injury has auxiliary curative effect, removes free radical.In addition, lotus seed protein oligopeptide can be applicable to multiple industries, before market
Scape is boundless.
Documents 1: application No. is " CN201310407100.1 ", entitled " lotus seed protein powder and its production method " is raw
Producing step is: 1. lotus seeds are checked and accepted: picking seven to medium well fresh shell lotus；2. crushing；3. salt proposes the heavy centrifugation of acid: after extraction
Mixed liquor carries out the adjusting of pH value with HCl, NaOH, and adjusting its pH value is 4.2.4. stirring, centrifugal filtration；5. ultrafiltration: by filtrate
It is concentrated by ultrafiltration, obtains the protein of high concentration；6. drying: being dried to obtain finished product albumen powder with spray dryer；⑦
Packaging: vacuum packaging.Lotus seed protein powder of the invention and preparation method thereof compared with prior art, generated beneficial effect
Be: 1) lotus seed protein powder be sufficiently reserved lotus seeds fragrant, white to it is light yellow, reconstituted with hot water after for homogeneous paste, mouth
Sense is fine and smooth.2) simple production process of the lotus seed protein powder is convenient for large-scale production, packaging and transport, sale.3) convenient for guarantor
It deposits, the room temperature lower shelf-life was up to 12 months.However the lotus seed protein of the case extracts and uses the recovery rate for subtracting salt-extracting method not high；
And the protein extracted, without desalting, mouthfeel is bad.
Documents 2: document " lotus seed protein matter Study on extraction~(9) Zhang Yu: Food Science .2007,28: 144~
147 " the techniques use alkali extraction and acid precipitation, solid-liquid ratio 1:25 (lotus nut starch: water), pH11 (alkali carries), extract 3h, protein at 40 DEG C
Recovery rate is up to 86.15%, however this method takes highly basic to extract, and it is deeper to obtain protein color, and poor taste simultaneously generates a large amount of useless
Liquid, post processing cost are high.
The lotus seed protein extraction process of CN101215316A and CN101215318A, takes microwave and ultrasound auxiliary salt to mention
CO2Or HCl acid sinks to obtain lotus seed protein, these technique extraction times are short, low energy consumption, yield height, but are difficult to industrialized production at present,
And the high requirements on the equipment, it puts into larger.The lotus seed protein peptide preparation process of CN101497913A, takes Ca (OH)2Instead of NaOH
Alkali carries technique, lack the processing step of removal foreign ion, CaCl can be remained in obtained protein peptides2, will affect product matter
Amount and mouthfeel.
In conclusion it is domestic at present seldom about the report of lotus seed protein oligopeptide preparation process and patent/patent application,
It is most of all to there is a problem of that yield is not high and impurity is more.
Summary of the invention
It is an object of the invention to overcome the defect of the prior art, a kind of lotus seed protein oligopeptide and its preparation side are provided
Method, the oligomeric peptide yield of the lotus seed protein that the present invention is prepared is up to 78.6%, and appearance luster and mouthfeel quality are all good.
The present invention is implemented as follows:
The present invention provides a kind of preparation method of lotus seed protein oligopeptide, includes the following steps:
Step 1, immersion, defibrination: it will go heart lotus seeds after warm water immersion, be mixed with water in flour mill and carry out wet-milling
It is broken, grind slurry sieving；
Step 2, salt extraction: it is that 1:10~20 adds NaCl to 0.2~0.5mol/L by feed liquid mass volume ratio, adjusts molten
Liquid pH value extracts lotus nut starch 2h~4h, filters and be centrifuged to obtain supernatant to 8~10,30 DEG C~45 DEG C of Extracting temperature；
Step 3, acid are heavy: the HCl adjusting supernatant pH value that addition concentration is 0.25~0.5mol/L to 4.3~
4.7, solution left standstill precipitating after centrifugation, obtains lotus seed protein sediment；
Step 4, enzymatic hydrolysis: it is 8%~16% that concentration of substrate will be added with water in the lotus seed protein sediment, temperature setting
At 35 DEG C~45 DEG C, PH is adjusted to 3~8, and protease is added, and the protease is neutral proteinase and papain, the egg
The total addition level of white enzyme is the 1%~5% of the quality of the lotus seed protein sediment, and the neutral proteinase and the wood
The mass ratio of melon protease is 1:1.5~2.5, digests 2~4h, obtains lotus seed protein peptide liquid；
Step 5, enzyme deactivation: the lotus seed protein polypeptide liquid is heated, and enzyme deactivation is living；
Step 6, dialysis: lotus seed protein polypeptide liquid is dialysed, and the lotus seed protein peptide liquid that desalts is obtained；
Step 7, film filtering: retention degree is set in 1500Da, and protein peptides liquid is obtained by filtration and carries out being concentrated to give concentrate, gained
The concentration of concentrate is 25%~45%；
The concentrate is dried step 8, obtains lotus seed protein oligopeptide.Purity is obtained greater than 97%, average molecular
Quality is less than the lotus seed protein oligopeptide of 1500Da.
The invention has the following advantages:
1, the preparation method of a kind of lotus seed protein oligopeptide provided by the invention by adding NaCl, and adjusts solution PH
Value is 8~10 to extract lotus seed protein, greatly reduces influence of the highly basic to lotus seed protein, reduces protein degradation and coagulates
It is poly-, the denaturation of lotus seed starch in the process is reduced, gained protein appearance luster, mouthfeel quality are all good, while remaining shallow lake
Powder can the higher product of continual exploitation added value；Specifically:
The applicant is by a large amount of innovative exploration discovery, when changing in the range of 5.0~13.0 with pH value, lotus
Sub- extraction rate of protein is in rising trend.In influence from pH to recovery rate, although recovery rate highest when pH is 13.0, with
Recovery rate alkalinity becomes strong, and lotus seed protein matter is easier to be precipitated, but the excessively high denaturation and degradation for easily causing protein of alkalinity, obtained lotus
Sub- albumen quality is poor.In view of application in the food industry, pH is excessively high to make its application have limitation (equipment and behaviour
Make), extract pH control 8~10, it is higher and more stable in this section recovery rate, equipment requirement is reduced, is reduced strong
The use of alkali.
2, the preparation method of a kind of lotus seed protein oligopeptide provided by the invention, by a large amount of innovative spy of the applicant
Suo Faxian is digested simultaneously using neutral proteinase and both enzymes of papain, and control the neutral proteinase with
The mass ratio of the papain is 1:1.5~2.5, and the total addition level of the protease is the lotus seed protein sediment
The 1%~5% of quality；First and most important, the degree of hydrolysis of albumen, the yield of polypeptide can be improved to the maximum extent.
Second, the scope of application of neutral proteinase and papain is wider, and enzymolysis process can be not required to adjust pH, simplifies hydrolysis process
Condition reduces production cost, eliminates neutralization procedure.Third is 8%~16% by the way that concentration of substrate is further arranged, temperature
Degree is set in 35 DEG C~45 DEG C, and PH is adjusted to 3~8, comes further accurate controlled enzymatic hydrolysis condition, hydrolysis and reaches the degree of requirement, both
Will not excessive hydrolysis generate small molecule bitter peptides, and final product quality can be improved.The molar mass average of product is controlled in 800D or so
(can detect 5000D or more molecular weight content≤2.9%), protein input-output ratio reaches 97% or more.
3, the present invention can remove the NaCl that extraction is added by dialysis, moreover it is possible to other ions brought into raw material are removed,
Improve product mouthfeel；The albumen required simultaneously by film filtering (retention degree is set in 1500Da) retention not up to hydrolysis, removal
Human body is nondigestible and the macro-molecular protein that absorbs, improves the purity of target protein peptide.
4, the oligomeric peptide yield of lotus seed protein that the present invention is prepared is up to 78.6%, and appearance luster and mouthfeel quality
It is all good.
Detailed description of the invention
Fig. 1 is the recovery rate of the lotus seed protein matter under different pH conditions.
Specific embodiment
Embodiment 1
Heart lotus seeds 3h is removed in the water immersion for being 35 DEG C with temperature, and lotus seeds and water quality ratio are 1:3, is uniformly added into fiberizer progress
Wet-milling, defibrination fineness are 60 mesh；Lotus nut starch is extracted with 0.3mol/LNaCl solution, solid-liquid ratio 1:10 adjusts pH to 10, extraction
2.5h, filters off screening, centrifugal filtrate, and centrifugation rate 2500rpm is centrifuged 25min, obtains supernatant；With the HCl of 0.25mol/L
Supernatant pH to 4.5 is adjusted, 1.0h, centrifugation rate 2500rpm are staticly settled, 15min is centrifuged, obtains lotus seed protein sediment；Lotus seeds
Albumen precipitation object is added with water to concentration 10%, adjusts pH to 6.0, and neutral proteinase and papain dosage are protein by weight
3.5%, and the mass ratio of the neutral proteinase and the papain is 1:2, hydrolysis temperature is 38 DEG C, digests 4h；It will
Polypeptide liquid is warming up to 85 DEG C, keeps 15min, albumen enzyme-deactivating；Polypeptide liquid is placed in circulation dialysis slot, set temperature
For 35 DEG C of dialysis 6h, dialysate flow rate 100mL/min, must desalt lotus seed protein peptide；Filter membrane retention degree is 1500Da, filtering
Pressure 0.1MPa, set temperature are 40 DEG C, obtain protein peptides liquid and are concentrated into concentration 30%；Feed liquid temperature is 55 DEG C, spraying import
125 DEG C of temperature, temperature is spray-dried under the conditions of being 75 DEG C, 80 DEG C of exhaust outlet temperature in tower, obtains the lotus seed protein that purity is 96%
Oligomeric Gly-His-Lys.Lotus seed protein oligopeptide (oligopeptide that molecular weight the is less than or equal to 1500Da) yield being prepared is 78.6%,
And appearance luster and mouthfeel quality are all good.
Embodiment 2
Heart lotus seeds 3.5h is removed in the water immersion for being 40 DEG C with temperature, and lotus seeds and water quality ratio are 1:3, be uniformly added into fiberizer into
Row wet-milling, defibrination fineness are 70 mesh；Lotus nut starch is extracted with 0.4mol/LNaCl solution, solid-liquid ratio 1:12 adjusts pH to 9, extraction
3h, filters off screening, centrifugal filtrate, and centrifugation rate 3000rpm is centrifuged 20min, obtains supernatant；With the HCl tune of 0.3mol/L
Supernatant pH to 4.4 staticly settles 1.0h, centrifugation rate 3000rpm, is centrifuged 10min, obtains lotus seed protein sediment；Lotus seeds egg
White sediment is added with water to concentration 12%, adjusts pH to 5.5, and neutral proteinase and papain dosage are protein by weight
4.5%, and the mass ratio of the neutral proteinase and the papain is 1:1.5, hydrolysis temperature is 42 DEG C, enzymatic hydrolysis
3.5h；Polypeptide liquid is warming up to 90 DEG C, keeps 10min, albumen enzyme-deactivating；Polypeptide liquid is placed in circulation dialysis slot,
Set temperature is 40 DEG C of dialysis 5h, dialysate flow rate 150mL/min, and must desalt lotus seed protein peptide；Filter membrane retention degree is
1500Da, filter pressure 0.15MPa, set temperature are 35 DEG C, obtain protein peptides liquid and are concentrated into concentration 40%；Feed liquid temperature is
60 DEG C, 130 DEG C of spray inlet temperature, temperature is spray-dried under the conditions of being 70 DEG C, 75 DEG C of exhaust outlet temperature in tower, and obtaining purity is
The oligomeric Gly-His-Lys of 97% lotus seed protein.The lotus seed protein oligopeptide (oligopeptide that molecular weight is less than or equal to 1500Da) being prepared
Yield is 78.2%, and appearance luster and mouthfeel quality are all good.
Embodiment 3
Heart lotus seeds 3h is removed in the water immersion for being 45 DEG C with temperature, and lotus seeds and water quality ratio are 1:5, is uniformly added into fiberizer progress
Wet-milling, defibrination fineness are 70 mesh；Lotus nut starch is extracted with 0.5mol/LNaCl solution, solid-liquid ratio 1:20 adjusts pH to 8, extraction
3h, filters off screening, centrifugal filtrate, and centrifugation rate 3000rpm is centrifuged 20min, obtains supernatant；With the HCl tune of 0.5mol/L
Supernatant pH to 4.7 staticly settles 1.0h, centrifugation rate 3000rpm, is centrifuged 10min, obtains lotus seed protein sediment；Lotus seeds egg
White sediment is added with water to concentration 16%, adjusts pH to 6.0, and neutral proteinase and papain dosage are protein by weight
5%, and the mass ratio of the neutral proteinase and the papain is 1:2.5, hydrolysis temperature is 45 DEG C, digests 3h；It will
Polypeptide liquid is warming up to 90 DEG C, keeps 10min, albumen enzyme-deactivating；Polypeptide liquid is placed in circulation dialysis slot, set temperature
For 40 DEG C of dialysis 5h, dialysate flow rate 150mL/min, must desalt lotus seed protein peptide；Filter membrane retention degree is 1500Da, filtering
Pressure 0.15MPa, set temperature are 35 DEG C, obtain protein peptides liquid and are concentrated into concentration 40%；Feed liquid temperature is 60 DEG C, spraying import
130 DEG C of temperature, temperature is spray-dried under the conditions of being 70 DEG C, 75 DEG C of exhaust outlet temperature in tower, obtains the lotus seed protein that purity is 98%
Oligomeric Gly-His-Lys.Lotus seed protein oligopeptide (oligopeptide that molecular weight the is less than or equal to 1500Da) yield being prepared is 77.8%,
And appearance luster and mouthfeel quality are all good.
Comparative example 1
Except enzymolysis process replaces with: lotus seed protein sediment is added with water to concentration 10%, adjusts pH to 6.0, neutral proteinase
It is the 3.5% of protein by weight with papain dosage, and the mass ratio of the neutral proteinase and the papain
For 1:1, hydrolysis temperature is 38 DEG C, digests 4h；Other steps are the same as embodiment 1.
Comparative example 2
Except enzymolysis process replaces with: lotus seed protein sediment is added with water to concentration 10%, adjusts pH to 6.0, neutral proteinase
It is the 3.5% of protein by weight with papain dosage, and the mass ratio of the neutral proteinase and the papain
For 1:3, hydrolysis temperature is 38 DEG C, digests 4h；Other steps are the same as embodiment 1.
Experimental example 1
It can be seen that by attached drawing 1, in the case that Extracting temperature, time, salinity are constant, as pH value is 5.0~13.0
In range when variation, lotus seed protein matter recovery rate is in rising trend.PH value 5.0 is that recovery rate is minimum, predicts the pH value and lotus seeds
The isoelectric point of protein is closer to, and it 5.0 is that recovery rate constantly increases that pH value, which is higher than, as pH13.0, recovery rate close to 80%, this
Illustrate that lotus seed protein matter is easier to be precipitated under alkaline condition, recovery rate is higher.
In influence from pH to recovery rate, although recovery rate highest when pH is 13.0, as recovery rate alkalinity becomes strong, lotus
Sub- protein is easier to be precipitated, but the excessively high denaturation and degradation for easily causing protein of alkalinity, obtained lotus seed protein are second-rate.It examines
Consider application in the food industry, pH is excessively high to make its application have limitation (equipment and operation).Extract pH control 8~
10, it is higher and more stable in this section recovery rate, equipment requirement is reduced, the use of highly basic is reduced.
Experimental example 2
The yield of the degree of hydrolysis of Statistics Implementation example 1~3 and comparative example 1~2, last resulting lotus seed protein oligopeptide, such as
The following table 1.
Table 1
| Group | Degree of hydrolysis (%) | Oligomeric peptide yield (%) | Mouthfeel |
| Comparative example 1 | 10.8 | 30.8 | Without bitter taste |
| Comparative example 2 | 9.3 | 36.2 | Without bitter taste |
| Embodiment 1 | 60.1 | 78.6 | Without bitter taste |
Shown by upper table 1 by a large amount of innovative exploration discovery of the applicant, using neutral proteinase and Papain
Both enzymes of enzyme are digested simultaneously, and the mass ratio for controlling the neutral proteinase and the papain be 1:1.5~
2.5, the yield of enough degree of hydrolysis for improving albumen to the maximum extent, polypeptide；The present invention by further be arranged concentration of substrate be
8%~16%, temperature is set in 35 DEG C~45 DEG C, and PH is adjusted to 3~8, comes further accurate controlled enzymatic hydrolysis condition, hydrolysis and reaches
It is required that degree, not only will not excessive hydrolysis generate small molecule bitter peptides, but also final product quality can be improved.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of preparation method of lotus seed protein oligopeptide, which comprises the steps of:
Step 1, immersion, defibrination: will go heart lotus seeds after warm water immersion, and the progress wet-milling in flour mill that is mixed with water is broken,
Slurry sieving is ground；
Step 2, salt extraction: it is that 1:10~20 adds NaCl to 0.2~0.5mol/L by feed liquid mass volume ratio, adjusts solution PH
Value extracts lotus nut starch 2h~4h, filters and be centrifuged to obtain supernatant to 8~10,30 DEG C~45 DEG C of Extracting temperature；
Step 3, acid are heavy: the HCl adjusting supernatant pH value that addition concentration is 0.25~0.5mol/L to 4.3~4.7, solution
It staticly settles, after centrifugation, obtains lotus seed protein sediment；
Step 4, enzymatic hydrolysis: it is 8%~16% that concentration of substrate will be added with water in the lotus seed protein sediment, and temperature is set in 35
DEG C~45 DEG C, PH is adjusted to 3~8, and protease is added, and the protease is neutral proteinase and papain, the protease
Total addition level be the lotus seed protein sediment quality 1%~5%, and the neutral proteinase and the Papain
The mass ratio of enzyme is 1:1.5~2.5, digests 2~4h, obtains lotus seed protein peptide liquid；
Step 5, enzyme deactivation: the lotus seed protein polypeptide liquid is heated, and enzyme deactivation is living；
Step 6, dialysis: lotus seed protein polypeptide liquid is dialysed, and the lotus seed protein peptide liquid that desalts is obtained；
Step 7, film filtering: retention degree is set in 1500Da, and protein peptides liquid is obtained by filtration and carries out being concentrated to give concentrate, gained concentration
The concentration of liquid is 25%~45%；
The concentrate is dried step 8, obtains lotus seed protein oligopeptide.
2. the preparation method of lotus seed protein oligopeptide as described in claim 1, which is characterized in that in the step 1, the temperature
The temperature that water impregnates is 25 DEG C~45 DEG C, and soaking time is 2h~4h；Lotus seeds are uniformly mixed with water according to the mass ratio of 1:2~5, and one
It plays that carry out wet-milling in addition flour mill broken, has ground and crossed 40~80 meshes after slurry.
3. the preparation method of lotus seed protein oligopeptide as described in claim 1, which is characterized in that in the step 2, centrifugation speed
Rate is 1500rpm~5000rpm, and centrifugation time is 20min~45m i n.
4. the preparation method of lotus seed protein oligopeptide as described in claim 1, which is characterized in that described quiet in the step 3
The time for setting precipitating is 0.5~2h；The centrifugation rate is 2000rpm~4000rpm, and centrifugation time is 15min~45min.
5. the preparation method of lotus seed protein oligopeptide as described in claim 1, which is characterized in that in the step 4, it is described in
Property protease and the papain mass ratio be 1:2.
6. the preparation method of lotus seed protein oligopeptide as described in claim 1, which is characterized in that described to add in the step 5
Hot temperature is 75 DEG C~95 DEG C, and heating time is 10min~20min.
7. the preparation method of lotus seed protein oligopeptide as described in claim 1, which is characterized in that described in the step 6
The specific steps of analysis are as follows: lotus seed protein polypeptide liquid is placed in circulation dialysis slot, temperature is set in 25 DEG C~45 DEG C, dialysis time 4
~8h, dialysate flow rate are set in 100~200mL/min.
8. the preparation method of lotus seed protein oligopeptide as described in claim 1, which is characterized in that in the step 7, the film
The pressure of filtering is 0.05MPa~0.25MPa, and temperature is 25 DEG C~45 DEG C.
9. the preparation method of lotus seed protein oligopeptide as described in claim 1, which is characterized in that described dry in the step 8
It is dry for spray drying, and the condition of the spray drying are as follows: 45 DEG C~75 DEG C of feed liquid temperature, spray inlet temperature 125 DEG C~130
DEG C, temperature is 65 DEG C~85 DEG C in tower, and exhaust outlet temperature is 70 DEG C~85 DEG C.
10. a kind of lotus seed protein oligopeptide that the method as described in claim 1~9 is any is prepared.
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| CN111607629A (en) * | 2020-06-05 | 2020-09-01 | 福建农林大学 | Lotus seed active polysaccharide, lotus seed active substance, extraction method and application thereof |
| CN114907623A (en) * | 2022-05-24 | 2022-08-16 | 华中农业大学 | Non-covalent crosslinking edible film of protein hydrolysate-polyphenol-esterified polysaccharide and preparation method thereof |
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