CN114921516B - Chinese lobster immunocompetent peptide, and preparation method and application thereof - Google Patents

Chinese lobster immunocompetent peptide, and preparation method and application thereof Download PDF

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CN114921516B
CN114921516B CN202210581153.4A CN202210581153A CN114921516B CN 114921516 B CN114921516 B CN 114921516B CN 202210581153 A CN202210581153 A CN 202210581153A CN 114921516 B CN114921516 B CN 114921516B
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李晓晖
杨志艳
李燕
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Shanghai Ocean University
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Abstract

The invention provides a Chinese lobster immune active peptide and a preparation method and application thereof, wherein the preparation method comprises the following steps: crushing Chinese lobster, homogenizing, adding distilled water, mixing, and inactivating enzyme in water bath; adding protease for enzymolysis, and inactivating enzyme in water bath again after the enzymolysis is finished; centrifuging or sieving the enzymolysis solution, collecting supernatant, separating by ceramic membrane, ultrafiltration and nanofiltration, and drying; the Chinese lobster immunocompetent peptide powder has important regulation function on immune system recovery of an immunosuppressive mouse, immune organ index, peripheral blood leukocyte number and serum immunoglobulin level of the immunosuppressive mouse are obviously increased, and NK cell killing activity is enhanced, so that the Chinese lobster peptide powder is capable of regulating cyclophosphamide-induced low immune function of the mouse, improving the immunity of the organism, mediating a liquid immune regulation mechanism, participating in inherent immunity and adaptive immunity of the organism, developing immunity-enhancing foods pertinently, and improving the immunity of people undergoing radiotherapy and chemotherapy.

Description

Chinese lobster immunocompetent peptide, and preparation method and application thereof
Technical Field
The invention belongs to the technical field of ocean functional food processing, and particularly relates to a Chinese lobster immunocompetent peptide, a preparation method and application thereof.
Background
The crayfish is small-sized planktonic shrimp of crayfish genus of Oriental cherry shrimp family, and has short life cycle, strong fertility, rapid generation update, high nutritive value, and unique flavor. The main varieties of the Penaeus vannamei Boone are Chinese Penaeus vannamei Boone (acetate Chinese) and Penaeus japonicus (Acetes japonicas Kishinouye) which occupy about 95% in the coastal areas of east China sea, bohai sea and the like, and the output of the Chinese Penaeus vannamei Boone in 2013-2020 is wholly improved according to the statistical annual-culture data of Chinese fishery, and reaches 630.73 ten thousand tons by 2020, and is increased by 3.94% in the same proportion. The shrimp species caught in the ocean in China are mainly Penaeus vannamei Boone, prawn, mantis shrimp and hawk claw shrimp. Wherein, the shrimp catching yield is highest, the yield in 2020 is 36.74 ten thousand tons, and the ratio of the shrimp catching yield to the total yield is 30.45 percent.
The Chinese lobster has high nutritive value, protein content up to 72.9% (dry basis), rich amino acid composition, rich trace elements such as calcium, phosphorus, magnesium, iron and the like required by human bodies, and high vitamin B5 and vitamin E content. The fresh shrimp shell is thin, small, the water content is above 80%, the fresh shrimp shell is difficult to process and easy to deteriorate, so that the traditional shrimp products supplied in the market mainly comprise dried shrimp shells, the water content of the dried shrimp is usually 25-40%, the water content is higher, and the problems of easy spoilage of microorganisms, standard exceeding of fat oxidation and color change exist in the purification process.
The Chinese shrimp is a marine low-value shrimp with extremely high nutritive value, and the unique amino acid composition of the Chinese shrimp has great development and utilization value in the aspects of taste, nutrition and health care. In recent years, domestic scholars have studied the process of preparing the polypeptides of penaeus monodon.
Researches on neutral and alkaline protease enzymolysis conditions of Chinese lobsters such as Pacific snow and the like show that the Chinese lobster polypeptides can obviously inhibit the activity of ACE in vitro, obviously reduce the arterial blood pressure of rats with renal vascular hypertension, have better antihypertensive effect, and have stronger antihypertensive effect than neutral enzymolysis polypeptides. The post Xue Changhu subject group also carries out separation and identification on the structure of the shrimp active peptide, the enzymolysis product of neutral protease 3942 obtains Ser-Pro (IC50=272 mu mol/L) and the enzymolysis product of flavobacterium low-temperature alkaline protease obtains Pro-Arg-Tyr (IC50=212 mu mol/L) two active peptide sequences. Cao Wengong and the like firstly study enzymolysis conditions and product functions of Chinese lobsters, and separate components obtained by enzymolysis of alkaline protease alcaise 2.4L, and the components with strongest scavenging activity on hydroxyl radicals are found to account for 21.4 percent, the molecular weight is mainly concentrated between 2259 Da and 869Da, the IC50 is 0.385mg/mL, and the peptide chain length is 22.4-7.7.
Chinese patent CN200610108297.9 (Chinese lobster protein antihypertensive peptide, preparation method and application thereof) separates and identifies 3 antihypertensive peptides with 6 amino acids from the proteolytic products of Chinese lobsters, has higher Angiotensin Converting Enzyme (ACE) inhibitory activity, and artificially synthesizes related 12 polypeptides.
The current patent and research mainly focus on the enzymolysis conditions of Chinese lobsters and the research on free radical removal and blood pressure reduction functions. The enzymes used are mainly neutral protease, alkaline protease and the like, so that more enzymolysis processes and physiological activities in the Chinese lobsters need to be researched, and the comprehensive utilization value of the Chinese lobsters is further improved.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a Chinese lobster immunocompetent peptide, and a preparation method and application thereof.
To achieve the above object, the solution of the present invention is:
in a first aspect, the invention provides a preparation method of Chinese lobster immunoactive peptide, which comprises the following steps:
(1) Crushing Chinese lobster, homogenizing, adding distilled water, mixing, and inactivating enzyme in water bath;
(2) Adding protease for enzymolysis, and inactivating enzyme in water bath again after the enzymolysis is finished;
(3) And cooling the enzymolysis liquid to normal temperature, centrifuging or sieving, and drying to obtain the Chinese lobster immunocompetent peptide.
Preferably, in step (1), when distilled water is added, the feed liquid ratio is 1:1 to 1:6, more preferably 1:2 to 1:4.
Preferably, in the step (1) and the step (2), the temperature of enzyme deactivation in the water bath is 90-95 ℃ and the time is 10-15min.
Preferably, in step (2), the protease is selected from more than one of bromelain, complex flavourzyme, papain, trypsin or neutral protease.
Preferably, in step (2), the temperature of the enzymatic hydrolysis is 50-55deg.C for 2-5 hours, preferably 2-4 hours.
Preferably, in step (3), the rotational speed of centrifugation is 8000-10000rpm for 15-20min.
Preferably, in step (3), the mesh number of the sieve is 100-200 mesh.
Preferably, in the step (3), the membrane separation operation is carried out after sieving, the membrane separation sequentially adopts ceramic microfiltration membranes with the membrane pore diameter of 0.2 mu m to clarify the fermentation liquor, namely bacteria and insoluble macromolecules with the size of about 1-22 mu m are removed, and the percolate is concentrated and separated by an ultrafiltration membrane with the molecular weight cutoff of 3000-5000Da, an RO reverse osmosis membrane or a nanofiltration membrane with the molecular weight cutoff of 150-500Da, so as to obtain concentrated trapped liquor.
Preferably, in the step (3), the trapped fluid can be subjected to freeze drying and spray drying to obtain the Chinese lobster immunoactive peptide. The spray drying condition is that the sample inlet temperature is 180 ℃ and the outlet temperature is 80-118 ℃.
In a second aspect, the invention provides a preparation method of Chinese lobster immunoactive peptide, which is obtained by the preparation method.
In a third aspect, the invention provides an application of an immunocompetent peptide such as Chinese lobster in the field of preparation of immunity-enhancing foods.
By adopting the scheme, the invention has the beneficial effects that:
the molecular weight of the Chinese lobster immune active peptide is more than 90 percent with the ratio of <1000 Da; the dry weight of the polypeptide powder is 50 percent of the dry weight of the penaeus monodon, and the protein content is more than 80 percent; the molecular weight content of small molecules <1000Da in the nanofiltration trapped fluid is increased to 91.49% from 54.82% of bromelain hydrolysate, and the salinity is reduced by 60.62% through a ceramic membrane, ultrafiltration and nanofiltration three-stage filtration system.
The Chinese lobster immunocompetent peptide powder has an important regulation effect on immune system recovery of an immunosuppressed mouse, immune organ index, peripheral blood leukocyte number and immunoglobulin (IgA, igG and IgM) level in serum of the immunosuppressed mouse are obviously increased, and cell factors (TNF-alpha, IL-2 and IL-6) are obviously increased, so that NK cell killing activity is enhanced. The Chinese lobster polypeptide powder is shown to be capable of regulating cyclophosphamide-induced low mouse immunity, improving the body immunity, mediating a liquid immunity regulating mechanism, participating in the inherent immunity and adaptive immunity of the body, and can pertinently develop immunity-enhancing foods and improve the immunity of people undergoing radiotherapy and chemotherapy.
Drawings
FIG. 1 is a graph showing the molecular weight distribution of the polypeptides of the present invention after enzymatic hydrolysis with different proteases.
FIG. 2 is a graph showing the effect of the powder of the immunocompetent peptide of the invention on the content of immunoglobulins in serum of immunosuppressive mice ((a) IgA, (b) IgG, (c) IgM; note that different letters represent significant differences between groups (p < 0.05)).
FIG. 3 is a graph showing the effect of the powder of the immunocompetent peptide of the invention on the cytokine content in serum of immunosuppressive mouse cells ((a) IL2, (b) IL6, (c) INF-alpha; note: different letters indicate significant differences between groups (p < 0.05)).
FIG. 4 is a graph of peripheral blood leukocyte counts for mice of each group of the present invention (note: different letters indicate significant differences between groups (p < 0.05)).
FIG. 5 is a graph showing the effect of the Chinese lobster immunocompetent peptide powder of the present invention on the activity of NK cells of immunosuppressed mice (note: different letters indicate significant differences between groups (p < 0.05)).
FIG. 6 is a graph of molecular weight distribution (a) and desalting effect of nanofiltration retentate of the present invention through a ceramic membrane, ultrafiltration and nanofiltration three stage filtration system.
Detailed Description
The invention provides a Chinese lobster immune active peptide and a preparation method and application thereof.
< preparation method of immunocompetent peptide of Chinese Mao shrimp)
The preparation method of the Chinese lobster immunoactive peptide comprises the following steps:
(1) Taking out frozen Chinese lobster (-80 ℃/-20 ℃ for preservation), naturally thawing, picking to remove impurities, and pulping; or crushing dried Chinese shrimp product (dried small shrimps), homogenizing, weighing minced shrimp, adding distilled water, mixing, and inactivating enzyme in water bath;
(2) Adding 1000-5000U/g (preferably 2000U/g) protease (as enzyme for hydrolysis) according to 1-2% of the weight of Chinese lobster, performing constant temperature enzymolysis in a magnetic stirrer for 2h, and inactivating enzyme in water bath again after enzymolysis;
(3) And cooling the enzymolysis liquid to normal temperature, centrifuging or sieving, and drying to obtain the Chinese lobster immunocompetent peptide.
Wherein, in the step (1), when distilled water is added, the feed liquid ratio (w/v) may be 1:1 to 1:6, preferably 1:2 to 1:4, more preferably 1:4.
In step (1) and step (2), the temperature of the water bath enzyme deactivation may be 90-95 ℃, preferably 90 ℃; the time may be 10-15min, preferably 15min.
In the step (2), the protease is selected from more than one of bromelain, compound flavor protease, papain, trypsin or neutral protease. Proteases are capable of producing immunologically active hydrolysates as shown in figure 1.
In step (2), the temperature of the enzymolysis may be 50-55deg.C, preferably 55deg.C; the time may be 2 to 5 hours, preferably 2 to 4 hours, and more preferably 3.5 hours.
In the step (3), the purpose of centrifugation is to remove the skin and shell of macromolecules and shrimps, and sieving only removes skin and slag, but not insoluble macromolecular substances, and then filtering with a ceramic membrane to remove further slag and macromolecular insoluble proteins. The active peptide with immune effect can be obtained by centrifugation or sieving, and the specific protein content and the molecular weight composition have different, but do not influence the immune activity and the product development and application.
In step (3), the rotational speed of centrifugation may be 8000-10000rpm, preferably 10000rpm; the time may be 15-20min, preferably 20min.
In step (3), the sieved screen is 100-200 mesh.
In the step (3), the membrane separation operation is carried out after sieving, the membrane separation sequentially adopts ceramic microfiltration membranes with the membrane pore diameter of 0.2 mu m to clarify the fermentation liquor, namely bacteria and insoluble macromolecules with the size of about 1-22 mu m are removed, and the percolate is concentrated and separated by ultrafiltration membranes with the molecular weight cutoff of 3000-5000Da, RO reverse osmosis membranes or nanofiltration membranes with the molecular weight cutoff of 150-500Da, so as to obtain concentrated trapped liquor.
As shown in fig. 6, the molecular weight of the nanofiltration trapped fluid, which is small molecules of <1000Da, is increased from 54.82% of bromelain hydrolysate to 91.49% and the salinity is reduced by 60.62% through a ceramic membrane, ultrafiltration and nanofiltration three-stage filtration system.
In the step (3), the trapped fluid can be subjected to freeze drying and spray drying to obtain the Chinese lobster immunoactive peptide. The spray drying condition is that the sample inlet temperature is 180 ℃ and the outlet temperature is 80-118 ℃.
In the step (3), the Chinese lobster polypeptide enzymolysis liquid can be directly dried to obtain the active peptide. Or further separation and purification are adopted, so that the molecular weight range of the target polypeptide is further reduced, and the content of the active peptide is higher.
< Chinese Mao shrimp immunocompetent peptide >
The Chinese lobster immunoactive peptide is obtained by the preparation method.
< application of immunocompetent peptide of Chinese Mao shrimp >
The Chinese lobster immunocompetent peptide is applied to the field of preparation of immunity enhancing foods.
The technical contents of the present invention are further described below with reference to examples. The following examples are illustrative, not limiting, and are not intended to limit the scope of the invention. The experimental methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1:
the preparation method of the Chinese lobster immunoactive peptide of the embodiment comprises the following steps:
(1) And taking out 50Kg of frozen Chinese lobsters, naturally thawing, crushing the frozen Chinese lobsters to minced shrimps, adding 200Kg of water according to the feed-liquid ratio of 1:4, uniformly mixing the mixture, and inactivating enzyme in a water bath at 90 ℃ for 15min.
(2) 500g bromelain is added, and the enzymolysis is carried out for 3.5 hours at 55 ℃ in a magnetic stirrer, and after the enzymolysis is finished, the enzyme is inactivated in a water bath at 90 ℃ for 15min.
(3) Cooling the enzymolysis liquid to normal temperature, sieving with a 200-mesh sieve, filtering to remove residues by a ceramic membrane of 0.2 mu m, ultrafiltering by a 3000Da ultrafilter membrane to obtain small molecular polypeptide, ultrafiltering by a 150Da nanofiltration membrane, and freeze-drying or spray-drying the retentate obtained by nanofiltration, wherein the inlet temperature of a spray dryer is 180 ℃, and the outlet temperature of the spray dryer is 118 ℃, so as to obtain the Chinese lobster immune active peptide.
Example 2:
the preparation method of the Chinese lobster immunoactive peptide of the embodiment comprises the following steps:
(1) And taking out 50Kg of frozen Chinese lobsters, naturally thawing, crushing the frozen Chinese lobsters to minced shrimps, adding 200Kg of water according to the feed-liquid ratio of 1:4, uniformly mixing the mixture, and inactivating enzyme in a water bath at 90 ℃ for 15min.
(2) 500g bromelain is added, and the enzymolysis is carried out for 3.5 hours at 55 ℃ in a magnetic stirrer, and after the enzymolysis is finished, the enzyme is inactivated in a water bath at 90 ℃ for 15min.
(3) Cooling the enzymolysis liquid to normal temperature, and centrifuging at high speed (the centrifugal speed is 10000rpm, and the time is 20 min); concentrating, spray drying, wherein the inlet temperature of the spray dryer is 180deg.C, the outlet temperature is 118 deg.C, and drying to obtain Chinese lobster immunocompetent peptide. The mass yield of the Chinese lobster immune active peptide reaches 40-50% of the dry weight of the lobster, and the protein content is 78-82%.
Example 3:
the preparation method of the Chinese lobster immunoactive peptide of the embodiment comprises the following steps:
(1) And taking out 50Kg of frozen Chinese lobsters, naturally thawing, crushing the frozen Chinese lobsters to minced shrimps, adding 200Kg of water according to the feed-liquid ratio of 1:4, uniformly mixing the mixture, and inactivating enzyme in a water bath at 90 ℃ for 15min.
(2) 500g bromelain is added, and the enzymolysis is carried out for 3.5 hours at 55 ℃ in a magnetic stirrer, and after the enzymolysis is finished, the enzyme is inactivated in a water bath at 90 ℃ for 15min.
(3) Cooling the enzymolysis liquid to normal temperature, filtering the enzymolysis liquid by a 100-mesh sieve to remove slag, and performing ultrafiltration operation. Filtering the residue-removing enzymolysis liquid by a 100-mesh sieve to remove residues and macromolecular insoluble substances by a 0.2 mu m ceramic membrane in sequence, ultrafiltering by an ultrafiltration membrane with a molecular cut-off of 3000Da to obtain micromolecular polypeptide with higher purity, and performing freeze drying or spray drying on a sample solution by an RO reverse osmosis membrane, wherein the inlet temperature of a spray dryer is 180 ℃, the outlet temperature is 118 ℃, and drying to obtain the Chinese lobster immunoactive peptide.
Example 4:
the preparation method of the Chinese lobster immunoactive peptide of the embodiment comprises the following steps:
(1) And taking out 50Kg of frozen Chinese lobsters, naturally thawing, crushing the frozen Chinese lobsters to minced shrimps, adding 200Kg of water according to the feed-liquid ratio of 1:4, uniformly mixing the mixture, and inactivating enzyme in a water bath at 90 ℃ for 15min.
(2) Adding 500g bromelain, performing enzymolysis at 55deg.C in 100L enzymolysis container for 3.5 hr, and inactivating enzyme in water bath at 90deg.C for 15min.
(3) Cooling the enzymolysis liquid to normal temperature, sieving with a 150-mesh sieve to remove slag, and performing ultrafiltration operation. Filtering the enzymatic hydrolysate after deslagging by a ceramic membrane with the thickness of 0.2 mu m to remove micro slag and macromolecular insoluble substances, ultrafiltering by an ultrafiltration membrane with the molecular interception of 3000Da, and then performing ultrafiltration by a nanofiltration membrane with the molecular interception of 500Da, and performing freeze drying or spray drying on the interception liquid obtained by nanofiltration, wherein the inlet temperature of a spray dryer is 180 ℃, and the outlet temperature of the spray dryer is 118 ℃ to obtain the Chinese lobster immunoactive peptide.
Experiment 1:
animal test immune function evaluation of Chinese lobster immunocompetent peptide powder on immunosuppressed mice:
in order to further analyze the immunoregulatory function of the active peptide of the Chinese lobster, the invention carries out a mouse test on the polypeptide powder of the lobster in the embodiment. In vivo, the immunomodulatory activity of the shrimp polypeptides was evaluated. The mice were divided into 3 low, medium and high dose groups of 0.25g/kg BW, 0.5g/kg BW and 1.0g/kg BW. And (3) researching indexes of the shrimp polypeptide powder on organ indexes, serum immunoglobulin and cytokine levels, NK cell activities, total peripheral blood white cells and the like of the immunosuppressive mice, and evaluating the immunoregulation effect of the shrimp polypeptide powder on the immunosuppressive mice.
The selected lobster polypeptide powder is prepared by enzymolysis of bromelain, centrifugation and freeze drying, and the total nitrogen content is more than 14%, and the content of the lobster oligopeptide is more than 55%.
Animal test study content:
a model of immunocompromised mice was established using Cyclophosphamide (CTX) induction. Mice given levamisole hydrochloride are used as positive control groups, and other normal control groups are established according to low, medium and high dose groups of the lobster polypeptide powder. Mice were randomly grouped, 12 per group.
After the mice are adapted to the environment 7d, the five groups except the blank control group are irrigated with 80g/kg CTX for 3d, and the four groups are irrigated with gastric test samples of 0.25g/kg, 0.5g/kg and 1.0g/kg which are low, medium and high doses every day, the blank group and the model group are irrigated with 0.9% Normal Saline (NS) in equal quantity, the positive control group is irrigated with 20mg/kg levamisole hydrochloride (LMS) for 15d. See tables 1 and 2. After 24 hours from the last administration, treatment and related index measurement were performed.
TABLE 1 Effect of shrimp active peptide on immunosuppressive mouse body weight and immune organ index
Note that: the different letters represent significant differences between groups (p < 0.05), the following.
TABLE 2 grouping and administration modes
The results show that the shrimp polypeptide powder promotes the increase of thymus index and spleen index of the immunized mice under the condition of each dosage group. Also significantly increases (p < 0.05) serum IgA, igG and IgM levels of mice (as shown in figure 2) and enhances the immune regulation function of organisms (as shown in figure 3). NK cell activity was significantly increased (p < 0.05) (as shown in fig. 5), peripheral blood leukocyte levels in mice were significantly increased (p < 0.05) (as shown in fig. 4), and levels were restored to the level of the placebo group. The results show that the shrimp bioactive peptide has a regulating effect on the immune function of the immunosuppressed mice.
Tumor Necrosis Factor (TNF) generally refers to TNF-alpha, which has a significant damaging effect on tumor cells; chemotherapy drugs cause adverse reactions such as leukopenia due to various degrees of myelosuppression. The shrimp polypeptide powder can raise tumor necrosis factor TNF-alpha obviously and raise the leukocyte number of immunosuppressive mouse. The immune globulin and the cell factor produced by the organism are increased, so that the method has good application value and development prospect in the aspects of tumor treatment and recovery after illness, and lays a foundation for developing the active peptide of the Chinese lobster and related products.
The Chinese lobster immunocompetent peptide powder has remarkable effects on improving the antibody level of mice, promoting the release of cytokines and increasing the leukocyte level, can pertinently develop food for enhancing the immunity, and improves the immunity of people subjected to radiotherapy and chemotherapy.
Implementation 2:
development of Chinese lobster immune active peptide powder enhanced immune food:
the investigation of the eplerian shows that among the 10 major functional demands of consumers, the demands related to intestinal health and improving immunity account for 3, wherein the demands for 'enhancing immunity' are only secondary to digestion, the attention is as high as 37%, and consumers are more concerned about the improvement of immunity brought by ingestion on the demands for enhancing immunity functionality.
Aiming at the nutrition characteristics of small molecular weight, easy absorption, high protein content, trace elements, calcium content and the like of the shrimp peptide powder, the shrimp peptide powder has certain immunity enhancing capability, and three high-quality nutrition foods are developed.
<1> development of immunocompetent peptide powder meringue food of China's palaemon
Mixing the weighed white granulated sugar and Chinese lobster immunocompetent peptide powder uniformly, firstly, low-speed stirring egg white, stirring until the egg white is in a fish eye foam state, then adding the uniformly mixed sugar for three times, stirring at high speed after all the sugar is added, stirring the egg white until the egg white is in a hard foam state, and lifting and stirring the egg white to see an upright sharp corner. And then filling the decorating heads into decorating bags, and filling the prepared proteins into the decorating bags. And extruding the pattern on a baking tray paved with oiled paper. Placing the baking tray into an oven, and setting the upper and lower fires at 80 ℃ for 60min. The baked protein sugar is hardened, can be easily taken off from the oiled paper, and is placed into a sealing bag after cooling.
Raw materials: egg white: white granulated sugar: the ratio of the Chinese lobster immune active peptide powder is optimally 9:5:3, and the lobster crisp product is shaped after baking, has a yellow to dark yellow color, is instant after being eaten, does not adhere to teeth, has the special taste of the lobster, and has a certain delicate flavor.
Any change in the ratio range of the formula falls into the protection scope of the invention.
<2> development of immunocompetent peptide milk bean food of Chinese penaeus monodon
Milk beans, also known as pineapple beans, are very popular as food for infants and elderly people because of their ready-to-eat nature. The main raw material is milk powder and sucrose. The milk beans have low water content, so the milk beans are suitable for long-term storage. The milk soybean milk powder contains abundant proteins, can absorb and mask peculiar smell, but the traditional milk soybean products have high sucrose content and high sweetness, are not beneficial to human health and have low product demand. In addition, the traditional milk bean ingredients are declared to be few in functional ingredients, so that the traditional milk bean ingredients cannot meet the consumption requirements of the young generation, and the sales of products are small.
The invention develops low-sugar shrimp peptide milk beans, adopts mannitol to replace sucrose, and adds a certain amount of Chinese shrimp immunocompetent peptide powder, improves the product formula, has the effect of enhancing health while being suitable for special people with diabetes and the like, and provides an innovative thought for polypeptide powder utilization and milk bean product innovation.
The ingredients provided by the invention solve the problems of single product category, heavy bitter taste and less other nutritional ingredients of the existing bioactive peptide products, improve the addition of low-calorie sugar sweeteners such as milk powder, mannose and the like, increase the overall nutritional value and functional propaganda of the products, meet the health requirements of new-generation consumers, and increase the added value of traditional products.
The optimal formula of the Chinese lobster immune active peptide milk bean product comprises 70-80g of milk powder, 10-20g of Chinese lobster immune active peptide powder, 30-40g of mannitol and 1.5-2.5g of citric acid, and the finally prepared milk bean has smooth appearance, fragrant and strong smell, moderate soft and hard taste, toughness, sour and sweet taste and proper salty and fragrant taste.
Wherein, the preferable formula comprises: 80g of milk powder, 20g of Chinese lobster immunocompetent peptide powder, 40g of mannitol and 2.1g of citric acid.
<3> development of immunocompetent peptide cookie food for chinese lobster
The cookies are optimal: 100g of low gluten flour, 75g of butter, 35g of sugar powder, 20g of egg liquid and 10g of Chinese lobster immunocompetent peptide powder.
Any change in the ratio range of the formula falls into the protection scope of the invention.
The preparation of the shrimp peptide cookies comprises the following steps:
softening butter at room temperature, adding white sugar for whitening;
scattering eggs for later use;
sieving flour, chinese lobster immunocompetent peptide powder and baking soda, and mixing;
adding the powder into the beaten butter for three times after mixing; after being kneaded into a round shape, the round shape is pressed and flattened to a bit; making a model;
preheating oven, and placing the batter model into oven at 150deg.C for 10-15min.
The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments. Those skilled in the art will appreciate that, in light of the principles of the present invention, improvements and modifications can be made without departing from the scope of the invention.

Claims (11)

1. A preparation method of Chinese lobster immunoactive peptide is characterized by comprising the following steps: the method comprises the following steps:
(1) Crushing Chinese lobster, homogenizing, adding distilled water, mixing, and inactivating enzyme in water bath;
(2) Adding protease for enzymolysis, and inactivating enzyme in water bath again after the enzymolysis is finished;
(3) Cooling the enzymolysis liquid to normal temperature, centrifuging or sieving, and drying to obtain the Chinese lobster immunocompetent peptide;
in the step (2), the protease is bromelain.
2. The method for preparing the Chinese lobster immunoactive peptide according to claim 1, wherein the method comprises the following steps: in the step (1), when distilled water is added, the feed-liquid ratio is 1:1-1:6.
3. The method for preparing the Chinese lobster immunoactive peptide according to claim 1, wherein the method comprises the following steps: in the step (1), when distilled water is added, the feed-liquid ratio is 1:2-1:4.
4. The method for preparing the Chinese lobster immunoactive peptide according to claim 1, wherein the method comprises the following steps: in the step (1) and the step (2), the temperature of the water bath enzyme deactivation is 90-95 ℃ and the time is 10-15min.
5. The method for preparing the Chinese lobster immunoactive peptide according to claim 1, wherein the method comprises the following steps: in the step (2), the enzymolysis temperature is 50-55 ℃ and the enzymolysis time is 2-5h.
6. The method for preparing the Chinese lobster immunoactive peptide according to claim 1, wherein the method comprises the following steps: in the step (3), the rotational speed of the centrifugation is 8000-10000rpm, and the time is 15-20min.
7. The method for preparing the Chinese lobster immunoactive peptide according to claim 1, wherein the method comprises the following steps: in the step (3), the mesh number of the sieving is 100-200 mesh.
8. The method for preparing the Chinese lobster immunoactive peptide according to claim 1, wherein the method comprises the following steps: in the step (3), the membrane separation operation is carried out after sieving, and the membrane separation sequentially adopts a ceramic micro-filtration membrane with the membrane aperture of 0.2 mu m, an ultrafiltration membrane with the molecular weight cutoff of 3000-5000Da, an RO reverse osmosis membrane or a nanofiltration membrane with the molecular weight cutoff of 150-500 Da.
9. The method for preparing the Chinese lobster immunoactive peptide according to claim 1, wherein the method comprises the following steps: in the step (3), the drying conditions are as follows: the temperature of the sample inlet is 180 ℃, and the temperature of the outlet is 80-118 ℃.
10. An immunocompetent peptide of Chinese lobster, which is characterized in that: obtained by the production process according to any one of claims 1 to 9.
11. The use of the immunocompetent peptide of chinese lobster of claim 10 in the field of preparation of immunoenhancing foods.
CN202210581153.4A 2022-05-26 2022-05-26 Chinese lobster immunocompetent peptide, and preparation method and application thereof Active CN114921516B (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1554257A (en) * 2003-12-24 2004-12-15 山东大学 Sea Aceta chinensis high value ecolugical utilizing process
CN1660888A (en) * 2004-12-29 2005-08-31 山东大学 Peptide of decreasing blood pressure of protein of acetes chinensis, preparing method and application
CN1982468A (en) * 2005-12-13 2007-06-20 王荣辉 Preparation of shrimp peptide powder
JP2011178750A (en) * 2010-03-03 2011-09-15 Ehime Univ Allergy inhibitor
CN106434814A (en) * 2016-11-27 2017-02-22 威海蓝印海洋生物科技有限公司 Method for preparing compounded antihypertensive peptide through marine organisms

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1554257A (en) * 2003-12-24 2004-12-15 山东大学 Sea Aceta chinensis high value ecolugical utilizing process
CN1660888A (en) * 2004-12-29 2005-08-31 山东大学 Peptide of decreasing blood pressure of protein of acetes chinensis, preparing method and application
CN1982468A (en) * 2005-12-13 2007-06-20 王荣辉 Preparation of shrimp peptide powder
JP2011178750A (en) * 2010-03-03 2011-09-15 Ehime Univ Allergy inhibitor
CN106434814A (en) * 2016-11-27 2017-02-22 威海蓝印海洋生物科技有限公司 Method for preparing compounded antihypertensive peptide through marine organisms

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