Sipunculus nudus myofibrillar protein composite nutrition powder and preparation method thereof

Abstract

The invention discloses sipunculus nudus myofibrillar protein composite nutrition powder and a preparation method thereof. The preparation method comprises the following steps: selecting fresh sipunculus nudus, and treating to obtain sipunculus nudus body wall homogenate; the sipunculus nudus body wall homogenate is prepared into myofibrillar protein freeze-dried powder; resuspending the myofibrillar protein freeze-dried powder in distilled water, adding procyanidine into the resuspended myofibrillar protein freeze-dried powder, and performing multi-mode ultrasonic treatment on the resuspended myofibrillar protein freeze-dried powder; performing freeze drying on the sipunculus nudus myofibrillar protein-procyanidine mixed solution subjected to the multi-mode ultrasonic treatment; the sipunculus nudus myofibrillar protein-procyanidine compound, soybean protein isolate powder, whey protein powder, trachinotus ovatus skin collagen peptide, fruit and vegetable powder, probiotics, compound vitamin powder and linoleic acid are uniformly mixed, dried, crushed and sieved by a sieve of 80-120 meshes, and the obtained powder is metered and packaged into a finished product. The sipunculus nudus myofibrillar protein composite nutrition powder is good in water solubility, high in antioxidant activity and easy to digest.

Description

Technical Field

[0001] The invention belongs to the technical field of marine product component processing, and particularly relates to a Sipunculus sphenotype myofibrillar protein composite nutrient powder and a preparation method thereof. Background Art

[0002] Animal protein has a complete range of essential amino acids and a reasonable ratio. It is easier to digest, absorb and utilize than general plant proteins, and its nutritional value is relatively high. Among them, the amino acid scores of aquatic proteins are all at the level of 0.9 to 1.0. It is not only an important source of protein in people's daily diet, but also a protein source for special groups of people. Sipunculus nudus, also known as naked star worms, commonly known as "sandworms", are mainly produced in coastal areas and are biological resources with important economic value in marine fishery resources. It is reported that the current breeding area of ​​Sipunculus nudus in China is about 45,000 mu, with an output of about 61,000 tons, which has a broad application prospect. Studies have shown that sandworms are rich in protein (85%, g / g, db), taste particularly delicious, have the effects of lowering blood pressure, treating dampness, and preventing cancer, and are known as the marine "cordyceps sinensis". Myofibrillar protein is the main component of sandworm carcass protein, which has an important influence on the processing characteristics of sandworms. However, the large processing characteristics of myofibrillar protein in aqueous solution or under low ionic strength conditions have seriously affected the widespread application of sandworm protein.

[0003] Physical modification, chemical modification, enzymatic modification and genetic engineering modification are all common protein modification treatment methods. However, compared with other modification methods, physical modification has the advantages of low energy consumption, high efficiency, safety and environmental protection, short action time and less impact on the nutritional properties of the substance. Among them, ultrasonic treatment can be used as a time-saving, efficient and reliable technology to change the properties of food proteins, and has the potential to contribute to the development of emerging foods. In recent years, ultrasonic treatment has become a common protein modification method. Multimodal ultrasonic treatment not only affects the solubility and structure of proteins, but also has a certain effect on the digestion characteristics of protein samples. However, physical modification alone is difficult to achieve the ideal effect. The introduction of phenolic compounds has aroused people's interest. Plant extracts rich in proanthocyanidins can interact with proteins in a complex manner during food processing and human ingestion and digestion. The interaction will have a certain impact on the functional properties and digestion characteristics of protein foods, as well as the health benefits of polyphenol foods. Therefore, how to achieve water solubility of myofibrillar protein and improve digestion characteristics has attracted much attention in the field of animal protein science, and conducting relevant research has become an urgent task. Summary of the invention

[0004] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a method for preparing a Sipuncula myofibrillar protein-proanthocyanidin complex by multi-mode ultrasonic treatment to improve the solubility and digestibility of myofibrillar protein; and to mix it with plant protein, fish skin collagen peptide and fruit and vegetable powder in a certain proportion to improve its antioxidant activity; thereby obtaining a Sipuncula myofibrillar protein composite nutritional powder with good water solubility, strong antioxidant activity and easy digestibility, and a preparation method thereof.

[0005] The technical solution adopted by the present invention to solve its technical problem is:

[0006] A Siegespitella myofibrillar protein composite nutritional powder comprises the following ingredients: Siegespitella myofibrillar protein-proanthocyanidin complex, isolated soy protein powder, whey protein powder, golden pomfret skin collagen peptide, fruit and vegetable powder, probiotics, composite vitamin powder and linoleic acid. The Siegespitella myofibrillar protein-proanthocyanidin complex is obtained by multi-mode ultrasonic treatment.

[0007] Preferably, the composition comprises the following ingredients in parts by weight: 20-40 parts of Sipunculus myofibrillar protein-proanthocyanidin complex, 10-20 parts of soy protein isolate powder, 5-15 parts of whey protein powder, 2-8 parts of golden pomfret skin collagen peptide, 2-10 parts of fruit and vegetable powder, 5-8 parts of probiotics, 0.5-2.0 parts of multivitamin powder and 1-5 parts of linoleic acid.

[0008] Preferably, the preparation method of the Sipunculus myofibrillar protein-proanthocyanidin complex is:

[0009] The lyophilized powder of Sipunculus square worm myofibrillar protein is resuspended in distilled water, and then proanthocyanidins with a mass percentage concentration of 0.5% to 2.5% are added thereto, and then multi-mode ultrasonic treatment is performed to obtain the product, wherein the ultrasonic rated power is 700-800W, the ultrasonic frequency is 20-35kHz, the ultrasonic amplitude is 20-100%, the ultrasonic time is 10-15min, and the ultrasonic treatment mode is on for 2.5-3.5s and off for 1.5-2.5s.

[0010] Preferably, the preparation method of the Sipunculus myofibrillar protein freeze-dried powder is:

[0011] The Sipunculus body wall homogenate is mixed with a phosphate buffer solution at a mass ratio of 1:2-4, stirred overnight, and the mixed solution is centrifuged at 3-5°C and 5000-8000 r / min for 15-25 min to obtain a precipitate; the centrifuged precipitate is then mixed with a 0.8M sodium chloride solution at a ratio of 1:2-4, and then stirred at 3-5°C for 22-24 h. The supernatant obtained by centrifugation is dialyzed in distilled water for 40-50 h using a dialysis bag with a molecular weight of 1-2 kDa; and finally the retained solution is freeze-dried to obtain the product.

[0012] Preferably, the preparation method of the Sipunculus body wall homogenate is:

[0013] Select fresh Sipuncula spp., remove the internal organs with a blunt-tipped bamboo stick or steel needle until there is no residue, and wash;

[0014] Chop the processed starfish into pieces using a meat grinder, select high speed gear and grind for 10-35 seconds with an interval of 4-6 seconds in between, repeat 3 times, and you have the finished product.

[0015] Another technical problem to be solved by the present invention is to provide a method for preparing Sipunculus myofibrillar protein composite nutritional powder, comprising the following steps:

[0016] Select fresh Sipuncula spp. and process them to obtain Sipuncula spp. body wall homogenate;

[0017] The body wall of Sipunculus quadratus was homogenized to prepare freeze-dried myofibrillar protein powder;

[0018] The myofibrillar protein freeze-dried powder is resuspended in distilled water, and then proanthocyanidins are added thereto, and then the powder is subjected to multi-mode ultrasonic treatment to obtain a Sipunculus myofibrillar protein-proanthocyanidin mixed solution subjected to multi-mode ultrasonic treatment;

[0019] The mixed solution of Siegespermum myofibrillar protein-proanthocyanidin treated with multi-mode ultrasound is freeze-dried to obtain Siegespermum myofibrillar protein-proanthocyanidin complex;

[0020] 20-40 parts of Sipunculus myofibrillar protein-proanthocyanidin complex, 10-20 parts of soy protein isolate powder, 5-15 parts of whey protein powder, 2-8 parts of golden pomfret skin collagen peptide, 2-10 parts of fruit and vegetable powder, 5-8 parts of probiotics, 0.5-2.0 parts of multivitamin powder and 1-5 parts of linoleic acid are mixed uniformly by weight, dried, crushed, sieved through 80-120 mesh, and the obtained powder is measured and packaged into a finished product.

[0021] Preferably, the method of selecting fresh Sipuncula spp. for processing to obtain Sipuncula spp. body wall homogenate is:

[0022] Select fresh Sipuncula worms, remove the internal organs of the sandworms with a blunt bamboo stick or steel needle until there is no residue, and wash them;

[0023] The processed Sipuncula were minced with a meat grinder, and the high speed gear was selected for 10-35s, with an interval of 4-6s in between, and the cycle was repeated 3 times to obtain a homogenate of the Sipuncula body wall.

[0024] Preferably, the method for preparing freeze-dried myofibrillar protein powder by homogenizing the body wall of Sipunculus sphenotype is as follows:

[0025] The Sipunculus body wall homogenate is mixed with a phosphate buffer solution at a mass ratio of 1:2-4, stirred overnight, and the mixed solution is centrifuged at 3-5°C and 5000-8000 r / min for 15-25 minutes to obtain a precipitate; the centrifuged precipitate is then mixed with a 0.8M sodium chloride solution at a ratio of 1:2-4, and then stirred at 3-5°C for 22-24 hours, and the supernatant obtained by centrifugation is dialyzed in distilled water for 40-50 hours using a dialysis bag with a molecular weight of 1-2 kDa; finally, the retained solution is lyophilized to obtain myofibrillar protein lyophilized powder.

[0026] Preferably, the preparation method of the Sipunculus myofibrillar protein-proanthocyanidin complex is:

[0027] The myofibrillar protein freeze-dried powder is resuspended in distilled water, and then 0.5% to 2.5% of proanthocyanidins are added thereto, and then multi-mode ultrasonic treatment is performed, wherein the ultrasonic rated power is 700-800W, the ultrasonic frequency is 20-35kHz, the ultrasonic amplitude is 20-100%, the ultrasonic time is 10-15min, and the off time is 1.5-2.5s, to obtain a Sipunculus myofibrillar protein-proanthocyanidin mixed solution treated with multi-mode ultrasonic treatment.

[0028] Preferably, the ultrasonic treatment mode is on for 2.5-3.5 s and off for 1.5-2.5 s.

[0029] The beneficial effects of the present invention are:

[0030] The present invention adopts multi-mode ultrasound technology to cooperate with proanthocyanidins to act on the myofibrillar protein of Sipunculus quadripunctatus, thereby significantly improving the physicochemical properties and digestion properties of the myofibrillar protein.

[0031] The present invention adds ingredients such as soy protein isolate, fish skin collagen, whey protein, and fruit and vegetable powder to the myofibrillar protein-proanthocyanidin complex, which can not only well complement the deficiencies of animal protein and plant protein, and be better absorbed by the human body, but also effectively improve its antioxidant and other functional activities;

[0032] The present invention improves the processing characteristics of myofibrillar protein by the interaction of physical modification and addition of polyphenol components, and has the characteristics of low energy consumption, high efficiency, safety and environmental protection, short action time, and little effect on the nutritional characteristics of the substance. It has good promotion and application prospects in production practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is a schematic diagram of the solubility of the composite nutritional powder prepared in the experimental example;

[0034] Figure 2 Schematic diagram of the degree of hydrolysis (DH%) of the digestive juice after digestion of the composite nutritional powder prepared in the experimental example;

[0035] Figure 3 Schematic diagram of the antioxidant activity of digestive juice after digestion of the composite nutritional powder prepared in the experimental example. DETAILED DESCRIPTION

[0036] The principles and features of the present invention are described below in conjunction with the accompanying drawings. The examples given are only used to explain the present invention and are not used to limit the scope of the present invention. The present invention is described in more detail by way of example with reference to the accompanying drawings in the following paragraphs. The advantages and features of the present invention will become clearer according to the following description and claims. It should be noted that the drawings are all in a very simplified form and are not in precise proportions, and are only used to conveniently and clearly assist in explaining the purpose of the embodiments of the present invention.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0038] Embodiment 1:

[0039] A method for preparing Sipunculus sphenanthera myofibrillar protein composite nutrient powder, the specific steps are as follows:

[0040] 1. Select fresh Sipuncula worms weighing 10g, remove the internal organs of the sandworms with a blunt bamboo stick or steel needle until there is no residue, and wash them;

[0041] 2. Crush the Sipunculi processed in step 1 with a meat grinder, set at high speed for 30 seconds, rest for 5 seconds in between, and repeat 3 times;

[0042] 3. The Sipunculus body wall homogenate in step 2 was mixed with phosphate buffer solution at a mass ratio of 1:2, stirred overnight, and the mixture was centrifuged at 4°C and 8000r / min for 20min to obtain a precipitate; the centrifuged precipitate was then mixed with 0.8M sodium chloride solution at a ratio of 1:2, and then stirred at 4°C for 24h. The supernatant obtained by centrifugation was dialyzed in distilled water with a dialysis bag with a molecular weight of 1kDa for 45h; finally, the retained solution was lyophilized for standby use;

[0043] 4. The myofibrillar protein freeze-dried powder obtained in step 3 was resuspended in distilled water, and then 0.5% of proanthocyanidins was added thereto, and then multi-mode ultrasonic treatment was performed, wherein the ultrasonic rated power was 750W, the ultrasonic frequency was 20kHz, the ultrasonic amplitude was 20%, and the ultrasonic time was 10min (on for 3s, off for 2s);

[0044] 5. freeze-drying the Sipunculus myofibrillar protein-proanthocyanidin mixed solution treated with multi-mode ultrasound obtained in step 4 to obtain a myofibrillar protein complex powder compositely modified by multi-mode ultrasound and proanthocyanidin;

[0045] 6. Mix 20 parts of the Sipunculus myofibrillar protein-proanthocyanidin complex in step 5 with 10 parts of soy protein isolate powder, 5 parts of whey protein powder, 2 parts of golden pomfret skin collagen peptide, 2 parts of fruit and vegetable powder, 5 parts of probiotics, 0.5 parts of multivitamin powder, and 1 part of linoleic acid, dry, crush, sieve fine powder of about 100 mesh, and measure and package into finished products;

[0046] 7. The solubility and digestibility of the prepared Sipunculus myofibrillar protein composite nutritional powder were tested.

[0047] Embodiment 2:

[0048] A method for preparing Sipunculus sphenanthera myofibrillar protein composite nutrient powder, the specific steps are as follows:

[0049] 1. Select fresh Sipuncula worms weighing 10g, remove the internal organs of the sandworms with a blunt bamboo stick or steel needle until there is no residue, and wash them;

[0050] 2. Crush the Sipunculi processed in step 1 with a meat grinder, set at high speed for 30 seconds, rest for 5 seconds in between, and repeat 3 times;

[0051] 3. The Sipunculus body wall homogenate in step 2 was mixed with a phosphate buffer solution at a mass ratio of 1:3, stirred overnight, and the mixture was centrifuged at 4°C and 6000r / min for 20min to obtain a precipitate; the centrifuged precipitate was then mixed with a 0.8M sodium chloride solution at a ratio of 1:3, and then stirred at 4°C for 24h. The supernatant obtained by centrifugation was dialyzed in distilled water with a dialysis bag with a molecular weight of 1kDa for 45h; finally, the retained solution was lyophilized for standby use;

[0052] 4. The myofibrillar protein freeze-dried powder obtained in step 3 was resuspended in distilled water, and then 0.5% of proanthocyanidins was added thereto, and then multi-mode ultrasonic treatment was performed, wherein the ultrasonic rated power was 750W, the ultrasonic frequency was 20kHz, the ultrasonic amplitude was 40%, and the ultrasonic time was 10min (on for 3s, off for 2s);

[0053] 5. freeze-drying the Sipunculus myofibrillar protein-proanthocyanidin mixed solution treated with multi-mode ultrasound obtained in step 4 to obtain a myofibrillar protein complex powder compositely modified by multi-mode ultrasound and proanthocyanidin;

[0054] 6. Mix 20 parts of the Sipunculus myofibrillar protein-proanthocyanidin complex in step 5 with 15 parts of soy protein isolate powder, 10 parts of whey protein powder, 2 parts of golden pomfret skin collagen peptide, 2 parts of fruit and vegetable powder, 6 parts of probiotics, 0.5 parts of multivitamin powder, and 1 part of linoleic acid, then dry, crush, sieve a fine powder of about 100 mesh, and measure and package it into a finished product;

[0055] 7. The solubility and digestibility of the prepared Sipunculus myofibrillar protein composite nutritional powder were tested.

[0056] Embodiment 3:

[0057] A method for preparing Sipunculus sphenanthera myofibrillar protein composite nutrient powder, the specific steps are as follows:

[0058] 1. Select fresh Sipuncula worms weighing 10g, remove the internal organs of the sandworms with a blunt bamboo stick or steel needle until there is no residue, and wash them;

[0059] 2. Crush the Sipunculi processed in step 1 with a meat grinder, set at high speed for 20 seconds, rest for 6 seconds, and repeat 3 times;

[0060] 3. The Sipunculus body wall homogenate in step 2 was mixed with phosphate buffer solution at a mass ratio of 1:4, stirred overnight, and the mixture was centrifuged at 4°C and 7000r / min for 20min to obtain a precipitate; the centrifuged precipitate was then mixed with 0.8M sodium chloride solution at a ratio of 1:2, and then stirred at 4°C for 24h. The supernatant obtained by centrifugation was dialyzed in distilled water with a dialysis bag with a molecular weight of 1kDa for 45h; finally, the retained solution was lyophilized for standby use;

[0061] 4. The myofibrillar protein freeze-dried powder obtained in step 3 was resuspended in distilled water, and then 0.5% of proanthocyanidins was added thereto, and then multi-mode ultrasonic treatment was performed, wherein the ultrasonic rated power was 750W, the ultrasonic frequency was 20kHz, the ultrasonic amplitude was 60%, and the ultrasonic time was 10min (on for 3s, off for 2s);

[0062] 5. freeze-drying the Sipunculus myofibrillar protein-proanthocyanidin mixed solution treated with multi-mode ultrasound obtained in step 4 to obtain a myofibrillar protein complex powder compositely modified by multi-mode ultrasound and proanthocyanidin;

[0063] 6. Mix 25 parts of the Sipunculus myofibrillar protein-proanthocyanidin complex in step 5, 15 parts of soy protein isolate powder, 10 parts of whey protein powder, 3 parts of golden pomfret skin collagen peptide, 5 parts of fruit and vegetable powder, 5 parts of probiotics, 1.0 part of multivitamin powder, and 2 parts of linoleic acid, dry, crush, sieve fine powder of about 100 mesh, and measure and package into finished products;

[0064] 7. The solubility and digestibility of the prepared Sipunculus myofibrillar protein composite nutritional powder were tested.

[0065] Embodiment 4:

[0066] A method for preparing Sipunculus sphenanthera myofibrillar protein composite nutrient powder, the specific steps are as follows:

[0067] 1. Select fresh Sipuncula worms weighing 10g, remove the internal organs of the sandworms with a blunt bamboo stick or steel needle until there is no residue, and wash them;

[0068] 2. Crush the Sipunculi processed in step 1 with a meat grinder, set at high speed for 10 seconds, rest for 5 seconds in between, and repeat for 3 times;

[0069] 3. The Sipunculus body wall homogenate in step 2 was mixed with phosphate buffer solution at a mass ratio of 1:5, stirred overnight, and the mixture was centrifuged at 4°C and 8000r / min for 20min to obtain a precipitate; the centrifuged precipitate was then mixed with 0.8M sodium chloride solution at a ratio of 1:3, and then stirred at 4°C for 24h. The supernatant obtained by centrifugation was dialyzed in distilled water with a dialysis bag with a molecular weight of 1kDa for 45h; finally, the retained solution was lyophilized for standby use;

[0070] 4. The myofibrillar protein freeze-dried powder obtained in step 3 was resuspended in distilled water, and then 1.0% proanthocyanidin was added thereto, and then multi-mode ultrasonic treatment was performed, wherein the ultrasonic rated power was 750 W, the ultrasonic frequency was 35 kHz, the ultrasonic amplitude was 80%, and the ultrasonic time was 12 min (on 3 s, off 2 s);

[0071] 5. freeze-drying the Sipunculus myofibrillar protein-proanthocyanidin mixed solution treated with multi-mode ultrasound obtained in step 4 to obtain a myofibrillar protein complex powder compositely modified by multi-mode ultrasound and proanthocyanidin;

[0072] 6. Mix 25 parts of the Sipunculus myofibrillar protein-proanthocyanidin complex in step 5 with 15 parts of soy protein isolate powder, 10 parts of whey protein powder, 6 parts of golden pomfret skin collagen peptide, 8 parts of fruit and vegetable powder, 6 parts of probiotics, 1.5 parts of multivitamin powder, and 3 parts of linoleic acid, dry, crush, sieve fine powder of about 100 mesh, and measure and package into finished products;

[0073] 7. The solubility and digestibility of the prepared Sipunculus myofibrillar protein composite nutritional powder were tested.

[0074] Embodiment 5:

[0075] A method for preparing Sipunculus sphenanthera myofibrillar protein composite nutrient powder, the specific steps are as follows:

[0076] 1. Select fresh Sipuncula worms weighing 10g, remove the internal organs of the sandworms with a blunt bamboo stick or steel needle until there is no residue, and wash them;

[0077] 2. Crush the Sipunculi processed in step 1 with a meat grinder, set at high speed for 10 seconds, rest for 5 seconds in between, and repeat for 3 times;

[0078] 3. The Sipunculus body wall homogenate in step 2 was mixed with phosphate buffer solution at a mass ratio of 1:5, stirred overnight, and the mixture was centrifuged at 4°C and 5000r / min for 20min to obtain a precipitate; the centrifuged precipitate was then mixed with 0.8M sodium chloride solution at a ratio of 1:4, and then stirred at 4°C for 22h. The supernatant obtained by centrifugation was dialyzed in distilled water with a dialysis bag with a molecular weight of 2kDa for 40h; finally, the retained solution was lyophilized for standby use;

[0079] 4. The myofibrillar protein freeze-dried powder obtained in step 3 was resuspended in distilled water, and then 2.5% of proanthocyanidins were added thereto, and then multi-mode ultrasonic treatment was performed, wherein the ultrasonic rated power was 750W, the ultrasonic frequency was 20kHz, the ultrasonic amplitude was 100%, and the ultrasonic time was 15min (on for 3s, off for 2s);

[0080] 5. freeze-drying the Sipunculus myofibrillar protein-proanthocyanidin mixed solution treated with multi-mode ultrasound obtained in step 4 to obtain a myofibrillar protein complex powder compositely modified by multi-mode ultrasound and proanthocyanidin;

[0081] 6. Mix 40 parts of the Sipunculus myofibrillar protein-proanthocyanidin complex in step 5, 20 parts of soy protein isolate powder, 15 parts of whey protein powder, 8 parts of golden pomfret skin collagen peptide, 10 parts of fruit and vegetable powder, 8 parts of probiotics, 2.0 parts of multivitamin powder, and 5 parts of linoleic acid, dry, crush, sieve fine powder of about 100 mesh, and measure and package into finished products;

[0082] 7. The solubility and digestibility of the prepared Sipunculus myofibrillar protein composite nutritional powder were tested.

[0083] Comparative Example 1

[0084] The same raw materials and method as those in Example 5 were used to prepare the Sipunculus myofibrillar protein composite nutrient powder, except that in step 4, the ultrasonic amplitude was 0%.

[0085] Comparative Example 2

[0086] The Sipunculus myofibrillar protein composite nutrient powder was prepared using the same raw materials and method as in Example 5, except that in step 4, the ultrasonic frequency was 35 kHz.

[0087] Comparative Example 3

[0088] The same raw materials and methods as those in Example 1 were used to prepare the Sipunculus myofibrillar protein composite nutritional powder, except that in step 6, no golden pomfret skin collagen peptide was added.

[0089] Experimental Example 1

[0090] Effect evaluation and performance testing:

[0091] Solubility determination

[0092] Test sample: composite modified myofibrillar protein composite nutritional powder prepared in Examples 1-5 and Comparative Examples 1-3

[0093] The solubility experiment scheme is as follows: accurately weigh 1 mg of the above sample, dissolve it in 1 mL of distilled water, shake for 30 seconds to mix it thoroughly, and then centrifuge the mixture for 20 minutes, remove 0.5 mL of the supernatant and mix it with 5 mL of Coomassie Brilliant Blue, let it stand for 10 minutes, and finally, use a UV spectrophotometer to measure the absorbance of the solution at 510 nm. The sample solubility can be expressed as the percentage of the protein content in the centrifuged supernatant to the total protein content in the protein suspension.

[0094] The solubility of the composite nutritional powder prepared by the present invention is as follows: Figure 1 As shown, the solubility of the myofibrillar protein composite nutritional powder prepared in Example 1 is 16.49%, the solubility of the myofibrillar protein composite nutritional powder prepared in Example 2 is 32.71%, the solubility of the myofibrillar protein composite nutritional powder prepared in Example 3 is 34.40%, the solubility of the myofibrillar protein composite nutritional powder prepared in Example 4 is 37.59%, and the solubility of the myofibrillar protein composite nutritional powder prepared in Example 5 is 37. .58%, indicating that the solubility of the nutrient powder increases with the increase of ultrasonic amplitude, indicating that the solubility of the star worm composite nutrient powder is effectively improved; the solubility of the nutrient powder prepared in comparative examples 1-2 of the present invention is significantly lower than that of the nutrient powder obtained in Example 5, indicating that the ultrasonic amplitude has a greater influence on the solubility of the nutrient powder; there is no significant change in the solubility of the nutrient powder prepared in comparative example 3 and the nutrient powder obtained in Example 1, indicating that after adding golden pomfret skin collagen peptide to the star worm myofibrillar protein composite nutrient powder, the solubility will not change significantly.

[0095] The solubility of the Sipunculus myofibrillar protein composite nutrient powder prepared by the invention is increased, and the solubility is 16% to 37%.

[0096] Experimental Example 2

[0097] Digestive properties test

[0098] Test sample: composite modified myofibrillar protein composite nutritional powder prepared in Examples 1-5 and Comparative Examples 1-3

[0099] In vitro digestion protocol: A three-stage simulated gastrointestinal tract (GIT) model was used. The digestion process consists of an oral phase, a gastric phase, and an intestinal phase. First, in the oral phase, 25 mL of sample solution (10 mg / mL) was mixed with 25 mL of preheated simulated saliva (SSF), the pH was adjusted to 6.8 with HCl, and then incubated at 37°C for 5 min. Then, gastric digestion was performed by mixing 30 mL of simulated gastric fluid (SGF, 2.27 mg / L pepsin) with the oral phase (30 mL). The mixture was adjusted to 2.5 and incubated at 37°C for 2 h. Finally, the intestinal digestion phase was started by adjusting 40 mL of the mixture from the gastric phase to 6.8 and adding 2 mL of simulated intestinal fluid (SIF) and 4.7 mL of bile salt solution. Afterwards, the pH was adjusted back to 7.0 with NaOH, and 3.3 mL of pancreatic enzyme solution (36 mg / mL) was added. The digestate samples (1 mL) were sampled after 120 min of gastric digestion and after 120 min of intestinal digestion. Sample digestion was terminated by raising the pH to 8.0 to completely inactivate enzyme activity. Blank digestion was performed by replacing the sample with distilled water and mixing it with mock digestion solution and the same concentrations of pancreatin and bile.

[0100] Experimental Example 3

[0101] Digestive juice hydrolysis degree test (DH%)

[0102] Hydrolysis degree experimental protocol: The degree of protein hydrolysis was determined by o-phthalaldehyde (OPA) spectrophotometry. First, an OPA solution was prepared by dissolving 7.620 g of disodium tetraborate decahydrate in 150 mL of distilled water, adding o-phthalaldehyde (160 mg / 4 mL of ethanol), and dispersing 200 mg of sodium dodecyl sulfate (SDS) and 176 mg of 99% dithiothreitol (DTT) in the solution, and then bringing the volume of the mixture solution to 200 mL. The OPA solution was stored away from light to prevent discoloration. 3 mL of fresh OPA reagent was added to 400 μL of sample solution and mixed with a vortex mixer for 5 s. After standing at room temperature for 2 min, the absorbance of the reaction mixture was measured at 340 nm using a UV-visible spectrophotometer. A calibration curve was determined using L-serine (1.95-500 g / mL), which was used to determine the free amino group concentration of in vitro digestion and acid hydrolysis samples.

[0103] The solubility of the composite nutritional powder prepared by the present invention is as follows: Figure 2As shown, the hydrolysis degrees of the myofibrillar protein composite nutritional powder prepared in Example 1 in gastric juice and small intestinal juice after digestion are 1.19% and 83.68% respectively, the hydrolysis degrees of the myofibrillar protein composite nutritional powder prepared in Example 2 in gastric juice and small intestinal juice after digestion are 2.30% and 81.63% respectively, the hydrolysis degrees of the myofibrillar protein composite nutritional powder prepared in Example 3 in gastric juice and small intestinal juice after digestion are 3.49% and 83.52% respectively, the hydrolysis degrees of the myofibrillar protein composite nutritional powder prepared in Example 4 in gastric juice and small intestinal juice after digestion are 3.89% and 89.70% respectively, the myofibrillar protein composite nutritional powder prepared in Example 5 in gastric juice and small intestinal juice after digestion are 3.49% and 83.52% respectively. The hydrolysis degrees of gastric juice and small intestinal juice are 3.89% and 89.71% respectively, indicating that the hydrolysis degrees of gastric juice and small intestinal juice after digestion of the nutrient powder increase with the increase of ultrasonic amplitude, which is consistent with the solubility effect. The hydrolysis degrees of gastric juice and small intestinal juice after digestion of the nutrient powder prepared in comparative examples 1-2 of the present invention and the nutrient powder obtained in Example 5 are significantly reduced, indicating that the ultrasonic amplitude has a promoting effect on the digestion and absorption of the nutrient powder in the small intestine, indicating that the digestion characteristics of the Sipunculus worm composite nutrient powder are effectively improved; there is no significant change in the hydrolysis degree of protein after digestion between the nutrient powder prepared in comparative example 3 and the nutrient powder obtained in Example 1, indicating that after adding golden pomfret skin collagen peptide to the Sipunculus worm myofibrillar protein composite nutrient powder, the digestion characteristics will not change significantly.

[0104] Experimental Example 4

[0105] Digestive juice antioxidant test

[0106] Antioxidant experimental scheme: In the ABTS+ free radical scavenging experiment, 0.0384g ABTS+ and 0.0065g potassium persulfate were dispersed in 10mL deionized water as a stock solution and allowed to stand for 12-16h in the dark. The stock solution was then diluted with ethanol to an absorbance value of 0.700 (±0.001). The sample was then added to the ABTS+ mixture and the absorbance was measured at 734nm after 30min. The standard curve was determined using Trolox as the standard (y=0.1063x+0.0008, R 2 =0.990).

[0107] The solubility of the composite nutritional powder prepared by the present invention is as follows: Figure 3As shown, the oxidation removal rates of the gastric juice and small intestinal juice after digestion of the Sipuncula myofibrillar protein composite nutritional powder prepared in Example 1 were 16.06% and 161.89% respectively, the oxidation removal rates of the gastric juice and small intestinal juice after digestion of the Sipuncula myofibrillar protein composite nutritional powder prepared in Example 2 were 16.18% and 144.78% respectively, the oxidation removal rates of the gastric juice and small intestinal juice after digestion of the Sipuncula myofibrillar protein composite nutritional powder prepared in Example 3 were 12.99% and 153.45% respectively, the oxidation removal rates of the gastric juice and small intestinal juice after digestion of the Sipuncula myofibrillar protein composite nutritional powder prepared in Example 4 were 16.35% and 169.89% respectively, and the oxidation removal rates of the gastric juice and small intestinal juice after digestion of the Sipuncula myofibrillar protein composite nutritional powder prepared in Example 5 were 7. 56% and 154.51%, respectively, indicating that the oxidative clearance rates of gastric juice and small intestinal juice after digestion of the nutrient powder first increase and then decrease with the increase of ultrasonic amplitude, indicating that appropriate ultrasonic amplitude treatment can also enhance the antioxidant activity of the myofibrillar protein-proanthocyanidin complex; the oxidative clearance rates of the small intestinal juice after digestion of the nutrient powder prepared in Comparative Examples 1-2 of the present invention and the nutrient powder obtained in Example 5 are significantly reduced, indicating that the ultrasonic amplitude has a promoting effect on the digestion and absorption of the nutrient powder in the small intestine, indicating that the digestion characteristics of the Siegespreichs composite nutrient powder are effectively improved; the oxidative clearance rates of the small intestinal juice after digestion of the nutrient powder prepared in Comparative Example 3 and the nutrient powder obtained in Example 1 are significantly increased, indicating that the antioxidant activity of the composite nutrient powder can be greatly increased by adding golden pomfret skin collagen peptide to the Siegespreichs myofibrillar protein composite nutrient powder.

[0108] The above embodiments of the present invention are not intended to limit the protection scope of the present invention, and the implementation modes of the present invention are not limited thereto. All other modifications, replacements or changes made to the above structures of the present invention based on the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, should fall within the protection scope of the present invention.

Claims

1. A Sipunculus myofibrillar protein composite nutritional powder, characterized in that: The invention comprises the following ingredients: a myofibrillar protein-proanthocyanidin complex of Siegesporium quadrilateral, isolated soy protein powder, whey protein powder, golden pomfret skin collagen peptide, fruit and vegetable powder, probiotics, multivitamin powder and linoleic acid. The myofibrillar protein-proanthocyanidin complex of Siegesporium quadrilateral is obtained by multi-mode ultrasonic treatment.

2. The Sipunculus myofibrillar protein composite nutritional powder according to claim 1, characterized in that: The invention comprises the following ingredients in parts by weight: 20-40 parts of Sipunculus truncatus myofibrillar protein-proanthocyanidin complex, 10-20 parts of isolated soy protein powder, 5-15 parts of whey protein powder, 2-8 parts of golden pomfret skin collagen peptide, 2-10 parts of fruit and vegetable powder, 5-8 parts of probiotics, 0.5-2.0 parts of complex vitamin powder and 1-5 parts of linoleic acid.

3. The Sipunculus myofibrillar protein composite nutritional powder according to claim 1, characterized in that: The preparation method of the Sipunculus myofibrillar protein-proanthocyanidin complex is as follows: The lyophilized powder of Sipunculus square worm myofibrillar protein is resuspended in distilled water, and then proanthocyanidins with a mass percentage concentration of 0.5% to 2.5% are added thereto, and then multi-mode ultrasonic treatment is performed to obtain the product, wherein the ultrasonic rated power is 700-800W, the ultrasonic frequency is 20-35kHz, the ultrasonic amplitude is 20-100%, the ultrasonic time is 10-15min, and the ultrasonic treatment mode is on for 2.5-3.5s and off for 1.5-2.5s.

4. The Sipunculus myofibrillar protein composite nutritional powder according to claim 1, characterized in that: The preparation method of the Sipunculus myofibrillar protein freeze-dried powder is as follows: The Sipunculus body wall homogenate is mixed with a phosphate buffer solution at a mass ratio of 1:2-4, stirred overnight, and the mixed solution is centrifuged at 3-5°C and 5000-8000 r / min for 15-25 min to obtain a precipitate; the centrifuged precipitate is then mixed with a 0.8M sodium chloride solution at a ratio of 1:2-4, and then stirred at 3-5°C for 22-24 h. The supernatant obtained by centrifugation is dialyzed in distilled water for 40-50 h using a dialysis bag with a molecular weight of 1-2 kDa; and finally the retained solution is freeze-dried to obtain the product.

5. The Sipunculus myofibrillar protein composite nutritional powder according to claim 1, characterized in that: The preparation method of the Sipunculus body wall homogenate is as follows: Select fresh Sipuncula spp., remove the internal organs with a blunt-tipped bamboo stick or steel needle until there is no residue, and wash; Chop the processed starfish into pieces using a meat grinder, select high speed gear and grind for 10-35 seconds with an interval of 4-6 seconds in between, repeat 3 times, and you have the finished product.

6. A method for preparing Sipunculus sphenanthera myofibrillar protein composite nutrient powder, characterized in that: The following steps are involved: Select fresh Sipuncula spp. and process them to obtain Sipuncula spp. body wall homogenate; The body wall of Sipunculus quadratus was homogenized to prepare freeze-dried myofibrillar protein powder; The myofibrillar protein freeze-dried powder is resuspended in distilled water, and then proanthocyanidins are added thereto, and then the powder is subjected to multi-mode ultrasonic treatment to obtain a Sipunculus myofibrillar protein-proanthocyanidin mixed solution subjected to multi-mode ultrasonic treatment; The mixed solution of Siegespermum myofibrillar protein-proanthocyanidin treated with multi-mode ultrasound is freeze-dried to obtain Siegespermum myofibrillar protein-proanthocyanidin complex; 20-40 parts of Sipunculus myofibrillar protein-proanthocyanidin complex, 10-20 parts of soy protein isolate powder, 5-15 parts of whey protein powder, 2-8 parts of golden pomfret skin collagen peptide, 2-10 parts of fruit and vegetable powder, 5-8 parts of probiotics, 0.5-2.0 parts of multivitamin powder and 1-5 parts of linoleic acid are mixed uniformly by weight, dried, crushed, sieved through 80-120 mesh, and the obtained powder is measured and packaged into a finished product.

7. The method for preparing the Sipunculus myofibrillar protein composite nutritional powder according to claim 6, characterized in that: The method for selecting fresh Sipuncula for processing to obtain Sipuncula body wall homogenate is as follows: Select fresh Sipuncula worms, remove the internal organs of the sandworms with a blunt bamboo stick or steel needle until there is no residue, and wash them; The processed Sipuncula were minced with a meat grinder, and the high speed gear was selected for 10-35s, with an interval of 4-6s in between, and the cycle was repeated 3 times to obtain a homogenate of the Sipuncula body wall.

8. The method for preparing the Sipunculus myofibrillar protein composite nutritional powder according to claim 6, characterized in that: The method for preparing freeze-dried myofibrillar protein powder from the homogenate of Sipunculus sphenotype body wall is as follows: The Sipunculus body wall homogenate is mixed with a phosphate buffer solution at a mass ratio of 1:2-4, stirred overnight, and the mixed solution is centrifuged at 3-5°C and 5000-8000 r / min for 15-25 minutes to obtain a precipitate; the centrifuged precipitate is then mixed with a 0.8M sodium chloride solution at a ratio of 1:2-4, and then stirred at 3-5°C for 22-24 hours, and the supernatant obtained by centrifugation is dialyzed in distilled water for 40-50 hours using a dialysis bag with a molecular weight of 1-2 kDa; finally, the retained solution is lyophilized to obtain myofibrillar protein lyophilized powder.

9. The method for preparing the Sipunculus sphenanthera myofibrillar protein composite nutritional powder according to claim 6, characterized in that: The preparation method of the Sipunculus myofibrillar protein-proanthocyanidin complex is as follows: The myofibrillar protein freeze-dried powder is resuspended in distilled water, and then 0.5% to 2.5% of proanthocyanidins are added thereto, and then multi-mode ultrasonic treatment is performed, wherein the ultrasonic rated power is 700-800W, the ultrasonic frequency is 20-35kHz, the ultrasonic amplitude is 20-100%, the ultrasonic time is 10-15min, and the off time is 1.5-2.5s, to obtain a Sipunculus myofibrillar protein-proanthocyanidin mixed solution treated with multi-mode ultrasonic treatment.

10. The method for preparing the Sipunculus sphenanthera myofibrillar protein composite nutritional powder according to claim 7, characterized in that: The ultrasonic treatment mode was on for 2.5-3.5s and off for 1.5-2.5s.

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