阅读说明：本技术 贝类肽的制备方法 (Method for preparing beta-peptide ) 是由 陈体昌 许劲 文志州 谢小俊 于 2021-07-13 设计创作，主要内容包括：本发明公开了一种贝类肽的制备方法,包括以下步骤：a、对贝类原料进行蛋白质变性,使蛋白质中的肽键水解；b、将蛋白质变性后的贝类原料与复合酶液混合后进行酶解反应,所述复合酶液包括蛋白质和淀粉质混合原料接种霉菌得到的曲料；c、将步骤b得到的酶解反应的产物与还原性单糖进行热反应。本发明还公开了一种上述贝类肽的制备方法制备得到的贝类肽。本发明还公开了一种调味产品,包括上述的贝类肽。(The invention discloses a preparation method of a shellfish peptide, which comprises the following steps: a. performing protein denaturation on the shellfish raw material to hydrolyze peptide bonds in the protein; b. mixing the shellfish raw material after protein denaturation with compound enzyme liquid, and then carrying out enzymolysis reaction, wherein the compound enzyme liquid comprises yeast material obtained by inoculating mould on the mixed raw material of protein and starchiness; c. and c, carrying out thermal reaction on the product of the enzymolysis reaction obtained in the step b and reducing monosaccharide. The invention also discloses the shellfish peptide prepared by the preparation method of the shellfish peptide. The invention also discloses a seasoning product which comprises the beta-peptide.)

1. A method for preparing a shellfish peptide, comprising the steps of:

a. performing protein denaturation on the shellfish raw material to hydrolyze peptide bonds in the protein;

b. mixing the shellfish raw material after protein denaturation with compound enzyme liquid, and then carrying out enzymolysis reaction, wherein the compound enzyme liquid comprises yeast material obtained by inoculating mould on the mixed raw material of protein and starchiness;

c. and c, carrying out thermal reaction on the product of the enzymolysis reaction obtained in the step b and reducing monosaccharide.

2. The method for preparing the beta-peptide according to claim 1, wherein the enzymolysis reaction in the step b is carried out at 45-65 ℃ for 3-6 hours.

3. The method for preparing shellfish peptide according to claim 2, wherein the temperature of the enzymatic reaction is increased with time gradient.

4. The method for preparing shellfish peptide as claimed in claim 2, wherein step b further comprises the step of treating the enzymolysis product at 85-120 ℃ after the enzymolysis reaction to inactivate enzyme.

5. The method for preparing the beta-peptide according to claim 1, wherein the temperature of the thermal reaction in the step c is 100 to 120 ℃ and the reaction time is 30 to 120 min.

6. The method for preparing the beta-peptide according to claim 1, wherein the protein denaturation in the step a is wet heat denaturation, and the protein denaturation is carried out for 60-70 min at 95-120 ℃.

7. The method for producing the shellfish peptide according to any one of claims 1 to 6, wherein the shellfish material is dried shellfish.

8. The shellfish peptide produced by the method for producing a shellfish peptide according to any one of claims 1 to 7, wherein the shellfish peptide contains a dipeptide.

9. The shellfish peptide of claim 8, wherein said dipeptide comprises one or more of Glu-Asp, Glu-Glu, Glu-Ser, and Glu-Thr.

10. A flavored product comprising the shellfish peptide of any of claims 8 to 9.

Technical Field

The invention relates to the technical field of seasoning processing, in particular to a method for preparing shellfish peptides.

Background

Shellfish is a marine product of the order of Pteriales of the order of Mollusca. There are more than 60 scallops in the world. There are four types of scallops distributed in our country, namely chlamys farreri, chlamys nobilis, bay scallop and Patinopecten yessoensis. At present, the deep development of scallops is gradually increased, the protein content of scallop meat is high, the amino acid composition is complete, and the comprehensive development and utilization potential is huge. The seafood flavor peptide prepared by using an enzyme engineering technology of protease hydrolysis can be used for further enhancing the delicate flavor of the seasoning.

According to the theory of the multilevel structure of proteins, peptides are intermediate structures from proteins to amino acids. The peptides can be divided into polypeptides and small molecule peptides according to the number of molecular weights and the combination mode of peptide bonds, wherein the small molecule peptides are peptides formed by dehydrating 2-10 amino acid molecules and are called small molecule peptides, and the polypeptides are compounds formed by dehydrating and condensing 10-100 amino acid molecules. According to the composition, the molecular weight of the polypeptide is 1000-3000Da, the molecular weight of the small molecular peptide is below 1000Da, and the small molecular peptide has better delicate flavor.

The preparation method of common fish and shellfish flavoring agent comprises cleaning fish and shellfish with sauce, crushing, performing enzymolysis, separating the enzymolysis solution, filtering, and drying to obtain flavoring agent. For example, CN201610317506.X adopts ultrasonic enzymolysis and ultrafiltration membrane filtration to produce seafood flavoring food rich in flavor amino acids and flavor peptides; and for example, 201610001743.X uses fresh oyster meat as raw material, and is prepared by crushing, performing enzymolysis with neutral protease, filtering, and freeze drying. However, the seasoning prepared by the methods is polypeptide with multiple amino acid numbers, and the dipeptide with delicate flavor is difficult to directionally obtain.

Disclosure of Invention

Accordingly, there is a need for a method for producing a mussel peptide from which a dipeptide can be obtained.

One of the purposes of the invention is to provide a preparation method of a shellfish peptide, which comprises the following steps:

a. performing protein denaturation on the shellfish raw material to hydrolyze peptide bonds in the protein;

b. mixing the shellfish raw material after protein denaturation with compound enzyme liquid, and then carrying out enzymolysis reaction, wherein the compound enzyme liquid comprises yeast material obtained by inoculating mould on the mixed raw material of protein and starchiness;

c. and c, carrying out thermal reaction on the product of the enzymolysis reaction obtained in the step b and reducing monosaccharide.

In some embodiments, the enzymolysis reaction in step b is carried out at 45-65 ℃ for 3-6 hours.

In some embodiments, the temperature of the enzymatic reaction is in a time gradient.

In some embodiments, step b further comprises the step of treating the enzymolysis product at 85-120 ℃ after the enzymolysis reaction to inactivate the enzyme.

In some embodiments, the temperature of the thermal reaction of step c is from 100 ℃ to 120 ℃ and the reaction time is from 30min to 120 min.

In some embodiments, the protein denaturation of step a is wet heat denaturation, and the treatment is carried out at 95-120 ℃ for 60-70 min.

In some embodiments, the shellfish material is dried shellfish.

The invention also aims to provide the shellfish peptide prepared by the preparation method of the shellfish peptide of any embodiment, wherein the shellfish peptide contains dipeptide.

In some embodiments, the dipeptide includes one or more of Glu-Asp, Glu-Glu, Glu-Ser, and Glu-Thr.

It is another object of the present invention to provide a flavored product comprising the above mentioned shellfish peptide.

The invention is obtained by performing protein denaturation, enzymolysis and reductive monosaccharide thermal reaction on shellfish raw materials. The method is characterized in that a compound enzyme solution from plants is adopted to carry out enzymolysis on moderately denatured animal protein, and the compound enzyme solution is obtained by inoculating mould to a mixed raw material of protein and starchiness for yeast making. The preparation method of the invention obtains the shellfish like peptide with the content of four delicious dipeptides Glu-Asp, Glu-Glu, Glu-Ser and Glu-Thr of more than 30 percent, and the dipeptide belongs to active peptide components, can be matched with nutrient components and has potential efficacies of regulating immunity, resisting virus, resisting oxidation and the like. The dipeptide is applied to seasoning products such as soy sauce, oyster sauce, soybean sauce and the like, and has obvious effects of improving freshness, thickening and marine product flavor.

Detailed Description

In order that the invention may be more fully understood, reference will now be made to the following description. The present invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The embodiment of the invention provides a preparation method of a shellfish peptide, which comprises the following steps:

a. performing protein denaturation on the shellfish raw material to hydrolyze peptide bonds in the protein;

b. mixing the shellfish raw material after protein denaturation with compound enzyme liquid, and then carrying out enzymolysis reaction, wherein the compound enzyme liquid comprises yeast material obtained by inoculating mould on the mixed raw material of protein and starchiness;

c. and c, carrying out thermal reaction on the product of the enzymolysis reaction obtained in the step b and reducing monosaccharide.

The invention is obtained by performing protein denaturation, enzymolysis and reductive monosaccharide thermal reaction on shellfish raw materials. The method is characterized in that a compound enzyme solution from plants is adopted to carry out enzymolysis on moderately denatured animal protein, and the compound enzyme solution is obtained by inoculating mould to a mixed raw material of protein and starchiness for yeast making. The preparation method of the invention obtains the shellfish like peptide with the content of four delicious dipeptides Glu-Asp, Glu-Glu, Glu-Ser and Glu-Thr of more than 30 percent, and the dipeptide belongs to active peptide components, can be matched with nutrient components and has potential efficacies of regulating immunity, resisting virus, resisting oxidation and the like. The dipeptide is applied to seasoning products such as soy sauce, oyster sauce, soybean sauce and the like, and has obvious effects of improving freshness, thickening and marine product flavor.

The shellfish raw material used in the invention is marine shellfish, belonging to the phylum mollusca, class Haliotis, class Pinctada, family scallops. The present invention is not limited to the kind of the raw material of the shellfish, and may be selected from any one or more of economic scallop varieties, including, for example, Chlamys farre, Argopecten irradians, Patinopecten yessoensis and Chlamys nobilis.

In some embodiments, the shellfish material used in the present invention is in the form of dried shellfish. Dried shellfish is a dried product of shellfish.

There are many factors causing protein denaturation, and physical factors include high temperature, ultraviolet rays, X-rays, ultrasonic waves, high pressure, vigorous stirring, shaking, and the like. The chemical factors include strong acid, strong base, urea, guanidine salt, detergent, and heavy metal salt (such as Hg)²⁺、Ag⁺、Pb²⁺Etc.), trichloroacetic acid, concentrated ethanol, etc. Different proteins are sensitive to various factors to different degrees, and different methods can change many properties of proteins after denaturation.

Preferably, the protein denaturation condition adopted in the step a of the preparation method is that the protein is treated for 60-70 min under the condition of moist heat denaturation at 95-120 ℃. Specifically, the temperature of the wet-heat denaturation can be 95 deg.C, 96 deg.C, 97 deg.C, 98 deg.C, 99 deg.C, 100 deg.C, 101 deg.C, 102 deg.C, 103 deg.C, 104 deg.C, 105 deg.C, 106 deg.C, 107 deg.C, 108 deg.C, 109 deg.C, 110 deg.C, 111 deg.C, 112 deg.C, 113 deg.C, 114 deg.C, 115 deg.C, 116 deg.C, 117 deg.C, 118 deg.C, 119 deg.C, and 120 deg.C. The treatment time can be 60min, 61min, 62min, 63min, 64min, 65min, 66min, 67min, 68min, 69min, and 70 min.

The raw material for preparing the compound enzyme liquid in the step b can be any one or more of soybean, soybean meal and other protein raw materials. The soybean can be soybean, green soybean and black soybean. The soybean meal is a product obtained by extracting oil with a soybean solvent. Other proteinaceous materials may include beans, peas, mung beans, and the like.

The starchy material may be selected from any one or more of wheat, bran and other starch materials. The wheat includes red skin wheat and white skin wheat, and the plasmid may be soft, hard and medium wheat. Other starch materials may include sweet potato, corn, barley, sorghum, millet, rice bran, and the like. The starchy material may be in powder form.

The mould is used as a mold starter to ferment protein and starchy raw materials. The mould may be selected from any one or more of aspergillus oryzae, rhizopus, trichome and aspergillus niger. Aspergillus oryzae and Mucor have strong protein decomposition and saccharification ability. Aspergillus niger and Rhizopus have various enzyme systems such as amylase, saccharifying enzyme, acid protease and cellulase, and are used for liquefying and saccharifying starch. Preferably, the mold comprises at least Aspergillus oryzae. More preferably, the mould is selected from aspergillus oryzae.

In some embodiments, the enzymolysis reaction in step b is carried out at 45-65 ℃ for 3-6 hours. Specifically, the enzymolysis temperature can be 45 deg.C, 50 deg.C, 55 deg.C, 60 deg.C, 65 deg.C.

In some embodiments, the temperature of the enzymatic reaction is in a time gradient.

In some embodiments, step b further comprises the step of treating the enzymatic hydrolysis product at 85 ℃ to 120 ℃ after the enzymatic hydrolysis reaction to inactivate the enzyme. Specifically, the enzyme deactivation temperature may be 85 deg.C, 86 deg.C, 87 deg.C, 88 deg.C, 89 deg.C, 90 deg.C, 91 deg.C, 92 deg.C, 93 deg.C, 94 deg.C, 95 deg.C, 96 deg.C, 97 deg.C, 98 deg.C, 99 deg.C, 100 deg.C, 101 deg.C, 102 deg.C, 103 deg.C, 104 deg.C, 105 deg.C, 106 deg.C, 107 deg.C, 108 deg.C, 109 deg.C, 110 deg.C, 111 deg.C, 112 deg.C, 114 deg.C, 115 deg.C, 116 deg.C, 117 deg.C, 118 deg.C, 119 deg.C, and 120 deg.C.

In some embodiments, the temperature of the thermal reaction of step c is from 100 ℃ to 120 ℃ and the reaction time is from 30min to 120 min.

In some embodiments, the reducing monosaccharide can be selected from any one or more of xylose, glucose, fructose syrup, galactose, ribose, deoxyribose, and maltose.

In some embodiments, the reaction ratio of step c is 80 to 90 parts by weight of the scallop enzymatic hydrolysate and 5 to 15 parts by weight of the reducing monosaccharide.

The embodiment of the invention also provides the shellfish peptide prepared by the preparation method of the shellfish peptide in any embodiment, wherein the shellfish peptide contains dipeptide.

In some embodiments, the dipeptide includes one or more of Glu-Asp, Glu-Glu, Glu-Ser, and Glu-Thr.

The embodiment of the invention also provides a seasoning product which comprises the beta-peptide.

The seasoning product provided by the invention can be a product with a seasoning effect, such as sauces (barbeque sauce, fermented soybean chili sauce, plum sauce, xo sauce and the like), sauces (light soy sauce, fresh shrimp sauce, fermented soybean sauce, straw mushroom dark soy sauce and the like), juice (barbecue juice, brine juice, juice, OK juice and the like), gourmet powder (pepper powder, sand ginger powder, garlic powder, chicken powder and the like) and the like. Specifically, the flavoring product can be scallop deep-processing product. The scallop deep-processed product includes scallop sauce, scallop soy sauce, scallop seasoning juice, etc.

The prepared product is detected by GPC determination or mass spectrum identification, and the result shows that the peptide content can be improved from 40-45% (calculated by a desalting dry basis) to 60-70% (calculated by a desalting dry basis) after enzymolysis, particularly the content of four delicious dipeptides Glu-Asp, Glu-Glu, Glu-Ser and Glu-Thr can reach more than 30% of the total peptide content, and the molecular weight is concentrated between 50-700 Da. Concretely, Glu-Asp (accounting for 1-15 percent of the total peptide), Glu-Glu (accounting for 1-15 percent of the total peptide), Glu-Ser (accounting for 1-10 percent of the total peptide) and Glu-Thr (accounting for 1-10 percent of the total peptide). The prepared shellfish peptide is added into seasoning products such as soy sauce, oyster sauce, soybean sauce and the like according to the mass fraction of 0.1-0.5%, so that the shellfish peptide has obvious fresh-keeping and thick-concentrating effects and is rich in marine product flavor.

The following are specific examples.

Example 1:

1. preparing a compound enzyme solution, and detecting for later use, wherein the activity of compound protease in the compound enzyme solution reaches 5000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling soybean and wheat flour to obtain starter propagation raw material; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 100 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 100 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 85 parts of scallop enzymatic hydrolysate, 10 parts of glucose and 5 parts of tap water, and carrying out thermal reaction at 115 ℃ for 45min to obtain a scallop peptide finished product.

The scallop peptide produced in this example had a total peptide content of 62% (on a dry desalted basis), a dipeptide content of 25% (on a dry desalted basis), a Glu-Asp content of 7% (on a dry desalted basis), a Glu-Glu content of 6% (on a dry desalted basis), a Glu-Ser content of 3% (on a dry desalted basis) and a Glu-Thr content of 4% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Example 2:

1. preparing a compound enzyme solution, and detecting for later use; the activity of the compound protease in the compound enzyme solution reaches 6000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling the soybean meal and the roasted wheat to obtain a starter propagation raw material; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 95 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 2 hours, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 85 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 90 parts of scallop enzymatic hydrolysate, 8 parts of fructose and 2 parts of tap water, and carrying out thermal reaction at 120 ℃ for 30min to obtain a scallop peptide finished product.

The scallop peptide produced in this example had a total peptide content of 65% (on a dry desalted basis), a dipeptide content of 26% (on a dry desalted basis), a Glu-Asp content of 8% (on a dry desalted basis), a Glu-Glu content of 7% (on a dry desalted basis), a Glu-Ser content of 4% (on a dry desalted basis) and a Glu-Thr content of 4% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Example 3:

1. preparing a compound enzyme solution, and detecting for later use; the activity of the compound protease in the compound enzyme solution reaches 8000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling soybean and wheat flour to obtain starter propagation raw materials; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 120 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 2 hours, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 110 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 80 parts of dried scallop enzymatic hydrolysate, 5 parts of ribose and 15 parts of tap water, and carrying out thermal reaction at 100 ℃ for 120min to obtain a finished product of dried scallop peptide.

The scallop peptide produced in this example had a total peptide content of 60% (on a dry desalted basis), a dipeptide content of 24% (on a dry desalted basis), a Glu-Asp content of 7% (on a dry desalted basis), a Glu-Glu content of 6% (on a dry desalted basis), a Glu-Ser content of 3% (on a dry desalted basis) and a Glu-Thr content of 3% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Example 4:

1. preparing a composite enzyme solution, and detecting for later use, wherein the activity of the composite protease in the composite enzyme solution reaches 7000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming black beans and rice flour to obtain starter propagation raw materials; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of dried scallop raw material at 110 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 50 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 120 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 85 parts of scallop enzymatic hydrolysate, 10 parts of glucose and 5 parts of tap water, and carrying out thermal reaction at 115 ℃ for 45min to obtain a scallop peptide finished product.

The scallop peptide produced in this example had a total peptide content of 66% (on a dry desalted basis), a dipeptide content of 27% (on a dry desalted basis), a Glu-Asp content of 8% (on a dry desalted basis), a Glu-Glu content of 7% (on a dry desalted basis), a Glu-Ser content of 5% (on a dry desalted basis) and a Glu-Thr content of 3% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Example 5:

1. preparing a compound enzyme solution, and detecting for later use, wherein the activity of the compound protease in the compound enzyme solution reaches 8000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling soybean and wheat flour to obtain starter propagation raw material; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:3, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 100 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:90, stirring uniformly at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 110 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 85 parts of scallop enzymatic hydrolysate, 10 parts of glucose and 5 parts of tap water, and carrying out thermal reaction at 115 ℃ for 45min to obtain a scallop peptide finished product.

The scallop peptide produced in this example had a total peptide content of 70% (on a dry desalted basis), a dipeptide content of 28% (on a dry desalted basis), a Glu-Asp content of 8% (on a dry desalted basis), a Glu-Glu content of 8% (on a dry desalted basis), a Glu-Ser content of 4% (on a dry desalted basis), and a Glu-Thr content of 5% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Example 6:

1. preparing a compound enzyme solution, and detecting for later use, wherein the activity of compound protease in the compound enzyme solution reaches 5000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling soybeans and rice flour to obtain a starter propagation raw material; 2) carrying out amplification culture on aspergillus niger to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 100 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 100 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 85 parts of scallop enzymatic hydrolysate, 10 parts of glucose and 5 parts of tap water, and carrying out thermal reaction at 115 ℃ for 45min to obtain a scallop peptide finished product.

The scallop peptide produced in this example had a total peptide content of 61% (on a dry desalted basis), a dipeptide content of 24% (on a dry desalted basis), a Glu-Asp content of 7% (on a dry desalted basis), a Glu-Glu content of 9% (on a dry desalted basis), a Glu-Ser content of 3% (on a dry desalted basis) and a Glu-Thr content of 3% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Comparative example 1:

1. preparing a compound enzyme solution, and detecting for later use, wherein the activity of compound protease in the compound enzyme solution reaches 5000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling soybeans to obtain a starter propagation raw material;

2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 100 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 100 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 85 parts of scallop enzymatic hydrolysate, 10 parts of glucose and 5 parts of tap water, and carrying out thermal reaction at 115 ℃ for 45min to obtain a scallop peptide finished product.

The scallop peptide produced in this example had a total peptide content of 60% (on a dry desalted basis), a dipeptide content of 15% (on a dry desalted basis), a Glu-Asp content of 5% (on a dry desalted basis), a Glu-Glu content of 3% (on a dry desalted basis), a Glu-Ser content of 4% (on a dry desalted basis), and a Glu-Thr content of 2% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Comparative example 2:

1. preparing a compound enzyme solution, and detecting for later use, wherein the activity of compound protease in the compound enzyme solution reaches 2000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming wheat flour to obtain starter propagation raw material; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 100 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 100 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 85 parts of scallop enzymatic hydrolysate, 10 parts of glucose and 5 parts of tap water, and carrying out thermal reaction at 115 ℃ for 45min to obtain a scallop peptide finished product. The scallop peptide produced in this example had a total peptide content of 15% (on a dry desalted basis), a dipeptide content of 8% (on a dry desalted basis), a Glu-Asp content of 1% (on a dry desalted basis), a Glu-Glu content of 4% (on a dry desalted basis), a Glu-Ser content of 1% (on a dry desalted basis), and a Glu-Thr content of 1% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Comparative example 3:

1. preparing a compound enzyme solution, and detecting for later use, wherein the activity of compound protease in the compound enzyme solution reaches 5000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling soybean and wheat flour to obtain starter propagation raw material; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 100 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis, inactivating enzyme at 100 deg.C for 20min, centrifuging at 4000rpm for 10min, collecting supernatant, and freeze drying.

The scallop peptide produced in this example had a total peptide content of 63% (on a dry desalted basis), a dipeptide content of 15% (on a dry desalted basis), a Glu-Asp content of 4% (on a dry desalted basis), a Glu-Glu content of 3% (on a dry desalted basis), a Glu-Ser content of 2% (on a dry desalted basis) and a Glu-Thr content of 2% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Comparative example 4:

1. preparing a compound enzyme solution, and detecting for later use, wherein the activity of compound protease in the compound enzyme solution reaches 5000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling soybean and wheat flour to obtain starter propagation raw material; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 100 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 100 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 85 parts of scallop enzymatic hydrolysate, 10 parts of glucose and 5 parts of tap water, and carrying out thermal reaction at 85 ℃ for 45min to obtain a scallop peptide finished product. The scallop peptide produced in this example had a total peptide content of 63% (on a dry desalted basis), a dipeptide content of 18% (on a dry desalted basis), a Glu-Asp content of 5% (on a dry desalted basis), a Glu-Glu content of 4% (on a dry desalted basis), a Glu-Ser content of 2% (on a dry desalted basis) and a Glu-Thr content of 3% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

Comparative example 5:

1. preparing a compound enzyme solution, and detecting for later use, wherein the activity of compound protease in the compound enzyme solution reaches 5000U/mL;

the preparation method of the compound enzyme solution comprises the following steps: 1) steaming and boiling soybean and wheat flour to obtain starter propagation raw material; 2) carrying out expanded culture on aspergillus oryzae to obtain a seed starter; 3) inoculating a starter into a starter-making raw material to prepare a starter material; 4) uniformly mixing the yeast material and water according to the mass ratio of 1:2, and centrifuging at 4000rpm for 10min to obtain supernatant, namely the compound enzyme solution;

2. wet denaturation of scallop raw material at 100 deg.C for 60 min;

3. mixing the compound enzyme solution and the dried scallop according to the mass ratio of 2:100, uniformly stirring at normal temperature, performing enzymolysis at 45 ℃ for 1 hour, performing enzymolysis at 55 ℃ for 2 hours, and performing enzymolysis at 65 ℃ for 2 hours;

4. after enzymolysis is finished, carrying out enzyme deactivation treatment at 100 ℃ for 20min, then centrifuging at 4000rpm for 10min, taking supernatant, and detecting for later use;

5. taking 85 parts of scallop enzymatic hydrolysate, 10 parts of glucose and 5 parts of tap water, and carrying out thermal reaction at 150 ℃ for 45min to obtain a scallop peptide finished product.

The scallop peptide produced in this example had a total peptide content of 65% (on a dry desalted basis), a dipeptide content of 6% (on a dry desalted basis), a Glu-Asp content of 2% (on a dry desalted basis), a Glu-Glu content of 1% (on a dry desalted basis), a Glu-Ser content of 1% (on a dry desalted basis), and a Glu-Thr content of 0% (on a dry desalted basis), as determined by GPC measurements or mass spectrometry.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, so as to understand the technical solutions of the present invention specifically and in detail, but not to be understood as the limitation of the patent protection scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the patent of the invention is subject to the appended claims, and the description can be used for explaining the contents of the claims.