CN113424941A

CN113424941A - Scallop flavor peptide product and preparation method thereof

Info

Publication number: CN113424941A
Application number: CN202110674952.1A
Authority: CN
Inventors: 邓丽; 侯杰; 桂军强; 王洪江; 吴明亮; 薛忠玉; 陈周
Original assignee: Haitian Oyster Sauce Jiangsu Food Co ltd; Haitian Oyster Sauce Nanning Food Co ltd; Haitian Oyster Sauce Tianjin Group Co ltd; Foshan Haitian Flavoring and Food Co Ltd; Foshan Haitian Gaoming Flavoring and Food Co Ltd
Current assignee: Xianzhiran Biotechnology Group Co.,Ltd.; Xianzhiran Guangdong Biotechnology Co ltd; Xianzhiran Tianjin Biotechnology Co ltd; Xianzhiran Tianjin Biotechnology Group Co ltd
Priority date: 2021-06-17
Filing date: 2021-06-17
Publication date: 2021-09-24
Anticipated expiration: 2041-06-17
Also published as: CN113424941B

Abstract

The invention relates to a scallop fresh peptide product and a preparation method thereof. The preparation method combines an ultrahigh pressure homogenization technology and an ultrasonic technology to carry out pretreatment on scallop protein, and improves the targeted extraction of scallop flavor peptides through segmented enzymolysis, a synergistic effect on hydrolysis control can be formed between the pretreatment and the segmented enzymolysis, the content of the flavor peptides in an enzymolysis product is increased on the basis of improving the protein hydrolysis rate, and the utilization degree of the scallop flavor substances is improved; the Mallotus oblongifolius muell-Arg extract can be introduced in the scallop protein hydrolysis process, fat oxidation is inhibited while hydrolysis is carried out, and compared with an after-treatment fishy smell removing mode such as activated carbon adsorption, the method is simpler and more effective. The preparation method of the scallop flavor peptide product provided by the invention can effectively solve the problems of low hydrolysis rate of scallop protein and insufficient utilization of scallop flavor substances, and further can simply and effectively inhibit the generation of fishy smell substances from the source.

Description

Scallop flavor peptide product and preparation method thereof

Technical Field

The invention relates to the technical field of biosynthesis and the technical field of seasonings, in particular to a scallop flavor peptide product and a preparation method thereof.

Background

The scallop has the characteristics of high protein, low fat and low cholesterol, and is delicious in taste and deeply favored by consumers. The scallop is rich in protein, can be hydrolyzed into delicious components such as polypeptide, amino acid and the like, so that the seasoning can be prepared by using the delicious substances extracted from the scallop hydrolysate.

The scallop is generally hydrolyzed by adopting an enzymolysis mode. Due to high aggregation of scallop protein molecules and high degree of molecule helicization, the hydrolysis degree of the scallop protein molecules is not high when the scallop protein molecules are subjected to enzymolysis treatment, and a considerable part of macromolecular peptides are not hydrolyzed into micromolecular umami peptides and free amino acids, so that the utilization rate of umami substances in the scallops is low, and the waste of raw materials is caused. At present, the protein hydrolysis rate is mainly improved by pretreatment of scallop materials in modes of high-temperature treatment and the like, but can be generally improved to 35-45 percent or even lower, and the improvement of the protein hydrolysis rate is limited in general.

In addition, the scallop protein is easy to generate fat oxidation in the enzymolysis process, so that fishy smell substances are generated, and the scallop protein is suitable for eating after the fishy smell of the enzymolysis products is removed. At present, most of methods for removing fishy smell of scallop enzymatic hydrolysate focus on removing fishy smell by means of Maillard reaction, activated carbon adsorption and the like after the enzymolysis is finished, but the fishy smell removing effect is not ideal.

At present, most researches on the enzymolysis of the scallop focus on the extraction of free amino acid, the amino acid content is taken as an important evaluation index of the content of the umami substances, and the utilization mode of the umami substance components is single.

Therefore, there is a need for a process that better utilizes scallop-derived umami taste substances.

Disclosure of Invention

Based on the technical problems, the invention provides a preparation method of a scallop fresh peptide product, which comprises the steps of firstly adopting an ultrahigh pressure homogenization technology and then adopting an ultrasonic technology to carry out pretreatment on scallop protein, obviously improving the hydrolysis rate of the scallop protein through the synergistic effect between the two treatment processes, and improving the utilization rate of raw materials.

The preparation method of the scallop fresh taste peptide product comprises the following steps:

providing raw materials: providing a scallop protein raw material;

pretreatment: performing pretreatment on the scallop protein raw material to prepare scallop protein liquid, wherein the pretreatment comprises ultrahigh pressure homogenization treatment and then ultrasonic treatment; wherein the pressure of the ultrahigh-pressure homogenizing treatment is preferably not less than 100 MPa;

segmented enzymolysis: carrying out segmented enzymolysis on the scallop protein liquid to prepare scallop protein enzymolysis liquid; the segmented enzymolysis comprises a first-stage enzymolysis by adopting endoprotease and a second-stage enzymolysis by adopting exoprotease;

and (3) post-treatment: and carrying out solid-liquid separation on the scallop protein enzymatic hydrolysate, and collecting clear liquid.

In some embodiments of the present invention, the step of performing the staged enzymolysis further comprises a step of adding an antioxidant to the scallop protein liquid. The antioxidant (such as the Mallotus furetianus extract) is introduced in the scallop protein hydrolysis process, fat oxidation can be inhibited during hydrolysis, fishy smell generation can be inhibited from the source, and therefore the content of fishy smell substances in the scallop fresh taste peptide product is effectively reduced, and compared with a post-treatment fishy smell removal mode such as activated carbon adsorption and the like, the method is simpler and more effective.

In some preferred embodiments of the present invention, the antioxidant comprises mallotus oblongifolius extract.

In some embodiments of the present invention, the antioxidant is selected from any one of antioxidants selected from mallotus oblongifolius extract, yeast extract (containing antioxidant peptides), chrysanthemum tea, and the like, or any combination thereof.

In some preferred embodiments of the present invention, the step of performing the staged enzymolysis further comprises a step of adding an antioxidant to the scallop protein liquid, wherein the antioxidant is partridge tea extract. Further, in some more preferred embodiments of the present invention, the addition amount of the mallotus oblongifolius extract is 0.5 to 2% of the mass of the scallop protein liquid.

In some embodiments of the invention, the scallop flavor peptide product has a scallop flavor peptide content of greater than or equal to 50% by weight of total peptides.

In some embodiments of the invention, in the scallop delicious peptide product, the mass content of the polypeptide with the molecular weight of 200 Da-1000 Da in the total peptide is more than or equal to 50 percent; and/or in the dry basis of the scallop fresh taste peptide product, the mass content of the polypeptide with the molecular weight of 200 Da-1000 Da is 30-45%.

In some embodiments of the present invention, the step of performing the ultra-high pressure homogenization treatment and the ultrasonic treatment further comprises performing a colloid mill treatment on the scallop protein raw material. In some embodiments of the invention, the water adding amount of the colloid mill is 2-5 times of the mass of the scallop protein raw material. In some embodiments of the invention, the colloid mill treatment is performed at least 1 time.

In some embodiments of the present invention, the pressure of the ultra-high pressure homogenization treatment is 100MPa to 150 MPa.

The ultrahigh pressure homogenization treatment in the invention can be carried out at normal temperature. The normal temperature is, for example, 15 ℃ to 30 ℃.

In some embodiments of the present invention, the time of the ultra-high pressure homogenization treatment is 10min to 20 min.

In some embodiments of the present invention, the number of times of performing the ultra-high pressure homogenization treatment is at least 1.

In some embodiments of the present invention, the power of the ultrasonic treatment is 1000W to 2000W, and the time is 15min to 30 min.

In some embodiments of the invention, the pressure of the ultrahigh-pressure homogenization treatment is 100 to 150 MPa; the power of ultrasonic treatment is 1000W-2000W, and the time is 15 min-30 min.

In some embodiments of the invention, the pretreatment step further comprises a step of performing a colloid mill treatment on the scallop protein raw material before performing the ultrahigh-pressure homogenization treatment and the ultrasonic treatment; the pretreatment step comprises colloid mill treatment, ultrahigh pressure homogenization treatment and ultrasonic treatment.

Further, in some preferred embodiments of the invention, the amount of water added in the colloid mill is 2-5 times of the mass of the scallop protein raw material; the pressure of the ultrahigh-pressure homogenization treatment is 100 to 150MPa (for example, 1 time, 2 times or 3 times); the power of ultrasonic treatment is 1000W-2000W, and the time is 15 min-30 min.

In some embodiments of the invention, the endoprotease comprises trypsin, and/or the exoprotease comprises flavourzyme.

In some embodiments of the invention, the endoprotease is any one of trypsin, papain, neutral protease, pepsin, or any combination thereof; and/or the exoprotease is any one of flavor protease and compound protease or the combination thereof.

In some preferred embodiments of the invention, the endoprotease is trypsin; and/or the exoprotease is flavourzyme.

In some embodiments of the invention, the addition amount of the endoprotease in the first stage of enzymolysis is 0.1-0.5% of the weight of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-10 h.

In some embodiments of the present invention, during the first stage of enzymolysis (endo-enzymolysis), NaOH solution is used to regulate pH.

In some embodiments of the present invention, the first stage enzyme hydrolysis step further comprises a first enzyme deactivation step. Further, in some embodiments of the present invention, the temperature of the first inactivation is 90 ℃ to 100 ℃ and the time is 10min to 20 min.

In some embodiments of the invention, the addition amount of the exoprotease for the second stage of enzymolysis is 0.1-0.5% of the weight of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-8 h.

In some embodiments of the present invention, the second stage enzymolysis (exo-enzymolysis) is finished and then a second stage enzyme deactivation step is included. Further, in some embodiments of the present invention, the temperature of the second inactivation is 90 ℃ to 100 ℃, and the time is 10min to 20 min.

In some embodiments of the invention, the stepwise enzymolysis comprises: adding endoprotease for carrying out first-stage enzymolysis (namely, endoprotease), wherein the addition amount of the endoprotease is 0.1-0.5% of the mass of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-10 h; then the first-stage enzyme deactivation is carried out at the temperature of 90-100 ℃ for 10-20 min; adding exoproteinase for second-stage enzymolysis (namely exozymolysis), wherein the addition amount of the exoproteinase is 0.1-0.5% of the mass of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-8 h; and after the second-stage enzymolysis is finished, performing second-stage enzyme deactivation at the temperature of 90-100 ℃ for 10-20 min. Examples of the endo-protease and the exo-protease and preferred modes thereof include, but are not limited to, those described above and below. Wherein, in some preferred embodiments of the present invention, the endoprotease is trypsin and the exoprotease is flavourzyme.

In some embodiments of the invention, the solid-liquid separation means in the post-treatment step is centrifugation. Centrifuging and collecting supernatant to obtain solution containing scallop flavor peptide. Further, in some embodiments of the present invention, the rotation speed of the centrifugation is 8000rpm to 10000rpm, and the centrifugation time is 5min to 15 min.

In some embodiments of the invention, the post-processing step is: centrifuging and collecting supernatant; and/or, one or two of the following steps are also included after the post-processing step: purifying and drying scallop peptide.

The second aspect of the invention provides another preparation method of the scallop fresh-taste peptide product, which utilizes a segmented enzymolysis technology to improve the content of the fresh-taste peptide substances in scallop enzymolysis liquid, and simultaneously introduces the partridge tea extract in the hydrolysis process of scallop protein, thereby realizing the effective fishy smell removal and the improvement of the content of the fresh-taste peptide of the scallop fresh-taste peptide product in a simple and convenient way.

The preparation method of the scallop umami peptide product in the second aspect of the invention comprises the following steps:

providing raw materials: providing a scallop protein raw material;

pretreatment: carrying out ultrahigh pressure homogenization treatment or ultrasonic treatment on the scallop protein raw material, or firstly carrying out ultrasonic treatment and then carrying out ultrahigh pressure homogenization treatment to prepare scallop protein liquid; wherein the pressure of the ultrahigh-pressure homogenizing treatment is preferably not less than 100 MPa; a step of performing or not performing colloid mill treatment on the scallop protein raw material before performing the ultrahigh pressure homogenization treatment or the ultrasonic treatment;

adding an antioxidant: adding a partridge tea extract into the scallop protein liquid;

carrying out segmented enzymolysis: performing segmented enzymolysis, namely performing first-stage enzymolysis by adopting trypsin, and performing second-stage enzymolysis by adopting flavourzyme to prepare scallop protein enzymolysis liquid;

carrying out post-treatment: carrying out solid-liquid separation on the scallop protein enzymatic hydrolysate, and collecting clear liquid;

optionally, the post-processing step may further include one or both of the following steps: purifying and drying scallop peptide.

The third aspect of the invention provides another preparation method of the scallop fresh taste peptide product, the preparation method combines an ultrahigh pressure homogenization technology and an ultrasonic technology to treat scallop protein, the synergistic effect between the two processes is utilized to improve the hydrolysis rate of the scallop, and the extract of the partridge tea is introduced in the hydrolysis process of the scallop protein, so that the effective fishy smell removal of the scallop fresh taste peptide product and the utilization rate of raw materials can be simultaneously realized in a simple and convenient manner.

providing raw materials: providing a scallop protein raw material;

pretreatment: carrying out ultrahigh pressure homogenization treatment on the scallop protein raw material, and then carrying out ultrasonic treatment to prepare scallop protein liquid; wherein the pressure of the ultrahigh-pressure homogenizing treatment is preferably not less than 100 MPa;

a step of performing or not performing colloid mill treatment on the scallop protein raw material before performing the ultrahigh-pressure homogenization treatment and the ultrasonic treatment;

carrying out enzymolysis: carrying out enzymolysis, wherein the used enzymes comprise endoprotease and exoprotease to prepare scallop protein enzymolysis liquid;

optionally, the post-processing step further comprises one or two of the following steps: purifying and drying scallop peptide.

The fourth aspect of the invention provides the scallop umami peptide product prepared by the preparation method of the first aspect, the second aspect or the third aspect. The scallop fresh taste peptide product can be in a liquid state or a dry state.

Compared with the prior art, each technical scheme of the invention has at least one of the following beneficial effects:

1. the scallop protein is processed by combining an ultrahigh pressure homogenization technology and an ultrasonic technology, wherein the enzyme binding sites of the protein can be exposed from the inside by processing the scallop protein by the ultrahigh pressure homogenization technology; the scallop protein suspension is processed by adopting an ultrasonic technology, so that the protein can be changed into a hollow sphere structure and a dispersed coil, the protein is favorably dispersed in water, the internal active site of the protein is favorably exposed, and the combination of the protein and enzyme is promoted. By combining the two technologies, ultrahigh pressure homogenization treatment is firstly carried out, then ultrasonic treatment is carried out, and a synergistic effect is generated between the two processes, so that the hydrolysis rate of the scallop protein can be effectively improved, and the contents of small molecular flavor peptides and free amino acids in a hydrolysate can be improved, thereby improving the utilization rate of the scallop protein and reducing the waste of raw materials.

2. The traditional research uses the proteolysis rate as an index to research the extraction of the umami substances of the scallops, focuses on the extraction of free amino acids in the scallops, and fails to perform selective extraction on the umami peptides in the scallops. The invention improves the pertinence extraction of the scallop fresh taste peptide by using the endoprotease and the flavourzyme to carry out the segmented extraction through the segmented enzymolysis technology, thereby more fully utilizing the scallop-derived fresh taste peptide.

Specifically, when segmented enzymolysis is carried out, the first segment of enzymolysis can be utilized for carrying out incision enzymolysis to extract macromolecular peptides, then the second segment of enzymolysis is utilized for carrying out exo-enzymolysis to extract small molecular peptides, the proper hydrolysis time of the second segment of enzymolysis can also be controlled, the proportion of the umami peptides in the polypeptides obtained by enzymolysis is improved to the maximum extent, bitter peptides cannot be generated due to insufficient hydrolysis degree, most of macromolecular peptides are hydrolyzed into free amino acids due to excessive hydrolysis, and the extraction of the umami peptide components in the scallops is facilitated to the maximum extent.

3. The mallotus oblongifolius extract is introduced in the scallop protein hydrolysis process, and the fat oxidation is inhibited by utilizing the antioxidation of polyphenol and polysaccharide substances in the mallotus oblongifolius extract, so that the fishy smell in the scallop fresh taste peptide product can be effectively removed; compared with the post-treatment fishy smell removal modes such as activated carbon adsorption and Maillard reaction in the traditional technology, the technical scheme of the invention allows hydrolysis and fishy smell generation inhibition, and the method is simple and effective, can simplify the process, and can effectively inhibit fishy smell generation from the source.

4. On the basis of processing the scallop protein by combining the ultrahigh-pressure homogenization technology and the ultrasonic technology, the targeted extraction of the umami peptide is further carried out by segmented enzymolysis (for example, the segmented enzymolysis is carried out by respectively adopting endoprotease and flavourzyme), and the umami peptide component in the scallop can be extracted in a targeted manner with high yield through the synergistic effect on hydrolysis control in the pretreatment process and the enzymolysis process. Can effectively solve the problems of low hydrolysis rate of scallop protein and insufficient utilization of scallop delicate flavor substances.

5. The method is characterized in that when sectional enzymolysis (for example, sectional enzymolysis by adopting endoprotease and flavourzyme) is adopted to carry out targeted extraction on the umami peptide, the Mallotus furetianus extract is introduced in the enzymolysis process, and the generation of fishy smell substances can be effectively inhibited by utilizing the fat oxidation inhibiting effect of the Mallotus furetianus extract, so that the effective fishy smell removal of the scallop umami peptide product and the improvement of the content of the umami peptide are simultaneously realized in a simple fishy smell removal mode.

6. On the basis of processing scallop protein by combining an ultrahigh pressure homogenization technology and an ultrasonic technology, the mallotus oblongifolius muell-Arg extract is introduced in the enzymolysis process, and the effect of inhibiting fat oxidation is utilized, so that fishy smell substances can be effectively inhibited from being generated, the effective fishy smell removal of scallop delicate flavor peptide products can be realized, the protein hydrolysis rate can be improved, and the utilization rate of raw materials can be improved.

Drawings

FIG. 1 is a schematic flow chart of one embodiment of the method for preparing the scallop umami peptide product of the invention.

FIG. 2 is a schematic flow chart of one embodiment of the method for preparing the scallop umami peptide product of the present invention.

Detailed Description

The scallop flavor peptide product and the preparation method thereof according to the present invention will be further described in detail with reference to the following embodiments and examples. It should be understood that the present invention may be embodied in many different forms and is not limited to the embodiments and specific examples described herein. Rather, these embodiments and examples 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 herein in the description of the invention is for the purpose of describing particular embodiments and examples only and is not intended to be limiting of the invention.

Term(s) for

Unless otherwise stated or contradicted, terms or phrases used herein have the following meanings:

the term "and/or", "and/or" as used herein is intended to be inclusive of any one of the two or more items listed in association, and also to include any and all combinations of the items listed in association, including any two or more of the items listed in association, any more of the items listed in association, or all combinations of the items listed in association.

As used herein, "a combination thereof," "any combination thereof," and the like, includes all suitable combinations of any two or more of the listed items.

In the present specification, the term "suitable" in "a suitable combination, a suitable manner," any suitable manner "and the like shall be construed to mean that the technical solution of the present invention can be implemented, the technical problem of the present invention can be solved, and the technical effect of the present invention can be achieved.

Herein, "preferred" merely describes a more effective embodiment or example, and it should be understood that the scope of the present invention is not limited thereto.

In the present invention, "optionally" means optionally, that is, means selected from any one of two juxtapositions "present" or "absent". If multiple optional parts appear in one technical scheme, if no special description exists, and no contradiction or mutual constraint exists, each optional part is independent.

In the present invention, the terms "first stage enzymolysis", "second stage enzymolysis", "first stage inactivating enzyme activity", "second stage inactivating enzyme activity", "first aspect", "second aspect", "third aspect", "fourth aspect" and the like are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or quantity, nor as implying an importance or quantity indicating a technical feature. Also, "first," "second," "third," "fourth," etc. are used for non-exhaustive enumeration of description purposes only and should not be construed as a closed limitation to the number.

In the present invention, the technical features described in the open type include a closed technical solution composed of the listed features, and also include an open technical solution including the listed features.

In the present invention, "polypeptide" and "peptide" have the same meaning and are used interchangeably and refer to amino acid chains having a molecular weight of greater than 200 Da.

As used herein, "polypeptide" refers to a polypeptide and its derivatives.

In the present invention, "macromolecular peptide" refers to a peptide having a molecular weight of more than 1000Da, and "small-molecular peptide" refers to a peptide having a molecular weight of not more than 1000 Da. In the present invention, the umami peptide is mainly a small molecule peptide with the molecular weight not more than 1000 Da.

In the invention, the small-molecular flavor peptide refers to flavor peptide with the molecular weight not more than 1000 Da.

In the present invention, the "scallop peptide" refers to a peptide derived from scallop, which comprises at least two amino acid units.

In the present invention, "amino acid" includes amino acids and derivatives thereof, unless otherwise specified.

In the invention, the scallop flavor peptide refers to a polypeptide flavor substance which is obtained by taking scallop as a raw material and has the molecular weight of mainly 200 Da-1000 Da. It will be appreciated that small amounts of higher molecular weight polypeptide umami materials are also permitted. For example, the peptide content in the molecular weight region of 200Da to 1000Da is not less than 60%, not less than 70%, not less than 80%, not less than 90%, or not less than 95% by mass of all the molecular weight peptides, and the like.

In the invention, the "scallop flavor peptide product" refers to a product containing scallop flavor peptides, which can be in a liquid state or a dry state, wherein the flavor components can only comprise peptide components, and can also comprise other flavor components (such as amino acid components). Preferably, the scallop flavor peptide product contains 30-45% of scallop flavor peptide by mass (calculated on a dry basis). More preferably, the mass content of the polypeptide with the molecular weight of 200 Da-1000 Da in the dry basis of the scallop umami peptide product is 30-45%.

In the present invention, the numerical range is defined to include both end points of the numerical range unless otherwise specified.

The ultra-high pressure homogenization treatment is a physical means which is emerging in the present year, and the pressure for carrying out the homogenization treatment is generally not lower than 100 MPa. The ultrahigh pressure homogenization treatment can effectively improve the emulsifying property and the texture property of the food. The protein structure can be fully changed by utilizing the shearing force, the high-speed fluid collision action and the vortex action in the ultrahigh-pressure homogenization process, so that protein particles are broken, action bonds are broken, and the protein solubility is improved, so that more protease binding sites are exposed from the inside, the hydrolysis of protein is accelerated, and the hydrolysis degree of the protein is improved.

Ultrasonic treatment is a technical means for effectively improving the structure and function of protein. The spatial structure of the substrate protein can be changed into a stretched and loose state by heating, cavitation, turbulence, shear stress and dynamic stirring in the ultrasonic treatment process, so that the internal active sites of the protein are exposed, and the hydrolysis of the substrate protein by protease can be promoted.

Colloid mill treatment, which is a technique for pulverizing materials. The colloid mill treatment can be carried out to the raw materials for preliminary fining, which is beneficial to the more thorough treatment of the raw materials in the subsequent homogenization or/and ultrasonic treatment process.

Fishy smell substances: the scallop protein is easy to generate fat oxidation in the enzymolysis process, which causes fishy smell. Wherein the fishy substances having large influence on umami sensory comprise hexanal, nonanal, decanal, 1-octen-3-ol, 1-penten-3-ol, 2, 4-heptadienal and 2, 4-decadienal. Research has shown that hexanal, nonanal, decanal, 2, 4-heptadienal, 2, 4-decadienal are typical fat oxidation products and contribute significantly to the production of fishy smell, while 1-octen-3-ol and 1-penten-3-ol are produced mainly by the oxidation of arachidonic acid.

Mallotus oblongifolius extract, which can be prepared by the following method: adding water into Mallotus furetianus, heating and extracting, and filtering to obtain Mallotus furetianus extract. Mallotus oblongifolius extract contains a large amount of antioxidant components, such as tea polyphenols such as catechin, anthocyanin, flavone, flavonol, phenolic acid, etc., and tea polysaccharides formed by polymerization of glucose, xylose, fucose, arabinose, etc. In the invention, the Mallotus furetianus extract is applied to proteolysis, and the oxidation of fat can be effectively inhibited by utilizing the antioxidation effect of polyphenol and polysaccharide substances in the Mallotus furetianus extract, so that the generation of fishy smell substances is inhibited.

The segmented enzymolysis in the invention comprises first-stage enzymolysis and second-stage enzymolysis. In the first stage of the enzymatic hydrolysis, endoprotease is used for enzymatic hydrolysis, and therefore, the method is also called endoproteolysis. In the second stage, the enzymatic hydrolysis is carried out using exoproteases and is therefore also referred to as exoenzymatic hydrolysis. The first section of enzymolysis hydrolyzes protein into macromolecular peptide from the interior of molecules, the second section of enzymolysis hydrolyzes the macromolecular peptide into micromolecular peptide and amino acid, and the two enzymolysis steps are cooperated with each other, so that high hydrolysis rate is realized, and extraction of the micromolecular peptide to the maximum extent is also realized.

The percentage contents referred to in the present invention mean, unless otherwise specified, mass percentages for solid-liquid mixing and solid-solid phase mixing, and volume percentages for liquid-liquid phase mixing.

The antioxidant addition amount, the mallotus oblongifolius tea addition amount and the protease addition amount in the enzymolysis stage in the invention are mass percentages relative to the scallop protein liquid, unless otherwise specified. Examples of descriptive means include, but are not limited to: the addition amount is 0.3 percent of the weight of the scallop protein liquid, the addition amount is 0.3 percent of the weight percentage of the scallop protein liquid, and the like.

The temperature parameter in the present invention is not particularly limited, and is allowed to be constant or to fluctuate within a certain temperature range. The constant temperature process allows for fluctuations within the accuracy of instrument control.

The invention provides a preparation method of a scallop fresh taste peptide product, which comprises the following steps:

providing raw materials: providing a scallop protein raw material;

segmented enzymolysis: carrying out segmented enzymolysis on the scallop protein liquid to prepare scallop protein enzymolysis liquid; the segmented enzymolysis comprises a first-stage enzymolysis (also called as endo enzymolysis) by adopting endoprotease, and a second-stage enzymolysis (also called as exo-enzymolysis) by adopting exoproteinase;

According to the preparation method provided by the first aspect of the invention, the scallop protein is treated by adopting an ultrahigh pressure homogenization technology and then an ultrasonic technology, the hydrolysis rate of the scallop can be obviously improved and the utilization rate of raw materials can be improved by utilizing the synergistic effect of the two treatment processes, and the targeted extraction of the scallop fresh taste peptide is improved by segmented enzymolysis; the pretreatment step and the segmented enzymolysis step form a synergistic effect on hydrolysis control, the protein hydrolysis rate is improved, and the content of the umami peptide in an enzymolysis product is improved, so that the scallop umami peptide can be more fully utilized.

The type of scallop of the scallop protein raw material is not particularly limited, and includes, but is not limited to, bay scallop, Japanese scallop, and chlamys farreri, as examples.

The source of the scallop protein material includes, but is not limited to, scallop meat, scallop skirt, scallop mantle, etc.

The scallop protein raw material can be obtained by the following steps: selecting fresh scallop and removing shell or removing shell of thawed scallop, washing with running water to obtain scallop protein raw material.

In some embodiments of the invention, the scallop flavor peptide product has a scallop flavor peptide content of greater than or equal to 50% by weight of the total peptides.

In some embodiments of the invention, the mass content of the polypeptides with the molecular weight of 200 Da-1000 Da in the scallop delicious peptide product in the total peptides is more than or equal to 50%. In some preferred embodiments of the invention, the mass content of the polypeptides with molecular weight of 200Da to 1000Da in the scallop umami peptide preparation in the total peptides is 50% to 65%, and specific examples include but are not limited to 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 62%, 63%, 64%, 65%.

In some embodiments of the invention, the mass content of the polypeptide with the molecular weight of 200 Da-1000 Da in the dry basis of the scallop fresh taste peptide preparation is 30-45%, and specific examples include but are not limited to 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%.

In some embodiments of the invention, the scallop protein is treated by combining an ultrahigh pressure homogenization technology and an ultrasonic technology, so that the hydrolysis rate of the protein can be effectively improved to be not less than 50 percent, even more than 70 percent. In some preferred embodiments of the present invention, the proteolysis rate is 55%, 60%, 65%, 70%, etc. In some preferred embodiments of the invention, the proteolysis rate is 55%, 60%, 65%, 68%, 70%, 72%, etc.

In some embodiments of the present invention, the step of performing the ultra-high pressure homogenization treatment and the ultrasonic treatment further comprises performing a colloid mill treatment on the scallop protein raw material. In some embodiments of the invention, the water adding amount of the colloid mill treatment is 2-5 times of the mass of the scallop protein raw material; in one preferred scheme, the water adding amount is 2 times, 2.5 times, 3 times, 3.5 times, 4 times, 4.5 times or 5 times of the mass of the scallop protein raw material. In some embodiments of the invention, the colloid mill treatment is performed at least 1 time; one of the preferred embodiments is to perform the colloid mill treatment 1, 2, 3 or 4 times. The colloid mill treatment can be carried out to the raw materials for preliminary fining, which is beneficial to the more thorough treatment of the raw materials in the subsequent homogenization or/and ultrasonic treatment process.

In the pretreatment step, the sequence of the ultrahigh-pressure homogenization treatment and the ultrasonic treatment has a great influence on the enzymolysis effect. The ultrahigh-pressure homogenization treatment is carried out firstly, then the ultrasonic treatment is carried out, the two treatment processes can play a synergistic role, and the functions of cavitation, shearing stress and the like can be played in the ultrasonic process after the ultrahigh-pressure homogenization. The inventor finds through experimental investigation that the sequence of the ultrahigh-pressure homogenization treatment and the ultrasonic treatment is adjusted, and the ultrahigh-pressure homogenization treatment is performed after the ultrasonic treatment, so that not only is the proteolysis rate seriously reduced, but also the synergistic effect between the two treatment modes is basically eliminated, and refers to example 4.

In some embodiments of the present invention, in the pre-treatment step, an ultra-high pressure homogenization treatment is performed first, and then an ultrasonic treatment is performed. The method comprises the steps of firstly adopting an ultrahigh pressure homogenization technology to process scallop protein to expose enzyme binding sites of the protein from the inside to obtain scallop protein suspension, and then further adopting an ultrasonic technology to process the scallop protein suspension to enable the protein to be changed into a hollow sphere structure and a dispersed coil, so that the protein is favorably dispersed in water, the combination of the protein and the enzyme is promoted, the hydrolysis rate of the scallop protein is effectively improved, and the utilization rate of the fresh taste substances of the scallop is improved.

In some embodiments of the present invention, the pressure of the ultra-high pressure homogenization treatment is 100MPa to 150 MPa. The inventor finds out through experimental exploration that if the pressure is too high (relative to 100 MPa-150 MPa), the hydrolysis degree of the scallop protein is not obviously influenced; if the pressure is too low, the degree of hydrolysis of the scallop protein cannot be significantly increased.

In some embodiments of the invention, the pressure of the ultra-high pressure homogenization treatment is 100MPa, 110MPa, 120MPa, 125MPa, 130MPa, 140MPa, 150 MPa.

The ultrahigh pressure homogenization treatment in the invention can be carried out at normal temperature. The normal temperature is, for example, 15 to 30 ℃. More specific examples are any one of the following temperatures or a temperature interval between any two temperatures: 15 ℃, 16 ℃, 17 ℃, 18 ℃, 19 ℃,20 ℃, 21 ℃, 22 ℃, 23 ℃, 24 ℃, 25 ℃, 26 ℃, 27 ℃, 28 ℃, 29 ℃ and 30 ℃. The temperature range between any two temperatures includes, for example, 15 ℃ to 24 ℃,20 ℃ to 30 ℃, 16 ℃ to 20 ℃, 16 ℃ to 26 ℃, 18 ℃ to 26 ℃ and the like.

In some embodiments of the present invention, the number of times of performing the ultra-high pressure homogenization treatment is at least 1. One of the preferred embodiments is to perform the ultra-high pressure homogenization treatment 1 time, 2 times or 3 times.

In some embodiments of the present invention, the number of times of performing the ultra-high pressure homogenization treatment is more than 1. One of the preferred embodiments is to perform the ultra-high pressure homogenization treatment 2 or 3 times.

In some embodiments of the present invention, the time for performing the ultra-high pressure homogenization treatment is 2min to 10 min. The inventor finds out through experimental exploration that if the homogenization treatment time is too short, the molecular structure of the protein is not fully changed, the hydrolysis rate of the scallop protein cannot be obviously improved, and if the homogenization treatment time is too long, the improvement on the hydrolysis rate of the scallop protein is not obvious.

Examples of the time for performing the ultrahigh-pressure homogenization treatment are 2min, 3min, 4min, 5min, 6min, 7min, 8min, 9min, and 10 min.

In some embodiments of the present invention, the power of the ultrasonic treatment is 1000W to 2000W, and the time is 15min to 30 min. The power of the ultrasonic treatment is, for example, 1000W, 1200W, 1400W, 1500W, 1600W, 1800W, 2000W. Examples of the time of the ultrasonic treatment include 15min, 20min, 25min, and 30 min. The inventor finds that the improvement on the hydrolysis rate of the scallop protein is not obvious if the power is too high through experimental exploration; if the power is too low, the protein molecule space structure is expanded to a small extent, which is not beneficial to enzymolysis. The inventor also discovers through experimental exploration that if the ultrasonic treatment time is too short, the effects of a cavity effect, shearing stress, dynamic stirring and the like cannot be completely generated, the spatial structure of the protein is not completely changed, and the purpose of improving the proteolysis rate cannot be achieved; if the ultrasonic treatment time is too long, the temperature of the scallop protein is too high, so that the protein molecules are excessively denatured and aggregated, and the proteolysis is not facilitated.

In some embodiments of the invention, the power of the ultrasonic treatment is 1600W and the time is 15min, 20min, 25min or 30 min.

In some embodiments of the present invention, the pre-treatment step comprises subjecting the scallop protein material to a colloid mill treatment, followed by an ultra-high pressure homogenization treatment, and then an ultrasonic treatment. In some preferred embodiments of the invention, the amount of water added in the colloid mill is 2-5 times of the mass of the scallop protein raw material; the pressure of the ultrahigh-pressure homogenization treatment is 100 to 150MPa (further, for example, 1 time, 2 times, or 3 times); the power of ultrasonic treatment is 1000W-2000W, and the time is 15 min-30 min.

In the step of segmented enzymolysis, incision enzymolysis is carried out in the first stage enzymolysis process, and excision enzymolysis is carried out in the second stage enzymolysis process. Experimental research shows that the order of the incision enzymolysis and the excision enzymolysis has great influence on the enzymolysis effect, and the incision enzymolysis and then the excision enzymolysis (the segmented enzymolysis mode of the first inside and the second outside) is superior to the segmented enzymolysis mode of the first outside and the second inside). In the first stage of enzymolysis process, firstly, endoprotease is used for hydrolyzing protein from the interior of molecules into macromolecular peptides, then in the second stage of enzymolysis process, exoprotease is used for continuously hydrolyzing the macromolecular peptides into small molecular peptides and free amino acids, and the scallop delicate flavor peptide content of the enzymolysis products can be improved in a targeted manner. The inventor finds out through experimental exploration that if the order of endoprotease and exoprotease is adjusted, the enzymolysis cannot be fully carried out under the same condition under other experimental conditions, and the content of the scallop flavor peptide in the scallop flavor peptide solution obtained after solid-liquid separation is tested, wherein the 'first outer and then inner' sectional enzymolysis mode is obviously lower than the 'first inner and then outer' sectional enzymolysis mode; and when the segmented enzymolysis mode of 'first outside then inside' is adopted, the enzymolysis time needs to be prolonged to reach the proper content of the umami peptide.

In some embodiments of the invention, the endoprotease is any one of trypsin, papain, neutrase, pepsin, and the like, or any combination thereof.

In some embodiments of the invention, the endoprotease comprises trypsin.

In some preferred embodiments of the invention, the endoprotease is trypsin. The inventor finds through experimental investigation that compared with other endoproteases, the proteolysis rate is higher when the endoprotease is used for endoprotease, excessive pH adjustment is not needed in the hydrolysis process, and the operation is simpler. According to the comparative experiment of example 1, under the same experimental conditions, the hydrolysis rates of trypsin, papain and neutral protease were 45%, 37% and 30%, respectively, and the mass contents of small-molecule umami peptide (200 Da-1000 Da) in the total peptide were 30%, 20% and 17%, respectively. Therefore, a synergistic effect is generated between the endo-enzymolysis (first-stage enzymolysis) and the exo-enzymolysis (second-stage enzymolysis), and the content of the micromolecule delicious peptide is improved by adopting trypsin for the endo-enzymolysis in the segmented enzymolysis.

In some embodiments of the invention, the exoprotease is any one of, or a combination of, flavourzyme, complex protease, and the like.

In some embodiments of the invention, the exoprotease comprises a flavourzyme.

In some preferred embodiments of the invention, the exoprotease is a flavourzyme. The inventor finds that compared with other exoproteases such as compound protease, the bitter taste of an enzymolysis product obtained by performing exo-enzymolysis (second stage enzymolysis) by using flavourzyme is the weakest.

In some embodiments of the invention, the endoprotease comprises trypsin and the exoprotease comprises flavourzyme.

The enzymolysis temperature and pH of the protease used in the invention are preferably the temperature and pH which can better exert the enzyme activity. The temperature and pH of the enzymatic hydrolysis can be controlled according to the instructions for the commercial product.

Further, the content of the delicious peptide in the enzymolysis product can be maximally increased by reasonably controlling the enzymolysis time. In some embodiments of the invention, the time for the first stage of the enzymatic hydrolysis using endoprotease is 6 to 10 hours and the time for the second stage of the enzymatic hydrolysis using exoprotease is 6 to 8 hours. The control of the hydrolysis time of the protease is very important. If the treatment time of the endoprotease in the first stage of enzymolysis is too short, a large amount of protein which is not hydrolyzed into macromolecular peptide exists, so that the content of the umami substances in the enzymolysis products is too low; if the endoprotease treatment time is too long, excessive proteolysis may occur, which may result in substantial hydrolysis of the macromolecular peptides to free amino acids, resulting in an underdetermined umami peptide content, and thus a reasonable control of the first enzymatic hydrolysis time is required. If the exoprotease treatment time of the second stage of enzymolysis is too short, the macromolecular peptide generated by the first stage of enzymolysis cannot be subjected to full enzymolysis, and most of unhydrolyzed macromolecular peptide still exists in the system; if the exoprotease treatment time is too long, the conversion of most of the small-molecule peptides to free amino acids is excessive, and the content ratio of the umami peptide in the protein hydrolysate is reduced; therefore, the second stage enzymolysis time needs to be reasonably controlled to maximize the fresh flavor peptide ratio. The protein hydrolysate is mainly hydrolyzed polypeptide and free amino acid.

In some embodiments of the invention, the addition amount (relative to the mass percentage of the scallop protein liquid) of the endoprotease in the first stage of enzymolysis is 0.1-0.5%, the enzymolysis temperature is 40-65 ℃, the pH of an enzymolysis system is 7-10, and the enzymolysis time is 6-10 h. Examples of the amount of the endoprotease include 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, and 0.5%. Examples of the temperature of the enzymatic hydrolysis include 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃ and 65 ℃. Examples of the pH of the enzymatic hydrolysis system include pH7, pH7.2, pH7.4, pH7.2 to pH7.6, pH7.8, pH8.0, pH8.5, pH9, pH9.5, and pH 10. Examples of the enzymolysis time include 6h, 6.5h, 7h, 7.5h, 8h, 8.5h, 9h, 9.5h and 10 h.

In some embodiments of the invention, the addition amount (relative to the mass percentage of the scallop protein liquid) of the exoprotease in the second stage of enzymolysis is 0.1-0.5%, the enzymolysis temperature is 40-65 ℃, the pH of the enzymolysis system is 7-10, and the enzymolysis time is 6-8 h. Examples of the addition amount of the exoprotease include 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, and 0.5%. Examples of the temperature of the enzymatic hydrolysis include 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃ and 65 ℃. Examples of the pH of the enzymatic hydrolysis system include pH7, pH7.2, pH7.4, pH7.2 to pH7.6, pH7.8, pH8.0, pH8.5, pH9, pH9.5, and pH 10. Examples of the enzymolysis time are 6h, 6.5h, 7h, 7.5h and 8 h.

The enzyme deactivation is carried out after each section of enzymolysis is finished, so that excessive enzymolysis can be prevented, and a certain sterilization effect can be achieved.

In some embodiments of the invention, the staged enzymatic hydrolysis is carried out in an aqueous solvent. In some embodiments of the invention, the solvent in which the staged enzymatic hydrolysis is carried out is water.

In some embodiments of the invention, the stepwise enzymolysis comprises: adding endoprotease for carrying out first-stage enzymolysis (i.e. endoprotease), wherein the addition amount of the endoprotease is 0.1-0.5% of the weight of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-10 h; then the first-stage enzyme deactivation is carried out at the temperature of 90-100 ℃ for 10-20 min; adding exoproteinase for second-stage enzymolysis (namely exoenzymolysis), wherein the addition amount of the exoproteinase is 0.1-0.5% of the mass of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-8 h; and after the second-stage enzymolysis is finished, performing second-stage enzyme deactivation at the temperature of 90-100 ℃ for 10-20 min. Examples of the endo-protease and the exo-protease and preferred modes thereof include, but are not limited to, those described above and below. Wherein, in some preferred embodiments of the present invention, the endoprotease is trypsin and the exoprotease is flavourzyme.

In some embodiments of the invention, the francolin tea extract is present in the hydrolysis process, and the fat oxidation inhibition effect of the francolin tea is utilized to achieve effective inhibition of fishy smell generation from the source during hydrolysis. In some preferred embodiments of the invention, the mallotus oblongifolius extract is added after pre-treatment and before staged enzymatic hydrolysis.

In some embodiments of the present invention, the step of performing the staged enzymolysis further comprises a step of adding an antioxidant to the scallop protein liquid. The antioxidant (such as the Mallotus furetianus extract) is introduced in the scallop protein hydrolysis process, fat oxidation can be inhibited during hydrolysis, fishy smell generation can be effectively inhibited from the source, the content of fishy smell substances in an enzymolysis product is effectively reduced, and compared with a post-treatment fishy smell removing mode such as activated carbon adsorption and the like, the method is simpler and more effective.

Further, in some embodiments of the invention, the antioxidant comprises mallotus oblongifolius extract. Further, in some preferred embodiments of the present invention, the addition amount of the mallotus oblongifolius extract is 0.5 to 2% of the mass of the scallop protein liquid. The addition amount of the mallotus oblongifolius extract is, for example, 0.5%, 1.0%, 1.5%, 2.0% by mass of the scallop protein solution.

In some embodiments of the present invention, the antioxidant is selected from any one of antioxidants of mallotus oblongifolius extract, yeast extract (containing antioxidant peptides), chrysanthemum tea, or any combination thereof.

In some preferred embodiments of the present invention, the step of performing the staged enzymolysis further comprises a step of adding an antioxidant to the scallop protein liquid, wherein the antioxidant is partridge tea extract. In some more preferred embodiments of the invention, the addition amount of the mallotus oblongifolius extract is 0.5-2% of the mass of the scallop protein liquid. The addition amount of the mallotus oblongifolius extract is, for example, 0.5%, 1.0%, 1.5%, 2.0% by mass of the scallop protein solution.

In other embodiments of the present invention, the antioxidant comprises a second antioxidant, which is not mallotus oblongifolius extract and is selected from any one or any combination of antioxidants from yeast extract (containing antioxidant peptides), chrysanthemum tea, and the like.

In other embodiments of the present invention, the antioxidant comprises mallotus oblongifolius extract and a second antioxidant.

In some embodiments of the invention, a second antioxidant is present during hydrolysis. The second antioxidant may be any one selected from yeast extract (containing antioxidant peptide), chrysanthemum tea, and the like, or any combination thereof. In some preferred embodiments of the present invention, the second antioxidant is added after the pretreatment and before the staged enzymolysis.

The invention firstly uses the Mallotus oblongifolius muell-Arg extract to extract the fresh taste of the scallop. In addition, the mallotus oblongifolius extract as an antioxidant has the following advantages compared with the antioxidant such as yeast extract (containing antioxidant peptide) and chrysanthemum tea: besides good fat oxidation inhibition effect, the antibacterial agent also has good antibacterial effect.

In some embodiments of the invention, mallotus oblongifolius extract and a second antioxidant are present during the hydrolysis process. Further, in some embodiments of the invention, the addition time of the partridge tea extract and the second antioxidant is both after pretreatment and before staged enzymolysis; the two can be added into scallop protein solution together or separately; when added separately, the order of addition is not particularly limited.

In some embodiments of the invention, the solid-liquid separation means in the post-treatment step is centrifugation. Centrifuging, collecting supernatant, and preparing solution containing scallop flavor peptide. Further, in some embodiments of the present invention, the rotation speed of the centrifugation is 8000rpm to 10000rpm, and the centrifugation time is 5min to 15 min. Examples of such rotational speeds include, but are not limited to, 8000rpm, 8500rpm, 9000rpm, 9500rpm, 10000 rpm. Examples of the centrifugation time include, but are not limited to, 5min, 6min, 7min, 8min, 9min, 10min, 11min, 12min, 13min, 14min, 15 min.

In some embodiments of the invention, the centrifugation is performed at 8000rpm for 10 min.

In some embodiments of the invention, the peptide distribution in the supernatant after hydrolysis is determined and peptides with molecular weights of 200Da to 1000Da are found to account for more than 55% by mass of the total peptides.

In some embodiments of the invention, the post-treatment step is followed by one or both of the following steps: purifying and drying scallop peptide.

The centrifugal supernatant containing the scallop flavor peptide is subjected to scallop peptide purification by adopting purification modes including but not limited to column chromatography, membrane filtration, ultrafiltration and the like, so that the scallop peptide component with specific molecular weight is collected to obtain a purified scallop flavor peptide product. The purification means may employ techniques including, but not limited to, conventional means for obtaining purified peptides. The purified product may be in the form of a solution or dried. The peptide component in the purified scallop fresh taste peptide product can be a single component or a combination of a plurality of peptide combinations.

Drying the supernatant (such as centrifugal supernatant) containing the scallop flavor peptide obtained by post-treatment to obtain a dry product of the scallop flavor peptide product. The fresh liquid containing the scallop flavor peptide can be purified by the scallop peptide and then dried to obtain a dry product of the scallop flavor peptide product. The drying method includes, but is not limited to, drying, freeze drying, spray drying, etc., and is preferably not harmful to the umami effect.

In some embodiments of the present invention, the method for preparing the scallop flavor peptide product adopts the process shown in fig. 1, and comprises the following steps: providing a scallop protein raw material, carrying out ultrahigh-pressure homogenization treatment and ultrasonic treatment in sequence, adding or not adding a Mallotus furetianus extract, carrying out first-stage enzymolysis (endo-enzymolysis) and second-stage enzymolysis (exo-enzymolysis) in sequence, carrying out solid-liquid separation (for example, centrifugation), and collecting clear liquid to obtain the scallop delicate flavor peptide solution.

In some embodiments of the present invention, the method for preparing the scallop umami peptide product adopts the process shown in fig. 2, and comprises the following steps: providing a scallop protein raw material, carrying out colloid mill treatment, carrying out ultrahigh pressure homogenization treatment and ultrasonic treatment sequentially, adding an antioxidant (including but not limited to a partridge tea extract), carrying out endo-enzymolysis and exo-enzymolysis (for example, firstly carrying out endo-enzymolysis by trypsin, namely first stage enzymolysis, and then carrying out exo-enzymolysis by flavourzyme, namely second stage enzymolysis), carrying out solid-liquid separation (for example, centrifugation), collecting clear liquid, obtaining scallop flavor peptide solution (liquid scallop flavor peptide product), optionally carrying out purification (liquid or dry scallop flavor product can be obtained), and drying (dry scallop flavor peptide product can be obtained). Can effectively solve the problems of low hydrolysis rate of scallop protein and insufficient utilization of scallop delicate flavor substances, and can simply and effectively inhibit the generation of fishy flavor substances from the source.

In some embodiments of the invention, the method for preparing the scallop umami peptide product comprises the following steps:

firstly, providing a scallop protein raw material;

secondly, preprocessing the scallop protein raw material to prepare scallop protein solution; wherein the pretreatment comprises ultrahigh pressure homogenization treatment and then ultrasonic treatment; before the ultrahigh pressure homogenization treatment and the ultrasonic treatment, the scallop protein raw material can be subjected to colloid mill treatment or subjected to colloid mill treatment;

thirdly, adding the partridge tea extract into the scallop protein liquid; a second antioxidant selected from any one of yeast extract (containing antioxidant peptide), chrysanthemum tea, and the like or any combination thereof may be added or not added;

fourthly, carrying out segmented enzymolysis to prepare scallop protein enzymolysis liquid; the segmented enzymolysis comprises the steps of firstly carrying out first-stage enzymolysis by adopting endoprotease and then carrying out second-stage enzymolysis by adopting exoprotease;

examples of the endo-protease and the exo-protease and preferred modes thereof include, but are not limited to, those described above and below;

fifthly, centrifuging the scallop protease hydrolysate and collecting supernatant.

The implementation modes and the preferred modes of colloid mill treatment, ultrahigh pressure homogenization treatment, ultrasonic treatment, staged enzymolysis treatment and centrifugation treatment include but are not limited to the above or below, and different technical characteristics can be combined in any suitable mode.

firstly, providing a scallop protein raw material;

secondly, preprocessing the scallop protein raw material to prepare scallop protein solution; wherein the pretreatment comprises colloid milling, ultrahigh pressure homogenization and ultrasonic treatment;

the water adding amount of the colloid mill is 2-5 times of the mass of the scallop protein raw material;

the pressure of the ultrahigh-pressure homogenizing treatment is 100-150 MPa, and the ultrahigh-pressure homogenizing treatment can be carried out for 1 time, 2 times or 3 times;

the power of ultrasonic treatment is 1000W-2000W, and the time is 15 min-30 min;

thirdly, adding a partridge tea extract into the scallop protein liquid, wherein the addition amount of the partridge tea extract is 0.5-2% of the mass of the scallop protein liquid;

a second antioxidant can be added or not added, wherein the second antioxidant is selected from any antioxidant in yeast extract (containing antioxidant peptide) and chrysanthemum tea or any combination mode of the antioxidants, and the total adding amount of the antioxidants is 0.5-2% of the weight of the scallop protein liquid;

fourthly, carrying out segmented enzymolysis to prepare scallop protein enzymolysis liquid; the segmented enzymolysis comprises the steps of firstly carrying out first-stage enzymolysis by adopting endoprotease, then carrying out first-stage enzyme inactivation treatment, then carrying out second-stage enzymolysis by adopting exoprotease, and then carrying out second-stage enzyme inactivation treatment;

the addition amount of the endoprotease is 0.1-0.5% of the weight of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-10 h;

the addition amount of the exoprotease is 0.1-0.5% of the weight of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-8 h;

fifthly, centrifuging the scallop protease hydrolysate, and collecting supernatant;

one of the preferable modes of centrifugation is 8000 rpm-10000 rpm, and the centrifugation time is 5 min-15 min.

In some embodiments of the present invention, the preparation method of the scallop flavor peptide product can be realized by the following steps: adding 2-5 times of water by mass into the scallop protein raw material, and carrying out colloid mill treatment for 3 times; then carrying out ultrahigh pressure homogenization treatment under the pressure of 100-150 MPa for 1, 2 or 3 times to obtain a homogenized solution; carrying out ultrasonic treatment on the homogeneous liquid, wherein the power is 1000W-2000W, and the ultrasonic time is 15 min-30 min, so as to obtain scallop protein liquid; adding the Mallotus oblongifolius muell-Arg extract into the scallop protein liquid, wherein the addition amount is 0.5-2% of the weight of the scallop protein liquid; carrying out segmented enzymolysis so as to obtain the delicious peptide to the maximum extent by controlling the enzymolysis process: firstly, adding endoprotease (such as trypsin) for carrying out first-stage enzymolysis, wherein the addition amount of the endoprotease is 0.1-0.5% of the mass of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, and NaOH solution is adopted to regulate and control the pH in the enzymolysis process so that the pH is controlled at 7-10, and the enzymolysis time is 6-10 h; then inactivating enzyme at 90-100 ℃ for 10-20 min; then adding flavourzyme for second-stage enzymolysis, wherein the addition amount of the flavourzyme is 0.1-0.5% of the mass of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH is controlled to be 7-10, and the enzymolysis time is 6-8 h so as to obtain the high umami peptide content; inactivating the enzyme at 90-100 ℃ for 10-20 min after the second stage of enzymolysis; then centrifuging for 5 min-15 min under the condition of 8000 rpm-10000 rpm, and collecting supernatant fluid to obtain the scallop flavor peptide solution.

The second aspect of the invention provides another preparation method of scallop delicious peptide products, which utilizes a segmented enzymolysis technology to improve the content of the delicious peptide substances in scallop enzymolysis liquid, and simultaneously introduces the Mallotus oblongifolius muell-Arg extract in the hydrolysis process of scallop protein, thereby realizing the improvement of the content of the delicious peptide while simply and effectively inhibiting the generation of fishy smell.

providing raw materials: providing a scallop protein raw material;

pretreatment: carrying out ultrahigh pressure homogenization treatment or ultrasonic treatment on the scallop protein raw material, or firstly carrying out ultrasonic treatment and then carrying out ultrahigh pressure homogenization treatment to prepare scallop protein liquid; wherein the pressure of the ultrahigh-pressure homogenizing treatment is preferably not less than 100 MPa;

the scallop protein raw material can be subjected to or not subjected to colloid mill treatment before the ultrahigh-pressure homogenization treatment or the ultrasonic treatment;

segmented enzymolysis: performing segmented enzymolysis, namely performing first-stage enzymolysis by adopting trypsin, and performing second-stage enzymolysis by adopting flavourzyme to prepare scallop protein enzymolysis liquid;

and (3) post-treatment: carrying out solid-liquid separation on the scallop protein enzymatic hydrolysate, and collecting clear liquid;

The technical characteristics of the preparation method of the scallop flavor peptide product according to the second aspect can be exemplified and optimized without contradiction, and the characteristics of the preparation method can be exemplified and optimized without limitation in the first aspect and the following specific examples.

The third aspect of the invention provides another preparation method of a scallop fresh taste peptide product, which combines an ultrahigh pressure homogenization technology and an ultrasonic technology to treat scallop protein to improve the hydrolysis rate of the scallop, and introduces a pintada extract in the hydrolysis process of the scallop protein, so that the yield and the hydrolysis rate of the protein can be improved while the generation of fishy smell can be simply and effectively inhibited.

providing raw materials: providing a scallop protein raw material;

pretreatment: carrying out ultrahigh pressure homogenization treatment on the scallop protein raw material, and then carrying out ultrasonic treatment to prepare scallop protein liquid; wherein the pressure of the ultrahigh-pressure homogenizing treatment is preferably not less than 100 MPa; the scallop protein raw material can be subjected to or not subjected to colloid mill treatment before the ultrahigh-pressure homogenization treatment and the ultrasonic treatment;

enzymolysis: carrying out enzymolysis to prepare scallop protein enzymolysis liquid; wherein the enzyme comprises endoprotease and exoprotease;

The technical characteristics of the preparation method of the scallop delicious peptide product can be exemplified and optimized in the aspects including but not limited to the characteristics of the first aspect and the following specific examples without contradiction.

The fourth aspect of the invention provides the scallop umami peptide product obtained by the preparation method of the first aspect, the second aspect or the third aspect. The obtained product can be in liquid state or dry state, and can be a composition of two or more peptide components or a single peptide component.

Evaluation of enzymolysis effect and related detection method

1. Determination of the proteolytic Rate

The utilization degree of the scallop protein can be measured by the proteolysis rate, and the calculation formula of the proteolysis rate is as follows:

proteolysis ratio (%) × 100 (ammonia ground state nitrogen content in the enzymatic hydrolysate/total nitrogen content in the enzymatic hydrolysate)

Wherein, the measurement of the amino nitrogen content and the total nitrogen content in the scallop enzymolysis liquid is known to the technicians in the field. For example, the total nitrogen content can be measured by the Kjeldahl method, and the amino nitrogen content can be measured by the formaldehyde titration method (GB 5009.235-2016).

Under the background technology that the current scallop protein is generally not utilized sufficiently, the improvement of the protein hydrolysis rate is beneficial to the improvement of the utilization rate of raw materials.

2. Method for measuring different components

Protein assays can be performed according to GB 5009.5-2016, amino acid assays can be performed according to GB 5009.124-2016, and fat assays can be performed according to GB/T5009.6-2003.

3. Peptide distribution assay

Peptide distribution detection refers to the detection of peptide content distribution of different molecular weights, which can be analyzed using mass content. Peptide distribution in the clear solution containing scallop umami peptide can be tested by referring to GB/T22492-2008 appendix A GPC/UV.

4. Sensory analysis

Those skilled in the art know how to perform organoleptic evaluation analysis of umami taste, such as scoring tests, brief description tests, and the like.

For example, polypeptides in a relatively large molecular weight range, including but not limited to 200Da to 1000Da, 1000Da to 5000Da, can be determined by the sensory analysis method for polypeptide umami substances described directly or/and indirectly in the references "M Ogasawara, T Katsuma, M Egi. step properties of Maillard reaction products prepared from 1000 to 5000Da Peptide [ J ]. Food Chemistry,2006,99(3):600- & 604.".

Sensory analysis of umami, fishy, bitter, etc. may also be performed using scoring methods including, but not limited to, quintile and heptatile.

5. Determination of the content of volatile odorous substances

The content of the volatile odorous substances can be measured by a Solid Phase Micro-extraction (SPME) method, and the relative content of the odorous substances is calculated from the ratio of the peak areas of the odorous compound and the internal standard substance.

The following are specific examples.

Experimental parameters not described in the following specific examples are preferably referred to the guidelines given in the present application, and may be referred to experimental manuals in the art or other experimental methods known in the art, or to experimental conditions recommended by the manufacturer.

The starting materials and reagents mentioned in the following specific examples are commercially available or can be easily obtained or prepared by those skilled in the art.

It should be noted that bay scallops are used in the following specific examples to describe the technical solution of the present invention in more detail. It should be understood that the scallop variety of the present invention is not limited to the specific scallop variety in the following examples, and may be replaced or supplemented with other varieties of scallops such as Japanese scallop, chlamys farreri, etc. The raw material of scallop protein may not be limited to one scallop source, allowing for two or more scallop sources.

Test method

1. Determination of the proteolytic Rate

The calculation formula of the proteolysis rate is as follows:

proteolysis ratio (%). ratio (amino nitrogen content in hydrolysate/total nitrogen content in hydrolysate). times.100

The total nitrogen content in the protein hydrolysate is determined by adopting a Kjeldahl method, and the amino nitrogen content in the protein hydrolysate is determined by adopting a formaldehyde titration method (GB 5009.235-2016).

2. Peptide distribution assay

The peptide distribution of the scallop fresh taste peptide in solution is tested by referring to GB/T22492-.

3. Content analysis of umami peptide

According to the peptide distribution detection result, the mass percentage of peptides with the molecular weight of 200 Da-1000 Da in the total peptides is calculated and is marked as the ratio of the umami peptides (200 Da-1000 Da), so that the content of the umami peptides in the centrifugal supernatant of the enzymolysis final product is measured.

4. Determination of the content of volatile odorous substances

The content of volatile odorous substances is determined by a Solid Phase Microextraction (SPME) method, and the relative content of the odorous substances is calculated by the ratio of the peak areas of the odorous compounds and the internal standard substance. The method mainly detects volatile fishy substances such as hexanal, nonanal, decanal, 1-octen-3-ol, 1-penten-3-ol, 2, 4-heptadienal, 2, 4-decadienal and the like. The specific test method is as follows:

SPME sample treatment: 2.5g of the enzymatic hydrolysate was put into a 20mL headspace bottle, 2.5mL of NaCl solution (0.18g/mL) was added, and 100. mu.L of o-dichlorobenzene solution (3.03. mu.g/mL) was added as an internal standard.

SPME test: the samples were SPME extracted using a 2cm tip of DVB/CAR/PDMS (Divinylbenzene/Car/Polydimethylsiloxane, Divinylbenzone/Carboxen/polydimethysiloxane) fiber. Extracting at 50 deg.C for 20min for 50min, and rapidly desorbing in GC sample inlet for 5 min. GC-MS conditions: the GC injection port temperature was 250 ℃ and the desorption time was 5 min. The odorous substances were separated using a DB-wax chromatography column (30 m.times.0.25 mm inner diameter. times.0.25 μm). The temperature rising procedure is as follows: keeping the temperature at 40 ℃ for 4min, heating to 230 ℃ at 4 ℃/min, and keeping the temperature for 5 min. The carrier gas was nitrogen and the flow rate was 1 mL/min. Electron Impact (EI) energy was 70eV, ion source temperature was 230 ℃, full scan mode was used.

5. Sensory evaluation

And performing sensory analysis on the fishy smell, the fresh taste and the bitter taste by a grading method. The scoring criteria for the three senses are shown in table 1.

TABLE 1 sensory analysis scoring criteria

Flavor (I) and flavor (II)	1 minute (1)	2 is divided into	3 points of
				Fishy smell	Obvious fishy smell	Slightly fishy	No fishy smell
Delicate flavour	Rich delicate flavor	The delicate flavor is general	Light flavor
				Bitter taste	Heavy bitter taste	Slightly bitter	Has no bitter taste

Raw material preparation

Mallotus oblongifolius extract: adding water into Mallotus furetianus tea in a mass ratio of 1:20, namely adding water with the mass of 20 times of Mallotus furetianus tea, extracting for 1h at 80 ℃, and filtering to obtain Mallotus furetianus tea extract.

Example 1 examination of the Effect of different endoproteases on fragmentation

The following stepwise enzymolysis experiments were performed using trypsin, papain, and neutral protease as endoproteases, and flavourzyme as exoproteases, respectively. The respective optimum enzymatic pH is controlled according to the instructions for the use of the protease.

Adding water with the mass of 3 times of that of 200g of the shelled scallop, pulping and crushing to obtain scallop protein liquid. Adding endoprotease, wherein the addition amount of the endoprotease is 0.3 percent of the mass of the scallop protein liquid, the enzymolysis temperature is 50 ℃, the pH is regulated and controlled by NaOH in the enzymolysis process, and the first enzymolysis time is 8 hours; then inactivating enzyme for 15min at 90-100 ℃; adding exoprotease (specifically flavourzyme), wherein the addition amount of the exoprotease is 0.3 percent of the mass of the scallop protein liquid, the enzymolysis temperature is 50 ℃, and the second enzymolysis time is 6 hours; and (3) inactivating the enzyme for 15min at 90-100 ℃ after the second-stage enzymolysis is finished. Centrifuging at 8000rpm for 10min, collecting supernatant, and collecting scallop umami peptide solution.

The results of the proteolysis rate determination and the umami peptide content analysis are shown in Table 2. Both the proteolysis rate and the content ratio of the small molecular flavor peptide, the trypsin is far superior to the papain and the neutral protease. In addition, the content ratio of the small-molecule umami peptide (200 Da-1000 Da) is different, which shows that the first-stage enzymolysis (endo enzymolysis) and the second-stage enzymolysis (exo-enzymolysis) have a synergistic effect, and the trypsin is more favorable for promoting the extraction of the small-molecule umami peptide. Therefore, when the present invention is applied to the staged enzymolysis, trypsin is more preferably used in the first stage (endo-enzymolysis) step.

TABLE 2 examination of the enzymatic Effect of different types of endoproteases

Numbering	Class of endoproteases	Rate of proteolysis	Fresh taste peptide ratio (200 Da-1000 Da)
				E1-1	Trypsin	45％	30％
E1-2	Papain	37％	20％
				E1-3	Neutral protease	30％	17％

Example 2 examination of the Effect of enzymatic hydrolysis time on staged enzymatic hydrolysis

Adding water with the mass of 3 times of that of 200g of the shelled scallop, pulping and crushing to obtain scallop protein liquid. Adding endoprotease (specifically trypsin), wherein the addition amount of the endoprotease is 0.3% of the weight of the scallop protein liquid, the enzymolysis temperature is 50 ℃, pH (pH7) is regulated by NaOH in the enzymolysis process, and the first enzymolysis time is 8 h; then enzyme deactivation is carried out for 15min at the temperature of 90-100 ℃; adding exoprotease (specifically flavourzyme), wherein the addition amount of the exoprotease is 0.3% of the mass of the scallop protein liquid, the enzymolysis temperature is 50 ℃, the pH value is 7, and the second-stage enzymolysis time is controlled to be 2h, 4h, 6h, 8h and 10h respectively by different samples; and (3) inactivating the enzyme for 15min at 90-100 ℃ after the second-stage enzymolysis is finished. Centrifuging at 8000rpm for 10min, collecting supernatant, and collecting scallop umami peptide solution.

The results of the measurement of the proteolysis ratio, the analysis of the content of amino acid nitrogen, and the analysis of the content of umami peptide are shown in Table 3.

TABLE 3 examination of the enzymatic effectiveness of the second stage of the enzymatic hydrolysis with different exoproteases

As can be seen from the experimental results in Table 3, the hydrolysis rate of the scallop protein gradually increases and the content of the amino acid nitrogen gradually increases with the extension of the second stage enzymolysis time, but the content of the umami peptide (200 Da-1000 Da) shows a trend of increasing first and then decreasing. When the second stage enzymolysis time is 6h, the fresh taste peptide ratio reaches the highest (30%). The experimental result shows that when the second-stage enzymolysis time is less than 6 hours, the macromolecular peptide generated by the first-stage enzymolysis cannot be subjected to full enzymolysis, and most of unhydrolyzed macromolecular peptide still exists in the solution; when the enzymolysis time is continued to be prolonged, most of small molecular peptides in the hydrolysate are converted into free amino acids, and the content ratio of the umami peptides is reduced, so that the second stage of enzymolysis time needs to be reasonably controlled so as to maximize the ratio of the umami peptides.

Example 3 Effect of alternative use and combination of ultra-high pressure homogenization and ultrasonic treatment on the enzymatic hydrolysis

3.1. Investigation of different ultra-high pressure homogenization conditions and ultrasonic treatment conditions

3.1.1. Preparing scallop protein liquid: adding water with the mass of 3 times into 200g of shelled scallops, and carrying out colloid mill treatment for 3 times; then respectively and independently carrying out ultrahigh pressure homogenization treatment and ultrasonic treatment, and combining the ultrahigh pressure homogenization treatment (firstly) and the ultrasonic treatment (secondly) to obtain the scallop protein liquid. The conditions of the ultra-high pressure homogenization treatment and the ultrasonic treatment are as follows, and table 4 is also referred to.

Ultrahigh pressure homogenization treatment: homogenizing for 1, 2 or 3 times under 150MPa and at room temperature (15-30 deg.C); the time for carrying out the ultrahigh pressure homogenization treatment for one time is 2min to 10 min.

Ultrasonic treatment: the power is 1600W, and the ultrasonic time is 15min, 20min, 25min or 30 min.

3.1.2. Preparing scallop protease hydrolysate: adding the Mallotus oblongifolius muell-Arg extract into the scallop protein liquid, wherein the addition amount is 0.5 percent of the mass of the scallop protein liquid, and then carrying out sectional enzymolysis.

The process of segmented enzymolysis is as follows: adding endoprotease (specifically trypsin) for first stage enzymolysis, wherein the addition amount of the endoprotease is 0.3% of the weight of the scallop protein solution, the enzymolysis temperature is 50 ℃, and NaOH solution is adopted to regulate and control the pH in the enzymolysis process, so that the pH is controlled at 7, and the enzymolysis time is 8 h; then inactivating enzyme for 15min at 90-100 ℃; adding exoprotease (specifically flavourzyme) for second-stage enzymolysis, wherein the addition amount of the exoprotease is 0.3% of the mass of the scallop protein liquid, the enzymolysis temperature is 50 ℃, the pH value is controlled at 7, and the enzymolysis time is 6 h; and after the second stage of enzymolysis is finished, inactivating the enzyme at 90-100 ℃ for 15min to obtain scallop protein enzymolysis liquid.

Post-processing to prepare the scallop delicate flavor peptide product: centrifuging at 8000rpm for 10min, and collecting supernatant to obtain scallop umami peptide solution.

3.2. Comparative example: adding water with the mass of 3 times of that of 200g of the shelled scallop, pulping and crushing to obtain scallop protein liquid. Adding trypsin, wherein the adding amount of the trypsin is 0.3 percent of the weight of the scallop protein liquid, the enzymolysis temperature is 50 ℃, and the pH is regulated and controlled by NaOH in the enzymolysis process to be 7, and the enzymolysis time is 8 hours; then inactivating enzyme for 15min at 90-100 ℃. Adding flavourzyme, wherein the addition amount of the flavourzyme is 0.3 percent of the mass of the scallop protein liquid, the enzymolysis temperature is 50 ℃, the pH value is controlled to be 7, and the enzymolysis time is 6 hours; and (3) inactivating the enzyme for 15min at 90-100 ℃ after the enzymolysis is finished. Then centrifuging for 10min at 8000rpm, and collecting supernatant to obtain scallop umami peptide solution.

3.3. Analysis of results

3.3.1. The results of the determination of the proteolysis rate, the analysis of the content of umami peptide, and the sensory evaluation of fishy smell, umami and bitter taste of the scallop umami peptide solution are shown in table 5.

(1) The method only adopts ultrahigh pressure homogenization treatment instead of ultrasonic treatment, can obviously improve the hydrolysis rate of scallop protein (can reach more than 50 percent and contains the content), and can obviously improve the content ratio of the umami peptide (can reach more than 40 percent and contains the content); when the homogenization is carried out for 2 times under the ultrahigh pressure, the proteolysis rate can be obviously improved to 55 percent, and the promotion of the proteolysis rate is slowed down by continuously increasing the homogenization times.

(2) The proportion of the hydrolysis rate of scallop protein and the content of the umami peptide can be slightly improved by only adopting ultrasonic treatment instead of ultrahigh pressure homogenization treatment.

(3) The ultrahigh pressure homogenization treatment and the ultrasonic treatment are jointly adopted, compared with the single pretreatment mode, the combined treatment mode has more obvious improvement on the proteolysis rate, and the proteolysis rate can be improved to more than 60 percent (including the cost). Moreover, changing the ultrasonic treatment time also has a significant effect on the proteolysis rate during the combined treatment, which is just the manifestation of the synergistic effect between the ultra-high pressure homogenization treatment and the ultrasonic treatment. Under the condition of ultrahigh pressure homogenization treatment for 2 times, the effect of improving the proteolysis rate tends to be stable when the ultrasonic treatment time is 20min, and can reach about 70%.

(4) The ultrahigh pressure homogenization treatment and the ultrasonic treatment are combined, so that the content of the umami peptide is improved more obviously, and can be improved to about 50 percent, even about 60 percent. Moreover, the change of the ultrasonic treatment time also has a significant influence on the content ratio of the umami peptide in the combined treatment, which is the embodiment of the synergistic effect between the pretreatment (ultrahigh pressure homogenization combined with ultrasonic treatment) and the subsequent enzymolysis (segmented enzymolysis). Under the condition of ultrahigh pressure homogenization treatment for 2 times, the improvement effect of the content ratio of the delicious peptide tends to be stable when the ultrasonic treatment time is 20min, and can reach about 60%.

(5) The results in table 5 also show that the addition of mallotus oblongifolius extract can effectively suppress the generation of fishy smell substances.

Table 4 experimental conditions for different experimental examples and comparative examples in example 3

Experiment number	Conditions of the experiment	Number of times of ultra-high pressure homogenization	Duration of ultrasonic treatment
				Comparative example C1	Inhomogeneous, non-ultrasonic	0 time	0min
E3-1	Homogenizing for 1 time	Homogenizing1 time of	0min
				E3-2	Homogenizing for 2 times	Homogenizing for 2 times	0min
E3-3	Homogenizing for 3 times	Homogenizing for 3 times	0min
				E3-4	Ultrasonic for 15min	0 time	Ultrasonic for 15min
E3-5	Ultrasonic treatment for 20min	0 time	Ultrasonic treatment for 20min
				E3-6	Ultrasonic for 25min	0 time	Ultrasonic for 25min
E3-7	Ultrasonic treatment for 30min	0 time	Ultrasonic treatment for 30min
				E3-8	Homogenizing for 2 times, and performing ultrasonic treatment for 15min	Homogenizing for 2 times	Ultrasonic for 15min
E3-9	Homogenizing for 2 times, and performing ultrasonic treatment for 20min	Homogenizing for 2 times	Ultrasonic treatment for 20min
				E3-10	Homogenizing for 2 times, and performing ultrasonic treatment for 25min	Homogenizing for 2 times	Ultrasonic for 25min
E3-11	Homogenizing for 2 times, and performing ultrasonic treatment for 30min	Homogenizing for 2 times	Ultrasonic treatment for 30min

TABLE 5 influence of different conditions of ultra-high pressure homogenization treatment and/or ultrasonic treatment on the enzymolysis effect in example 3

3.3.2. Measuring the content of fishy smell substances

The SPME method is adopted to measure the contents of different fishy smell components in the scallop flavor peptide solution, and the contents are expressed as microgram of the fishy smell components contained in each 1Kg of scallop flavor peptide solution by relative contents (microgram/Kg). The results are shown in Table 6. The results in table 6 show that, compared with comparative example C1, after the partridge tea extract is added, the content of the fishy smell components, such as hexanal, nonanal, decanal, 1-octen-3-ol, 1-penten-3-ol, 2, 4-heptadienal and 2, 4-decadienal, related to fat oxidation is significantly reduced, which indicates that the partridge tea extract has a good inhibition effect on fat oxidation during scallop enzymolysis, can effectively inhibit the generation of fishy smell substances, and achieves an excellent level of no fishy smell in sensory evaluation.

Influence of extract of Mallotus oblongifolius on content of fishy smell substances in scallop flavor peptide solution

Note: "-" indicates no detection in the sample.

Example 4 examination of the Effect of the sequence of the ultra-high pressure homogenization treatment and the ultrasonic treatment on the proteolysis of scallop (E4)

Adding water with the mass of 3 times into the scallop after shelling, and processing for 3 times by a colloid mill. Carrying out ultrasonic treatment with ultrasonic power of 1600W for 20 min. Then homogenizing for 2 times under 150 MPa. Obtaining scallop protein liquid. And then adding the mallotus oblongifolius muell-Arg extract which accounts for 1.0 percent of the mass of the scallop protein liquid into the scallop protein liquid.

Then carrying out segmented enzymolysis: adding endoprotease (specifically trypsin), wherein the addition amount of the endoprotease is 0.3% of the weight of the scallop protein liquid, the enzymolysis temperature is 50 ℃, and the pH is controlled by NaOH solution in the enzymolysis process to be 7, and the enzymolysis time is 8 h; then inactivating enzyme for 15min at 90-100 ℃; adding exoprotease (specifically flavourzyme), wherein the addition amount of the exoprotease is 0.3 percent of the mass of the scallop protein liquid, the enzymolysis temperature is 50 ℃, the pH value is controlled at 7, and the enzymolysis time is 6 hours; inactivating the enzyme for 15min at 90-100 ℃ after the second stage of enzymolysis is finished; then centrifuging at 8000rpm for 10min, collecting supernatant, and obtaining scallop umami peptide solution.

And (4) analyzing results: the results of the measurement of the proteolysis ratio, the analysis of the umami peptide content, and the sensory evaluation of the fishy smell, umami taste and bitter taste are shown in table 7. From the results in table 7, it can be seen that the enzymolysis effect of the ultra-high pressure homogenization followed by the ultrasonic treatment (E3-9) is significantly better than that of the reverse sequence treatment (E4, ultrasonic treatment followed by ultra-high pressure homogenization), and the proteolysis rate and the umami peptide ratio are both significantly improved. In addition, the proteolysis rate (E4) of the combined treatment after the pre-treatment and the ultrahigh-pressure homogenization treatment is not much different from the proteolysis rate (E3-2) of the single ultrahigh-pressure homogenization treatment; the reason is that the ultrahigh pressure homogenization treatment and the ultrasonic treatment have a synergistic effect only in a specific sequence, the ultrahigh pressure homogenization treatment is firstly carried out to uniformly disperse the protein, and the subsequent ultrasonic treatment can play roles of cavitation, shear stress and the like.

TABLE 7 influence of different sequences of ultra-high pressure homogenization and ultrasonic treatment on the enzymolysis effect

Example 5 examination of the Effect of Partridge tea extract dosage on scallop proteolysis

5.1. Experimental example: adding different concentrations of Mallotus oblongifolius extract

Adding water in an amount which is 3 times the weight of the scallop after the shell removal, and processing the scallop for 3 times by a colloid mill; homogenizing at 150MPa for 2 times to obtain homogenized solution; and then carrying out ultrasonic treatment on the homogeneous liquid with the ultrasonic power of 1600W for 20min to obtain the scallop protein liquid.

Adding different dosages of Mallotus furetianus extract into scallop protein liquid, wherein the adding amounts are respectively 0.25%, 0.5%, 0.75% and 1.0% by mass percent relative to the scallop protein liquid.

Then carrying out segmented enzymolysis: adding endoprotease (specifically trypsin), wherein the addition amount of the endoprotease is 0.3% (relative to the mass percentage of the scallop protein liquid), the enzymolysis temperature is 50 ℃, and the pH is controlled by NaOH solution in the enzymolysis process to be 7, and the enzymolysis time is 8 h; then inactivating enzyme for 15min at 90-100 ℃; adding exoprotease (specifically flavourzyme), wherein the addition amount of the exoprotease is 0.3% (relative to the mass percentage of the scallop protein liquid), the enzymolysis temperature is 50 ℃, the pH value is controlled to be 7, and the enzymolysis time is 6 hours; inactivating the enzyme for 15min at 90-100 ℃ after the enzymolysis is finished; then centrifugating at 8000rpm for 10min, collecting supernatant to obtain scallop umami peptide solution.

5.2. Comparative example: the Mallotus oblongifolius extract was not added, and the remaining parameters were consistent with those in the experimental example of 5.1.

5.3. And (4) testing results: sensory evaluation of fishy smell, umami taste and bitter taste was performed, and the results are shown in table 8. As can be seen from the results in table 8, the fishy smell of the scallop enzymatic hydrolysate was significantly reduced by adding the partridge tea extract. When the adding amount is 0.5 percent (relative to the mass percent of the scallop protein liquid), no obvious fishy smell exists.

TABLE 8 influence of Partridge tea extract addition on scallop enzymatic hydrolysate flavor

The technical features of the embodiments and examples described above can be combined in any suitable manner, and for the sake of brevity, all possible combinations of the technical features of the embodiments and examples described above are not described, but should be considered within the scope of the present disclosure 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 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. It should be understood that the technical solutions provided by the present invention, which are obtained by logical analysis, reasoning or limited experiments, are within the scope of the appended claims. Therefore, the protection scope of the present invention should be subject to the content of the appended claims, and the description and the drawings can be used for explaining the content of the claims.

Claims

1. A preparation method of a scallop fresh taste peptide product is characterized by comprising the following steps:

providing raw materials: providing a scallop protein raw material;

pretreatment: performing pretreatment on the scallop protein raw material to prepare scallop protein liquid, wherein the pretreatment comprises ultrahigh pressure homogenization treatment and then ultrasonic treatment; wherein the pressure of the ultrahigh-pressure homogenizing treatment is more than or equal to 100 MPa;

2. The method for preparing the fresh taste peptide product of scallop as claimed in claim 1, further comprising a step of adding an antioxidant into the scallop protein liquid before the step of performing the step of segmented enzymolysis, wherein the antioxidant is an extract of Mallotus furetianus, and the addition amount of the extract of Mallotus furetianus is 0.5-2% of the mass of the scallop protein liquid.

3. The method for preparing the scallop flavor peptide product of claim 1, wherein in the scallop flavor peptide product, the mass content of polypeptides with molecular weight of 200 Da-1000 Da in the total peptides is more than or equal to 50%; and/or in the dry basis of the scallop fresh taste peptide product, the mass content of the polypeptide with the molecular weight of 200 Da-1000 Da is 30-45%.

4. The method for preparing the scallop delicious peptide product according to the claim 1, characterized in that the pretreatment step further comprises the step of colloid milling the scallop protein raw material before the ultrahigh pressure homogenization treatment and the ultrasonic treatment;

the pretreatment step comprises the steps of firstly carrying out colloid mill treatment, then carrying out ultrahigh pressure homogenization treatment and then carrying out ultrasonic treatment;

the pressure of the ultrahigh-pressure homogenizing treatment is 100-150 MPa;

the power of ultrasonic treatment is 1000W-2000W, and the time is 15 min-30 min.

5. The method for preparing the scallop delicious peptide product according to the claim 1, characterized in that the endoprotease is any one of trypsin, papain, neutral protease and pepsin or any combination thereof; and/or the exoprotease is any one of flavor protease and compound protease or the combination thereof.

6. The method for preparing the scallop flavor peptide product of any one of claims 1-5, wherein the step-wise enzymatic hydrolysis comprises: adding the endoprotease to carry out the first-stage enzymolysis, wherein the addition amount of the endoprotease is 0.1-0.5% of the mass of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-10 h; then carrying out first-stage enzyme deactivation at the temperature of 90-100 ℃ for 10-20 min; then adding the exoprotease for the second-stage enzymolysis, wherein the addition amount of the exoprotease is 0.1-0.5% of the mass of the scallop protein liquid, the enzymolysis temperature is 40-65 ℃, the pH value is 7-10, and the enzymolysis time is 6-8 h; and after the second-stage enzymolysis is finished, performing second-stage enzyme deactivation at the temperature of 90-100 ℃ for 10-20 min.

7. The method for preparing the scallop umami peptide product of claim 1, wherein the post-processing steps comprise: centrifuging at 8000-10000 rpm for 5-15 min, and collecting supernatant; and/or the presence of a catalyst in the reaction mixture,

the post-treatment step further comprises one or two of the following steps: purifying and drying scallop peptide.

8. A preparation method of a scallop fresh taste peptide product is characterized by comprising the following steps:

providing raw materials: providing a scallop protein raw material;

pretreatment: carrying out ultrahigh pressure homogenization treatment or ultrasonic treatment on the scallop protein raw material, or firstly carrying out ultrasonic treatment and then carrying out ultrahigh pressure homogenization treatment to prepare scallop protein liquid; wherein the pressure of the ultrahigh-pressure homogenizing treatment is more than or equal to 100 MPa;

9. A preparation method of a scallop fresh taste peptide product is characterized by comprising the following steps:

providing raw materials: providing a scallop protein raw material;

pretreatment: performing pretreatment on the scallop protein raw material, performing ultrahigh-pressure homogenization treatment, and then performing ultrasonic treatment to prepare scallop protein liquid; wherein the pressure of the ultrahigh-pressure homogenizing treatment is more than or equal to 100 MPa;

enzymolysis: carrying out enzymolysis, wherein the used enzymes comprise endoprotease and exoprotease to prepare scallop protein enzymolysis liquid;

10. A scallop umami peptide product prepared according to the preparation method of any one of claims 1-9.