Method, device, electronic device and storage medium for determining enzymatic hydrolysis conditions

Abstract

The present invention provides a method, device, electronic device and storage medium for determining enzymatic hydrolysis conditions, which relate to the field of biochemical technology, wherein the method comprises: matching a plurality of peptide segments to be studied with the sequences of enzymatic hydrolysis product peptide spectra in a database, respectively, and determining a plurality of target peptide spectra containing the peptide segments to be studied from the database; marking the peptide segments to be studied in the sequences of each target peptide spectra based on a target color set, and determining the optimal enzymatic hydrolysis conditions for obtaining the plurality of peptide segments to be studied based on the coincident colors in the target peptide spectra. The optimal enzymatic hydrolysis conditions are determined based on the recognition of the coincident colors. Based on the database screening and the coincident color recognition screening method, the rapid determination process of the optimal enzymatic hydrolysis conditions can be realized, and the efficiency of determining the optimal enzymatic hydrolysis conditions is improved.

Description

Technical Field

[0001] The present invention relates to the field of biochemical technology, and in particular to a method, device, electronic equipment and storage medium for determining enzymatic hydrolysis conditions. Background Art

[0002] After the peptide to be studied is determined, the enzymatic hydrolysis conditions for obtaining the peptide to be studied need to be studied to determine the optimal enzymatic hydrolysis conditions for obtaining the peptide to be studied.

[0003] The existing method determines the optimal enzymatic conditions for the peptide to be studied by enzymatically hydrolyzing the corresponding protein under different enzymatic conditions and analyzing the products of each enzymatic hydrolysis to determine the optimal enzymatic conditions. However, when there are multiple peptides to be studied, the analysis of multiple conditions one by one is relatively complicated and inefficient. Summary of the invention

[0004] The present invention provides a method, device, electronic device and storage medium for determining enzymatic hydrolysis conditions, which are used to improve the efficiency of determining the optimal enzymatic hydrolysis conditions for a peptide segment to be studied.

[0005] The present invention provides a method for determining enzymatic hydrolysis conditions, comprising the following steps:

[0006] Matching the multiple peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectra in the database respectively, and determining multiple target peptide spectra containing the peptide segments to be studied from the database;

[0007] Based on the target color set, the peptides to be studied are marked in the sequence of each target peptide spectrum, and based on the overlapping colors in each target peptide spectrum, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied are determined, and the color categories in the target color set correspond to each peptide to be studied one by one;

[0008] The database construction process includes:

[0009] Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time;

[0010] The database is constructed based on the peptide spectra of enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, and the peptide spectra of enzymatic hydrolysis products are obtained based on the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions marked in the amino acid sequence of the target protein.

[0011] According to a method for determining enzymatic hydrolysis conditions provided by the present invention, when the multiple peptides to be studied are different forms of peptides with the same function, determining the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the overlapping colors in the spectra of each target peptide comprises:

[0012] Based on the overlapping colors in the spectra of each target peptide, determining a plurality of overlapping peptide segments;

[0013] Based on the strength of the functional activity of each overlapping peptide segment, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptide segments to be studied are determined.

[0014] According to a method for determining enzymatic hydrolysis conditions provided by the present invention, after marking the peptide segments to be studied in the sequence of each target peptide spectrum based on the target color set, the method further comprises:

[0015] In the case where the multiple peptides to be studied are different functional peptides, overlapping colors in the spectra of the target peptides are screened to obtain overlapping peptide sequences;

[0016] Based on the overlapping colors, determining the overlapping amino acids in the overlapping peptide sequences and the different amino acids in the overlapping peptide sequences;

[0017] Based on the overlapping amino acids and the different amino acids, the interaction relationship between the function and the amino acid target is determined.

[0018] A method for determining enzymatic hydrolysis conditions provided by the present invention further comprises:

[0019] Based on the preset combination conditions, the multiple peptides to be studied are combined to obtain a target peptide;

[0020] Based on the overlapping colors in the target peptide spectra, the combined enzymatic hydrolysis conditions for obtaining the target peptide segment are determined.

[0021] According to a method for determining enzymatic hydrolysis conditions provided by the present invention, the peptide segments to be studied are marked in the sequence of each target peptide spectrum based on the target color set, further comprising:

[0022] Based on the number of peptides to be studied contained in the target peptide spectrum, determine the depth of the annotation color corresponding to the peptides to be studied;

[0023] Based on the target color set and the depth of the labeled color corresponding to the peptide segment to be studied, the peptide segment to be studied in the sequence of each target peptide spectrum is marked.

[0024] According to a method for determining enzymatic hydrolysis conditions provided by the present invention, the optimal enzymatic hydrolysis conditions for obtaining the plurality of peptide segments to be studied are determined based on the overlapping colors in the spectra of the target peptides, including:

[0025] Based on the depth of the overlapping colors in the target peptide spectra, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptide segments to be studied are determined.

[0026] According to a method for determining enzymatic hydrolysis conditions provided by the present invention, after determining the overlapping colors in the spectra of each target peptide, the method further comprises:

[0027] Based on the overlapping colors in the target peptide spectra, the changing pattern of the peptide segment to be studied with the enzymatic hydrolysis conditions is determined.

[0028] The present invention also provides a device for determining enzymatic hydrolysis conditions, comprising the following modules:

[0029] A peptide spectrum screening module is used to match the multiple peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectra in the database, and determine multiple target peptide spectra containing the peptide segments to be studied from the database;

[0030] An enzymatic hydrolysis condition determination module is used to mark the peptides to be studied in the sequence of each target peptide spectrum based on a target color set, and determine the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the overlapping colors in each target peptide spectrum, wherein the color categories in the target color set correspond one-to-one to each peptide to be studied;

[0031] The database construction process includes:

[0032] Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time;

[0033] The database is constructed based on the peptide spectra of enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, and the peptide spectra of enzymatic hydrolysis products are obtained based on the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions marked in the amino acid sequence of the target protein.

[0034] The present invention also provides an electronic device, comprising a memory, a processor, and a computer program stored in the memory and running on the processor, wherein when the processor executes the computer program, the method for determining the enzymatic hydrolysis conditions as described above is implemented.

[0035] The present invention also provides a non-transitory computer-readable storage medium having a computer program stored thereon, and when the computer program is executed by a processor, the method for determining the enzymatic hydrolysis conditions as described in any one of the above is implemented.

[0036] The method, device, electronic device and storage medium for determining enzymatic hydrolysis conditions provided by the present invention pre-construct a database containing enzymatic hydrolysis product peptide spectra corresponding to each enzymatic hydrolysis condition, and determine multiple target peptide spectra from the database, thereby realizing a screening process for enzymatic hydrolysis product peptide spectra corresponding to the peptide segments to be studied. Based on marking the peptide segments to be studied in the sequence of each target peptide spectrum, and based on the recognition of overlapping colors, the optimal enzymatic hydrolysis conditions are determined. Based on the database screening and the overlapping color recognition screening method, the rapid determination process of the optimal enzymatic hydrolysis conditions can be realized, and the efficiency of determining the optimal enzymatic hydrolysis conditions is improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0038] Figure 1 It is a schematic flow chart of the method for determining the enzymatic hydrolysis conditions provided by the present invention.

[0039] Figure 2 It is the peptide spectrum of the enzymatic hydrolysis product provided by the present invention.

[0040] Figure 3 It is a schematic diagram of the structure of the device for determining enzymatic hydrolysis conditions provided by the present invention.

[0041] Figure 4 It is a structural schematic diagram of the electronic device provided by the present invention. DETAILED DESCRIPTION

[0042] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0043] Figure 1 It is a schematic diagram of the process for determining the enzymatic hydrolysis conditions provided by the present invention, such as Figure 1 As shown, the method includes the following:

[0044] Step 110, matching the multiple peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectra in the database, and determining multiple target peptide spectra containing the peptide segments to be studied from the database;

[0045] Step 120, based on the target color set, marking the peptides to be studied in the sequence of each target peptide spectrum, and determining the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the overlapping colors in each target peptide spectrum, and the color categories in the target color set correspond one-to-one to each peptide to be studied;

[0046] The database construction process includes:

[0047] Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time;

[0048] The database is constructed based on the peptide spectra of enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, and the peptide spectra of enzymatic hydrolysis products are obtained based on the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions marked in the amino acid sequence of the target protein.

[0049] The execution subject of the method for determining the enzymolysis conditions provided by the present invention can be an electronic device, a component in an electronic device, an integrated circuit, or a chip. The electronic device can be a mobile electronic device or a non-mobile electronic device. Exemplarily, the mobile electronic device can be a mobile phone, a tablet computer, a notebook computer, a palmtop computer, an ultra-mobile personal computer (ultra-mobile personal computer, UMPC), a netbook or a personal digital assistant (personal digital assistant, PDA), etc., and the non-mobile electronic device can be a server, a network attached storage (Network Attached Storage, NAS) or a personal computer (personal computer, PC), etc., which is not specifically limited by the present invention.

[0050] The following takes the method for determining the enzymatic hydrolysis conditions provided by the present invention executed by a computer as an example to describe the technical solution of the present invention in detail.

[0051] In step 110, a database containing a plurality of peptide spectra of enzymatic hydrolysis products is pre-constructed.

[0052] Determine different enzymatic hydrolysis conditions. The enzymatic hydrolysis conditions can be obtained by combining different enzyme types, enzyme addition amounts, pH values, temperatures, and enzymatic hydrolysis times.

[0053] Based on the determined different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions.

[0054] Different enzymatic hydrolysis conditions are optimized based on a combination of a series of key factors. Specifically, these conditions include the type of enzyme, the amount of enzyme added, pH value, temperature and enzymatic hydrolysis time, which have a significant impact on the efficiency of the enzymatic hydrolysis process, the generation of products and the final peptide spectrum results.

[0055] Based on the determination of the enzymatic hydrolysis conditions of a combination of multiple factors, and based on the peptide spectra of the enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, a more comprehensive database can be constructed, which facilitates the subsequent research process of multiple peptide segments to be studied.

[0056] It should be noted that the target protein can be different types of proteins. Based on different enzymatic hydrolysis conditions, the target protein may be hydrolyzed to obtain a hydrolysis product containing one peptide to be studied, a hydrolysis product containing multiple peptides to be studied, or a hydrolysis product containing no peptide to be studied.

[0057] Based on the enzymatic hydrolysis products obtained under each enzymatic hydrolysis condition, a peptide spectrum of the enzymatic hydrolysis products is constructed. The peptide spectrum of the enzymatic hydrolysis products is obtained based on the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions marked in the amino acid sequence of the target protein. Figure 2 The enzymatic hydrolysis product peptide spectrum provided by the present invention is shown in FIG. Wherein, each segment under the amino acid sequence represents a sequence of an enzymatic hydrolysis product.

[0058] For each enzymatic hydrolysis condition, a peptide spectrum of the enzymatic hydrolysis product is constructed. For multiple enzymatic hydrolysis conditions, multiple peptide spectra of the enzymatic hydrolysis product are obtained. The multiple constructed peptide spectra of the enzymatic hydrolysis product are summarized to build a database containing multiple peptide spectra of the enzymatic hydrolysis product.

[0059] After the database is constructed, multiple peptides to be studied are matched with the sequences of the enzymatic hydrolysis product peptide spectra in the database respectively to screen the enzymatic hydrolysis product peptide spectra in the database and determine multiple target peptide spectra in the database that contain the peptides to be studied.

[0060] Specifically, each peptide to be studied can be matched with the sequence of the enzymatic hydrolysis product peptide spectrum in the database. As long as the sequence has a matching peptide to be studied, the enzymatic hydrolysis product peptide spectrum corresponding to the sequence is determined as the target peptide spectrum.

[0061] Optionally, peptide matching can be based on some specific algorithms, such as KMP algorithm (Knuth-Morris-Pratt algorithm), etc. These algorithms are usually efficient and accurate, and can quickly find matching sequences in large-scale data sets.

[0062] In step 120, based on the target color set, the peptide segments to be studied are marked in the sequence of each target peptide spectrum.

[0063] The target color set includes colors of various color categories, such as red, yellow, green, etc. Among them, each color in the target color set corresponds to each peptide to be studied. For example, if the multiple peptides to be studied are the first peptide to be studied, the second peptide to be studied, and the third peptide to be studied. The first peptide to be studied corresponds to red, the second peptide to be studied corresponds to yellow, and the third peptide to be studied corresponds to green.

[0064] It is understandable that each target peptide spectrum contains at least one peptide segment to be studied. Based on the color corresponding to each peptide segment to be studied, the peptide segments to be studied in the sequence in the target peptide spectrum are marked to obtain a color-marked target peptide spectrum.

[0065] By analyzing the overlapping colors in each target peptide spectrum, the optimal enzymatic hydrolysis conditions for multiple peptides to be studied can be determined. Among them, the optimal enzymatic hydrolysis conditions are preset conditions corresponding to preset requirements, which can be adjusted according to the requirements and are the optimal conditions that meet the corresponding requirements.

[0066] It can be understood that the new color obtained after the overlap of different colors is different from the colors in the target color set. A new color can be obtained, which does not belong to the target color set and can be identified from the color-marked target peptide spectrum based on computer vision to achieve a rapid identification process of overlapping colors.

[0067] It is understandable that the overlapping colors in the target peptide spectrum are the results of the overlap of at least two peptides to be studied. The overlapping colors are the colors corresponding to the peptides to be studied. The color overlap indicates that the sequences of at least two peptides to be studied have overlapped.

[0068] It is understandable that the optimal enzymatic hydrolysis condition does not only include a single enzymatic hydrolysis condition, but may also be a condition obtained by combining different enzymatic hydrolysis conditions. For multiple peptides to be studied, it may be impossible to obtain all the peptides to be studied based on one enzymatic hydrolysis condition. Based on the overlapping colors, multiple enzymatic hydrolysis conditions can be combined to determine the optimal enzymatic hydrolysis condition.

[0069] At the same time, for the study of overlapping colors, a sequence containing two peptides to be studied can be obtained with only one enzyme digestion process. Therefore, the more overlapping colors there are, the higher the efficiency. The higher the efficiency, the better the enzymatic hydrolysis conditions, and thus the optimal enzymatic hydrolysis conditions can be determined based on this.

[0070] For example, if multiple peptides to be studied have different functions, after overlapping, a sequence with two functions can be obtained. When determining the optimal enzymatic hydrolysis conditions, it may be desirable to have as many peptides with multiple functions as possible. Based on the overlapping colors, an optimal enzymatic hydrolysis condition can be determined to achieve an efficient screening process for such peptides.

[0071] Based on the identification of overlapping colors in the target peptide spectra, multiple target peptide spectra can be screened to determine which target peptide spectra have the most overlapping colors or are most suitable for the needs. For example, if you want to hydrolyze the target protein and obtain the largest number of multiple peptides to be studied at the same time, you can analyze the number of overlapping colors in each target peptide spectrum and determine that the enzymatic hydrolysis conditions corresponding to the target peptide spectrum with the largest number of overlapping colors are the optimal enzymatic hydrolysis conditions for obtaining multiple peptides to be studied. Similarly, the optimal enzymatic hydrolysis conditions can also be determined based on other measurement factors, such as output efficiency.

[0072] It should be noted that the multiple peptides to be studied may also be peptides corresponding to multiple functional peptides with different functions. For example, the multiple peptides to be studied may be immunomodulatory functional peptides, blood pressure lowering peptides, and antioxidant peptides. In actual research, not only functional peptides with a single function are studied, but also the combination of multiple functional peptides needs to be studied. That is, it is necessary to determine peptides that have both immunomodulatory function and blood pressure lowering function, or blood pressure lowering function and antioxidant function based on multiple single functional peptides.

[0073] For example, after screening the peptide segment with the sequence "ypfpgpi" based on the overlapping colors, its "ypfpgp" sequence part was determined to be a blood pressure lowering active peptide (yellow mark), and its "ypfpgpi" sequence was an immunomodulatory active peptide (green mark). A new color is obtained by mixing yellow and green. Based on the recognition of this new color, the screening process of the peptide segment with the sequence "ypfpgpi" can be quickly realized.

[0074] In summary, researchers can easily and quickly screen out active peptide combinations with "ypfpgpi" as the dominant product by enzymatic hydrolysis under specified conditions. This enzymatic hydrolysis product has a strong immunomodulatory function. Since the dominant product "ypfpgpi" contains "ypfpgp" which is predicted to have antihypertensive activity, future research can focus on observing the content of this sequence polypeptide in the enzymatic hydrolysis product and whether the enzymatic hydrolysis product also has antihypertensive activity.

[0075] After determining the required overlapping colors, the optimal enzymatic hydrolysis conditions for multiple peptides to be studied are determined based on the identification of the overlapping colors in each target peptide spectrum. Among them, the number of overlapping colors contained in the identified target peptide spectrum (the number of peptides is fed back) and the specific implementation difficulty of the conditions in the identified target peptide spectrum containing overlapping colors can be screened to determine the optimal enzymatic hydrolysis conditions. Specifically, multiple target peptide spectra can be sorted according to the number of overlapping colors, and based on the difficulty of implementing the corresponding enzymatic hydrolysis conditions of each target peptide spectrum, the most suitable target peptide spectrum for enzymatic hydrolysis can be determined from the sorted target peptide spectra. The enzymatic hydrolysis conditions corresponding to the target peptide spectrum that is most suitable for enzymatic hydrolysis are the optimal enzymatic hydrolysis conditions.

[0076] It is understandable that the overlapping colors can intuitively reflect the specific position of the amino acid sequence of the target protein. That is, the peptide to be studied corresponding to the overlapping colors is at the specific position of the amino acid sequence of the target protein. For the amino acid sequence of the target protein, the difficulty of obtaining the peptide to be studied at different positions is different. Therefore, the auxiliary determination process of the optimal enzymatic hydrolysis conditions can also be realized based on the position.

[0077] The method for determining enzymatic hydrolysis conditions provided by the present invention realizes the screening process of enzymatic hydrolysis product peptide spectra corresponding to the peptide segments to be studied by pre-constructing a database containing enzymatic hydrolysis product peptide spectra corresponding to each enzymatic hydrolysis condition and determining multiple target peptide spectra from the database. Based on marking the peptide segments to be studied in the sequence of each target peptide spectrum and based on the recognition of overlapping colors, the optimal enzymatic hydrolysis conditions are determined. Based on the database screening and the overlapping color recognition screening method, the rapid determination process of the optimal enzymatic hydrolysis conditions can be realized, and the efficiency of determining the optimal enzymatic hydrolysis conditions is improved.

[0078] In one embodiment, when the multiple peptides to be studied are different forms of peptides with the same function, determining the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the overlapping colors in the target peptide spectra includes: determining multiple overlapping peptides based on the overlapping colors in the target peptide spectra; and determining the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the strength of the functional activity of each overlapping peptide.

[0079] Different forms of peptides with the same function refer to multiple peptides with the same function but different peptide sequences. For example, multiple peptides are all peptides with blood pressure lowering function, but the amino acid sequences of the peptides are different.

[0080] The overlapping colors reflect that there are overlapping parts in the sequences of at least two peptides to be studied with the same function, and the sequences corresponding to these peptides to be studied are similar.

[0081] In the case of similar sequences and the same functions, since the target is unique, biological experiments and digital simulations can be used to determine the strength of the functional activity of overlapping peptides, their interactions and binding modes with the target, thereby finding the key factors for the active peptides to exert their effects, thereby further assisting in the process of determining the optimal enzymatic hydrolysis conditions.

[0082] For example, under the premise of obtaining more highly active blood pressure-lowering functional peptides, the enzymatic hydrolysis conditions are screened based on the number of peptides with strong functional activity in the overlapping colors to determine the optimal enzymatic hydrolysis conditions.

[0083] In one embodiment, after marking the peptide to be studied in the sequence of each target peptide spectrum based on the target color set, it also includes: when the multiple peptides to be studied are peptides of different functions, the overlapping colors in each target peptide spectrum are screened to obtain overlapping peptide sequences; based on the overlapping colors, the overlapping amino acids in the overlapping peptide sequences and the differential amino acids in the overlapping peptide sequences are determined; based on the overlapping amino acids and the differential amino acids, the interaction relationship between function and amino acid target is determined.

[0084] It is understandable that overlapping colors reflect the similarity of the sequences of different peptides to be studied. For example, if the middle core amino acid sequence is the same, but the amino acid sequences at the two ends are different, you may get:

[0085] Peptides with similar amino acid sequences but different functions;

[0086] Peptides with similar amino acid sequences but the same functions.

[0087] In both cases, a third overlapping color will appear. The appearance of this color is of great significance for studying the relationship between the amino acid sequence, structure and function of the peptide. For active peptides with similar amino acid sequences but different functions, researchers can study the interaction between functional peptides and target sites by studying the different amino acids.

[0088] The multiple peptides to be studied are peptides with different functions, which means that the functions of the peptides to be studied are different. For example, the functions of the multiple peptides to be studied are respectively blood pressure lowering function, immune regulation function, and antioxidant function.

[0089] The overlapping peptide sequence corresponding to the obtained overlapping colors contains overlapping amino acids and differential amino acids. For example, if the determined overlapping peptide sequence is "ypfpgpi". This overlapping peptide sequence contains the "ypfpgp" peptide to be studied with blood pressure lowering function and "ypfpgpi" with immune regulation function. The differential amino acid is "i" and the overlapping amino acid is "ypfpgp".

[0090] For example, when the difference at the end of the functional peptide lies in a branched-chain amino acid or an amino acid with a benzene ring, this branched-chain amino acid or amino acid with a benzene ring is likely to be a key structure for forming hydrogen bonds and other interactions with the target. By understanding the role of these key amino acids, such key amino acids can be added to the ends of the active peptides that have been discovered and have these functions, which may greatly enhance the function of the original active peptides.

[0091] Similarly, for active peptides with similar amino acid sequences but different functions, different amino acids are crucial to revealing the relationship between the sequence, structure and function of active peptides. Researchers can also use the method of adding or subtracting amino acids based on the core amino acid sequence to observe and study the changes in activity and the interaction with the target, thereby further revealing and predicting the relationship between the amino acid sequence and function of active peptides.

[0092] In one embodiment, the method further includes: combining the multiple peptides to be studied based on preset combination conditions to obtain a target peptide; and determining the combined enzymatic hydrolysis conditions for obtaining the target peptide based on the overlapping colors in the spectra of the target peptides.

[0093] It is understandable that, for the research process of multiple peptides to be studied, sometimes one wants to implement a prediction process, that is, to determine the research process of an unknown combination of peptides based on multiple known peptides to be studied.

[0094] The preset combination conditions can be determined based on the combination of multiple peptides to be studied. For example, for the peptides to be studied "ypfpgpi" and the peptides to be studied "ipnslpq", a new target peptide "ypfpgpipnslpq" can be obtained after combination. In the actual research process, it is often necessary to realize the research process of obtaining a new target peptide based on the combination of some known peptides to be studied.

[0095] If you want to obtain an active peptide with the sequence "ypfpgpipnslpq", you can choose a combination of different enzymes to overlap. The database itself contains the peptide sequences produced by the enzymatic hydrolysis of two enzymes. When the two overlap, many new peptides will appear that are not produced by a single enzyme during the enzymatic hydrolysis process. The peptides hydrolyzed by the two enzymes can also be represented by different colors. The new peptides produced when the two overlap will appear in a third color. If the target peptide you want to select appears in the third color after the overlap, scientific researchers can choose these two enzymes to perform enzymatic digestion, and finally analyze the enzymatic hydrolysis results to see if the target peptide appears.

[0096] This prediction-assisted selection can be two different enzymes, different enzymatic hydrolysis conditions of the same enzyme, or a combination of different enzymes and different enzymatic hydrolysis conditions.

[0097] Based on the overlapping colors in the target peptide spectra, the combined enzymatic hydrolysis conditions for obtaining the target peptide segment are determined. For example, when the target peptide segment is a polypeptide "ypfpgp" with antihypertensive activity. Enzymatic hydrolysis by protease A can obtain a polypeptide with a sequence of "ypfpgpi", and enzymatic hydrolysis by protease B can obtain a polypeptide with a sequence of "ipnslpq". When the enzymatic hydrolysis product of protease A coincides with the enzymatic hydrolysis product of protease B, the superimposed part is "ypfpgpipnslpq" as the required target sequence active peptide. In actual experiments, scientific researchers select a combination of protease A + protease B to obtain the target active peptide "ypfpgp". In the screening process, the process of determining the enzymatic hydrolysis conditions can be realized based on the overlapping colors.

[0098] In one embodiment, based on the target color set, marking the peptides to be studied in the sequence of each target peptide spectrum further includes: determining the depth of the marked color corresponding to the peptides to be studied based on the number of peptides to be studied contained in the target peptide spectrum; marking the peptides to be studied in the sequence of each target peptide spectrum based on the target color set and the depth of the marked color corresponding to the peptides to be studied.

[0099] Based on the depth of the overlapping colors in the target peptide spectra, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptide segments to be studied are determined.

[0100] In the target peptide spectrum, the number of peptides to be studied usually reflects the abundance and quality of the peptides generated under specific enzymatic conditions. For ease of analysis, the number of these peptides can be associated with the depth of color to establish an intuitive visual labeling relationship.

[0101] Specifically, the number of peptides can be mapped to the depth of color: the more peptides there are, the darker the color; the fewer peptides there are, the lighter the color.

[0102] Based on the number of peptides to be studied contained in the target peptide spectrum, the depth of the annotation color corresponding to the peptides to be studied is determined.

[0103] Specifically, the color depth of each color category in the target color set can be graded. For example, brown in the target color set corresponds to the first peptide to be studied. The brown in the target color set is graded, and the levels can be set to light brown and dark brown.

[0104] In the process of marking based on the target color set, a marking process of different brown levels is performed according to the number of the first peptide segments to be studied. For example, when the number of all the first peptide segments to be studied is greater than a preset number threshold, dark brown is used for marking, and when it is less than or equal to the preset number threshold, light brown is used for marking.

[0105] The depth of the overlapping color that can be obtained can be used to determine the number of the first peptide fragment to be studied obtained by enzyme cleavage.

[0106] According to the quantity requirement of the first peptide to be studied by the scientific research staff, the optimal enzymatic hydrolysis conditions of the first peptide to be studied can be screened and determined based on the depth of the color marked on the first peptide to be studied, so as to quickly obtain the first peptide to be studied in the quantity corresponding to the requirement.

[0107] In one embodiment, after determining the overlapping colors in the target peptide spectra, the method further includes: determining the variation pattern of the peptide segments to be studied with the enzymatic hydrolysis conditions based on the overlapping colors in the target peptide spectra.

[0108] The most important thing in choosing enzymatic hydrolysis conditions is whether a sufficient number of target functional peptides can be produced under these conditions. Different enzymatic hydrolysis products can be produced by choosing different enzymes and different enzymatic hydrolysis conditions (such as temperature, pH value, reaction time, enzyme addition amount, etc.). Through peptidomics analysis, the composition and relative abundance of peptides in these enzymatic hydrolysis products can be obtained, thereby establishing a database of peptide distribution of enzymatic hydrolysis products. However, such a database cannot include all enzymatic hydrolysis conditions. For example, it is impossible to take enzymatic hydrolysis products for peptidomics analysis every minute. The enzymatic hydrolysis conditions in the database must be composed of many discontinuous points. By overlapping different functional peptide databases with the peptidomics database obtained by enzymatic hydrolysis, the number and relative abundance of target functional peptides contained in a certain point in the peptide database (characterized by a third color and color depth) can be known. By comparing multiple peptidomics database points, researchers can analyze which point, that is, which enzymatic hydrolysis condition, can produce the most target functional peptides. By comparing and analyzing multiple points in such a large number of points, the conditions that produce the most and deepest target colors can be found, and then this condition can be selected for enzymatic hydrolysis.

[0109] The overlapping colors in each target peptide spectrum reflect which enzymatic products contain or are rich in this peptide sequence. Based on these enzymatic products, the corresponding enzymes and enzymatic conditions can be found, as well as the change pattern of the produced peptides with the enzymatic conditions. Researchers can quickly screen out the corresponding enzymes and enzymatic conditions for the target active peptides they want to obtain.

[0110] The following is a description of the device for determining enzymatic hydrolysis conditions provided by the present invention. The device for determining enzymatic hydrolysis conditions described below and the method for determining enzymatic hydrolysis conditions described above can be referred to each other.

[0111] like Figure 3 As shown, the device comprises:

[0112] The peptide spectrum screening module 310 is used to match the multiple peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectrum in the database, and determine multiple target peptide spectra containing the peptide segments to be studied from the database;

[0113] The enzymatic hydrolysis condition determination module 320 is used to mark the peptides to be studied in the sequence of each target peptide spectrum based on the target color set, and determine the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the overlapping colors in each target peptide spectrum, and the color categories in the target color set correspond to each peptide to be studied one by one;

[0114] The database construction process includes:

[0115] Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time;

[0116] The database is constructed based on the peptide spectra of enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, and the peptide spectra of enzymatic hydrolysis products are obtained based on the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions marked in the amino acid sequence of the target protein.

[0117] The device for determining enzymatic hydrolysis conditions provided by the present invention realizes the screening process of enzymatic hydrolysis product peptide spectra corresponding to the peptide segments to be studied by pre-constructing a database containing enzymatic hydrolysis product peptide spectra corresponding to each enzymatic hydrolysis condition and determining multiple target peptide spectra from the database. Based on marking the peptide segments to be studied in the sequence of each target peptide spectrum and based on the recognition of overlapping colors, the optimal enzymatic hydrolysis conditions are determined. Based on the database screening and the overlapping color recognition screening method, the rapid determination process of the optimal enzymatic hydrolysis conditions can be realized, and the efficiency of determining the optimal enzymatic hydrolysis conditions is improved.

[0118] In one embodiment, the enzymatic hydrolysis condition determination module 320 is specifically used to:

[0119] In the case where the multiple peptides to be studied are different forms of peptides with the same function, determining the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the overlapping colors in the target peptide spectra includes:

[0120] Based on the overlapping colors in the spectra of each target peptide, determining a plurality of overlapping peptide segments;

[0121] Based on the strength of the functional activity of each overlapping peptide segment, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptide segments to be studied are determined.

[0122] In one embodiment, the enzymolysis condition determination module 320 is further specifically used for:

[0123] After marking the peptide segments to be studied in the sequence of each target peptide spectrum based on the target color set, the method further includes:

[0124] In the case where the multiple peptides to be studied are different functional peptides, overlapping colors in the spectra of the target peptides are screened to obtain overlapping peptide sequences;

[0125] Based on the overlapping colors, determining the overlapping amino acids in the overlapping peptide sequences and the different amino acids in the overlapping peptide sequences;

[0126] Based on the overlapping amino acids and the different amino acids, the interaction relationship between the function and the amino acid target is determined.

[0127] In one embodiment, the enzymolysis condition determination module 320 is further specifically used for:

[0128] Based on the preset combination conditions, the multiple peptides to be studied are combined to obtain a target peptide;

[0129] Based on the overlapping colors in the target peptide spectra, the combined enzymatic hydrolysis conditions for obtaining the target peptide segment are determined.

[0130] In one embodiment, the enzymolysis condition determination module 320 is further specifically used for:

[0131] The step of marking the peptide segments to be studied in the sequence of each target peptide spectrum based on the target color set further includes:

[0132] Based on the number of peptides to be studied contained in the target peptide spectrum, determine the depth of the annotation color corresponding to the peptides to be studied;

[0133] Based on the target color set and the depth of the labeled color corresponding to the peptide segment to be studied, the peptide segment to be studied in the sequence of each target peptide spectrum is marked.

[0134] In one embodiment, the enzymolysis condition determination module 320 is further specifically used for:

[0135] Based on the overlapping colors in the target peptide spectra, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptide segments to be studied are determined, including:

[0136] Based on the depth of the overlapping colors in the target peptide spectra, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptide segments to be studied are determined.

[0137] In one embodiment, the enzymolysis condition determination module 320 is further specifically used for:

[0138] After determining the overlapping colors in the target peptide spectra, the method further includes:

[0139] Based on the overlapping colors in the target peptide spectra, the changing pattern of the peptide segment to be studied with the enzymatic hydrolysis conditions is determined.

[0140] Figure 4An example of a physical structure diagram of an electronic device is shown in FIG. Figure 4 As shown, the electronic device may include: a processor 410, a communications interface 420, a memory 430 and a communications bus 440, wherein the processor 410, the communications interface 420 and the memory 430 communicate with each other through the communications bus 440. The processor 410 may call the logic instructions in the memory 430 to execute a method for determining enzymatic hydrolysis conditions, the method comprising: matching a plurality of peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectra in the database respectively, and determining a plurality of target peptide spectra containing the peptide segments to be studied from the database;

[0141] Based on the target color set, the peptides to be studied are marked in the sequence of each target peptide spectrum, and based on the overlapping colors in each target peptide spectrum, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied are determined, and the color categories in the target color set correspond to each peptide to be studied one by one;

[0142] The database construction process includes:

[0143] Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time;

[0144] The database is constructed based on the peptide spectra of enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, and the peptide spectra of enzymatic hydrolysis products are obtained based on the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions marked in the amino acid sequence of the target protein.

[0145] In addition, the logic instructions in the above-mentioned memory 430 can be implemented in the form of a software functional unit and can be stored in a computer-readable storage medium when it is sold or used as an independent product. Based on this understanding, the technical solution of the present invention, in essence, or the part that contributes to the prior art or the part of the technical solution, can be embodied in the form of a software product, and the computer software product is stored in a storage medium, including a number of instructions for a computer device (which can be a personal computer, a server, or a network device, etc.) to perform all or part of the steps of the method described in each embodiment of the present invention. The aforementioned storage medium includes: U disk, mobile hard disk, read-only memory (ROM, Read-Only Memory), random access memory (RAM, Random Access Memory), disk or optical disk, etc. Various media that can store program codes.

[0146] On the other hand, the present invention also provides a computer program product, which includes a computer program, which can be stored on a non-transitory computer-readable storage medium. When the computer program is executed by a processor, the computer can execute the method for determining the enzymatic hydrolysis conditions provided by the above methods, which method includes: matching a plurality of peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectra in the database, respectively, and determining a plurality of target peptide spectra containing the peptide segments to be studied from the database;

[0147] Based on the target color set, the peptides to be studied are marked in the sequence of each target peptide spectrum, and based on the overlapping colors in each target peptide spectrum, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied are determined, and the color categories in the target color set correspond to each peptide to be studied one by one;

[0148] The database construction process includes:

[0149] Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time;

[0150] The database is constructed based on the peptide spectra of enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, and the peptide spectra of enzymatic hydrolysis products are obtained based on the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions marked in the amino acid sequence of the target protein.

[0151] In another aspect, the present invention further provides a non-transitory computer-readable storage medium having a computer program stored thereon, which is implemented when the computer program is executed by a processor to perform the method for determining the enzymatic hydrolysis conditions provided by the above methods, the method comprising: matching a plurality of peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectra in a database, respectively, and determining a plurality of target peptide spectra containing the peptide segments to be studied from the database;

[0152] Based on the target color set, the peptides to be studied are marked in the sequence of each target peptide spectrum, and based on the overlapping colors in each target peptide spectrum, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied are determined, and the color categories in the target color set correspond to each peptide to be studied one by one;

[0153] The database construction process includes:

[0154] Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time;

[0155] The database is constructed based on the peptide spectra of enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, and the peptide spectra of enzymatic hydrolysis products are obtained based on the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions marked in the amino acid sequence of the target protein.

[0156] The device embodiments described above are merely illustrative, wherein the units described as separate components may or may not be physically separated, and the components displayed as units may or may not be physical units, that is, they may be located in one place, or they may be distributed on multiple network units. Some or all of the modules may be selected according to actual needs to achieve the purpose of the scheme of this embodiment. Ordinary technicians in this field can understand and implement it without paying creative labor.

[0157] Through the description of the above implementation methods, those skilled in the art can clearly understand that each implementation method can be implemented by means of software plus a necessary general hardware platform, and of course, can also be implemented by hardware. Based on this understanding, the above technical solution is essentially or the part that contributes to the prior art can be embodied in the form of a software product, and the computer software product can be stored in a computer-readable storage medium, such as ROM / RAM, a disk, an optical disk, etc., including a number of instructions for a computer device (which can be a personal computer, a server, or a network device, etc.) to execute the methods described in each embodiment or some parts of the embodiments.

[0158] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for determining enzymatic hydrolysis conditions, characterized in that: include: Matching the multiple peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectra in the database respectively, and determining multiple target peptide spectra containing the peptide segments to be studied from the database; Based on the target color set, the peptides to be studied are marked in the sequence of each target peptide spectrum, and based on the overlapping colors in each target peptide spectrum, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied are determined, and the color categories in the target color set correspond to each peptide to be studied one by one; The database construction process includes: Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time; The database is constructed based on the peptide spectra of the enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, wherein the peptide spectra of the enzymatic hydrolysis products are obtained by marking the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions in the amino acid sequence of the target protein; Also includes: Based on the preset combination conditions, the multiple peptides to be studied are combined to obtain a target peptide; Based on the overlapping colors in the target peptide spectra, the combined enzymatic hydrolysis conditions for obtaining the target peptide segment are determined.

2. The method for determining enzymolysis conditions according to claim 1, characterized in that: In the case where the multiple peptides to be studied are different forms of peptides with the same function, determining the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the overlapping colors in the target peptide spectra includes: Based on the overlapping colors in the spectra of each target peptide, determining a plurality of overlapping peptide segments; Based on the strength of the functional activity of each overlapping peptide segment, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptide segments to be studied are determined.

3. The method for determining enzymolysis conditions according to claim 1, characterized in that: After marking the peptide segments to be studied in the sequence of each target peptide spectrum based on the target color set, the method further includes: In the case where the multiple peptides to be studied are different functional peptides, overlapping colors in the spectra of the target peptides are screened to obtain overlapping peptide sequences; Based on the overlapping colors, determining the overlapping amino acids in the overlapping peptide sequences and the different amino acids in the overlapping peptide sequences; Based on the overlapping amino acids and the different amino acids, the interaction relationship between the function and the amino acid target is determined.

4. The method for determining enzymolysis conditions according to claim 1, characterized in that: The step of marking the peptide segments to be studied in the sequence of each target peptide spectrum based on the target color set further includes: Based on the number of peptides to be studied contained in the target peptide spectrum, determine the depth of the annotation color corresponding to the peptides to be studied; Based on the target color set and the depth of the labeled color corresponding to the peptide segment to be studied, the peptide segment to be studied in the sequence of each target peptide spectrum is marked.

5. The method for determining enzymolysis conditions according to claim 4, characterized in that: The step of determining the optimal enzymatic hydrolysis conditions for obtaining the plurality of peptide segments to be studied based on the overlapping colors in the target peptide spectra includes: Based on the depth of the overlapping colors in the target peptide spectra, the optimal enzymatic hydrolysis conditions for obtaining the multiple peptide segments to be studied are determined.

6. The method for determining enzymolysis conditions according to claim 1, characterized in that: After determining the overlapping colors in the target peptide spectra, the method further includes: Based on the overlapping colors in the target peptide spectra, the variation pattern of the peptide to be studied with the enzymatic hydrolysis conditions is determined.

7. A device for determining enzymatic hydrolysis conditions, characterized in that: include: A peptide spectrum screening module is used to match the multiple peptide segments to be studied with the sequences of the enzymatic hydrolysis product peptide spectra in the database, and determine multiple target peptide spectra containing the peptide segments to be studied from the database; An enzymatic hydrolysis condition determination module is used to mark the peptides to be studied in the sequence of each target peptide spectrum based on a target color set, and determine the optimal enzymatic hydrolysis conditions for obtaining the multiple peptides to be studied based on the overlapping colors in each target peptide spectrum, wherein the color categories in the target color set correspond one-to-one to each peptide to be studied; The database construction process includes: Based on different enzymatic hydrolysis conditions, the target protein is enzymatically hydrolyzed to obtain enzymatic hydrolysis products corresponding to the different enzymatic hydrolysis conditions, wherein the different enzymatic hydrolysis conditions are determined based on the type of enzyme, the amount of enzyme added, the pH value, the temperature and the enzymatic hydrolysis time; The database is constructed based on the peptide spectra of the enzymatic hydrolysis products corresponding to each enzymatic hydrolysis condition, wherein the peptide spectra of the enzymatic hydrolysis products are obtained by marking the enzymatic hydrolysis products corresponding to the enzymatic hydrolysis conditions in the amino acid sequence of the target protein; Also includes: Based on the preset combination conditions, the multiple peptides to be studied are combined to obtain a target peptide; Based on the overlapping colors in the target peptide spectra, the combined enzymatic hydrolysis conditions for obtaining the target peptide segment are determined.

8. An electronic device comprising a memory, a processor, and a computer program stored in the memory and running on the processor, characterized in that: When the processor executes the computer program, the method for determining the enzymatic hydrolysis conditions according to any one of claims 1 to 6 is implemented.

9. A non-transitory computer-readable storage medium having a computer program stored thereon, characterized in that: When the computer program is executed by a processor, the method for determining the enzymatic hydrolysis conditions according to any one of claims 1 to 6 is implemented.

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