Stropharia rugoso-annulata characteristic oligopeptide database and construction method and application thereof

Abstract

The invention discloses a Stropharia rugoso-annulata characteristic oligopeptide database and a construction method and application thereof, and belongs to the technical field of Stropharia rugoso-annulata characteristic oligopeptide database construction. A stropharia rugoso-annulata peptide molecule sequence is determined from stropharia rugoso-annulata fermentation mycelium, a stropharia rugoso-annulata sporocarp ultrasonic extract and a stropharia rugoso-annulata sporocarp enzymolysis product by utilizing a peptidomics technology, based on a fingerprint spectrum of peptide molecule mass and fragments and utilizing a bioinformatics tool, and a stropharia rugoso-annulata characteristic oligopeptide database is constructed. The construction method can comprehensively and quickly obtain the sequence information of the characteristic peptide molecules in the stropharia rugoso-annulata, has potential application value in the development of food, health care products and medicines, and provides powerful technical support for the development of food, health care products and medicines.

Description

Technical Field

[0001] The present invention belongs to the technical field of the construction of a characteristic oligopeptide database of Stropharia rugosoannulata, and particularly relates to a characteristic oligopeptide database of Stropharia rugosoannulata, a construction method thereof, and an application thereof. Background Art

[0002] Stropharia rugosoannulata is an edible mushroom with rich nutrition and unique taste. Its fruiting bodies and mycelia contain abundant proteins and peptide molecules. The peptide molecules of Stropharia rugosoannulata have various biological activities, such as blood pressure lowering, blood sugar lowering, antioxidant, anti-tumor, etc., and have a unique umami taste property. However, there is currently a lack of a characteristic peptide molecule database of Stropharia rugosoannulata, and effective quality control of Stropharia rugosoannulata peptide products cannot be achieved. Therefore, constructing a characteristic peptide database of Stropharia rugosoannulata and using the characteristic peptide sequences of Stropharia rugosoannulata as the quality control standards for peptide products are of great significance in the development of foods, health products, and drugs with Stropharia rugosoannulata peptides as the main nutritional and functional components. Summary of the Invention

[0003] In view of this, the purpose of the present invention is to provide a characteristic oligopeptide database of Stropharia rugosoannulata, which accurately and comprehensively contains the sequence information of characteristic peptide molecules in Stropharia rugosoannulata.

[0004] Another purpose of the present invention is to provide a construction method of the characteristic oligopeptide database of Stropharia rugosoannulata.

[0005] Another purpose of the present invention is to provide an application of the characteristic oligopeptide database of Stropharia rugosoannulata or the characteristic oligopeptide database of Stropharia rugosoannulata constructed by the construction method in the identification or quality control of Stropharia rugosoannulata products.

[0006] Another purpose of the present invention is to provide a method for identifying a product containing Stropharia rugosoannulata peptides.

[0007] In order to achieve the above invention purposes, the present invention provides the following technical solutions:

[0008] The present invention provides a characteristic oligopeptide database of Stropharia rugosoannulata, and the database contains 60 oligopeptide molecules shown in the following table:

[0009]

[0010]

[0011] The present invention also provides a construction method of the characteristic oligopeptide database of Stropharia rugosoannulata, and the construction method includes the following steps:

[0012] Perform mass spectrometry identification on the peptide sequences in the fermented mycelium extract of Stropharia rugoso-annulata, the ultrasonic extract of the fruiting body of Stropharia rugoso-annulata, and the enzymolysis product of the fruiting body of Stropharia rugoso-annulata to obtain the peptide spectra of each sample; screen the peptide segments with the number of occurrences ≥ 2 times and high confidence in each peptide spectrum; after merging the screened peptide segments, remove the repeatedly occurring peptide segments to obtain the characteristic oligopeptide database of Stropharia rugoso-annulata.

[0013] Preferably, the ultrasonic extract of the fruiting body of Stropharia rugoso-annulata includes a polyenergy ultrasonic extract and a flat plate divergent ultrasonic extract.

[0014] Preferably, the enzymolysis product of the fruiting body of Stropharia rugoso-annulata includes a product obtained by pretreatment with a complex enzyme system combined with moderate enzymolysis by an endonuclease and a product obtained by multi-mode synchronous ultrasonic-assisted directional enzymolysis.

[0015] Preferably, the mass spectrometry identification is performed by liquid chromatography-tandem mass spectrometry.

[0016] Preferably, the high confidence means that the peptide confidence score -10lgP ≥ 15.

[0017] Preferably, the peptide segment is a peptide molecule containing 7-9 amino acids.

[0018] The present invention also provides an application of the characteristic oligopeptide database of Stropharia rugoso-annulata or the characteristic oligopeptide database of Stropharia rugoso-annulata constructed by the construction method in the identification or quality control of Stropharia rugoso-annulata products.

[0019] Preferably, the Stropharia rugoso-annulata products include products containing peptides of Stropharia rugoso-annulata.

[0020] The present invention also provides a method for identifying a product containing peptides of Stropharia rugoso-annulata, and the method includes the following steps:

[0021] Perform mass spectrometry identification on the sample to be tested to obtain the peptide spectrum of the sample to be tested; compare the peptide segments in the peptide spectrum of the sample to be tested with the characteristic oligopeptide database of Stropharia rugoso-annulata or the characteristic oligopeptide database of Stropharia rugoso-annulata constructed by the construction method; if it contains the peptide segments in the characteristic oligopeptide database of Stropharia rugoso-annulata, it means that the sample to be tested contains peptides of Stropharia rugoso-annulata.

[0022] The beneficial effects of the present invention:

[0023] The characteristic oligopeptide database of Stropharia rugosoannulata in the present invention contains 60 characteristic oligopeptide molecules of Stropharia rugosoannulata, accurately and comprehensively including the sequence information of the characteristic peptide molecules in Stropharia rugosoannulata. Applying the characteristic oligopeptide database of Stropharia rugosoannulata in the present invention to identify the characteristic peptide molecules of Stropharia rugosoannulata in foods, health products, and drugs with Stropharia rugosoannulata peptides as the main nutritional and functional components can reflect the peptide molecule characteristics and product quality in the products, effectively distinguish them from other non-Stropharia rugosoannulata peptide products, and provide strong support for the precise identification of Stropharia rugosoannulata peptide products.

[0024] The present invention provides a construction method for efficiently and accurately constructing a characteristic oligopeptide database of Stropharia rugosoannulata based on peptide molecular fingerprinting, which can comprehensively and rapidly obtain the sequence information of characteristic peptide molecules in Stropharia rugosoannulata, has potential application value in the development of foods, health products, and drugs, and provides strong technical support for the development of foods, health products, and drugs. Description of the Drawings

[0025] Figure 1 It is the peptide fingerprint of the extract of the mycelium of Stropharia rugosoannulata fermented in Example 1;

[0026] Figure 2 It is the distribution diagram of the peptide segment scores matched by the peptide fingerprint of the extract of the mycelium of Stropharia rugosoannulata fermented in Example 1;

[0027] Figure 3 It is the peptide fingerprint of the extract of the fruiting body of Stropharia rugosoannulata by focused ultrasound in Example 1;

[0028] Figure 4 It is the distribution diagram of the peptide segment scores matched by the peptide fingerprint of the extract of the fruiting body of Stropharia rugosoannulata by focused ultrasound in Example 1;

[0029] Figure 5 It is the peptide fingerprint of the extract of the fruiting body of Stropharia rugosoannulata by plate divergent ultrasound in Example 1;

[0030] Figure 6 It is the distribution diagram of the peptide segment scores matched by the peptide fingerprint of the extract of the fruiting body of Stropharia rugosoannulata by plate divergent ultrasound in Example 1;

[0031] Figure 7 It is the peptide fingerprint of the enzymolysis product of the fruiting body of Stropharia rugosoannulata prepared by the pretreatment of complex enzyme system combined with moderate enzymolysis by endonuclease in Example 1;

[0032] Figure 8 It is the distribution diagram of the peptide segment scores matched by the peptide fingerprint of the enzymolysis product of the fruiting body of Stropharia rugosoannulata prepared by the pretreatment of complex enzyme system combined with moderate enzymolysis by endonuclease in Example 1;

[0033] Figure 9For the peptide fingerprint map of the enzymolysis product of the fruiting body of Stropharia rugoso-annulata prepared by multi-mode synchronous ultrasonic-assisted directional enzymolysis in Example 1;

[0034] Figure 10 For the peptide segment score distribution map matched with the peptide fingerprint map of the enzymolysis product of the fruiting body of Stropharia rugoso-annulata prepared by multi-mode synchronous ultrasonic-assisted directional enzymolysis in Example 1;

[0035] Figure 11 For the ion map of the KPPPPAP peptide segment in Example 1;

[0036] Figure 12 For the ion map of the HVVVGKN peptide segment in Example 1;

[0037] Figure 13 For the ion map of the DVKVPEGD peptide segment in Example 1;

[0038] Figure 14 For the ion map of the VVNPITSKL peptide segment in Example 1;

[0039] Figure 15 For the ion map of the PLVPVNH peptide segment in Example 1;

[0040] Figure 16 For the ion map of the DTVFGTH peptide segment in Example 1;

[0041] Figure 17 For the ion map of the SKVPWIL peptide segment in Example 1;

[0042] Figure 18 For the ion map of the RAFNDLL peptide segment in Example 1;

[0043] Figure 19 For the ion map of the WNPWIGK peptide segment in Example 1;

[0044] Figure 20 For the ion map of the VVVVPLPK peptide segment in Example 1;

[0045] Figure 21 For the ion map of the SHAVKIPGL peptide segment in Example 1;

[0046] Figure 22 For the ion map of the VVGTPGRVF peptide segment in Example 1;

[0047] Figure 23 For the ion map of the VNLIPVVAK peptide segment in Example 1;

[0048] Figure 24 For the ion map of the LVRVPPVGQ peptide segment in Example 1;

[0049] Figure 25Ion chromatogram of IGLPPIINF peptide in Example 1;

[0050] Figure 26 Ion chromatogram of IIEDQVRFK peptide in Example 1;

[0051] Figure 27 Ion chromatogram of ILEDQVRFK peptide in Example 1;

[0052] Figure 28 Ion chromatogram of KSWDDFFTR peptide in Example 1;

[0053] Figure 29 Peptide fingerprint of enzymatically hydrolyzed extract of Lentinula edodes in Test Example 1;

[0054] Figure 30 Peptide fingerprint of enzymatically hydrolyzed extract of Morchella esculenta in Test Example 1. Detailed implementation mode

[0055] The present invention provides a characteristic oligopeptide database of Stropharia rugoso-annulata, and the database contains 60 oligopeptide molecules shown in Table 1:

[0056] Table 1 Characteristic peptide molecule database of Stropharia rugoso-annulata

[0057]

[0058]

[0059] The present invention also provides a construction method of the characteristic oligopeptide database of Stropharia rugoso-annulata, and the construction method includes the following steps:

[0060] Perform mass spectrometry identification on the peptide sequences in the extracts of Stropharia rugoso-annulata fermented mycelium, the ultrasonic extract of Stropharia rugoso-annulata fruiting body, and the enzymatic hydrolysis product of Stropharia rugoso-annulata fruiting body respectively to obtain the peptide spectra of each sample; screen the peptide segments with the appearance times ≥ 2 times and high confidence in each peptide spectrum; after combining the screened peptide segments, remove the repeatedly appearing peptide segments to obtain the characteristic oligopeptide database of Stropharia rugoso-annulata.

[0061] In the present invention, the extract of Stropharia rugoso-annulata fermented mycelium is an extract obtained by extracting with Stropharia rugoso-annulata fermented mycelium as the raw material, and the Stropharia rugoso-annulata fermented mycelium is obtained by fermenting with the fermentation strain isolated from the Stropharia rugoso-annulata fruiting body.

[0062] As an alternative implementation mode, the preparation method of the extract of Stropharia rugoso-annulata fermented mycelium preferably includes the following steps: isolate the fermentation strain from the Stropharia rugoso-annulata fruiting body, inoculate it into the medium for liquid fermentation; centrifuge to collect the mycelium; use the high-pressure extraction method to extract the peptides in the mycelium to obtain the extract of Stropharia rugoso-annulata fermented mycelium.

[0063] In the present invention, there are no special limitations on the type and source of the fruiting bodies of Stropharia rugosoannulata, and preferably the fruiting bodies of Stropharia rugosoannulata Shenqiu No. 2 are used. The method for separating and purifying the fermentation strain can be conventionally selected according to actual needs. There are no special limitations on the fermentation strain, and preferably the strain with the NCBI strain release number SRR14469700 is used.

[0064] In the present invention, the fermentation strain is inoculated into a medium for liquid fermentation. The medium is preferably a PDB medium; the inoculation amount of the fermentation strain is preferably 5% - 15% (v / v), more preferably 7.5% - 12.5% or 10%; the temperature of the liquid fermentation is preferably 20 - 30 °C, more preferably 22 - 28 °C or 26 °C, the stirring speed is preferably 50 - 150 r / min, more preferably 75 - 125 r / min or 100 r / min, the ventilation volume is preferably 20 - 30 L / min, more preferably 22 - 27 L / min or 25 L / min, and the pH is preferably the natural pH.

[0065] In the present invention, the rotation speed of the centrifugation is preferably 3000 - 5000 r / min, more preferably 3500 - 4500 r / min or 4000 r / min, and the time of the centrifugation is preferably 5 - 15 min, more preferably 7 - 12 min or 10 min.

[0066] In the present invention, after centrifugally collecting the mycelium, it preferably further includes washing and drying the mycelium. The methods of washing and drying can be conventionally selected according to actual needs. As an implementable method, it is preferably to wash the mycelium with distilled water and then freeze-dry it. The temperature of the freeze-drying is preferably -80 to -60 °C, more preferably -75 to -65 °C or -70 °C; the time of the freeze-drying is preferably 36 - 60 h, more preferably 40 - 54 h or 48 h.

[0067] In the present invention, the material-liquid ratio of the high-pressure extraction method is preferably 1:10 - 30 (g:mL), more preferably 1:15 - 25 (g:mL) or 1:20 (g:mL); the temperature is preferably 110 - 130 °C, more preferably 115 - 125 °C or 121 °C; the pressure is preferably 0.05 - 0.5 MPa, more preferably 0.08 - 0.2 MPa or 0.1 MPa; the extraction time is preferably 90 - 150 min, more preferably 110 - 130 min or 120 min.

[0068] In the present invention, after leaching, the supernatant is collected by centrifugation, and the fermented mycelium extract of Stropharia rugoso-annulata is obtained by drying. The rotation speed of the centrifugation is preferably 6000-10000 r / min, more preferably 7000-9000 r / min or 8000 r / min; the time of the centrifugation is preferably 10-30 min, more preferably 15-25 min or 20 min. The drying is preferably freeze-drying, the temperature of the freeze-drying is preferably -80 to -60 °C, more preferably -75 to -65 °C or -70 °C, and the time of the freeze-drying is preferably 36-60 h, more preferably 40-54 h or 48 h.

[0069] In the present invention, the peptide content in the fermented mycelium extract of Stropharia rugoso-annulata is preferably 40-60 mg / g dry weight, more preferably 45-55 mg / g dry weight or 53.43 mg / g dry weight.

[0070] In the present invention, the ultrasonic extract of Stropharia rugoso-annulata fruiting body is an extract obtained by using the ultrasonic extraction method with the Stropharia rugoso-annulata fruiting body as the raw material, preferably including the poly-energy ultrasonic extract and the flat-divergent ultrasonic extract. The type and source of the Stropharia rugoso-annulata fruiting body are not particularly limited, and preferably the fruiting body of Stropharia rugoso-annulata Shenqiu No. 2.

[0071] As an alternative embodiment, the preparation method of the poly-energy ultrasonic extract preferably includes: extracting the free peptides of the Stropharia rugoso-annulata fruiting body by using the poly-energy ultrasonic mode. The material-liquid ratio for extraction is preferably 1:10-30 (g:mL), more preferably 1:15-25 (g:mL) or 1:20 (g:mL); the ultrasonic power density of the poly-energy ultrasonic mode is preferably 200-400 W / L, more preferably 250-350 W / L or 300 W / L; the ultrasonic frequency mode is preferably synchronous dual-frequency, and the frequencies of the synchronous dual-frequency are preferably 20 kHz and 28 kHz; the intermittent ratio is preferably 4-6 s / 4-6 s, more preferably 5 s / 5 s; the extraction time is preferably 10-30 min, more preferably 15-25 min or 20 min.

[0072] In the present invention, after ultrasonic extraction, centrifugation is carried out to collect the supernatant, and the poly-energy ultrasonic extract is obtained by drying. The rotation speed of the centrifugation is preferably 6000-10000 r / min, more preferably 7000-9000 r / min or 8000 r / min; the time of the centrifugation is preferably 10-30 min, more preferably 15-25 min or 20 min. The drying is preferably freeze-drying, the temperature of the freeze-drying is preferably -80 to -60 °C, more preferably -75 to -65 °C or -70 °C, and the time of the freeze-drying is preferably 36-60 h, more preferably 40-54 h or 48 h.

[0073] In the present invention, the peptide content in the converging ultrasound extract is preferably 300-500 mg / g dry weight, more preferably 350-400 mg / g dry weight or 385.29 mg / g dry weight.

[0074] As an alternative embodiment, the preparation method of the plate divergent ultrasound extract preferably includes: extracting the free peptides from the fruiting bodies of Stropharia rugoso-annulata by using the plate divergent ultrasound mode. The material-liquid ratio for extraction is preferably 1:10-30 (g:mL), more preferably 1:15-25 (g:mL) or 1:20 (g:mL); the ultrasound power density of the plate divergent ultrasound mode is preferably 200-400 W / L, more preferably 250-350 W / L or 300 W / L; the ultrasound frequency mode is preferably synchronous dual-frequency, and the frequencies of the synchronous dual-frequency are preferably 20 kHz and 28 kHz; the intermittent ratio is preferably 4-6 s / 4-6 s, more preferably 5 s / 5 s; the extraction time is preferably 10-30 min, more preferably 15-25 min or 20 min.

[0075] In the present invention, after ultrasonic extraction, centrifugation is carried out, the supernatant is collected, and the plate divergent ultrasound extract is obtained by drying. The rotation speed for centrifugation is preferably 6000-10000 r / min, more preferably 7000-9000 r / min or 8000 r / min; the centrifugation time is preferably 10-30 min, more preferably 15-25 min or 20 min. The drying is preferably freeze-drying, the temperature for freeze-drying is preferably -80 to -60 °C, more preferably -75 to -65 °C or -70 °C, and the freeze-drying time is preferably 36-60 h, more preferably 40-54 h or 48 h.

[0076] In the present invention, the peptide content in the plate divergent ultrasound extract is preferably 250-450 mg / g dry weight, more preferably 300-400 mg / g dry weight or 343.52 mg / g dry weight.

[0077] In the present invention, the enzymatic hydrolysate of Stropharia rugoso-annulata fruit bodies is an enzymatic hydrolysate obtained by subjecting Stropharia rugoso-annulata fruit bodies as raw materials to enzymatic hydrolysis by an enzymatic hydrolysis method, preferably including a product obtained by combined pretreatment with a complex enzyme system and moderate enzymatic hydrolysis with an endonuclease, and a product obtained by multi-mode synchronous ultrasonic-assisted directional enzymatic hydrolysis. Cellulase is a complex enzyme composed of multiple enzyme systems, and it has a relatively high ability to degrade mushroom cellulose, which is beneficial to the release of mushroom proteins cross-linked with cellulose; while alkaline protease, as an endonuclease, has strong proteolytic ability, and its ability to produce peptides by hydrolysis is significantly higher than that of flavor protease, trypsin, etc. The energy generated by the poly-energy ultrasound instantaneously is relatively high, and there are effects such as shear force generated by ultrasonic cavitation, instantaneous high pressure and high temperature of cavitation bubbles, and micro-streaming. The raw materials are pretreated by using the physical and cavitation equivalent effects generated by ultrasound, so that the protein structure changes and the tissue becomes loose, which is beneficial to the release of peptides and improves the conversion rate of the product. Therefore, cellulase and poly-energy ultrasound are selected to pretreat the raw materials respectively, and alkaline protease is used as the protease for enzymatic hydrolysis to obtain two enzymatic hydrolysates of Stropharia rugoso-annulata fruit bodies.

[0078] In the present invention, there are no special limitations on the type and source of the Stropharia rugoso-annulata fruit bodies, and preferably the fruit bodies of Stropharia rugoso-annulata Shenqiu No. 2 are used.

[0079] As an alternative embodiment, the preparation method of the product obtained by combined pretreatment with a complex enzyme system and moderate enzymatic hydrolysis with an endonuclease preferably includes: dissolving the powder of Stropharia rugoso-annulata fruit bodies in water, adding cellulase for pretreatment; after the pretreatment is completed, adding alkaline protease for enzymatic hydrolysis to obtain the product obtained by combined pretreatment with a complex enzyme system and moderate enzymatic hydrolysis with an endonuclease.

[0080] In the present invention, when the powder of Stropharia rugoso-annulata fruit bodies is dissolved in water, the concentration of the obtained feed liquid is preferably 30 - 60 g / L, more preferably 40 - 55 g / L or 48 g / L; the enzyme activity of the cellulase is preferably 100000 - 1000000 U / g, more preferably 250000 - 750000 U / g or 500000 U / g; the addition amount of the cellulase is preferably 0.5% - 1.5% (w / w), more preferably 0.6% - 1% or 0.8%; the temperature for cellulase pretreatment is preferably 50 - 70 °C, more preferably 55 - 65 °C or 60 °C; the pretreatment time is preferably 90 - 150 min, more preferably 110 - 130 min or 120 min.

[0081] In the present invention, the enzyme activity of the alkaline protease is preferably 100,000 - 500,000 U / g, more preferably 150,000 - 300,000 U / g or 200,000 U / g; the addition amount of the alkaline protease is preferably 0.5% - 1.5% (w / w), more preferably 0.7% - 1.2% or 1%; the temperature for enzymatic hydrolysis with the alkaline protease is preferably 35 - 50 °C, more preferably 40 - 45 °C or 42 °C; the time for enzymatic hydrolysis is preferably 40 - 60 min, more preferably 45 - 55 min or 51 min.

[0082] In the present invention, after enzymatic hydrolysis, it preferably further includes inactivating the enzyme, centrifuging, collecting the supernatant, and drying to obtain the product of combined pretreatment with complex enzyme system and moderate enzymatic hydrolysis with endonuclease.

[0083] In the present invention, the enzyme inactivation preferably adopts boiling water bath for enzyme inactivation, and the time for enzyme inactivation is preferably 5 - 15 min, more preferably 7 - 12 min or 10 min. The rotation speed for centrifuging is preferably 6000 - 10000 r / min, more preferably 7000 - 9000 r / min or 8000 r / min; the time for centrifuging is preferably 10 - 30 min, more preferably 15 - 25 min or 20 min. The drying is preferably freeze-drying, the temperature for freeze-drying is preferably -80 - -60 °C, more preferably -75 - -65 °C or -70 °C, and the time for freeze-drying is preferably 36 - 60 h, more preferably 40 - 54 h or 48 h.

[0084] In the present invention, in the product of combined pretreatment with complex enzyme system and moderate enzymatic hydrolysis with endonuclease, the peptide content is preferably 400 - 550 mg / g dry weight, more preferably 450 - 500 mg / g dry weight or 461.31 mg / g dry weight.

[0085] As an alternative embodiment, the preparation method of the multi-mode synchronous ultrasonic-assisted directional enzymatic hydrolysis product preferably includes: dissolving the powder of fruiting bodies of Stropharia rugosoannulata in water, and performing ultrasonic extraction with energy-gathering ultrasound; adding alkaline protease and performing assisted enzymatic hydrolysis with divergent ultrasound to obtain the multi-mode synchronous ultrasonic-assisted directional enzymatic hydrolysis product.

[0086] In the present invention, when the powder of fruiting bodies of Stropharia rugosoannulata is dissolved in water, the obtained concentration of the feed liquid is preferably 30 - 60 g / L, more preferably 40 - 55 g / L or 48 g / L; the working frequency of the energy-gathering ultrasound is preferably 15 - 25 kHz, more preferably 20 kHz; the working intermittent ratio is preferably 4 - 6 s / 4 - 6 s, more preferably 5 s / 5 s; the power density of the energy-gathering ultrasound is preferably 200 - 400 W / L, more preferably 250 - 350 W / L or 300 W / L; the treatment time of the energy-gathering ultrasound is preferably 10 - 20 min, more preferably 12 - 17 min or 15 min.

[0087] In the present invention, the enzyme activity of the alkaline protease is preferably 100,000 - 500,000 U / g, more preferably 150,000 - 300,000 U / g or 200,000 U / g; the addition amount of the alkaline protease is preferably 0.5% - 1.5% (w / w), more preferably 0.7% - 1.2% or 1%; the enzymatic hydrolysis temperature is preferably 35 - 50 °C, more preferably 40 - 45 °C or 42 °C; the working frequency of the divergent ultrasound is preferably 15 - 25 kHz, more preferably 20 kHz; the working intermittent ratio is preferably 4 - 6 s / 4 - 6 s, more preferably 5 s / 5 s; the power density of the divergent ultrasound is preferably 100 - 150 W / L, more preferably 110 - 130 W / L or 120 W / L; the action time of the divergent ultrasound-assisted enzymatic hydrolysis is preferably 30 - 50 min, more preferably 35 - 45 min or 40 min.

[0088] In the present invention, after enzymatic hydrolysis, it preferably further includes inactivating the enzyme, centrifuging, collecting the supernatant, and drying to obtain a multi-mode synchronous ultrasound-assisted directional enzymatic hydrolysis product.

[0089] In the present invention, the enzyme inactivation preferably uses boiling water bath for enzyme inactivation, and the time of enzyme inactivation is preferably 5 - 15 min, more preferably 7 - 12 min or 10 min. The rotation speed of the centrifugation is preferably 6000 - 10000 r / min, more preferably 7000 - 9000 r / min or 8000 r / min; the time of centrifugation is preferably 10 - 30 min, more preferably 15 - 25 min or 20 min. The drying is preferably freeze-drying, the temperature of the freeze-drying is preferably -80 - -60 °C, more preferably -75 - -65 °C or -70 °C, and the time of the freeze-drying is preferably 36 - 60 h, more preferably 40 - 54 h or 48 h.

[0090] In the present invention, in the multi-mode synchronous ultrasound-assisted directional enzymatic hydrolysis product, the peptide content is preferably 400 - 600 mg / g dry weight, more preferably 450 - 550 mg / g dry weight or 492.87 mg / g dry weight.

[0091] In the present invention, after obtaining the peptide substrates by the above different methods, mass spectrometry identification is respectively carried out to obtain the peptide spectra of each peptide substrate sample. Before mass spectrometry identification, it is preferably to carry out desalting pretreatment on the peptide substrate, and preferably use a ZipTip C18 microchromatography column to carry out desalting pretreatment on the peptide substrate. The method of the desalting pretreatment can be conventionally selected according to actual needs.

[0092] In the present invention, the mass spectrometry identification is preferably carried out by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Preferably, the peptide base material is dissolved in a dissolution solution, vortexed, centrifuged, and the supernatant is collected as the injection solution. The dissolution solution is preferably a solution containing 0.1% formic acid and 5% acetonitrile. The temperature of the centrifugation is preferably 2-6 °C, more preferably 4 °C; the rotation speed of the centrifugation is preferably 10000-15000 r / min, more preferably 12000-14000 r / min or 13500 r / min; the time of the centrifugation is preferably 10-30 min, more preferably 15-25 min or 20 min.

[0093] In the present invention, the injection volume of the injection solution is preferably 5-10 μL, more preferably 6-9 μL or 8 μL. Mobile phase A is preferably 0.1% formic acid, and mobile phase B is preferably an aqueous solution containing 0.1% formic acid and 80% acetonitrile.

[0094] In the present invention, in the liquid chromatography-tandem mass spectrometry method, the liquid chromatography conditions preferably include: 0 min 97% A + 3% B, flow rate 400 nL / min; 3 min 97% A + 3% B, flow rate 400 nL / min; 7 min 92% A + 8% B, flow rate 400 nL / min; 46 min 68% A + 32% B, flow rate 400 nL / min; 51 min 56% A + 44% B, flow rate 400 nL / min; 56 min 1% A + 99% B, flow rate 400 nL / min; 60 min 1% A + 99% B, flow rate 400 nL / min; 60.1 min 97% A + 3% B, flow rate 300 nL / min; 70 min 97% A + 3% B, flow rate 400 nL / min.

[0095] In the present invention, in the liquid chromatography-tandem mass spectrometry method, the mass spectrometry conditions include: the resolution of the first-stage mass spectrometry is 120000, the automatic gain control is 4×10 5 , the maximum ion injection time is 50 ms, and the mass scanning range is 350-1550 m / z; the resolution of the second-stage mass spectrometry is 30000, the automatic gain control is 1×10 5 , the maximum ion injection time is 100 ms, the number of ions selected for second-stage fragmentation in the first-stage mass spectrometry is 20, and the collision energy in the NCE mode is 32.

[0096] In the present invention, after obtaining the peptide spectra of each peptide base material sample, peptide segments with the number of occurrences ≥ 2 times and high confidence in each peptide spectrum are screened; the high confidence means that the peptide confidence score - 10lgP ≥ 15; the peptide segment is preferably a peptide molecule containing 7-9 amino acids.

[0097] In the present invention, the screened peptide segments are combined, and the repeatedly occurring peptide segments are removed to obtain a characteristic oligopeptide database of Stropharia rugosoannulata.

[0098] The present invention also provides an application of the characteristic oligopeptide database of Stropharia rugosoannulata or the characteristic oligopeptide database of Stropharia rugosoannulata constructed by the construction method in the identification or quality control of Stropharia rugosoannulata products.

[0099] In the present invention, the Stropharia rugosoannulata products preferably include products containing Stropharia rugosoannulata peptides. There is no special limitation on the types of the products, including foods, drugs or health products.

[0100] The present invention also provides a method for identifying a product containing Stropharia rugosoannulata peptides, and the method includes the following steps:

[0101] Performing mass spectrometry identification on a sample to be tested to obtain a peptide spectrum of the sample to be tested; comparing the peptide segments in the peptide spectrum of the sample to be tested with the characteristic oligopeptide database of Stropharia rugosoannulata or the characteristic oligopeptide database of Stropharia rugosoannulata constructed by the construction method; if it contains the peptide segments in the characteristic oligopeptide database of Stropharia rugosoannulata, it indicates that the sample to be tested contains Stropharia rugosoannulata peptides.

[0102] In the present invention, the mass spectrometry identification method is preferably to perform identification by using the LC-MS / MS method; the steps and parameters of the LC-MS / MS method are preferably the same as those in the above-mentioned construction method of the characteristic oligopeptide database of Stropharia rugosoannulata.

[0103] In the present invention, from the fermented mycelium of Stropharia rugosoannulata, the ultrasonic extract of the fruiting body of Stropharia rugosoannulata, and the enzymatic hydrolysate of the fruiting body of Stropharia rugosoannulata, by using peptidomics technology, based on the fingerprint maps of peptide molecular weights and fragments, bioinformatics tools are used to determine the molecular sequences of Stropharia rugosoannulata peptides, and a characteristic oligopeptide database of Stropharia rugosoannulata is constructed. The characteristic oligopeptides in the characteristic oligopeptide database of Stropharia rugosoannulata of the present invention are peptide molecules containing 7-9 amino acids, belonging to oligopeptide molecules. As quality control molecules, due to their long molecular chains, stable structures, not easily degradable, and having specificity, in the application process, the non-specificity of short peptides (containing 2-3 amino acids) that can be detected in a variety of foods is avoided.

[0104] Using the Stropharia rugosoannulata characteristic oligopeptide database constructed by the present invention to screen a substrate rich in Stropharia rugosoannulata characteristic oligopeptides as a food ingredient or food additive can significantly improve the taste, flavor and nutritional value of food, and enhance the quality and market competitiveness of food. Using the Stropharia rugosoannulata characteristic oligopeptide database constructed by the present invention to screen Stropharia rugosoannulata characteristic peptide molecules as health product ingredients can prepare health products with various biological activities to meet people's needs for a healthy life. Using the Stropharia rugosoannulata characteristic oligopeptide database constructed by the present invention to screen Stropharia rugosoannulata characteristic peptide molecules as drug efficacy ingredients can develop drugs with specific pharmacological activities and provide new options for the treatment of diseases.

[0105] Applying the Stropharia rugosoannulata characteristic oligopeptide database constructed by the present invention to identify Stropharia rugosoannulata characteristic peptide molecules in foods, health products and drugs with Stropharia rugosoannulata peptides as the main nutritional and functional ingredients can reflect the peptide molecule characteristics and product quality of the products, effectively distinguish them from other non-Stropharia rugosoannulata peptide products, and provide strong support for the accurate identification of Stropharia rugosoannulata peptide products. Applying the Stropharia rugosoannulata characteristic oligopeptide database to the actual production of Stropharia rugosoannulata peptide substrates can ensure the consistency and stability of products by real-time monitoring of the changes in peptide molecules during the production process.

[0106] The technical solutions provided by the present invention will be described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0107] In the following embodiments, unless otherwise specified, all are conventional methods.

[0108] The materials, reagents, etc. used in the following embodiments, unless otherwise specified, can all be obtained from commercial channels.

[0109] Example 1

[0110] A method for constructing a Stropharia rugosoannulata characteristic oligopeptide database, and the preparation method steps are as follows:

[0111] (1) Extract of Stropharia rugosoannulata fermented mycelium: The fermented mycelium strain was isolated from the fruiting body of Stropharia rugosoannulata Shenqiu No. 2, and the NCBI strain release number is SRR14469700. Using the same strain as the fruiting body for liquid fermentation can ensure the consistency of the provenance of different morphological samples.

[0112] The fermentation conditions of the mycelium of Stropharia rugosoannulata are as follows: Liquid fermentation is carried out in a 30L fermenter with an inoculum size of 10% (v / v); among them, the fermenter is filled with 25L of PDB medium, the fermentation temperature is 26°C, the stirring speed is 100r / min, the ventilation volume is 25L / min, and the pH is the natural pH. After fermentation, the fermentation sample is centrifuged at 4000r / min for 10min to collect the mycelium. After the mycelium is thoroughly rinsed with distilled water, it is freeze-dried at -70°C for 48h to obtain freeze-dried mycelium.

[0113] The extraction conditions of the peptide base material from the extract of the fermented mycelium of Stropharia rugosoannulata are as follows: The solid-liquid ratio is 1:20 (g:mL), and it is extracted under high pressure at 121°C and 0.1MPa for 120min. After the extract is centrifuged at 8000r / min for 20min, the supernatant is collected and freeze-dried at -70°C for 48h to obtain the peptide base material from the extract of the fermented mycelium of Stropharia rugosoannulata.

[0114] (2) Ultrasonic extract of Stropharia rugosoannulata fruiting body: Two modes of ultrasonic energy concentration and flat-panel divergent ultrasound are used to extract free peptides from Stropharia rugosoannulata. The raw material used is the fruiting body of Stropharia rugosoannulata Shenqiu No. 2.

[0115] ① Conditions for extracting free peptides from Stropharia rugosoannulata by ultrasonic energy concentration: The ultrasonic power density is 300W / L, the ultrasonic frequency mode is synchronous dual frequency (20kHz / 28kHz), the intermittent ratio is 5s / 5s, the extraction time is 20min, and the solid-liquid ratio is 1:20 (g:mL). After the extract is centrifuged at 8000r / min for 20min, the supernatant is collected and freeze-dried at -70°C for 48h to obtain the peptide base material from the ultrasonic energy concentration extract of Stropharia rugosoannulata fruiting body.

[0116] ② Conditions for extracting free peptides from Stropharia rugosoannulata by flat-panel divergent ultrasound: The ultrasonic power density is 300W / L, the ultrasonic frequency mode is synchronous dual frequency (20kHz / 28KHz), the intermittent ratio is 5s / 5s, the extraction time is 20min, and the solid-liquid ratio is 1:20 (g:mL). After the extract is centrifuged at 8000r / min for 20min, the supernatant is collected and freeze-dried at -70°C for 48h to obtain the peptide base material from the flat-panel divergent ultrasound extract of Stropharia rugosoannulata fruiting body.

[0117] (3) Enzymatic hydrolysate of Stropharia rugosoannulata fruiting body: Two enzymatic hydrolysis techniques are used to prepare the peptide base material of the enzymatic hydrolysate of Stropharia rugosoannulata fruiting body, including the combined pretreatment of complex enzyme systems and moderate enzymatic hydrolysis with endonuclease, and the multi-mode synchronous ultrasonic-assisted directional enzymatic hydrolysis technique. The raw material used is the fruiting body of Stropharia rugosoannulata Shenqiu No. 2.

[0118] ① Preparation of peptide-based substrate of Stropharia rugosoannulata by pretreatment with composite enzyme system and moderate enzymatic hydrolysis by endoenzyme: The method for raw material pretreatment is as follows: the substrate concentration is 48 g / L, the dosage of cellulase (enzyme activity 500000 U / g) is 0.8% (w / w), the temperature is 60 °C, and the pretreatment time is 120 min. The process for preparing the peptide-based substrate is as follows: the dosage of alkaline protease (enzyme activity 200000 U / g) is 1% (w / w), the enzymatic hydrolysis temperature is 42 °C, and the enzymatic hydrolysis time is 51 min. After inactivating the enzyme in the enzymatic hydrolysate in a boiling water bath for 10 min, centrifuge at 8000 r / min for 20 min to collect the supernatant, and freeze-dry at -70 °C for 48 h to obtain the peptide-based substrate of the enzymatic hydrolysate of Stropharia rugosoannulata fruiting bodies.

[0119] ② Preparation of peptide-based substrate of Stropharia rugosoannulata by multi-mode synchronous ultrasonic-assisted directional enzymatic hydrolysis: The method for raw material pretreatment is as follows: the substrate concentration is 48 g / L, the working frequency of the polyenergy ultrasonic wave is 20 kHz, the working intermittent ratio is 5 s / 5 s, the power density of the polyenergy ultrasonic wave is 300 W / L, and the treatment time of the polyenergy ultrasonic wave is 15 min. The process for preparing the peptide-based substrate is as follows: the dosage of alkaline protease (enzyme activity 200000 U / g) is 1% (w / w), the enzymatic hydrolysis temperature is 42 °C, the working frequency of the divergent ultrasonic wave is 20 kHz, the working intermittent ratio is 5 s / 5 s, the power density of the divergent ultrasonic wave is 120 W / L, and the action time of the divergent ultrasonic wave-assisted enzymatic hydrolysis is 40 min. After inactivating the enzyme in the enzymatic hydrolysate in a boiling water bath for 10 min, centrifuge at 8000 r / min for 20 min to collect the supernatant, and freeze-dry at -70 °C for 48 h to obtain the peptide-based substrate of the enzymatic hydrolysate of Stropharia rugosoannulata fruiting bodies.

[0120] (4) After the above 5 kinds of peptide-based substrates are prepared, the peptide content in each peptide-based substrate is detected respectively.

[0121] The detection method is as follows: The analysis of the peptide content in the peptide-based substrate of Stropharia rugosoannulata adopts the kit method (Suzhou Mengxi Biopharmaceutical Technology Co., Ltd.). Use 10% trichloroacetic acid solution to precipitate and remove macromolecular proteins in the sample, and retain peptide molecules in the sample. The peptide bonds in the molecules reduce Cu 2+ to Cu + ; 2 molecules of BCA combine with Cu + to form a purple complex, which has an absorption peak at 540 - 595 nm, and the absorption peak is the strongest at 562 nm. Weigh 0.1 g of freeze-dried substrate, add 1 mL of the extraction solution of the kit (10% trichloroacetic acid), homogenize in an ice bath and let it stand for 30 min, centrifuge at 12000 r / min at 4 °C for 10 min, and then collect the supernatant. Take 10 μL of the supernatant sample, add 190 μL of the working solution of the kit, incubate in an oven at 60 °C for 30 min, and measure the absorbance value at 562 nm. The calculation of the peptide content refers to the method of the kit.

[0122] The results show that the peptide content in the above 5 kinds of peptide-based substrates is shown in Table 2.

[0123] Table 2 Peptide content in peptide-based substrates

[0124]

[0125] (5) Peptide sequence mass spectrometry identification method: A ZipTip C18 microchromatography column was used to perform desalting pretreatment on the freeze-dried substrate. The desalting method is as follows: Accurately weigh 10 mg of the freeze-dried sample, add 30 μL of 0.1% trifluoroacetic acid (TFA) to dissolve it; rinse the chromatography column 10 times with 50 μL of 60% acetonitrile (ACN) and 0.1% TFA; rinse the chromatography column 10 times with 10 μL of 0.1% TFA; aspirate and discharge the sample through the chromatography column 20 times; rinse the chromatography column 5 times with 10 μL of 0.1% TFA; elute the chromatography column with 10 μL of 60% ACN and 0.1% TFA, transfer the eluted peptide segments to a polypropylene centrifuge tube, and detect them on the machine after vacuum drying.

[0126] Dissolve the peptide segments with 20 μL of the dissolution solution (containing 0.1% formic acid and 5% ACN), vortex and oscillate, centrifuge at 13,500 r / min at 4 °C for 20 min, collect the supernatant and transfer it to the sample injection tube. The injection volume of the sample solution is 8 μL, and mass spectrometry peptide sequence analysis and identification are carried out. The mobile phase A of the liquid chromatography is 0.1% formic acid; the mobile phase B is an aqueous solution containing 0.1% formic acid and 80% ACN; the LC-MS / MS setting parameters are shown in Table 3. The PEAKS software was used for peptide sequence database retrieval. The peptide spectra of each sample obtained by mass spectrometry identification are as Figures 1 - 10 shown.

[0127] Table 3 LC-MS / MS parameter settings

[0128]

[0129] (6) Screen the effective peptide segments with the number of occurrences ≥ 2 times and high confidence (peptide confidence score -10lgP ≥ 15, protein confidence score -10lgP ≥ 20) in each peptide spectrum. The results are shown in Table 4 - Table 8. 16 oligopeptide molecules were identified from the extract of the fermented mycelium of Stropharia rugosoannulata (Table 4), 9 oligopeptide molecules were identified from the polyenergy ultrasonic extract of the fruiting body of Stropharia rugosoannulata (Table 5), 5 oligopeptide molecules were identified from the plate divergent ultrasonic extract of the fruiting body of Stropharia rugosoannulata (Table 6), 18 oligopeptide molecules were identified from the enzymolysis product of the fruiting body of Stropharia rugosoannulata prepared by the combined pretreatment of complex enzyme system and moderate enzymolysis with endonuclease (Table 7), and 25 oligopeptide molecules were identified from the enzymolysis product of the fruiting body of Stropharia rugosoannulata prepared by multi-mode synchronous ultrasonic-assisted directional enzymolysis (Table 8).

[0130] Table 4 Peptides in the extract of the fermented mycelium of Stropharia rugosoannulata obtained by mass spectrometry identification

[0131]

[0132]

[0133] Peptides in the poly-energetic ultrasonic extract of the fruiting body of Stropharia rugoso-annulata identified by mass spectrometry

[0134] Serial number Peptide sequence Molecular weight Chain length Source (NCBI database number of precursor protein) 1 PLVPVNH 774 7 A0A409XF43_PSICY 2 DTVFGTH 775 7 A0A409WID3_PSICY 3 AIEPPVRP 878 8 A0A0D2LMI9_HYPSF 4 LDYGDGKW 952 8 A0A409WQ59_PSICY 5 SDIKHFPF 989 8 A0A067TNR5_GALM3 6 ELPPTHPI 902 8 A0A0D2LNW4_HYPSF 7 AIEPPVRPS 965 9 A0A0D2LMI9_HYPSF 8 GPVSDFPTR 974 9 A0A409XLX0_PSICY 9 VVDEGILQH 1009 9 A0A0D2LLX1_HYPSF

[0135] Peptides in the plate divergence ultrasonic extract of the fruiting body of Stropharia rugoso-annulata identified by mass spectrometry

[0136] Serial number Peptide sequence Molecular weight Chain length Source (NCBI database number of precursor protein) 1 PLVPVNH 774 7 A0A409XF43_PSICY 2 DTVFGTH 775 7 A0A409WID3_PSICY 3 AIEPPVRP 878 8 D8WMA7_9AGAR 4 AIEPPVRPS 965 9 D8WMA7_9AGAR 5 VVDEGILQH 1009 9 A0A0D2LLX1_HYPSF

[0137] Peptides in the enzymolysis product of the fruiting body of Stropharia rugoso-annulata prepared by pretreatment with complex enzyme system combined with moderate enzymolysis by endonuclease identified by mass spectrometry

[0138]

[0139]

[0140] Peptides in the enzymolysis product of the fruiting body of Stropharia rugoso-annulata prepared by multi-mode synchronous ultrasonic-assisted directional enzymolysis identified by mass spectrometry

[0141]

[0142]

[0143] (7) After merging the effective peptide segments in Tables 4 - 8 and removing the repeatedly occurring peptide segments, a characteristic peptide molecular database of Stropharia rugoso-annulata was obtained, as shown in Table 9, with a total of 60 oligopeptide molecules.

[0144] Table 9 Characteristic peptide molecular database of Stropharia rugoso-annulata

[0145]

[0146]

[0147]

[0148] In Table 9, the peptide segment ion chromatograms marked with * are as Figures 11 - 28 shown.

[0149] Experimental Example 1

[0150] Peptides were extracted from Lentinula edodes and Morchella esculenta respectively by the method of biological enzyme hydrolysis.

[0151] (1) For the shiitake mushroom powder and water, the solid-liquid ratio is 1:30 (g:mL), the temperature is 50 °C, the pH is 7.0, add flavor protease with an enzyme activity of 1000 U / g (enzyme activity compared to the mass of mushroom powder) (enzyme activity 30000 U / g) and enzymatically hydrolyze for 45 min. Then add trypsin with an enzyme activity of 2000 U / g (enzyme activity compared to the mass of mushroom powder) (enzyme activity 250 U / mg), and enzymatically hydrolyze at 37 °C and pH 8.0 for another 45 min. Inactivate the enzyme in the enzymatic hydrolysate, collect the centrifuged supernatant, and dry it in the same steps as the extraction of the enzymatic hydrolysate of the fruiting body of Stropharia rugosoannulata in Example 1.

[0152] (2) For the morel powder and water, the solid-liquid ratio is 1:30 (g:mL), the addition amount of pectinase (enzyme activity 40 U / mg) is 7% (w / w), the temperature is 52 °C, the pH is 5.0, and the enzymatic hydrolysis time is 48 min. Inactivate the enzyme in the enzymatic hydrolysate, collect the centrifuged supernatant, and dry it in the same steps as the extraction of the enzymatic hydrolysate of the fruiting body of Stropharia rugosoannulata in Example 1.

[0153] Use the peptide sequence mass spectrometry identification method (the same as the (5) peptide sequence mass spectrometry identification method in Example 1) to perform mass spectrometry detection on the shiitake mushroom peptide extract and the morel peptide extract respectively. The results are as Figures 29 - 30 shown.

[0154] From Figure 29 and Figure 30 the results, it can be seen that the peptide sequences in the shiitake mushroom and morel extracts do not contain the peptide sequences in the characteristic oligopeptide database of Stropharia rugosoannulata of the present invention, indicating that neither of them is the peptide extract of Stropharia rugosoannulata.

[0155] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A database of characteristic oligopeptides of Stropharia rugosoannulata, characterized in that, The database contains 60 oligopeptide molecules as shown in the following table: 。 2. The method for constructing the characteristic oligopeptide database of Stropharia rugosoannulata according to claim 1, characterized in that The construction method includes the following steps: Perform mass spectrometry identification on the peptide sequences in the fermented mycelium extract of Stropharia rugosoannulata, the ultrasonic extract of the fruiting body of Stropharia rugosoannulata, and the enzymolysis product of the fruiting body of Stropharia rugosoannulata to obtain the peptide profiles of each sample; screen the peptide segments with the appearance times ≥ 2 and high confidence in each peptide profile; after combining the screened peptide segments, remove the repeatedly appearing peptide segments to obtain the characteristic oligopeptide database of Stropharia rugosoannulata.

3. The construction method according to claim 2, characterized in that, The ultrasonic extract of the fruiting body of Stropharia rugosoannulata includes a poly-energy ultrasonic extract and a flat-divergent ultrasonic extract.

4. The construction method according to claim 2, characterized in that The enzymolysis product of the fruiting body of Stropharia rugosoannulata includes a product obtained by pretreatment with a complex enzyme system combined with moderate enzymolysis by an endonuclease and a product obtained by multi-mode synchronous ultrasonic-assisted directional enzymolysis.

5. The construction method according to claim 2, characterized in that The mass spectrometry identification is performed by liquid chromatography-tandem mass spectrometry.

6. The construction method according to claim 2, characterized in that, The high confidence means that the peptide confidence score - 10lgP ≥ 15.

7. The construction method according to claim 2, characterized in that, The peptide segment is a peptide molecule containing 7 - 9 amino acids.

8. Application of the characteristic oligopeptide database of Stropharia rugosoannulata described in claim 1 or the characteristic oligopeptide database of Stropharia rugosoannulata constructed by the construction method described in any one of claims 2 - 7 in the identification or quality control of Stropharia rugosoannulata products.

9. The application according to claim 8, wherein The Stropharia rugosoannulata products include products containing peptides of Stropharia rugosoannulata.

10. A method for identifying a product containing stropharia rugosoannulata peptide, characterized in that, The method includes the following steps: Perform mass spectrometry identification on the test sample to obtain the peptide profile of the test sample; compare the peptide segments in the peptide profile of the test sample with the characteristic oligopeptide database of Stropharia rugosoannulata; if it contains the peptide segments in the characteristic oligopeptide database of Stropharia rugosoannulata, it means that the test sample contains peptides of Stropharia rugosoannulata. The characteristic oligopeptide database of Stropharia rugosoannulata is the characteristic oligopeptide database of Stropharia rugosoannulata described in claim 1 or the characteristic oligopeptide database of Stropharia rugosoannulata constructed by the construction method described in any one of claims 2 - 7.

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