Glycosylated fish maw peptide and instant coffee powder rich in fish maw glycopeptide

Abstract

The invention belongs to the technical field of compound coffee powder, and relates to glycosylated fish maw peptide and instant coffee powder rich in fish maw glycopeptide. A preparation method of the glycosylated fish maw peptide comprises the following steps: (1) pretreating fish maw, and adding protease for hydrolysis to obtain a fish maw hydrolysate; (2) mixing the lycium barbarum polysaccharide and cellulase, heating and reacting to obtain lycium barbarum polysaccharide hydrolysate; and (3) mixing the fish maw hydrolysate and the lycium barbarum polysaccharide hydrolysate according to a volume ratio, adjusting the pH value, and carrying out ultrasonic-assisted water bath heating reaction to obtain the glycosylated fish maw peptide. The invention also discloses instant coffee powder rich in fish maw glycopeptides, and a preparation method of the instant coffee powder comprises the following steps: mixing a coffee solution and a glycosylated fish maw peptide solution according to a volume ratio, adding an auxiliary agent, and treating through high-pressure micro-jet equipment to obtain a fish maw peptide coffee liquid; and freeze-drying the fish maw peptide coffee liquid to obtain the instant coffee powder rich in fish maw glycopeptides.

Description

Technical Field

[0001] The invention belongs to the technical field of composite coffee powder and relates to glycosylated fish maw peptides and instant coffee powder rich in fish maw glycopeptides. Background Art

[0002] As a material with a long history of both food and medicine, fish maw occupies an important position in the field of modern functional foods due to its unique nutritional components and multiple health benefits. The current traditional processing technology still faces significant challenges: the processing of raw materials requires a time-consuming drying and rehydration process, and there are technical bottlenecks in the extraction of core ingredients, which leads to extended product development cycles and increased production costs, making it difficult to adapt to the fast-moving consumer goods market's demand for efficient production. Especially in the field of deep processing, the industrial preparation of fish maw peptides is still limited by the traditional enzymatic hydrolysis process - the pretreatment of raw materials requires a rehydration process of more than 48 hours, which not only consumes a lot of water, but the final product also has obvious fishy smell residues and strong hygroscopicity. Technical difficulties directly affect the product's palatability and shelf stability. Therefore, breaking through high-efficiency and low-consumption preparation technology, establishing a flavor modification system, and improving bioavailability have become key technical paths to expand its application in emerging food fields.

[0003] As a representative example of a traditional medicinal and edible resource, goji berries have diversified from primary agricultural products to high-value-added products. Modern research reveals that their active ingredients, including characteristic goji berry polysaccharides, carotenoids, and phenolic compounds, form a unique network of benefits. These ingredients, along with a nutrient matrix composed of amino acids and trace elements, support their complex physiological functions, including anti-oxidative stress, regulation of glucose and lipid metabolism, and improvement of microcirculation.

[0004] Glycosylation is the process of modifying biomolecules by introducing sugars. This modification can significantly improve the solubility, stability, taste, and bioavailability of the target molecule. Glycosylation technology offers new avenues for modifying the aforementioned polysaccharides and peptides. Summary of the Invention

[0005] The purpose of the present invention is to address the above-mentioned problems existing in the prior art and propose a glycosylated fish maw peptide based on fish maw hydrolysate and wolfberry polysaccharide hydrolysate, which has good protein content, hypoglycemic activity and antioxidant activity.

[0006] One object of the present invention is achieved by the following technical solutions:

[0007] A glycosylated fish maw peptide, the preparation method of which comprises:

[0008] (1) pre-treating the fish maw, adding protease to hydrolyze it, and obtaining a fish maw hydrolysate;

[0009] (2) mixing wolfberry polysaccharide and cellulase and heating them to react to obtain wolfberry polysaccharide hydrolyzate;

[0010] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:(0.1-10), the pH was adjusted to 8-11, and the mixture was heated in an ultrasonic-assisted water bath at 60-80°C for 1-6 hours to obtain glycosylated fish maw peptides.

[0011] Preferably, the fish maw pretreatment in (1) comprises: stewing the dry fish maw and water at a high temperature and high pressure of 110-130° C. and 100-120 Pa for 0.1-5 hours to obtain the pretreated fish maw.

[0012] Preferably, the protease in (1) includes pepsin and trypsin.

[0013] Preferably, the preparation method of the fish maw hydrolyzate described in (1) comprises: stewing dry fish maw and water at a high temperature and high pressure of 110-130° C. and 100-120 Pa for 0.1-5 h to obtain pretreated fish maw, adding phosphate buffer for homogenization, adjusting the pH to 2-3, adding pepsin, and oscillating the reaction at 37° C. for 1-6 h, then adjusting the pH to 7-8, adding trypsin, and oscillating the reaction at 37° C. for 1-6 h, heating with boiling water to terminate the reaction, and freeze-drying the hydrolyzed fish maw peptide solution and storing it at -20° C. for later use.

[0014] More preferably, the mass ratio of the pretreated fish maw to pepsin in (1) is 1:(20-40).

[0015] More preferably, the mass ratio of the pretreated fish maw to trypsin in (1) is 1:(30-50).

[0016] Preferably, the method for preparing the Lycium barbarum polysaccharide hydrolyzate described in (2) comprises: adding Lycium barbarum polysaccharide to water, mixing, adjusting the pH to 4.5-6.0, adding cellulase and mixing evenly, and then shaking the mixture in a constant temperature water bath at 40-60°C for 1-12 hours. After the reaction is completed, adjusting the pH to neutral to terminate the enzymatic hydrolysis reaction; and filtering to remove impurities to obtain Lycium barbarum polysaccharide hydrolyzate.

[0017] More preferably, the mass fraction of the cellulase in (2) is 0.1 to 0.9% (w / v).

[0018] More preferably, the mass ratio of the wolfberry polysaccharide to the cellulase in (2) is (20-50):1.

[0019] Preferably, the volume ratio of the fish maw hydrolysate to the wolfberry polysaccharide hydrolysate in (3) is 1:(0.5-2).

[0020] More preferably, the volume ratio of the fish maw hydrolysate to the wolfberry polysaccharide hydrolysate in (3) is 1:1.

[0021] Preferably, the grafting degree of the glycosylated fish glue peptide is greater than 30%.

[0022] More preferably, the grafting degree of the glycosylated fish glue peptide is greater than 35%.

[0023] The second object of the present invention is achieved through the following technical solutions:

[0024] An instant coffee powder rich in fish maw glycopeptide, the preparation method of which comprises:

[0025] A coffee solution with a concentration of 1 to 10 wt% is mixed with a glycosylated fish gum peptide solution in a volume ratio of 1:(0.1 to 10), and then 0 to 2 wt% of an auxiliary agent is added. The mixture is processed 1 to 3 times through a high-pressure microfluidizer at 50 to 100 MPa to obtain fish gum peptide coffee liquid. The fish gum peptide coffee liquid is freeze-dried to obtain instant coffee powder rich in fish gum glycopeptides.

[0026] Preferably, the volume ratio of the 2 wt% coffee solution to the glycosylated fish peptide solution is 1:(0.5-2).

[0027] More preferably, the volume ratio of the 2 wt % coffee solution to the glycosylated fish peptide solution is 1:1.

[0028] Preferably, the auxiliary agent includes one or more of matcha powder, cinnamon powder, cocoa powder, turmeric powder, nutmeg powder, clove powder, and ginger powder.

[0029] Preferably, the preparation method of the instant coffee powder rich in fish maw glycopeptides comprises: mixing a coffee solution with a concentration of 2wt% and a glycosylated fish maw peptide solution in a volume ratio of 1:(0.5-2), adding 0.1-2wt% of an auxiliary agent, and treating the mixture 1-3 times with a high-pressure microfluidizer at 50-100MPa to obtain fish maw peptide coffee liquid; and freeze-drying the fish maw peptide coffee liquid to obtain instant coffee powder rich in fish maw glycopeptides.

[0030] Preferably, the protein content of the instant coffee powder rich in fish maw glycopeptides is greater than 40 mg / ml.

[0031] More preferably, the protein content of the instant coffee powder rich in fish maw glycopeptides is greater than 46 mg / ml.

[0032] Preferably, the blood sugar lowering activity of the instant coffee powder rich in fish maw glycopeptides is greater than 50%.

[0033] More preferably, the blood sugar lowering activity of the instant coffee powder rich in fish maw glycopeptides is greater than 60%.

[0034] Preferably, the antioxidant activity of the instant coffee powder rich in fish maw glycopeptides is ≥0.1.

[0035] More preferably, the antioxidant activity of the instant coffee powder rich in fish maw glycopeptides is ≥0.15.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] 1. The glycosylated fish maw peptide of the present invention is based on fish maw hydrolysate and wolfberry polysaccharide hydrolysate, and has good protein content, hypoglycemic activity and antioxidant activity.

[0038] 2. The glycosylated fish gelatin peptide of the present invention is applied to instant coffee powder, which can achieve the refreshing effect of traditional coffee while also having nutritional value.

[0039] 3. The instant coffee powder rich in fish maw glycopeptide of the present invention is added with adjuvants such as matcha powder and cinnamon powder to increase the flavor while enhancing the hypoglycemic activity and antioxidant activity. DETAILED DESCRIPTION

[0040] The technical solutions of the present invention are further described below through specific embodiments. It should be understood that the specific embodiments described herein are only used to help understand the present invention and are not used to specifically limit the present invention.

[0041] Unless otherwise specified, the raw materials used in the examples of the present invention are all commonly used raw materials in the art, and the methods used in the examples are all conventional methods in the art.

[0042] In this article, the test methods include:

[0043] (1) Grafting degree: Using L-serine as the standard curve, 200 μL of sample was mixed with 4 mL of OPA reagent (OPA reagent was prepared as follows: 200 mg of OPA was dissolved in 5 mL of ethanol solution, 9.53 g of sodium tetraborate, 0.25 g of sodium dodecyl sulfate (SDS), and 220 mg of dithiothreitol (DTT) were dissolved in 180 mL of deionized water, and the two solutions were mixed and diluted to 250 mL). The mixture was reacted at room temperature in the dark for 10 min. The absorbance at a wavelength of 340 nm was measured, and the absorbance values before and after glycosylation were recorded as A0 and A1. The grafting degree of the sample was calculated according to the following formula (1).

[0044]

[0045] (2) Protein content: The protein content of the sample was determined using a BCA kit using bovine serum albumin as the standard curve.

[0046] (3) Hypoglycemic activity (α-glucosidase inhibitory activity): 100 μL of sample solution was mixed with 100 μL of α-glucosidase solution (0.4 U / mL) and placed in a 37°C water bath for 10 minutes. 100 μL of PNPG (2.5 mmol / L) and 400 μL of phosphate buffer (0.1 M, pH 6.8) were added and the reaction was continued at 37°C for 30 minutes. After the reaction was completed, 1 mL of 1 M Na2CO3 solution was used to terminate the reaction and the absorbance value was measured at a wavelength of 403 nm and recorded as A1. Next, the α-glucosidase solution was replaced with phosphate buffer for measurement to obtain the absorbance value A2; the sample solution was replaced with phosphate buffer for measurement to obtain the absorbance value A0. Finally, the α-glucosidase inhibitory activity was calculated according to formula (2).

[0047]

[0048] (4) Antioxidant activity (total reducing power): 0.5 mL of sample, 2.5 mL of 0.2 M phosphate buffer (pH 6.6), and 2.5 mL of 1.0% potassium percyanate solution were thoroughly mixed and incubated in a 50°C water bath for 20 min. Then, 2.5 mL of 10% trichloroacetic acid (w / v) was added, and the mixture was centrifuged at 3000 rpm for 15 min at room temperature. 2.5 mL of the supernatant was thoroughly mixed with 2.5 mL of distilled water and 0.5 mL of 0.1% ferric chloride solution (w / v), incubated at room temperature for 10 min, and the absorbance at 700 nm was measured.

[0049] Example 1

[0050] (1) The cleaned dried fish maw and water were stewed at a ratio of 1:8.3 (dried fish maw: water, w / v) at high temperature and high pressure (121℃, 105pa) for 2h to obtain pretreated fish maw for later use. Weigh an appropriate amount of pretreated fish maw, add ten times the volume of phosphate buffer (PBS, 10mM, Na2HPO4-NaH2PO4, pH7.0), homogenize at 10000r / min for 60s, repeat twice, and set aside. Adjust the pH to 2-3 with 1M HCl, add pepsin (1:30, w / w), hydrolyze at 37℃, rotate at 150r / min, and after 2h, adjust the pH to 7-8 with 1M NaOH to inactivate pepsin. After the hydrolysis in the stomach is completed, add trypsin (1:40, w / w), shake under the same conditions as above for 2h, heat with boiling water for 5min to terminate the reaction, freeze-dry the hydrolyzed fish maw peptide solution, and store at -20℃ for later use.

[0051] (2) Lycium barbarum polysaccharide was mixed with ten times the volume of water, and the pH was adjusted to 5.0. 0.5% (w / v) cellulase was added and mixed evenly. The mixture was then shaken in a constant temperature water bath at 50°C for 4 hours at a speed of 150 rpm. After the reaction was completed, the pH was adjusted to neutral (pH = 7.0) with 1 M NaOH to terminate the enzymatic hydrolysis reaction. Impurities were removed by filtration through filter cloth to obtain Lycium barbarum polysaccharide hydrolyzate.

[0052] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 8, and the mixture was heated in an ultrasonic-assisted water bath at 60°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0053] (4) Prepare a 2% black coffee solution, add the glycosylated fish maw peptide solution in a volume ratio of 1:1, and mix thoroughly; add 0.4% cinnamon powder and 0.8% matcha powder to the mixture and stir evenly; treat the mixture three times with a 60MPa high-pressure microfluidizer, spray-dry the micro-jet-treated mixture, and immediately seal it in an aluminum foil composite film bag with a built-in deoxidizer, and store it in an environment below 25°C and with a relative humidity of ≤30%, thereby obtaining instant coffee powder rich in fish maw glycopeptides.

[0054] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0055] Example 2

[0056] (1-2) are the same as steps (1-2) in Example 1.

[0057] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 9, and the mixture was heated in an ultrasonic-assisted water bath at 60°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0058] (4) Same as step (4) in Example 1.

[0059] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0060] Example 3

[0061] (1-2) are the same as steps (1-2) in Example 1.

[0062] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 10, and the mixture was heated in an ultrasonic-assisted water bath at 60°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0063] (4) Same as step (4) in Example 1.

[0064] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0065] Example 4

[0066] (1-2) are the same as steps (1-2) in Example 1.

[0067] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 11, and the mixture was heated in an ultrasonic-assisted water bath at 60°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0068] (4) Same as step (4) in Example 1.

[0069] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0070] Example 5

[0071] (1-2) are the same as steps (1-2) in Example 1.

[0072] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 8, and the mixture was heated in an ultrasonic-assisted water bath at 70°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0073] (4) Same as step (4) in Example 1.

[0074] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0075] Example 6

[0076] (1-2) are the same as steps (1-2) in Example 1.

[0077] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 9, and the mixture was heated in an ultrasonic-assisted water bath at 70°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0078] (4) Same as step (4) in Example 1.

[0079] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0080] Example 7

[0081] (1-2) are the same as steps (1-2) in Example 1.

[0082] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 10, and the mixture was heated in an ultrasonic-assisted water bath at 70°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0083] (4) Same as step (4) in Example 1.

[0084] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0085] Example 8

[0086] (1-2) are the same as steps (1-2) in Example 1.

[0087] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 11, and the mixture was heated in an ultrasonic-assisted water bath at 70°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0088] (4) Same as step (4) in Example 1.

[0089] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0090] Example 9

[0091] (1-2) are the same as steps (1-2) in Example 1.

[0092] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 8, and the mixture was heated in an ultrasonic-assisted water bath at 80°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0093] (4) Same as step (4) in Example 1.

[0094] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0095] Example 10

[0096] (1-2) are the same as steps (1-2) in Example 1.

[0097] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 9, and the mixture was heated in an ultrasonic-assisted water bath at 80°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0098] (4) Same as step (4) in Example 1.

[0099] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0100] Example 11

[0101] (1-2) are the same as steps (1-2) in Example 1.

[0102] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 10, and the mixture was heated in an ultrasonic-assisted water bath at 80°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0103] (4) Same as step (4) in Example 1.

[0104] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0105] Example 12

[0106] (1-2) are the same as steps (1-2) in Example 1.

[0107] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:1, the pH was adjusted to 11, and the mixture was heated in an ultrasonic-assisted water bath at 80°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0108] (4) Same as step (4) in Example 1.

[0109] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0110] Example 13

[0111] (1-2) are the same as steps (1-2) in Example 1.

[0112] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:2, the pH was adjusted to 9, and the mixture was heated in an ultrasonic-assisted water bath at 70°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0113] (4) Same as step (4) in Example 1.

[0114] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0115] Example 14

[0116] (1-2) are the same as steps (1-2) in Example 1.

[0117] (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:0.5, the pH was adjusted to 9, and the mixture was heated in an ultrasonic-assisted water bath at 70°C for 2 h to obtain a glycosylated fish maw peptide solution. After the reaction, the mixture was cooled in an ice-water bath, freeze-dried, and stored in a -20°C refrigerator for later use.

[0118] (4) Same as step (4) in Example 1.

[0119] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0120] Example 15

[0121] (1-2) are the same as steps (1-2) in Example 6.

[0122] (3) Same as step (3) in Example 6.

[0123] (4) Prepare a 2% black coffee solution, add the glycosylated fish maw peptide solution in a volume ratio of 1:2, and mix thoroughly; add 0.4% cinnamon powder and 0.8% matcha powder to the mixture and stir evenly; process the mixture three times through a 60MPa high-pressure microfluidizer, spray-dry the microjet-treated mixture, and immediately seal it in an aluminum foil composite film bag with a built-in deoxidizer, and store it in an environment below 25°C and with a relative humidity of ≤30%, thereby obtaining instant coffee powder rich in fish maw glycopeptides.

[0124] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0125] Example 16

[0126] (1-2) are the same as steps (1-2) in Example 6.

[0127] (3) Same as step (3) in Example 6.

[0128] (4) Prepare a 2% black coffee solution, add the glycosylated fish maw peptide solution in a volume ratio of 1:0.5, and mix thoroughly; add 0.4% cinnamon powder and 0.8% matcha powder to the mixture and stir evenly; process the mixture three times through a 60MPa high-pressure microfluidizer, spray-dry the microjet-treated mixture, and immediately seal it in an aluminum foil composite film bag with a built-in deoxidizer, and store it in an environment below 25°C and with a relative humidity of ≤30% to obtain instant coffee powder rich in fish maw glycopeptides.

[0129] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0130] Example 17

[0131] (1-2) are the same as steps (1-2) in Example 6.

[0132] (3) Same as step (3) in Example 6.

[0133] (4) A 2% black coffee solution was prepared, and the glycosylated fish maw peptide solution was added in a volume ratio of 1:1 and mixed thoroughly; the mixed solution was treated three times by a 60 MPa high-pressure microfluidizer, and the microjet-treated mixed solution was immediately sealed in an aluminum foil composite film bag after spray drying, with a built-in deoxidizer, and stored in an environment below 25°C and a relative humidity of ≤30%, thereby obtaining instant coffee powder rich in fish maw glycopeptides.

[0134] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0135] Example 18

[0136] (1) The washed dried fish maw and water were mixed in a ratio of 1:8.3 (dried fish maw: water, w / v) and soaked for 24 hours to obtain pretreated fish maw for later use. An appropriate amount of pretreated fish maw was weighed and added with ten times the volume of phosphate buffer (PBS, 10mM, Na2HPO4-NaH2PO4, pH7.0), homogenized at 10000r / min for 60s, repeated twice and set aside. The pH was adjusted to 2-3 with 1M HCl, pepsin (1:30, w / w) was added, and hydrolyzed at 37℃ with a speed of 150r / min. After 2 hours, the pH was adjusted to 7-8 with 1M NaOH to inactivate pepsin. After the hydrolysis in the stomach was completed, trypsin (1:40, w / w) was added, and the mixture was shaken for 2 hours under the same conditions as above. The reaction was terminated by heating with boiling water for 5 minutes. The hydrolyzed fish maw peptide solution was freeze-dried and stored at -20℃ for later use.

[0137] (2-4) are the same as steps (2-4) in Example 6.

[0138] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0139] Example 19

[0140] (1-2) are the same as steps (1-2) in Example 6.

[0141] (3) Same as step (3) in Example 6.

[0142] (4) Prepare a 2% black coffee solution, add the glycosylated fish maw peptide solution in a volume ratio of 1:1, and mix thoroughly; add 1.2% cinnamon powder to the mixture and stir evenly; treat the mixture three times with a 60MPa high-pressure microfluidizer, spray-dry the microjet-treated mixture, and immediately seal it in an aluminum foil composite film bag with a built-in deoxidizer, and store it in an environment below 25°C and with a relative humidity of ≤30%, thereby obtaining instant coffee powder rich in fish maw glycopeptides.

[0143] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0144] Example 20

[0145] (1-2) are the same as steps (1-2) in Example 6.

[0146] (3) Same as step (3) in Example 6.

[0147] (4) Prepare a 2% black coffee solution, add the glycosylated fish maw peptide solution in a volume ratio of 1:1, and mix thoroughly; add 0.1% cinnamon powder and 1.1% matcha powder to the mixture and stir evenly; treat the mixture three times with a 60MPa high-pressure microfluidizer, spray-dry the microjet-treated mixture, and immediately seal it in an aluminum foil composite film bag with a built-in deoxidizer, and store it in an environment below 25°C and with a relative humidity of ≤30%, thereby obtaining instant coffee powder rich in fish maw glycopeptides.

[0148] The performance of the obtained instant coffee powder rich in fish maw glycopeptides was tested.

[0149] Comparative Example 1

[0150] (1-2) are the same as steps (1-2) in Example 6.

[0151] (3) A 2% black coffee solution was prepared, and fish maw hydrolysate and wolfberry polysaccharide hydrolysate were added in a volume ratio of 1:0.5:0.5, and the mixture was thoroughly mixed; 0.4% cinnamon powder and 0.8% matcha powder were added to the mixture and stirred evenly; the mixture was treated three times by a 60 MPa high-pressure microfluidizer, and the microjet-treated mixture was spray-dried and immediately sealed in an aluminum foil composite film bag with a built-in deoxidizer, and stored in an environment below 25°C and a relative humidity of ≤30% to obtain instant coffee powder.

[0152] The obtained instant coffee powder was subjected to performance tests.

[0153] Comparative Example 2

[0154] (1-3) are the same as steps (1-3) in Example 6.

[0155] (3) Adding glycosylated fish glue peptide solution to water in a volume ratio of 1:1 and mixing thoroughly; adding 0.4% cinnamon powder and 0.8% matcha powder to the mixture and stirring evenly; treating the mixed solution three times with a 60MPa high-pressure microfluidizer, spray-drying the microjet-treated mixed solution and immediately sealing it in an aluminum foil composite film bag with a built-in deoxidizer, and storing it in an environment below 25°C and a relative humidity of ≤30% to obtain instant powder.

[0156] The obtained instant powder was subjected to performance tests.

[0157] Comparative Example 3

[0158] (1) Same as step (1) in Example 6.

[0159] (2) A 2% black coffee solution was prepared, and fish maw hydrolyzate was added in a volume ratio of 1:1, and the mixture was thoroughly mixed; 0.4% cinnamon powder and 0.8% matcha powder were added to the mixture and stirred evenly; the mixture was treated three times by a 60 MPa high-pressure microfluidizer, and the microjet-treated mixture was spray-dried and immediately sealed in an aluminum foil composite film bag with a built-in deoxidizer, and stored in an environment below 25°C and a relative humidity of ≤30% to obtain an instant powder.

[0160] The obtained instant powder was subjected to performance tests.

[0161] Comparative Example 4

[0162] (1) Same as step (2) in Example 6.

[0163] (2) Prepare a 2% black coffee solution, add wolfberry polysaccharide hydrolyzate in a volume ratio of 1:1, and mix thoroughly; add 0.4% cinnamon powder and 0.8% matcha powder to the mixture and stir evenly; process the mixture three times through a 60 MPa high-pressure microfluidizer, spray-dry the microjet-treated mixture, and immediately seal it in an aluminum foil composite film bag with a built-in deoxidizer, and store it in an environment below 25°C and with a relative humidity of ≤30% to obtain an instant powder.

[0164] The obtained instant powder was subjected to performance tests.

[0165] Table 1. Performance of instant coffee powder, instant coffee powder and instant powder rich in fish maw glycopeptide

[0166]

[0167] According to Table 1, in Examples 1 to 12, when pH = 9 and the water bath temperature is 70° C., the grafting degree is the highest and the optimal protein content, hypoglycemic activity and antioxidant activity are exhibited.

[0168] In Comparative Example 1, the fish maw hydrolysate and wolfberry polysaccharide hydrolysate were added directly to the coffee solution without glycosylation, resulting in significantly lower performance. In Comparative Example 2, no coffee or additives were added, resulting in lower performance than Example 6. The fish maw hydrolysate and wolfberry polysaccharide hydrolysate alone in Comparative Examples 3 and 4 exhibited poor hypoglycemic and antioxidant activities.

[0169] In summary, the glycosylated fish maw peptides based on fish maw hydrolysate and wolfberry polysaccharide hydrolysate of the present invention have good protein content, hypoglycemic activity, and antioxidant activity; and can be applied to instant coffee to enhance its nutritional value.

[0170] The various aspects, embodiments, and features of the present invention should be considered in all respects as illustrative and not limiting, the scope of which is defined solely by the claims. Other embodiments, modifications, and uses will be apparent to those skilled in the art without departing from the spirit and scope of the invention as claimed.

[0171] In the preparation method of the present invention, the order of the steps is not limited to the order listed. Persons skilled in the art will appreciate that variations in the order of the steps are within the scope of the present invention without inventive effort. Furthermore, two or more steps or actions may be performed simultaneously.

[0172] Finally, it should be noted that the specific embodiments described herein are merely illustrative of the present invention and are not intended to limit its implementation. Persons skilled in the art may make various modifications, additions, or substitute similar methods for the described specific embodiments. It is not necessary and impossible to provide comprehensive examples of all implementations here. However, obvious variations or modifications arising from the essential spirit of the present invention remain within the scope of protection of the present invention, and interpreting them as any additional limitations would be contrary to the spirit of the present invention.

Claims

1. A glycosylated fish maw peptide, characterized in that: The preparation method thereof comprises: (1) pre-treating the fish maw, adding protease to hydrolyze it, and obtaining a fish maw hydrolysate; (2) mixing wolfberry polysaccharide and cellulase and heating them to react to obtain wolfberry polysaccharide hydrolyzate; (3) After mixing fish maw hydrolysate and wolfberry polysaccharide hydrolysate in a volume ratio of 1:(0.1-10), the pH was adjusted to 8-11, and the mixture was heated in an ultrasonic-assisted water bath at 60-80°C for 1-6 hours to obtain glycosylated fish maw peptides.

2. The glycosylated fish glue peptide according to claim 1, characterized in that: The fish maw pretreatment described in (1) includes: stewing the dry fish maw and water at a high temperature and high pressure of 110-130° C. and 100-120 Pa for 0.1-5 hours to obtain the pretreated fish maw.

3. The glycosylated fish glue peptide according to claim 1, characterized in that: The proteases mentioned in (1) include pepsin and trypsin.

4. The glycosylated fish glue peptide according to claim 1, characterized in that The preparation method of the fish maw hydrolyzate described in (1) comprises: stewing dry fish maw and water at a high temperature and high pressure of 110-130°C and 100-120 Pa for 0.1-5 hours to obtain pretreated fish maw, adding phosphate buffer to homogenize, adjusting the pH to 2-3, adding pepsin, shaking reaction at 37°C for 1-6 hours, then adjusting the pH to 7-8, adding trypsin, shaking reaction at 37°C for 1-6 hours, heating with boiling water to terminate the reaction, freeze-drying the hydrolyzed fish maw peptide solution, and storing it at -20°C for later use.

5. The glycosylated fish glue peptide according to claim 1, characterized in that: The method for preparing the Lycium barbarum polysaccharide hydrolyzate described in (2) comprises: adding Lycium barbarum polysaccharide to water, mixing the mixture, adjusting the pH to 4.5-6.0, adding cellulase, and mixing the mixture evenly, and then oscillating the mixture in a constant temperature water bath at 40-60° C. for 1-12 hours. After the reaction is completed, adjusting the pH to neutral to terminate the enzymatic hydrolysis reaction; and filtering to remove impurities to obtain the Lycium barbarum polysaccharide hydrolyzate.

6. The glycosylated fish glue peptide according to claim 1, characterized in that: The volume ratio of the fish maw hydrolysate to the wolfberry polysaccharide hydrolysate in (3) is 1:(0.5-2).

7. The glycosylated fish glue peptide according to claim 1, characterized in that: The grafting degree of the glycosylated fish glue peptide is greater than 30%.

8. An instant coffee powder rich in fish maw glycopeptide, characterized in that: The preparation method comprises: mixing a coffee solution with a concentration of 1 to 10 wt% and a glycosylated fish maw peptide solution in a volume ratio of 1:(0.1 to 10), adding 0 to 2 wt% of an auxiliary agent, and treating the mixture 1 to 3 times with a high-pressure microfluidizer at 50 to 100 MPa to obtain fish maw peptide coffee liquid; and freeze-drying the fish maw peptide coffee liquid to obtain instant coffee powder rich in fish maw glycopeptides.

9. The instant coffee powder rich in fish maw glycopeptide according to claim 1, characterized in that: The volume ratio of the coffee solution with a concentration of 2wt% to the glycosylated fish glue peptide solution is 1:(0.5-2).

10. The instant coffee powder rich in fish maw glycopeptide according to claim 1, characterized in that: The blood sugar lowering activity of the instant coffee powder rich in fish maw glycopeptide is greater than 50%; and the antioxidant activity of the instant coffee powder rich in fish maw glycopeptide is greater than or equal to 0.1.

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