CN115119929A - Color protection method for Chinese chestnut processing - Google Patents
Color protection method for Chinese chestnut processing Download PDFInfo
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- CN115119929A CN115119929A CN202210783169.3A CN202210783169A CN115119929A CN 115119929 A CN115119929 A CN 115119929A CN 202210783169 A CN202210783169 A CN 202210783169A CN 115119929 A CN115119929 A CN 115119929A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
- A23L25/20—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof consisting of whole seeds or seed fragments
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B9/00—Preservation of edible seeds, e.g. cereals
- A23B9/02—Preserving by heating
- A23B9/04—Preserving by heating by irradiation or electric treatment
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B9/00—Preservation of edible seeds, e.g. cereals
- A23B9/06—Preserving by irradiation or electric treatment without heating effect
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B9/00—Preservation of edible seeds, e.g. cereals
- A23B9/16—Preserving with chemicals
- A23B9/24—Preserving with chemicals in the form of liquids or solids
- A23B9/26—Organic compounds; Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/40—Colouring or decolouring of foods
- A23L5/41—Retaining or modifying natural colour by use of additives, e.g. optical brighteners
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
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- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
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Abstract
The invention belongs to the technical field of fruit and vegetable storage, and particularly relates to a color protection method for processing Chinese chestnuts. The color protection method for processing the Chinese chestnut comprises the following steps: picking and cleaning Chinese chestnut, cutting a cross at the top of the Chinese chestnut, curing, cooling, peeling to obtain Chinese chestnut kernel, soaking the Chinese chestnut kernel in polypeptide liquid of young grapefruit, taking out, draining, bagging, and performing microwave sterilization. The color protection method for Chinese chestnut processing adopts the bioactive peptide as the color fixative in the Chinese chestnut processing process, not only can achieve very good color protection effect, but also can avoid the side effect caused by organic matter residue in the existing color fixative, has the advantages of nature, safety and environmental protection, and is a relatively ideal Chinese chestnut color protection process.
Description
Technical Field
The invention belongs to the technical field of fruit and vegetable storage, and particularly relates to a color protection method for processing Chinese chestnuts.
Background
Chinese chestnut (Castanea mollissima) is a famous traditional agricultural and sideline product in China, is rich in nutrition, contains various nutritional ingredients such as fat, calcium, phosphorus, iron, various vitamins and trace elements, and is a treasure with high medicinal value and health care function. Chinese chestnut resources are rich in China, but fresh Chinese chestnuts are not durable to store, so deep processing of Chinese chestnuts becomes an important means for long-term storage of Chinese chestnuts.
The browning phenomenon often appears in the deep processing process of the Chinese chestnut, not only the taste of the Chinese chestnut is influenced, but also the appearance of the Chinese chestnut product is seriously influenced, and the quality of the product is greatly reduced, so the browning phenomenon is a technical problem which is solved firstly in the production process of the Chinese chestnut. The browning of the Chinese chestnut comprises enzymatic browning and non-enzymatic browning, and currently, a physical regulation and control method and a chemical color fixative are available for the browning, such as: a storage method for regulating and controlling the gas components, proportion and temperature and humidity of chestnut storage environment. The method has the advantages that the respiration of the Chinese chestnut is reduced under the environment of low temperature, high humidity, relative hypoxia and high carbon dioxide, the material consumption is reduced, and the growth and the propagation of microorganisms are inhibited, so that the Chinese chestnut is kept in a fresh state for a long time. The treatment of sulfurous acid and salts thereof can obviously control the enzymatic browning and non-enzymatic browning of Chinese chestnuts, but the use of sulfurous acid is controversial. Therefore, the color protection treatment in the deep processing process of Chinese chestnut is still the focus of current research.
Patent document CN109907280A discloses a color protection method for producing and processing chestnuts, which comprises the following steps: s1: picking and cleaning Chinese chestnuts, and cutting openings on the side surfaces and the top of the Chinese chestnuts; s2: vacuum permeating Vc color protecting liquid into Chinese chestnut; s3: the permeated Chinese chestnuts enter a tunnel type microwave channel to inactivate enzyme; s4: quickly putting the chestnuts with the enzyme inactivated by the microwave into the Vc color protection liquid for cooling; s5: removing shell of cooled Chinese chestnut, and removing Chinese chestnut with wormhole and black spot; s6: cooking the finished Chinese chestnuts by using sodium metabisulfite liquid; s7: and (3) putting the cooked chestnut kernels into the Vc color protection liquid for cooling, fishing out, putting into a plastic bag for food, carrying out vacuum packaging, and then putting into tunnel type microwave equipment for sterilization. The color protection process has the advantages of simple flow, good color protection effect, low production cost, small difficulty and easy mass production and use.
Patent document CN 111789215a discloses a color protection and sterilization process of chestnut kernels, which comprises the following specific processes: (1) cleaning the Chinese chestnut with water, and removing stains on the surface of the Chinese chestnut; (2) peeling off chestnut shells, taking out chestnut kernels, and soaking the chestnut kernels in color protection liquid for 10 min; (3) taking out the color-protected chestnut kernels; (4) mixing the chestnut kernels with subacid electrolyzed water at 40-50 ℃ according to a material-liquid ratio of 1:3-1:5g/L, soaking for 10min, and taking out the chestnut kernels; repeating the step twice; (5) draining liquid on the surface of the chestnut kernels, and then carrying out vacuum packaging on the chestnut kernels; (6) storing the packaged semen Castaneae at low temperature of 0-4 deg.C. The color protection process can fully maintain the color of the peeled Chinese chestnut kernels, reduce the quantity of mold on the surfaces of the Chinese chestnut kernels and effectively prolong the shelf life of the Chinese chestnut kernels.
However, in the prior art, the color protection treatment is performed by soaking the chestnut in the composite preservative solution prepared from citric acid, ascorbic acid, EDTA-2Na and the like, the browning degree is taken as an evaluation index, and the optimal formula of the composite preservative solution is screened out through an orthogonal test on the basis of a single-factor test. However, the chestnut processed by the compound preservative solution has the side effect of organic matter residue.
Disclosure of Invention
In order to solve the defects of the prior art, the invention provides a color protection method for Chinese chestnut processing, which mainly adopts bioactive peptide as a color protection agent in the Chinese chestnut processing process, not only can achieve very good color protection effect, but also can avoid side effect caused by organic matter residue in the existing color protection agent, and is a natural, safe and environment-friendly color protection process.
The invention provides a color protection method for Chinese chestnut processing, which comprises the following steps:
s1, selecting and cleaning Chinese chestnuts, cutting a cross cut at the tops of the Chinese chestnuts, and performing curing treatment, wherein the curing treatment comprises the following steps: frying the Chinese chestnut for 25-35 min or putting the Chinese chestnut into boiling water for boiling for 10-20 min;
s2, cooling the Chinese chestnuts processed in the step S1, and peeling the Chinese chestnuts to obtain Chinese chestnut kernels;
and S3, soaking the chestnut kernels prepared in the step S2 in the young pomelo fruit polypeptide liquid for 8-15 min, wherein the material-liquid ratio of the chestnut kernels to the young pomelo fruit polypeptide liquid is 1g: 20-1 g:50mL, taking out, draining, bagging and carrying out microwave sterilization, wherein the microwave sterilization conditions are as follows: the power density is 6-8W/g, and the treatment time is 80-90 s.
Further, the concentration of the grapefruit young fruit polypeptide liquid in the step S3 is 4-8 mg/mL.
Further, the concentration of the grapefruit young fruit polypeptide liquid in the step S3 is 5 mg/mL.
Further, the preparation method of the grapefruit young fruit polypeptide liquid in the step S3 includes:
adding distilled water into young pomelo fruit powder, uniformly stirring to obtain young pomelo fruit powder solution, then placing the young pomelo fruit powder solution in a water bath at the temperature of 90 ℃ for 8-12 min, cooling to 25 ℃, adding a pH regulator to regulate the pH of the solution to 8.5 to obtain reaction liquid, then adding neutral protease to carry out enzymolysis for 0.4-0.6 h in the water bath at the temperature of 55 ℃, adding phenylmethylsulfonyl fluoride after enzymolysis, and centrifuging for 8-12 min at the speed of 4000r/min to obtain the shaddock fruit powder.
The CAS number of the neutral protease is 9014-01-1 brand: solarbio cargo number: p9090.
Further, the concentration of the grapefruit young fruit powder solution is 48-52 mg/mL, and the concentration of the grapefruit young fruit powder solution is 50 mg/mL.
Further, the pH regulator is NaOH solution with the concentration of 0.1mol/L or HCL solution with the concentration of 0.1 mol/L.
Further, the amount of the neutral protease added was 0.5% w/v of the reaction solution, and the amount of the phenylmethylsulfonyl fluoride added was 0.02% w/v of the reaction solution.
Further, step S3 further includes a pulse processing process, which includes the specific steps of: and after soaking, performing pulse treatment, taking out, draining, and bagging to obtain the product.
Further, the conditions of the pulse processing are as follows: the pulse width pulse is 160-180 ns, the pulse rise time is 25-30 ns, the frequency is 60-70 Hz, the pulse amplitude is 15-18 Kv/cm, and the pulse time is 4-6 min.
Further, the conditions of the pulse processing are as follows: the pulse width pulse is 165ns, the pulse rise time is 28ns, the frequency is 66Hz, the pulse amplitude is 16Kv/cm, and the pulse time is 5 min.
The young pomelo fruit is the physiological fallen pomelo fruit in the growing period of pomelo, and has low cost, large yield and low utilization rate. The inventor is continuously dedicated to the research on the deep processing of agricultural and sideline products, and particularly has a plurality of results on the resource research of the young pomelo fruits. The inventor finds a shaddock young fruit enzymolysis polypeptide in the process of intensive research on shaddock young fruits, wherein the shaddock young fruit enzymolysis polypeptide is a shaddock young fruit polypeptide with the characteristic peptide of IVVPGIY, and the content of the shaddock young fruit polypeptide is not less than 1% of the total peptide. The inventor uses the shaddock young fruit polypeptide for color protection treatment in a Chinese chestnut processing process for the first time, and unexpectedly discovers that the shaddock young fruit polypeptide can remarkably delay a browning phenomenon in the Chinese chestnut processing process, is a natural, safe and nutritional bioactive peptide, solves the problem of side effect of residual organic matters in the existing color protection complex liquid, and is a natural, safe and environment-friendly Chinese chestnut processing color protection agent.
The characteristic peptide provided by the invention is that IVVPGIY shaddock young fruit polypeptide has a remarkable color protection effect on chestnut kernels, and the inventor conjectures that the principle is as follows: although browning of chestnuts in a processing process is mainly divided into enzymatic browning and non-enzymatic browning, the essence of the two is oxidation of phenolic substances in chestnuts, the phenolic substances in the chestnuts are oxidized to form quinone substances, and further, the quinone substances are polymerized to form brown substances, thereby causing browning. The invention unexpectedly discovers that the characteristic peptide obtained by the invention is IVVPGIY shaddock young fruit polypeptide which can be polymerized with quinone substances formed by oxidizing phenolic substances in Chinese chestnuts, so that the oxidation of the phenolic substances in the Chinese chestnuts is inhibited, and the effect of delaying browning is achieved. The grapefruit young fruit polypeptide liquid with the concentration of 4-8 mg/mL provided by the invention has a particularly remarkable polymerization effect on quinone substances formed by oxidizing phenolic substances in chestnuts, so that browning in the chestnut processing process is remarkably inhibited, and the quality of chestnut products is improved.
In addition, in order to prolong the shelf life of the Chinese chestnut products, the inventor adopts microwave sterilization, selects the microwave condition with the power density of 6-8W/g and the processing time of 80-90 s to process the Chinese chestnut kernels, and the microwave processing condition can not affect the browning of the Chinese chestnut kernels. However, the inventor researches the long-term stability test of the Chinese chestnut kernel and finds that the microwave sterilization does not affect the browning of the Chinese chestnut kernel, but the prepared Chinese chestnut product has poor stability. In order to pursue more perfect quality, the inventor continuously searches for innovation, and unexpectedly finds that the pulse treatment is combined with the mode of inhibiting the browning of the Chinese chestnut by adopting the bioactive polypeptide, so that the Chinese chestnut sterilizing agent not only can play a good sterilizing effect, but also can further delay the browning of the Chinese chestnut and improve the stability of products.
The invention provides a pulse condition that the pulse width pulse is 160-180 ns, the pulse rise time is 25-30 ns, the frequency is 60-70 Hz, the pulse amplitude is 15-18 Kv/cm, and the pulse time is 4-6 min, for treating the chestnut kernels soaked in the polypeptide liquid of the young pomelos, so that the sterilization effect can be achieved, and the stability of chestnut products can be improved. The specific principle is presumed as follows: under the action of the pulse electric field provided by the invention, bacteria and fungi are changed rapidly along with the frequency of the pulse electric field, are deformed continuously and are finally broken, so that the sterilization effect is achieved. Moreover, the pulse treatment can make polymers formed by the shaddock young fruit polypeptide liquid and quinone substances fall into the soaking liquid, and further inhibit the browning phenomenon. Meanwhile, the specific pulse treatment provided by the invention can also dissociate heavy metal ions in the Chinese chestnut, and the heavy metal ions are reduced and fall off under the pulse electric field, so that the phenomenon of non-enzymatic browning such as metal ion catalysis is reduced, and the browning of the Chinese chestnut is further delayed.
In a word, compared with the prior art, the color protection method for processing the Chinese chestnut provided by the invention has the following advantages:
(1) the color fixative in the color fixative method for processing the Chinese chestnut provided by the invention is the shaddock young fruit polypeptide, has rich and cheap raw materials, is safe and has no side effect, can obviously inhibit the brown stain of the Chinese chestnut, and can greatly improve the safety of the Chinese chestnut product;
(2) the Chinese chestnut product prepared by the color protection method for Chinese chestnut processing provided by the invention has bright color, rich taste and good stability, and is an ideal Chinese chestnut processing method.
Description of the drawings:
FIG. 1 is an amino acid sequence diagram of a polypeptide of young pomelo fruit obtained in example 1 of the present invention.
Detailed Description
The present invention is further described below by way of specific examples, but the present invention is not limited to only the following examples. Variations, combinations, or substitutions of the invention, which are within the scope of the invention or the spirit, scope of the invention, will be apparent to those of skill in the art and are within the scope of the invention.
Example 1 preparation of polypeptide solution of young pomelo fruit
Adding distilled water into the shaddock young fruit powder, and uniformly stirring to obtain a shaddock young fruit powder solution, wherein the concentration of the shaddock young fruit powder solution is 50 mg/mL; and then placing the shaddock young fruit powder solution in a water bath at the temperature of 90 ℃ for 10min, cooling to 25 ℃, adding a pH regulator to regulate the pH of the solution to 8.5, wherein the pH regulator is a NaOH solution with the concentration of 0.1mol/L or a HCL solution with the concentration of 0.1mol/L to obtain a reaction solution, then adding neutral protease to carry out enzymolysis for 0.5h under the water bath condition at the temperature of 55 ℃, the addition amount of the neutral protease is 0.5% w/v of the reaction solution, adding phenylmethylsulfonyl fluoride after the enzymolysis, the addition amount of the phenylmethylsulfonyl fluoride is 0.02% w/v of the reaction solution, and centrifuging for 10min under the condition of 4000r/min to obtain the shaddock powder.
Example 2 color protection method for chestnut processing
S1, picking and cleaning Chinese chestnuts, cutting a cross at the tops of the Chinese chestnuts, preheating for 30min at 220 ℃ by a frying machine, and frying for 30min by putting the Chinese chestnuts;
s2, cooling the Chinese chestnuts processed in the step S1, and peeling the Chinese chestnuts to obtain Chinese chestnut kernels;
step S3, soaking the chestnut kernels prepared in the step S2 in the grapefruit young fruit polypeptide liquid prepared in the example 1 for 10min, wherein the concentration of the grapefruit young fruit polypeptide liquid is 5mg/mL, the material-liquid ratio of the chestnut kernels to the grapefruit young fruit polypeptide liquid is 1g:20mL, taking out, draining, bagging and carrying out microwave sterilization, wherein the microwave sterilization conditions are as follows: the power density is 6W/g, and the processing time is 90 s.
Example 3 color protection method for chestnut processing
S1, picking and cleaning Chinese chestnuts, cutting a cross cut on the tops of the Chinese chestnuts, and then putting the Chinese chestnuts into boiling water to boil for 15 min;
s2, cooling the Chinese chestnuts processed in the step S1, and peeling the Chinese chestnuts to obtain Chinese chestnut kernels;
step S3, soaking the chestnut kernels prepared in the step S2 in the grapefruit young fruit polypeptide liquid prepared in the example 1 for 8min, wherein the concentration of the grapefruit young fruit polypeptide liquid is 6mg/mL, the material-liquid ratio of the chestnut kernels to the grapefruit young fruit polypeptide liquid is 1g:30mL, taking out, draining, bagging and carrying out microwave sterilization, wherein the microwave sterilization conditions are as follows: the power density is 8W/g, and the processing time is 80 s.
Example 4 color protection method for chestnut processing
S1, picking and cleaning Chinese chestnuts, cutting a cross at the tops of the Chinese chestnuts, preheating for 30min at 220 ℃ by a frying machine, and frying for 30min by putting the Chinese chestnuts;
s2, cooling the Chinese chestnuts processed in the step S1, and peeling the Chinese chestnuts to obtain Chinese chestnut kernels;
step S3, soaking the chestnut kernels obtained in step S2 in the polypeptide liquid of young fruit of pomelo obtained in example 1 for 10min, wherein the concentration of the polypeptide liquid of young fruit of pomelo is 5mg/mL, and the ratio of the chestnut kernels to the polypeptide liquid of young fruit of pomelo is 1g:20mL, and after the soaking, performing pulse treatment under the following conditions: the pulse width pulse is 165ns, the pulse rise time is 28ns, the frequency is 66Hz, the pulse amplitude is 16Kv/cm, the pulse time is 5min, and the product is obtained by taking out, draining and bagging.
Example 5 color protection method for chestnut processing
S1, picking and cleaning Chinese chestnuts, cutting a cross cut at the tops of the Chinese chestnuts, and then putting the Chinese chestnuts into boiling water to be boiled for 15 min;
s2, cooling the Chinese chestnuts processed in the step S1, and peeling the Chinese chestnuts to obtain Chinese chestnut kernels;
step S3, soaking the Chinese chestnut kernels prepared in the step S2 in the grapefruit young fruit polypeptide liquid prepared in the example 1 for 10min, wherein the concentration of the grapefruit young fruit polypeptide liquid is 5mg/mL, the material-liquid ratio of the Chinese chestnut kernels to the grapefruit young fruit polypeptide liquid is 1g:30mL, and after soaking, performing pulse treatment under the conditions that: the pulse width pulse is 160ns, the pulse rise time is 30ns, the frequency is 70Hz, the pulse amplitude is 18Kv/cm, the pulse time is 6min, and the product is obtained after taking out, draining and bagging.
Comparative example 1 color protection method for chestnut processing
The difference from example 2 is that: the soaking solution of the step S3 is: 1.0g of cod polypeptide powder was weighed, dissolved, and made up to 1000mL to prepare a cod polypeptide solution having a concentration of 0.1%, and the rest was similar to example 2.
The preparation method of the cod polypeptide powder comprises the following steps:
dissolving 50g of cod skin in 150mL of distilled water, stirring, adjusting the pH value to 9.0 by using 1mol/L NaOH, adding 0.6g of marine low-temperature alkaline protease, performing enzymolysis for 1h in a constant-temperature water bath at 40 ℃, continuously adjusting the pH value by using 1mol/L NaOH, maintaining the pH value at 9.0, inactivating the enzyme of the enzymolysis liquid through a boiling water bath for 8min, cooling the enzymolysis liquid to 10000r/min, centrifuging the enzymolysis liquid for 30min, taking the supernatant, namely the cod polypeptide solution, and then performing a series of decoloration, deodorization, concentration and drying treatment to obtain the final cod polypeptide powder. (refer to patent document CN 109122841A for cod polypeptide preparation)
Comparative example 2 color protection method for chestnut processing
The difference from example 2 is that: the soaking solution of the step S3 is: the process is similar to that of example 2 except that the process is carried out with a color protection solution consisting of 0.3 mass percent of ascorbic acid, 0.7 mass percent of citric acid, 0.3 mass percent of phytic acid, 1 mass percent of sodium chloride and the balance of water.
Test example I, test for testing polypeptide solution of young grapefruit
1. The test method comprises the following steps:
amino acid sequencing was performed on the grapefruit young fruit polypeptide liquid obtained in example 1. The specific determination method comprises the following steps:
(1) 100ug of the grapefruit young fruit polypeptide sample prepared in example 1 was dissolved in Washing buffer (0.1% FA, 2% ACN);
(2) transferring the grapefruit young fruit polypeptide sample solution into a 10KD ultrafiltration centrifugal tube, and centrifuging for 10min at 12000 g;
(3) desalting the solution after ultrafiltration by using a C18 desalting column;
(4) elute the sample with an elute buffer (0.1% FA, 60% ACN) and transfer the Elution solution to a new EP tube;
(5) and (4) carrying out centrifugal concentration and drying on the eluted sample, and carrying out mass spectrometry.
The desalted polypeptide sample was dried by centrifugation, re-dissolved in 100uL Nano-LC mobile phase A (0.1% formic acid/water), bottled and subjected to on-line LCMS analysis. The solubilized sample was loaded onto a nanobipe C18 pre-column in a volume of 2 μ L (3 μm,
) Then desalted by 20ul volume flush. The liquid phase is Easy nLC 1200 nanoliter liquid phase system (ThermoFisher, USA), the polypeptide sample of fructus Citri Grandis is desalted and retained on pre-column, and then separated by analytical column (Acclaim PepMap RSLC,75 μm × 25cm C18-2 μm)
) The gradient used in the experiment was an increase of mobile phase B (80% acetonitrile, 0.1% formic acid) from 5% to 38% within 30 min. Mass Spectrometry A ThermoFisher Q active system (ThermoFisher, USA) was used in combination with a nanoliter nebulizing Nano Flex ion source (ThermoFisher, USA), the nebulizing voltage was 1.9kV, and the ion transfer tube heating temperature was 275 ℃. The mass spectrum scanning mode is in an information-Dependent acquisition working mode (DDA, Data Dependent Analysis), the primary mass spectrum scanning resolution is 70000, the scanning range is 100-. Maximum collection of 20 charges of 1+ under each DDA cycleAnd (3) obtaining a secondary spectrum of 3+, wherein the maximum ion injection time of the secondary mass spectrum is 50 ms. The collision cell energy (high energy collision induced dissociation, HCD) was set to 28eV for all precursor ions and the dynamic exclusion was set to 6 seconds.
The raw profile file collected by mass spectrometry is processed and retrieved and analyzed by PEAKS Studio 8.5(Bioinformatics Solutions Inc. Waterloo, Canada) software, the database is a protein database of Citrus maxima species downloaded by NCBI, and the retrieval parameters are set as follows: the mass tolerance of the primary mass spectrum is 10ppm, and the mass tolerance of the secondary mass spectrum is 0.05 Da. The variable modifications are oxidation (M), Acetylation (Protein N-term), deamidation (NQ), Pyro-glu from E, Pyro-glu from Q.
2. Test results
2.1, the amino acid sequence of the grapefruit young fruit polypeptide prepared in the embodiment 1 of the invention is as follows: IVVPGIY.
2.2, the amino acid sequence diagram of the grapefruit young fruit polypeptide prepared in the embodiment 1 of the invention is shown in figure 1.
Second test example color difference value measurement test of chestnut kernels
1. Test subjects:
the chestnut kernels obtained in example 2 and example 3 were designated as example 2 group and example 3 group, respectively, and the chestnut kernels obtained in step 2 of example 2 were used as control group a, and the chestnut kernels obtained in step 2 of example 3 were used as control group B.
2. The test method comprises the following steps:
the color difference values of the chestnut kernels of the groups of example 2, example 3, control group A and control group B were measured by a KONICA MINOLTA CR-400 color difference meter.
(1) And (3) white board correction: pressing a correction key ON a measurement screen of the colorimeter, inputting white plate correction data, then vertically placing a probe in the middle of a white correction plate, and pressing the measurement key to finish correction when a ready lamp is confirmed to be ON;
(2) selecting a color space: pressing a color space key to select a color space as Hunter Lab;
(3) setting a color difference standard color: pressing a color difference standard color key on the measurement screen to enter a color difference standard color setting screen, pressing a color difference target color key to set the color difference target color No. T06, and pressing an enter key to return to the measurement interface;
(4) measuring chromatic aberration: and tightly attaching the probe to the surface of the Chinese chestnut, and pressing a measuring key ON the probe after the ready lamp is turned ON to finish the color difference value measurement of the Chinese chestnut.
(5) And (3) data analysis: analyzing the color difference value data (delta a, delta b, delta E and delta L) by an LAB system, and evaluating the color protection effect of the grapefruit young fruit polypeptide on chestnut kernels.
3. And (3) test results:
the test results are shown in table 1.
TABLE 1 color change during storage of chestnut kernels
Note: the same group is marked with different lower case letters to represent obvious difference; statistical analysis of experimental results one-way analysis of variance was used (P < 0.05).
In the present study, the color change of chestnut kernels was analyzed using the LAB system (Lab color space defined by L, a and b coordinate space). The greater the Δ E change, the more pronounced the color change.
As can be seen from Table 1, the browning degree of the group in example 2 (the polypeptide-treated group of young pomelo fruits, which is stir-fried Chinese chestnut) did not change much during storage, while the Δ E of the control group A increased significantly between 0 and 12 hours. During the storage period of 0-8h, the color change of the group in example 3 (the shaddock young fruit polypeptide treated group cooks the Chinese chestnut) is not obviously changed, while the color of the group B in the control group is obviously higher than that of the group in example 3.
When the yellow/blue and red/green deviation ratios were calculated, the colors of control a and control B shifted significantly toward cyan, and the brightness decreased with the increase in storage time. Unlike control group A and control group B, the changes of delta a, delta B and delta L of the chestnut kernels treated by the grapefruit young fruit polypeptides in the groups of example 2 and example 3 are relatively slow. Food color analysis shows that the changes of delta E, delta a, delta b and delta L represent browning degrees, and in the research of the invention, the shaddock young fruit polypeptide treated Chinese chestnut kernels of the groups of the example 2 and the group of the example 3 show slight changes of delta a, delta b and delta L, which indicates that the shaddock young fruit polypeptide prepared in the example 1 of the invention can inhibit the browning of the Chinese chestnut kernels.
Test example III stability test of chestnuts
1. Test subjects:
example 2, example 3, example 4, example 5, comparative example 1 and comparative example 2.
2. The test method comprises the following steps:
the packaged and sealed chestnut kernels obtained in example 2, example 3, example 4, example 5, comparative example 1 and comparative example 2 were left at room temperature, and color changes were observed at day 0, month 1, month 3, month 6, month 9 and month 12.
Wherein: the color grading standard and requirements are as follows: the scoring criteria are shown in table 2, and the scoring requirements are that 10 chestnut kernels are placed on A4 white paper for comparative evaluation at each evaluation, the scoring values are recorded, and the average value is taken as the result.
TABLE 2 sensory Scoring Standard Table
| Item | Score value |
| Has obvious gray, black and the like | 0～20 |
| Has light gray, black and the like | 20～40 |
| No gray color,Black, etc., but no chestnut kernel should have color and luster | 40～60 |
| The original color of the Chinese chestnut kernels is kept and is consistent with that of the freshly treated Chinese chestnut kernels | 80～100 |
3. Test results
The test results are shown in table 3:
TABLE 3 color protection score results for chestnut kernels
| Day 0 | |
Month 3 | Month 6 | Month 9 | Month 12 | |
| Example 2 | 100 | 99 | 95 | 91 | 85 | 82 |
| Example 3 | 100 | 98 | 93 | 90 | 84 | 80 |
| Example 4 | 100 | 100 | 98 | 96 | 94 | 93 |
| Example 5 | 100 | 99 | 97 | 95 | 91 | 90 |
| Comparative example 1 | 100 | 92 | 84 | 73 | 60 | 45 |
| Comparative example 2 | 100 | 94 | 88 | 76 | 64 | 52 |
As can be seen from table 3, the specific shaddock young fruit polypeptide prepared in example 1 provided by the present invention can effectively alleviate browning of the chestnut kernels, and the effect of alleviating browning of the chestnut kernels is better than that of the existing chestnut kernel color protection liquid, so that the quality of the chestnut kernel product can be effectively improved. Meanwhile, the specific pulse treatment provided by the invention is combined with the specific shaddock young fruit polypeptide prepared in the embodiment 1, so that a good sterilization effect can be achieved, the browning of the Chinese chestnuts can be further delayed, and the product stability is improved. Test example four product quality test of chestnut kernels
1. Test subjects:
chestnut kernels according to example 2, example 3, example 4 and example 5.
2. The test method comprises the following steps:
the chestnut kernels obtained in example 2, example 3, example 4 and example 5 were tested for total bacteria, coliform bacteria and pathogenic bacteria. Wherein: the total number of bacteria is detected by referring to a detection method of GB4789.2, escherichia coli is detected by referring to a detection method of GB4789.3, and pathogenic bacteria is detected by referring to a detection method of GB 4789.4.
3. And (3) test results:
the test results are shown in table 4.
TABLE 4 detection of the strains of chestnut kernels
| Total number of bacteria (number/g) | Escherichia coli (each/100 g) | Pathogenic bacteria | |
| Example 2 | 0 | 0 | Is free of |
| Example 3 | 0 | 0 | Is free of |
| Example 4 | 0 | 0 | Is composed of |
| Example 5 | 0 | 0 | Is free of |
As can be seen from Table 4, the microbial indicators of the chestnut kernels prepared by the method all meet the relevant requirements.
Claims (10)
1. The color protection method for processing the Chinese chestnuts is characterized by comprising the following steps of:
s1, selecting and cleaning Chinese chestnuts, cutting a cross cut at the tops of the Chinese chestnuts, and performing curing treatment, wherein the curing treatment comprises the following steps: frying the Chinese chestnut for 25-35 min or putting the Chinese chestnut into boiling water for boiling for 10-20 min;
s2, cooling the Chinese chestnuts processed in the step S1, and peeling the Chinese chestnuts to obtain Chinese chestnut kernels;
step S3, soaking the Chinese chestnut kernels prepared in the step S2 in the young grapefruit polypeptide liquid for 8-15 min, wherein the material-liquid ratio of the Chinese chestnut kernels to the young grapefruit polypeptide liquid is 1g:20 mL-1 g:50mL, taking out, draining, bagging and carrying out microwave sterilization, wherein the microwave sterilization conditions are as follows: the power density is 6-8W/g, and the processing time is 80-90 s.
2. The color protection method for chestnut processing according to claim 1, wherein the concentration of the grapefruit young fruit polypeptide liquid in the step S3 is 4-8 mg/mL.
3. The color protection method for chestnut processing according to claim 2, wherein the concentration of the grapefruit young fruit polypeptide liquid in the step S3 is 5 mg/mL.
4. The color protection method for chestnut processing according to claim 1, wherein the preparation method of the grapefruit young fruit polypeptide liquid in the step S3 comprises the following steps:
adding distilled water into grapefruit young fruit powder, uniformly stirring to obtain grapefruit young fruit powder solution, then placing the grapefruit young fruit powder solution in a water bath at 90 ℃ for 8-12 min, cooling to 25 ℃, adding a pH regulator to adjust the pH of the solution to 8.5 to obtain reaction liquid, then adding neutral protease to carry out enzymolysis for 0.4-0.6 h in the water bath at 55 ℃, adding phenylmethylsulfonyl fluoride after enzymolysis, and centrifuging at 4000r/min for 8-12 min to obtain the grapefruit fruit powder.
5. The color protection method for Chinese chestnut processing according to claim 4, wherein the concentration of the shaddock young fruit powder solution is 48-52 mg/mL.
6. The color protection method for chestnut processing according to claim 4, wherein the pH adjustor is NaOH solution having a concentration of 0.1mol/L or HCL solution having a concentration of 0.1 mol/L.
7. The color protection method for chestnut processing according to claim 4, wherein the addition amount of the neutral protease is 0.5% w/v of the reaction solution, and the addition amount of the phenylmethylsulfonyl fluoride is 0.02% w/v of the reaction solution.
8. The color protection method for chestnut processing according to claim 1, wherein the step S3 further includes a pulse processing, which includes the specific steps of: and after soaking, carrying out pulse treatment, taking out, draining and bagging to obtain the finished product.
9. The color protecting method for chestnut processing according to claim 8, wherein the pulse processing conditions are: the pulse width pulse is 160-180 ns, the pulse rise time is 25-30 ns, the frequency is 60-70 Hz, the pulse amplitude is 15-18 Kv/cm, and the pulse time is 4-6 min.
10. The color-protecting method for chestnut processing according to claim 9, wherein the pulse-processing conditions are: the pulse width pulse is 165ns, the pulse rise time is 28ns, the frequency is 66Hz, the pulse amplitude is 16Kv/cm, and the pulse time is 5 min.
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| CN109907280A (en) * | 2019-02-27 | 2019-06-21 | 陕西省生物农业研究所 | A kind of color protecting method for Chinese chestnut production and processing |
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