CN112425737A - Fishy smell-removed and decolored sea cucumber powder and preparation method thereof - Google Patents
Fishy smell-removed and decolored sea cucumber powder and preparation method thereof Download PDFInfo
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- CN112425737A CN112425737A CN202011235447.9A CN202011235447A CN112425737A CN 112425737 A CN112425737 A CN 112425737A CN 202011235447 A CN202011235447 A CN 202011235447A CN 112425737 A CN112425737 A CN 112425737A
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- sea cucumber
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- enzymolysis
- enzyme
- fishy smell
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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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/70—Comminuted, e.g. emulsified, fish products; Processed products therefrom such as pastes, reformed or compressed products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
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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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/65—Addition of, or treatment with, microorganisms or enzymes
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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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
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- 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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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
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- Zoology (AREA)
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- Oil, Petroleum & Natural Gas (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses fishy smell removing and discoloring sea cucumber powder and a preparation method thereof, and belongs to the field of aquatic product processing. The method comprises the following steps: (1) removing internal organs and gonads of fresh sea cucumbers, soaking and washing the inner and outer surfaces of the body walls of the sea cucumbers; (2) draining the body wall of the sea cucumber, shearing and then homogenizing; (3) adding water and a complex enzyme into the homogenate for enzymolysis, wherein the complex enzyme is prepared from the following special enzymes for aquatic products in a compounding ratio: flavourzyme 2-8: 2-8 (U); (4) inactivating enzyme, and cooling to room temperature; (5) centrifuging and reserving supernatant; (6) concentrating by rotary evaporation to 1/10 of the original volume; (7) storing the concentrated solution in a refrigerator at-80 deg.C for freezing; (8) freeze-drying at low temperature to obtain sea cucumber powder. The method of the invention has no addition of other chemical components, thereby not only reducing the time cost and the material cost, but also improving the safety of the product, and being suitable for enterprise production.
Description
Technical Field
The invention relates to fishy smell-removed and decolored sea cucumber powder and a preparation method thereof, belonging to the field of aquatic product processing.
Background
Sea cucumber belongs to the organisms of the order of the Holothuroidea of the class Holothuroidea of the phylum Echinodermata, and the types recorded in the world exceed 1100, wherein more than 140 sea cucumbers grow in China, and the quality of stichopus japonicus produced by Liaoning, Shandong and Fujian is the best. The sea cucumber is widely known from ancient times by using rare medicinal materials and nourishing food materials, the organic components of the wall of the dried sea cucumber body comprise 90% of protein, 4% of lipid and 6% of polysaccharide, and the sea cucumber body is also rich in various mineral substances and vitamins required by a human body and is a high-quality food material with high protein, low fat and low sugar. In addition, a large number of documents show that the sea cucumber also has various physiological functions of reducing blood pressure, reducing blood sugar, resisting fatigue, improving immunity, delaying senility and the like. In recent years, the market demand of sea cucumbers is continuously rising, the sea cucumber breeding industry is developed vigorously, but the sea cucumbers have different sizes and forms due to climate reasons and individual differences in the breeding process, so that a large amount of sea cucumbers with poor appearances are not fully utilized, and a large amount of economic loss and resource waste are caused.
The prior literature has demonstrated that the body wall of sea cucumber contains a variety of active ingredients, such as: the sea cucumber collagen and the peptides thereof, which are the largest organic matters in the body wall of the sea cucumber, have the physiological functions of resisting oxidation, resisting fatigue, reducing blood pressure, reducing blood fat, easing pain and the like; the sea cucumber polysaccharide comprises mucopolysaccharide and fucosan, and has physiological effects of resisting tumor, blood coagulation, radiation injury, oxidation and the like; stichopus japonicus saponins have physiological effects of regulating immunity, resisting bacteria, resisting cytotoxicity and relieving fatigue. In addition, the sea cucumber also contains various mineral substances, such as Fe, Ca, Mg and Zn, and further contains trace elements such as Se, Ge, Sr, Cu and the like which are beneficial to the human body, so that the sea cucumber is a marine product with extremely high nutritional value.
However, most of the existing researches on the biological activity of the sea cucumber are focused on a certain substance after being extracted, such as sea cucumber polysaccharide, sea cucumber saponin or sea cucumber collagen, and the research on the total nutritional functionality of the sea cucumber is less. In addition, the fine extraction process is not only tedious, but also causes a great loss of nutrient substances, which is not very practical for enterprises requiring simple and convenient process, low time consumption and low cost.
Disclosure of Invention
In order to solve at least one problem, the invention provides a method for preparing fishy smell-removed and decolored sea cucumber powder, which is obtained from fresh sea cucumbers by an enzymolysis technology and can solve the problems of sea cucumber resource waste and sea cucumber active substance extraction technology complexity.
The first purpose of the invention is to provide a method for preparing fishy smell-removed and decolored sea cucumber powder, which comprises the following steps:
adding complex enzyme into the homogenate of the body wall of the sea cucumber for enzymolysis, wherein the complex enzyme is a compound of special enzyme for aquatic products and flavor protease.
In one embodiment of the invention, the complex enzyme is compounded in a ratio of 2-8: 2-8 (U) of the special aquatic enzyme to 2-8: 2-8 (U) of flavourzyme, and is further optimized to 6-8: 2-4 (U), and is further optimized to 6: 4 (U).
In one embodiment of the invention, the aquatic-specific enzyme and the flavourzyme are obtained from bioscience, Inc. of Henghuadao, Dongning.
In one embodiment of the invention, the enzyme activity of the aquatic product specific enzyme is 300000U/g.
In one embodiment of the invention, the enzymatic activity of the flavourzyme is 30000U/g.
In one embodiment of the present invention, the conditions of the enzymatic hydrolysis are: the addition amount of the complex enzyme is 8000-20000U/g pro (pro is equal to the content of substrate protein), the enzymolysis temperature is 50-55 ℃, the pH value is 6.5-7.5, and the enzymolysis time is 10 min-6 h.
In one embodiment of the present invention, the conditions of the enzymatic hydrolysis are: the addition amount of the complex enzyme is 20000U/g pro, namely 14000U/20g of sea cucumber homogenate, the enzymolysis temperature is 50-55 ℃, the pH is 6.5-7.5, and the enzymolysis time is 30 min-6 h; more preferably, the enzymolysis time of the compound enzyme is 30 min.
In an embodiment of the present invention, the enzymatic hydrolysis conditions are: the addition amount of the complex enzyme is 20000U/g pro, namely 14000U/20g of sea cucumber homogenate, the enzymolysis temperature is 50-55 ℃, the pH value is 6.5-7.5, and the enzymolysis time is 1-6 h; more preferably, the enzymolysis time of the compound enzyme is 5 h.
In one embodiment of the present invention, a method for preparing a deodorized and decolorized sea cucumber powder comprises the following steps:
(1) pretreatment: cutting open fresh sea cucumber from abdomen, removing viscera and gonad, and only retaining the body wall part of sea cucumber; soaking the sea cucumber by clear water, and then cleaning the inner and outer surfaces of the body wall of the sea cucumber by running water;
(2) homogenizing: draining the body wall of the sea cucumber treated in the step (1), shearing and then homogenizing to obtain homogenate;
(3) enzymolysis: placing the homogenate prepared in the step (2) into a fermentation bottle, and adding water and complex enzyme for enzymolysis to obtain an enzymolysis liquid;
(4) enzyme deactivation: inactivating enzyme of the enzymatic hydrolysate treated in the step (3), and cooling to room temperature;
(5) centrifuging: centrifuging the enzymolysis liquid treated in the step (4), and reserving supernatant;
(6) and (3) rotary steaming: performing rotary evaporation on the supernatant collected in the step (5), and concentrating to 5-15% of the original volume;
(7) freezing and storing: storing the concentrated solution treated in the step (6) at-90 to-70 ℃ for freezing compaction;
(8) freeze-drying: and (4) freeze-drying, grinding and crushing the concentrated solution stored in the step (7) to obtain a finished product of the sea cucumber powder with fishy smell removed and decolored.
In one embodiment of the present invention, a method for preparing a deodorized and decolorized sea cucumber powder comprises the following steps:
(1) pretreatment: cutting open fresh sea cucumber from abdomen, removing viscera and gonad, and only retaining the body wall part of sea cucumber; soaking the sea cucumber in clear water for 3-5 min, and then cleaning the inner and outer surfaces of the body wall of the sea cucumber by running water;
(2) homogenizing: draining the body wall of the sea cucumber treated in the step (1), shearing and then placing the sea cucumber into a multifunctional stirrer for homogenate;
(3) enzymolysis: placing the homogenate prepared in the step (2) into a fermentation bottle, adding water and complex enzyme for enzymolysis, and carrying out enzymolysis on insoluble protein in the homogenate to obtain polypeptide with good solubility;
(4) enzyme deactivation: inactivating the enzyme of the enzymatic hydrolysate treated in the step (3), heating in a boiling water bath at 95-100 ℃ for 10-15 min, and then placing in cold water to cool to room temperature;
(5) centrifuging: centrifuging the enzymolysis liquid treated in the step (4), and reserving supernatant;
(6) and (3) rotary steaming: performing rotary evaporation on the supernatant collected in the step (5), and concentrating to 1/10 of the original volume;
(7) freezing and storing: storing and freezing the concentrated solution treated in the step (6) in an ultra-low temperature refrigerator at-80 ℃;
(8) freeze-drying: and (4) placing the concentrated solution stored in the step (7) into a freeze dryer, freeze-drying for 24-48 h at-80 ℃, and grinding and crushing to obtain a finished product of the fishy smell-removed and decolored sea cucumber powder.
In one embodiment of the present invention, the sea cucumber used in step (1) is one of apostichopus japonicus, thelenota ananas, holothuria leucospilota, holothuria miqueliana, stichopus japonicus selenka, and holothuria scabra.
In one embodiment of the present invention, the operation in step (2) is specifically: cutting the wall of the sea cucumber body into small pieces of 1-2 cm multiplied by 1-2 cm, placing the small pieces in a multifunctional stirrer, wherein the volume of the small pieces is not more than 80% of the volume of a container, and stirring the small pieces at low speed for 1min without adding water.
In one embodiment of the present invention, the mass ratio of the homogenate to the water in the step (3) is 1: 2 to 6(w/w), and more preferably 1: 4.
In one embodiment of the present invention, the centrifugation conditions in step (5) are: centrifuging at 8000-12000 rpm/min and 4 ℃ for 15-20 min, and further preferably: 10000rpm/min, 4 ℃, centrifuging for 20min, and the obtained enzymolysis liquid has the characteristics of degreasing and impurity removal.
In one embodiment of the present invention, the rotary evaporation conditions in the step (6) are as follows: the amount of the supernatant added into the rotary evaporation container is not more than 50% of the volume of the container, the rotating speed is set to be 45-60 rpm/min, the vacuum degree is 80-120 mbar, the temperature is 60-65 ℃, and particularly, the solution is preferably in a slightly boiling state.
In one embodiment of the present invention, the freezing operation in the step (7) is: and (3) subpackaging the collected concentrated solution into a container with a larger bottom surface area, wherein the thickness is not more than 1.5cm, sealing the container with a preservative film, placing the container in an ultralow temperature refrigerator at minus 80 ℃ for 12-24 hours, freezing the concentrated solution to be solid, and facilitating low-temperature freeze-drying.
The second purpose of the invention is the sea cucumber powder prepared by the method.
The third purpose of the invention is the application of the sea cucumber powder in the whole milk powder or the soybean milk powder.
The invention has the beneficial effects that:
(1) the preparation method provided by the invention is simple to operate, only needs enzymolysis and centrifugation technologies, removes impurity removal technologies such as acid precipitation, alkali precipitation, microfiltration and membrane filtration, and does not add other chemical components in the preparation process, thereby not only reducing the time cost and the material cost, but also improving the safety of the product, and being suitable for enterprise production;
(2) the preparation method provided by the invention uses the compound enzyme with a proper proportion, can reduce the fishy smell of the sea cucumber while carrying out high-efficiency enzymolysis, and simultaneously optimizes the color of the final product sea cucumber powder;
(3) the invention provides a new way for high-value utilization of the sea cucumber, provides guiding significance for the research of physiological functionality of the sea cucumber, and has important significance for the healthy sustainable development of the functional food industry.
(4) In the method, the hydrolysis degree of the enzymolysis liquid reaches more than 4 percent; the proportion of the sea cucumber powder with the molecular weight of less than 1000Da is more than 82%, the whole color is lightened, the proportion of ketone substances in volatile substances is greatly reduced, and typical fishy substances are reduced.
Drawings
FIG. 1 shows the degree of hydrolysis and molecular weight of the enzymatic hydrolysate under different conditions; wherein (A) is the hydrolysis degree of neutral protease, papain, aquatic product special enzyme and flavourzyme under the same conditions; (B) is the molecular weight distribution of the hydrolysate of neutral protease, papain, aquatic special enzyme and flavourzyme; (C) is the hydrolysis degree of the compound protease with different proportions under the same conditions; (D) is the molecular weight distribution of the hydrolysate of the compound protease with different proportions.
Fig. 2 is a radar chart (n-9) for sensory evaluation of hydrolysate of compound protease (aquatic product special enzyme: flavourzyme) in different proportions.
FIG. 3 is a diagram showing the color difference between the sea cucumber powder with different enzymolysis degrees and the raw material freeze-dried powder, wherein (A) is a color difference change diagram; (B) is a simulated color block diagram obtained by the values of L, a and b.
FIG. 4 shows the volatile flavor composition of the sea cucumber powder and the raw material freeze-dried powder with different enzymolysis degrees.
FIG. 5 shows the content changes of typical volatile fishy smell substances, i.e. n-hexanal, n-octanal and n-heptanal, in the sea cucumber powder and the raw material freeze-dried powder with different enzymolysis degrees.
FIG. 6 shows the content changes of typical volatile fishy smell substances of 3, 5-octadiene-2-one, 3, 5-nonadiene-2-one and 1-octen-3-ol in sea cucumber powder and raw material freeze-dried powder with different enzymolysis degrees.
Detailed Description
The following description of the preferred embodiments of the present invention is provided for the purpose of better illustrating the invention and is not intended to limit the invention thereto.
The test method comprises the following steps:
1. and (3) determining the degree of hydrolysis: the experiment adopts ninhydrin colorimetry to determine the degree of hydrolysis, and comprises the following specific steps:
(1) determination of detection wavelength: and (3) moderately diluting the standard glycine solution and the polypeptide hydrolysate to be used as sample liquid to be detected for later use. And (2) adding 2mL of sample liquid and 1mL of ninhydrin color developing agent into a 10mL colorimetric tube respectively, shaking and mixing uniformly, reacting in a boiling water bath for 15min, taking out, standing and cooling for 5-10 min, adding 5mL of ethanol with the volume fraction of 40%, shaking and mixing uniformly, standing for 15-20 min, detecting under an ultraviolet spectrophotometer, and selecting the optimal absorption wavelength, wherein the detection wavelength range is 500-800 nm. The results show that the optimal detection wavelength is 570 nm.
(2) Establishing a glycine standard curve: accurately weighing 10mg of standard glycine, dissolving in ultrapure water, fully dissolving, and metering to 100mL to obtain a standard solution with the concentration of 100 mu g/mL. The standard solution was diluted according to the concentration gradient to prepare glycine standard solutions of 50. mu.g/mL, 20. mu.g/mL, 10. mu.g/mL, 5. mu.g/mL and 2. mu.g/mL, respectively. And (3) taking 10mL of colorimetric tube, respectively adding 2mL of standard solution with each dilution and 1mL of ninhydrin color developing agent, oscillating and uniformly mixing, reacting in a boiling water bath for 15min, taking out, standing and cooling for 5-10 min, then adding 5mL of ethanol with the volume fraction of 40%, oscillating and uniformly mixing, standing for 15-20 min, measuring the absorbance under an ultraviolet spectrophotometer, wherein the detection wavelength is 570nm, and zeroing with ultrapure water. A glycine standard curve was established from the results, with absorbance on the ordinate and glycine concentration (μ g/mL) on the abscissa.
(3) And (3) determining the content of free amino nitrogen in the hydrolysate: 1mL of the sea cucumber enzymolysis liquid with different degrees of hydrolysis is respectively diluted by 50 times, 2mL of the diluted liquid is taken to be placed in a 10mL colorimetric tube, the subsequent steps are described in the establishment part of the standard curve, the absorbance is detected at the wavelength of 570nm, and the content (mu mol/mL) of free amino nitrogen in the sample is calculated according to the glycine standard curve.
(4) Determination of the free amino nitrogen content in the sample stock solution (not enzymolyzed): and (3) during enzymolysis, taking 1 part of the sea cucumber homogenate with equal mass into 1 fermentation bottle with the same specification, wherein the only difference is that no enzyme is added and other conditions are unchanged, then placing the mixture into a constant-temperature shaking incubator, taking out 10mL of the mixture in turn after 10min, 30min and 5h respectively, and the centrifugation and filtration conditions are also the same. Taking 1mL of filtrate, diluting by 50 times, taking 2mL of diluent in a 10mL colorimetric tube, detecting the absorbance at the wavelength of 570nm according to the subsequent steps as described in the establishment part of the standard curve, and calculating the content (mmol/L) of free amino nitrogen in the sample stock solution according to the glycine standard curve.
(5) Calculating the hydrolysis degree:
wherein: h is hydrolysateMillimoles, mmol of peptide bonds of (a); h istotThe number of millimoles and mmol of peptide bonds of each gram of sea cucumber polypeptide can be calculated according to the amino acid composition of the sea cucumber polypeptide, and h is obtained by calculationtot8.5 mmol/g; a is the free amino nitrogen content of the stock solution, mmol/g; b is the free amino nitrogen content of the hydrolysate, mu mol/mL; n is the total nitrogen content of the hydrolysate, mmol/L, and is measured by a Kjeldahl method.
2. Determination of molecular weight distribution:
fully dissolving the sea cucumber powder by using ultrapure water, and diluting into a clear and transparent solution with the protein concentration of 1 mg/mL. The molecular weight distribution was determined by Waters 1525HPLC (high Performance liquid chromatography) equipped with a 2487 UV detector and GPC analysis software in an Em-power workstation.
The set parameters of the chromatogram are as follows: column TSK GEL 2000SWXL (300 mm. times.7.8 mm); mobile phase: 40% acetonitrile (containing 0.1% trichloroacetic acid) + 60% ultrapure water (V/V); the detection wavelength is 220 nm; the flow rate is 0.5 mL/min; the column temperature is 30 ℃;
the sample injection volume is 10 mu L; the sample concentration is 1 mg/mL; and (4) standard product: cytochrome C (Mw ═ 12384Da), aprotinin (Mw ═ 6500Da), bacillase (Mw ═ 1422Da), ethionine-tyrosine-arginine (Mw ═ 451Da), and ethionine-ethionine (Mw ═ 189 Da);
3. and (3) color difference analysis: a sea cucumber powder sample with equal mass is filled in a completely transparent plastic package bag, is fully and uniformly mixed, 6 random points of the sample are measured by a colorimeter, and the values of L, a and b of the sample are measured, wherein L represents brightness, a represents red (+) green (-), and b represents yellow (+) blue (-).
4. Analysis of volatile flavor composition:
a sea cucumber powder sample with the equal mass (5g) is filled into a detection bottle, wherein the mass does not exceed 1/3 of the sample bottle, and the volatile flavor substances are measured by adopting a full two-dimensional GC x GC/TOF-MS (gas phase-high flux time-of-flight mass spectrometer).
GCxGC conditions: one-dimensional chromatography column TR-FFAP (30 m.times.0.25 mm. times.0.25 μm); two-dimensional chromatography column Rxi-17Sil ms (1 m.times.0.25 mm.times.0.25 μm); temperature rising procedure: the initial temperature is 45 ℃, the temperature is kept for 3min, the temperature is increased to 230 ℃ at the speed of 10 ℃/min, and the temperature is kept for 5 min; the temperature of a sample inlet is 250 ℃; sample introduction mode: split-flow sample injection, split-flow ratio 5: 1, the flow rate is 5 mL/min; the flow rate of the carrier gas (He) was 1 mL/min.
MS conditions: the ion source temperature is 210 ℃; an electron ionization source; electron energy 70 eV; emission current 1 mA; the mass scanning range is 33-400 mu; the acquisition frequency was 1 kHz.
5. Sensory evaluation analysis:
sensory evaluation criteria are shown in table 1:
TABLE 1 organoleptic evaluation criteria of enzymatic hydrolysate of Stichopus japonicus
The specific evaluation method comprises the following steps: in the environment of excluding other interference, the same amount of enzymolysis liquid is put into a disposable measuring cup with high transparency, and the visual inspection, the nasal smell, the stirring and other methods are carried out one by one according to the table above to observe the color, the transparency, the viscosity and the smell. Nine sensory evaluators form a sensory evaluation group, and sensory evaluation training is carried out on the enzymolysis liquid sample, wherein each index is fully divided into 5 points, and the total number is 20 points.
Example 1: preparation of low-enzymolysis-degree sea cucumber powder (LDH)
The preparation method of the low-enzymolysis-degree sea cucumber powder (LDH) comprises the following steps:
(1) cutting open fresh sea cucumber from the abdomen, removing internal organs and gonads, soaking in clear water for 3-5 min, and cleaning the inner and outer surfaces of the body wall of the sea cucumber with running water;
(2) draining the cleaned sea cucumber body wall, cutting into small pieces of 1-2 cm multiplied by 1-2 cm, placing the small pieces in a multifunctional stirrer, wherein the volume of the small pieces is not more than 80% of the volume of a container, and stirring the small pieces at low speed for 1min without adding water;
(3) adding 160g of homogenate into a fermentation bottle, adding 640g of water and 112000U of complex enzyme (67200U of aquatic special enzyme +44800U of flavourzyme), wherein the enzymolysis temperature is 50-55 ℃, the pH value is natural pH (6.5-7.5), and the enzymolysis time is 10 min;
(4) heating in a boiling water bath at 95-100 ℃ for 15min to inactivate enzyme;
(5) centrifuging at 10000rpm/min and 4 deg.C for 20min, and collecting supernatant;
(6) the supernatant is subjected to rotary evaporation at the rotating speed of 45-60 rpm/min, the vacuum degree of 80-120 mbar and the temperature of 60-65 ℃, and the solution is preferably in a slightly boiling state and is concentrated to 1/10 of the original volume;
(7) subpackaging the concentrated solution into large plates with thickness of about 1cm, sealing with preservative film, and storing in-80 deg.C ultra-low temperature refrigerator for 16 h;
(8) and (4) carrying out vacuum freeze drying on the completely frozen concentrated solution for 48h, and grinding and crushing to obtain a finished sea cucumber powder product.
Example 2: preparation of sea cucumber powder (MDH) with medium enzymolysis degree
The sea cucumber powder is prepared by the method of the embodiment 1, except that the enzymolysis time is 30min, and other steps are the same as the embodiment 1, so that a finished sea cucumber powder product is obtained.
Example 3: preparation of high-enzymolysis-degree sea cucumber powder (HDH)
The method of the embodiment 1 is adopted to prepare the sea cucumber powder, the difference is that the enzymolysis time is only 5 hours, and other steps are the same as the embodiment 1, namely the finished product of the sea cucumber powder.
Comparative example 1: preparation of enzymatic hydrolysate of different kinds of enzymes
(1) Cutting open fresh sea cucumber from the abdomen, removing internal organs and gonads, soaking in clear water for 3-5 min, and cleaning the inner and outer surfaces of the body wall of the sea cucumber with running water;
(2) draining the cleaned sea cucumber body wall, cutting into small pieces of 1-2 cm multiplied by 1-2 cm, placing the small pieces in a multifunctional stirrer, wherein the volume of the small pieces is not more than 80% of the volume of a container, and stirring the small pieces at low speed for 1min without adding water;
(3) adding 160g of homogenate into a fermentation bottle, adding 640g of water and 112000U of protease, wherein the enzymolysis temperature is 50-55 ℃, the pH is natural pH (6.5-7.5), and the enzymolysis time is 10 min;
(4) heating in a boiling water bath at 95-100 ℃ for 15min to inactivate enzyme;
(5) centrifuging at 10000rpm/min and 4 deg.C for 20min, and collecting supernatant; obtaining enzymolysis liquid;
wherein, the protease used in the step (3) is neutral protease, papain, aquatic special enzyme and flavourzyme respectively.
Comparative example 2: preparation of enzymolysis liquid of complex enzymes with different proportions
(1) Cutting open fresh sea cucumber from the abdomen, removing internal organs and gonads, soaking in clear water for 3-5 min, and cleaning the inner and outer surfaces of the body wall of the sea cucumber with running water;
(2) draining the cleaned sea cucumber body wall, cutting into small pieces of 1-2 cm multiplied by 1-2 cm, placing the small pieces in a multifunctional stirrer, wherein the volume of the small pieces is not more than 80% of the volume of a container, and stirring the small pieces at low speed for 1min without adding water;
(3) adding 160g of homogenate into a fermentation bottle, adding 640g of water and 112000U of compound protease, wherein the enzymolysis temperature is 50-55 ℃, the pH is natural pH (6.5-7.5), and the enzymolysis time is 10 min.
(4) Heating in a boiling water bath at 95-100 ℃ for 15min to inactivate enzyme;
(5) centrifuging at 10000rpm/min and 4 deg.C for 20min, and collecting supernatant; obtaining enzymolysis liquid;
wherein, the proportion of the compound protease used in the step (3) is respectively as follows: the ratio of the special enzyme for aquatic products to the flavourzyme is 0: 100; the ratio of the special enzyme to the flavourzyme for the aquatic products is 20: 80; the ratio of the special enzyme to the flavourzyme for the aquatic products is 40: 60; the ratio of the special enzyme to the flavourzyme for the aquatic products is 60: 40; the ratio of the special aquatic enzyme to the flavourzyme is 80: 20 and the ratio of the special aquatic enzyme to the flavourzyme is 100: 0.
Results analysis 1: selection of species and proportion of compound enzyme
The degree of hydrolysis of the enzymatic hydrolysate prepared in comparative examples 1 to 2 was measured according to the above-described method for measuring the degree of hydrolysis, and the results are shown in FIG. 1. Fig. 1 (a): enzymolysis is carried out at the optimum temperature and pH of four enzymes, the added enzyme activity, substrate concentration and enzymolysis time are consistent, the lowest papain hydrolysis degree and the highest flavourzyme hydrolysis degree can be found, and the difference between the hydrolysis degrees of the aquatic special enzyme and flavourzyme is small. FIG. 1 (B): comparing the molecular weight distribution with the enzymolysis degree curve, the finding shows that although the hydrolysis degree of the aquatic special enzyme is slightly lower than that of the flavourzyme, the small molecular peptide in the enzymolysis product of the aquatic special enzyme is more than that of the flavourzyme, so the aquatic special enzyme and the flavourzyme are selected as the best hydrolase for compounding. Fig. 1(C, D): the special enzyme for the aquatic products and the flavourzyme are compounded, the degree of hydrolysis is determined after 5 hours of enzymolysis under the same enzyme activity, substrate concentration, pH and temperature, and the hydrolysis effect is best when the ratio of the special enzyme for the aquatic products to the flavourzyme is 6: 4.
Meanwhile, sensory evaluation analysis is carried out on the enzymolysis liquid of the comparative example 2, and the result is shown in figure 2, so that when the compound enzyme is the aquatic special enzyme and the flavourzyme in the compounding ratio of 60: 40, the four sensory indexes of the enzymolysis liquid are all highest in score.
Results analysis 2: hydrolysis degree and molecular weight distribution of sea cucumber powder prepared in examples 1 to 3
The results of measuring the hydrolysis degrees of the enzymatic hydrolysates in the preparation processes of examples 1 to 3 according to the above-described hydrolysis degree measuring method are shown in table 2: the degree of hydrolysis of example 1 was 4.07 ± 0.09%; the degree of hydrolysis of example 2 was 4.90 ± 0.09%; the degree of hydrolysis of example 3 was 8.98. + -. 0.14%.
Because the organic components of the sea cucumber body wall are mainly protein, the percentage of the protein is up to 90 percent, and the rest is a small amount of fat and heteropolysaccharide. The sample is degreased and purified through the centrifugation step after enzymolysis in the embodiment 1-3, and the finally prepared sea cucumber powder mainly contains the organic components of polypeptide. The results of measuring the molecular weight distribution of the sea cucumber powder of examples 1 to 3 according to the above-mentioned method for measuring the molecular weight distribution are shown in Table 2: the proportion of the sea cucumber powder with the molecular weight of less than 1000Da in the embodiment 1 is more than 82 percent; the proportion of the sea cucumber powder with the molecular weight of less than 1000Da in the embodiment 2 is more than 88 percent; the sea cucumber powder of example 3, with a molecular weight of less than 1000Da, accounts for more than 96%, and can be regarded as oligopeptide. The sea cucumber powder prepared by the method provided by the invention has good solubility, and insoluble collagen in the wall of the sea cucumber body is almost completely hydrolyzed into water-soluble small molecular polypeptide.
TABLE 2 hydrolysis degree of Stichopus japonicus powder with different enzymolysis degree and molecular weight distribution of its polypeptide
Results analysis 3: color and flavor composition of sea cucumber powder
The sea cucumber has dark color and peculiar fishy smell of marine products, so that the organoleptic properties of products prepared from the sea cucumber are poor, and the product forms of the sea cucumber are limited.
According to the determination methods of the color difference analysis and the volatile flavor composition analysis, the appearance, color and volatile flavor substances of the sea cucumber powder prepared in the examples 1 to 3 are determined. As can be seen from table 3 and fig. 3(a), L of the enzymatically hydrolyzed sea cucumber powder is significantly increased, a and b are significantly decreased, and the overall color is characterized by being faded; the simulated color block diagram of fig. 3(B) also visually shows that the color of the sea cucumber powder after enzymolysis is obviously lighter than that of the sea cucumber powder without enzymolysis.
Analysis of the volatile materials of examples 1-3 by GC-TOF-MS detected nearly a hundred flavor species, mainly hydrocarbons, ketones, alcohols, aldehydes and acids, as shown in FIG. 4. A large amount of existing literature proves that the fishy smell of marine products mainly comes from ketones and aldehydes, and as can be seen from fig. 4, the proportion of ketones in the volatile substances of the sea cucumber powder in examples 1 to 3 is greatly reduced, and the proportion of aldehydes in the sea cucumber powder in example 3 is also greatly reduced.
The existing literature researches more on volatile fishy smell substances of marine products, and summarizes ten volatile substances with typical fishy smell, six types of flavor substances with typical fishy smell are screened according to the literature, wherein the six types of flavor substances are n-hexanal, n-octanal, n-heptanal, 3, 5-octadien-2-one, 3, 5-nonadien-2-one and 1-octen-3-ol, and the contents of the flavor substances are compared. As can be seen from fig. 5 and 6, the content of the typical fishy smell substances in the sea cucumber powder of examples 1 to 3 is reduced, especially the content of the sea cucumber powder of example 3 is reduced most significantly.
As can be seen from the results of table 3, fig. 4, fig. 5 and fig. 6, the color and flavor of the sea cucumber powder prepared in examples 1 to 3 are optimized to a certain extent, and especially, the sea cucumber powder in example 3 is optimized, so that the method of the present invention has the function of removing fishy smell and decoloring the sea cucumber powder.
TABLE 3 color difference values of Stichopus japonicus powder with different enzymolysis degrees and raw materials without enzymolysis
| Sample (I) | L* | a* | b* |
| Sea cucumber raw material freeze-dried powder | 60.56±1.04c | 2.77±0.04a | 9.66±0.25a |
| Example 1 sea cucumber powder with Low enzymatic hydrolysis degree | 72.72±2.02a | 2.26±0.15b | 6.38±0.14b |
| Example 2 sea cucumber powder with medium enzymolysis degree | 72.34±1.04a | 1.83±0.01c | 5.90±0.09c |
| Example 3 sea cucumber powder with high enzymatic hydrolysis degree | 69.03±0.98b | 1.83±0.05c | 5.96±0.15c |
Note: different letters in the same column represent significant differences, P <0.05
Example 4 comparison of the application of sea cucumber powder in milk powder
The sea cucumber powder of example 1 was mixed with commercially available whole milk powder (brand: Anjia) uniformly, wherein the contents of the sea cucumber powder and the milk powder were 10% and 90%, respectively.
Comparative example 3
Mixing commercially available Stichopus japonicus powder (Dalianfei biological industry Co., Ltd.) and commercially available whole milk powder (brand: Anjia) uniformly, wherein the content of Stichopus japonicus powder is 10% and the content of milk powder is 90%.
Example 5
The sea cucumber powder of example 1 was mixed with commercially available whole milk powder (brand: Anjia) uniformly, wherein the contents of the sea cucumber powder and the milk powder were 20% and 80%, respectively.
Comparative example 4
Mixing commercially available Stichopus japonicus powder (Dalianfei biological industry Co., Ltd.) and commercially available whole milk powder (brand: Anjia) uniformly, wherein the content of Stichopus japonicus powder is 20% and the content of milk powder is 80%.
Example 6 comparison of application of sea cucumber powder in Bean flour
The sea cucumber powder of example 1 was uniformly mixed with commercially available soybean milk powder (brand: Bingquan), wherein the content of the sea cucumber powder was 10% and the content of the commercially available soybean milk powder (brand: Bingquan) was 90%.
Comparative example 5
Mixing commercially available Stichopus japonicus powder (Dalianfei bio-industry Co., Ltd.) and commercially available soybean milk powder (brand: Bingquan) uniformly, wherein the content of Stichopus japonicus powder is 10% and the content of commercially available soybean milk powder (brand: Bingquan) is 90%.
Example 7
The sea cucumber powder of example 1 was uniformly mixed with commercially available soybean milk powder (brand: Bingquan), wherein the content of the sea cucumber powder was 20% and the content of the commercially available soybean milk powder (brand: Bingquan) was 80%.
Comparative example 6
Mixing commercially available Stichopus japonicus powder (Dalianfei bio-industry Co., Ltd.) and commercially available soybean milk powder (brand: Bingquan) uniformly, wherein the content of Stichopus japonicus powder is 20% and the content of commercially available soybean milk powder (brand: Bingquan) is 80%.
Results analysis 4: application comparison of sea cucumber powder
The samples prepared in comparative examples 3 to 6 and examples 4 to 7 were subjected to color difference measurement, wherein L represents brightness, and a smaller value of L represents a darker color of the sample. As shown in table 4, it can be found that the final sample of the sea cucumber powder prepared in the example has a lighter color than a sample mixed with a certain commercially available sea cucumber powder when the sea cucumber powder is mixed with milk powder and soybean milk powder, which indicates that the sample prepared in the example has a smaller influence on the color of the final product in application, and the application range of the sea cucumber powder is expanded.
TABLE 4 color difference values of the sea cucumber powder and the whole milk powder mixture sample
| Example (b) | Sample (I) | L* |
| Example 4 | Example 1 Whole milk powder 1: 9 | 85.51±0.47A |
| Comparative example 3 | Control sample to whole milk powder 1: 9 | 77.76±0.29a |
| Example 5 | Example 1 Whole milk powder 2: 8 | 82.37±0.33B |
| Comparative example 4 | Control sample to whole milk powder 2: 8 | 73.63±0.27b |
| Example 6 | Example 1 soymilk powder 1: 9 | 74.99±0.59C |
| Comparative example 5 | Control sample soybean milk powder is 1: 9 | 78.23±0.25c |
| Example 7 | Example 1 soymilk powder 2: 8 | 81.13±0.09D |
| Comparative example 6 | Control sample soybean milk powder 2: 8 | 83.44±0.51d |
Note: the case between the same letters in the table represents a significant difference, P < 0.05; the control sample represents sea cucumber powder of Dalian biological industries, Ltd.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A method for preparing fishy smell-removed and decolored sea cucumber powder is characterized by comprising the following steps:
adding complex enzyme into the homogenate of the body wall of the sea cucumber for enzymolysis, wherein the complex enzyme is a compound of special enzyme for aquatic products and flavor protease.
2. The method according to claim 1, wherein the enzyme activity ratio of the complex enzyme compound is that of the aquatic special enzyme: the flavor protease is 2-8: 2-8.
3. The method according to claim 1 or 2, wherein the conditions of the enzymatic hydrolysis are: the addition amount of the complex enzyme is 8000-20000U/g pro, the enzymolysis temperature is 50-55 ℃, the pH value is 6.5-7.5, and the enzymolysis time is 10 min-6 h.
4. A method according to any one of claims 1 to 3, comprising the steps of:
(1) pretreatment: cutting open fresh sea cucumber from abdomen, removing viscera and gonad, and only retaining the body wall part of sea cucumber; soaking the sea cucumber by clear water, and then cleaning the inner and outer surfaces of the body wall of the sea cucumber by running water;
(2) homogenizing: draining the body wall of the sea cucumber treated in the step (1), shearing and then homogenizing to obtain homogenate;
(3) enzymolysis: placing the homogenate prepared in the step (2) into a fermentation bottle, and adding water and complex enzyme for enzymolysis to obtain an enzymolysis liquid;
(4) enzyme deactivation: inactivating enzyme of the enzymatic hydrolysate treated in the step (3), and cooling to room temperature;
(5) centrifuging: centrifuging the enzymolysis liquid treated in the step (4), and reserving supernatant;
(6) and (3) rotary steaming: performing rotary evaporation on the supernatant collected in the step (5), and concentrating to 5-15% of the original volume;
(7) freezing and storing: storing the concentrated solution treated in the step (6) at-90 to-70 ℃ for freezing compaction;
(8) freeze-drying: and (4) freeze-drying, grinding and crushing the concentrated solution stored in the step (7) to obtain a finished product of the sea cucumber powder with fishy smell removed and decolored.
5. The method of claim 4, wherein the mass ratio of the homogenate to the water in the step (3) is 1: 2-6.
6. The method according to claim 4 or 5, wherein the centrifugation conditions in step (5) are: centrifuging at 8000-12000 rpm/min for 15-20 min.
7. The method according to any one of claims 4 to 6, wherein the sea cucumber used in step (1) is one of Apostichopus japonicus selenka, Thelenota ananas, Holothuria miq, Stichopus japonicus selenka, and Holothuria scabra.
8. The method according to any one of claims 4 to 7, wherein the rotary evaporation conditions in the step (6) are as follows: the amount of the supernatant added into the rotary evaporation container is not more than 50% of the volume of the container, the rotating speed is set to be 45-60 rpm/min, the vacuum degree is 80-120 mbar, and the temperature is 60-65 ℃.
9. A fishy smell removed and decolored sea cucumber powder prepared by the method of any one of claims 1 to 8.
10. Use of the fishy smell removed and decolored sea cucumber powder of claim 9 in whole milk powder or soybean milk powder.
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Cited By (4)
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| CN114287609A (en) * | 2021-12-29 | 2022-04-08 | 好当家集团有限公司 | Processing method of 3D printed instant sea cucumbers stored at normal temperature |
| CN114343139A (en) * | 2021-12-29 | 2022-04-15 | 好当家集团有限公司 | Preparation method of artificial sea cucumber |
| CN114522183A (en) * | 2022-01-24 | 2022-05-24 | 江南大学 | Sea cucumber extract for increasing bone mineral density |
| CN116491632A (en) * | 2023-01-31 | 2023-07-28 | 大连工业大学 | Preparation method of low-fishy low-salt sea cucumber powder |
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2020
- 2020-11-05 CN CN202011235447.9A patent/CN112425737A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114287609A (en) * | 2021-12-29 | 2022-04-08 | 好当家集团有限公司 | Processing method of 3D printed instant sea cucumbers stored at normal temperature |
| CN114343139A (en) * | 2021-12-29 | 2022-04-15 | 好当家集团有限公司 | Preparation method of artificial sea cucumber |
| CN114522183A (en) * | 2022-01-24 | 2022-05-24 | 江南大学 | Sea cucumber extract for increasing bone mineral density |
| CN116491632A (en) * | 2023-01-31 | 2023-07-28 | 大连工业大学 | Preparation method of low-fishy low-salt sea cucumber powder |
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