CN109430514B - Method for preparing tilapia mossambica-soybean meal coprecipitation protein - Google Patents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
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- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/001—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste
- A23J1/005—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste from vegetable waste materials
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Abstract
The invention provides a method for preparing tilapia mossambica-soybean meal coprecipitation protein, which is characterized by comprising the following steps: s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 1:5-5:1 (dry basis), and adding ice distilled water into the mixture according to the mass ratio of water =1:5-15(g: mL); s2.10000-15000r/min for homogenizing for 1-5 min; s3, adjusting the pH value of the homogeneous mixture to 3.0-8.0, and standing for 5 min; s4, adding a cosolvent with the mass ratio of 2%, stirring at room temperature and dissolving for 5-30 min; s5.4 ℃, 8000 plus 11000r/min for 10-20 min; s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5; and S7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and performing vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitated protein. The invention prepares a novel double-protein product to improve fish protein solubility, emulsibility, gel property and the like and develop a novel double-protein product combining animal and plant proteins.
Description
Technical Field
The invention belongs to the technical field of high-value utilization of aquatic protein, and particularly relates to a method for preparing tilapia mossambica-soybean meal coprecipitation protein.
Background
China is a big producing country, a big trading country and a big consuming country of tilapia, the export of tilapia is mainly sold fresh, and the products mainly comprise frozen tilapia fillets and frozen whole tilapia. The primary processing of tilapia mossambica processing products accounts for the main part, the products are single, and the profits are low, so the development of the tilapia mossambica industry is restricted. Aiming at the problems, the tilapia mossambica is subjected to fine and deep processing to improve the added value and profit margin of the tilapia mossambica.
Different types of proteins generally have different nutritional characteristics and functional characteristics, and the utilization of high-protein raw materials can be improved by mixing the proteins from different sources, so that the problems of insufficient content of essential amino acids in the protein from a single source, low nutritional value, poor functional characteristics and the like are solved. Mixing of proteins mainly includes direct mixing (physical mixing) and chemical mixing of protein materials. Co-precipitated protein (Co-PP) refers to the use of isoelectric precipitation, acid-heat precipitation, or acid-heat induction with the aid of a precipitating agent such as CaCl 2 And the like, a mixed protein obtained from one or more raw materials.
The coprecipitation protein prepared by the proteins from different sources not only can realize the complementation of amino acids nutritionally, but also has good functional characteristics and ideal organoleptic characteristics. The above studies indicate that co-precipitated proteins have a great potential application in the food industry.
At present, tilapia is used as a raw material, and novel products on the market comprise minced fillet products, pickled products, frozen products and the like. Tilapia protein is unstable, easy to change and poor in solubility in the actual production and processing process, and further the structural and functional characteristics of the protein are changed. The tilapia mossambica protein has low gel property and emulsifying property, influences the quality of products and limits the application of the tilapia mossambica protein in the field of foods. At present, a method for improving the quality of tilapia products, expanding the application of tilapia products in the food field and improving the overall profit margin of tilapia is urgently needed.
Disclosure of Invention
In view of this, the present invention utilizes the nutritional advantages of tilapia mossambica protein and the functional characteristic advantages of soybean protein, uses tilapia mossambica meat and defatted soybean meal as raw materials, prepares coprecipitated protein by a pH regulation method, prepares a novel 'double-protein' product, so as to improve fish protein solubility, emulsibility, gel characteristics, etc., and develops a novel double-protein product combining animal and plant proteins.
The technical scheme of the invention is as follows: a method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia mossambica meat and defatted soybean meal powder in a mass ratio of 1:5-5:1 (dry basis), and adding ice distilled water into the mixture according to the mass ratio of water =1:5-15(g: mL);
s2.10000-15000r/min for homogenizing for 1-5 min;
s3, adjusting the pH value of the homogeneous mixture to 3.0-8.0, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, stirring at room temperature and dissolving for 5-30 min; the mass ratio is the ratio of the cosolvent to the sum of the mass of the added tilapia mossambica and soybean meal.
S5.4 ℃, 8000 plus 11000r/min for 10-20 min;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
s7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and carrying out vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitation protein.
Further, in step S3, the homogeneous mixture is brought to a pH of 4.0 to 6.0.
Further, in step S6, the pH is adjusted to 4.0 to 6.0.
By finding that the soluble protein yield shows an upward trend in the acidic and alkaline pH ranges deviating from the isoelectric point, and the dissolution yield reaches the maximum under the conditions of extreme acid-base pH values of 2.0, 3.0, 11.0 and 12.0. This is because in the acidic range, the negative charges of the branched carboxyl groups of the residues of partially acidic amino acids are neutralized, so that the protein molecules have net positive charges; within the range of the basic strip, the basic groups of partial amino acids undergo deprotonation reaction, so that protein molecules carry net negative charges, the hydration of the protein is enhanced, and the solubility of the protein is increased. Through a large number of creative tests, soluble protein solutions extracted by mixing tilapia meat and defatted soybean meal in a mass ratio (on a dry basis) are dissolved at pH 2.0, 3.0, 11.0 and 12.0, precipitate at pH4.0-6.0, and the precipitation yield is 88% -95%, preferably, the yield is the highest at pH 4.5, so that the optimal isoelectric point of the tilapia-soybean meal acid/alkali soluble protein is determined to be 4.5. The pH value of the acid-precipitated isoelectric point of the vegetable protein is generally relatively low, and after the tilapia mossambica is mixed with the soybean meal, the pH value of the acid-precipitated isoelectric point is biased to the isoelectric point of the soybean meal. Thus, it can be determined that the precipitation pH of acid/alkali soluble protein of tilapia mossambica-soybean meal is 4.0-6.0, preferably pH 4.5.
Further, in step S1, tilapia meat and defatted soybean meal powder are mixed (on a dry basis) at a mass ratio of 1:2-2:1, and ice-cold distilled water is added according to the ratio of material: water =1:7-10(g: mL). According to the invention, through a large number of creative experiments, the characteristics of two different proteins are combined, the optimal mixing ratio and the material-water ratio during dissolution are obtained, and the optimal preparation condition of the co-precipitated protein is obtained.
Further, the cosolvent comprises the following components in parts by weight: 1-10 parts of lecithin, 12-19 parts of mannose, 3-9 parts of silicon dioxide, 0.2-0.9 part of lysozyme, 11-18 parts of lysine, 14-22 parts of arginine and 7-16 parts of glutamic acid. In the invention, the added cosolvent has the function of enhancing the interaction of the protein near the isoelectric point in the process of forming the tilapia-soybean meal coprecipitation protein, so that the tilapia protein and the soybean meal protein are further combined to form a stable coprecipitation protein structure, and the yield of the coprecipitation protein is improved.
Specifically, under the coexistence environment of lysozyme, lysine, arginine and glutamic acid, the method is favorable for protein renaturation, promotes the protein to be correctly folded into a natural active state, avoids the protein from generating irreversible aggregation and precipitation before the protein is formed into coprecipitation protein, and ensures the interaction of tilapia protein and soybean meal protein at the isoelectric point; meanwhile, the lecithin, mannose and silicon dioxide can improve the uniformity and stability of the whole reaction system, so that the tilapia mossambica protein and the bean pulp protein do not have undesirable reaction near the isoelectric point.
Further, the cosolvent comprises the following components in parts by weight: 3-7 parts of lecithin, 14-16 parts of mannose, 4-7 parts of silicon dioxide, 0.3-0.6 part of lysozyme, 13-17 parts of lysine, 15-20 parts of arginine and 9-14 parts of glutamic acid.
Further, the cosolvent comprises the following components in parts by weight: lecithin 5, mannose 15, silicon dioxide 5.5, lysozyme 0.4, lysine 14, arginine 17 and glutamic acid 11. In particular, the enzyme activity of the lysozyme is 100-300U/g, and preferably, the enzyme activity of the lysozyme is 150U/g.
The innovation points of the invention are as follows: the tilapia protein is unstable, easy to change and poor in solubility in the actual production and processing process, so that the structural and functional characteristics of the protein are changed, and the gel characteristic and the emulsibility of the tilapia protein are low. The tilapia-soybean meal coprecipitation protein is developed and prepared by combining the advantages of better solubility, emulsification stability, foamability, good sensory characteristics and the like of the soybean protein isolate, so that the solubility, the emulsibility, the gel characteristics and the like of fish protein are well improved, and a novel animal and plant protein combined double-protein product is developed. The tilapia-soybean meal coprecipitation protein prepared by the method has high yield, is safe and stable, and provides an effective way for improving the quality of tilapia products, expanding the application method of the tilapia products in the food field and improving the overall profit margin of tilapia.
Detailed Description
The technical solutions of the present invention will be described below clearly and completely in conjunction with the embodiments, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
Example 1
A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia mossambica meat and defatted soybean meal powder in a mass ratio of 1:5 (dry basis), and adding ice distilled water into the mixture according to the mass ratio of water =1: 5(g: mL);
s2.10000r/min for homogenizing for 1 min;
s3, adjusting the pH value of the homogeneous mixture to 3.0, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, stirring at room temperature and dissolving for 5 min;
s centrifuging at 8000 r/min for 10 min at 5.4 deg.C;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
s7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and carrying out vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitation protein.
Further, in step S6, the pH is adjusted to 4.5.
Further, the cosolvent comprises the following components in parts by weight: lecithin 1, mannose 12, silicon dioxide 3, lysozyme 0.2 at 150U/g, lysine 11, arginine 14 and glutamic acid 7. In the invention, the added cosolvent has the function of enhancing the interaction of the protein near the isoelectric point in the process of forming the tilapia-soybean meal coprecipitated protein, so that the tilapia protein and the soybean meal protein are further combined to form a stable coprecipitated protein structure, and the yield of the coprecipitated protein is improved.
Example 2
A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 5:1 (dry basis), and adding ice distilled water according to the mass ratio of water =1: 15(g: mL);
s2.15000r/min homogenizing for 5 min;
s3, adjusting the pH value of the homogeneous mixture to 8.0, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, stirring and dissolving for 30min at room temperature;
s centrifuging at 11000r/min for 20min at 5.4 ℃;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
s7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and carrying out vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitation protein.
Further, in step S6, the pH is adjusted to 4.5.
Further, the cosolvent comprises the following components in parts by weight: lecithin 10, mannose 19, silicon dioxide 9, 150U/g lysozyme 0.9, lysine 18, arginine 22 and glutamic acid 16. In the invention, the added cosolvent has the function of enhancing the interaction of the protein near the isoelectric point in the process of forming the tilapia-soybean meal coprecipitated protein, so that the tilapia protein and the soybean meal protein are further combined to form a stable coprecipitated protein structure, and the yield of the coprecipitated protein is improved.
Example 3
A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 1:2 (dry basis), and adding ice distilled water according to the ratio of material: water =1:7 (g: mL);
s2.10000r/min for homogenizing for 2.5 min;
s3, adjusting the pH value of the homogeneous mixture to 4.0, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, and stirring and dissolving for 10 min at room temperature;
s, centrifuging at 9000 r/min for 12min at 5.4 ℃;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
s7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and carrying out vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitation protein.
Further, in step S6, the pH was adjusted to 4.0.
Further, the cosolvent comprises the following components in parts by weight: lecithin 3, mannose 14, silicon dioxide 4, 150U/g lysozyme 0.3, lysine 13, arginine 15 and glutamic acid 9.
Example 4
A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 2:1 (dry basis), and adding ice distilled water according to the ratio of material: water =1: 10(g: mL);
s2.13000r/min for 3.5 min;
s3, adjusting the pH value of the homogeneous mixture to 6.0, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, stirring and dissolving at room temperature for 25 min;
s, centrifuging the mixture for 17 min at the temperature of 5.4 ℃ and the speed of 10000 r/min;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
and S7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and performing vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitated protein.
Further, in step S6, the pH was adjusted to 6.0.
Further, the cosolvent comprises the following components in parts by weight: lecithin 7, mannose 16, silicon dioxide 7, 150U/g lysozyme 0.6, lysine 17, arginine 20 and glutamic acid 14.
Example 5
A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 2:3 (dry basis), and adding ice distilled water according to the mass ratio of water =1:9(g: mL);
s2.12000r/min homogenizing for 2 min;
s3, adjusting the pH value of the homogeneous mixture to 4.2, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, and stirring and dissolving for 20min at room temperature;
s5.4 ℃, centrifuging for 15 min at 9500 r/min;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
s7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and carrying out vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitation protein.
Further, in step S6, the pH is adjusted to 4.5.
Further, the cosolvent comprises the following components in parts by weight: lecithin 5, mannose 15, silicon dioxide 5.5, 150U/g lysozyme 0.4, lysine 14, arginine 17 and glutamic acid 11.
Comparative example 1
A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 2:3 (dry basis), and adding ice distilled water according to the mass ratio of water =1:9(g: mL);
s2.12000r/min homogenizing for 2 min;
s3, adjusting the pH value of the homogeneous mixture to 4.2, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, stirring at room temperature and dissolving for 20 min;
s5.4 ℃, centrifuging for 15 min at 9500 r/min;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
and S7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and performing vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitated protein.
Further, in step S6, the pH is adjusted to 4.5.
Further, the cosolvent comprises the following components in parts by weight: lecithin 5, mannose 15 and silicon dioxide 5.5.
Comparative example 2
A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 2:3 (dry basis), and adding ice distilled water into the mixture according to the ratio of material: water =1:9(g: mL);
s2.12000r/min homogenizing for 2 min;
s3, adjusting the pH value of the homogeneous mixture to 4.2, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, stirring at room temperature and dissolving for 20 min;
s, centrifuging for 15 min at 5.4 ℃ and 9500 r/min;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
and S7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and performing vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitated protein.
Further, in step S6, the pH is adjusted to 4.5.
Further, the cosolvent comprises the following components in parts by weight: 150U/g lysozyme 0.4, lysine 14, arginine 17, glutamic acid 11.
Comparative example 3
A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 2:3 (dry basis), and adding ice distilled water into the mixture according to the ratio of material: water =1:9(g: mL);
s2.12000r/min homogenizing for 2 min;
s3, adjusting the pH value of the homogeneous mixture to 4.2, and standing for 5 min;
s4, stirring and dissolving for 20min at room temperature;
s5.4 ℃, centrifuging for 15 min at 9500 r/min;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
s7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and carrying out vacuum freeze drying to obtain the tilapia mossambica-soybean meal coprecipitation protein.
Further, in step S6, the pH is adjusted to 4.5.
Experimental testing
Yield of coprecipitated protein
SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) analysis is carried out on the supernatant of the example 1-5 and the supernatant of the comparative example 1-3 to obtain the content of soluble protein, and the yield of the precipitated protein, namely the crude yield of the coprecipitated protein, is obtained by deducting the content of the soluble protein. SDS-PAGE electrophoresis was performed using a separation gel concentration of 12% and a concentration gel concentration of 5%. Mixing the sample supernatant with SDS-PAGE sample buffer (4: 1, v/v), boiling for 6 min before adding into the electrophoresis tank, wherein the volume is equal to 10 μ L, staining with R-250 Coomassie brilliant blue, decolorizing with acetic acid solution of high methanol, namely adding 75 mL acetic acid and 50 mL methanol, and adding water to volume of 1L. The results are shown in the following table:
emulsification Performance test of coprecipitated proteins
The concentration of the prepared protein is 0.5%, and the turbidity is adoptedThe method measures the Emulsifying Activity (EA) and the Emulsifying Stability (ES) of the protein. 6 mL of sample was put into a 50 mL small beaker, and 2 mL of first-class soybean oil of Syngnathus was added thereto, followed by homogenization for 1 min with a homogenizer (fixed rotation speed 12000 r/min). Using UV light at 500 nm with 0.1% SDS as blank, rapidly taking 20 from the bottom of the beakerμThe L emulsion was dissolved in 4 mL of 0.1% SDS solution, shaken, and the absorbance value at 0min was measured (A 0 ) (ii) a Standing for 10 min, and taking 20μL emulsion determination of the absorbance value (A 10 ). EA is expressed as the Emulsion Activity Index (EAI) and ES is expressed as the Emulsion Stability Index (ESI). The formula is as follows:
wherein:Dd is 200 for dilution factor;Cin terms of the concentration of the protein,C=0.5%（g/mL）；Φis the number of oil volume fractions,Φ=1/4；Lis the thickness of the cuvette and is,L=1 cm；△tand 10 min. The results are shown in the following table:
it will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present specification describes embodiments, not every embodiment includes only a single embodiment, and such description is for clarity purposes only, and it is to be understood that all embodiments may be combined as appropriate by one of ordinary skill in the art to form other embodiments as will be apparent to those of skill in the art from the description herein. It should be noted that the technical features not described in detail in the present invention can be implemented by any prior art in the field.
Claims (4)
1. A method for preparing tilapia mossambica-soybean meal coprecipitation protein is characterized by comprising the following steps:
s1, mixing tilapia meat and defatted soybean meal powder in a mass ratio of 1:5-5:1 (dry basis), and adding ice distilled water into the mixture according to the mass ratio of water =1:5-15(g: mL);
s2.10000-15000r/min for homogenizing for 1-5 min;
s3, adjusting the pH value of the homogeneous mixture to be 4.0-6.0, and standing for 5 min;
s4, adding a cosolvent with the mass ratio of 2%, stirring at room temperature and dissolving for 5-30 min; the cosolvent comprises the following components in parts by weight: 1-10 parts of lecithin, 12-19 parts of mannose, 3-9 parts of silicon dioxide, 0.2-0.9 part of lysozyme, 11-18 parts of lysine, 14-22 parts of arginine and 7-16 parts of glutamic acid;
s5.4 ℃, 8000-;
s6, taking the supernatant, adjusting the pH value to enable the supernatant to precipitate, and repeating the step S5;
s7, collecting the precipitates obtained after centrifugation in the steps S5 and S6, and carrying out vacuum freeze drying to obtain tilapia mossambica-soybean meal coprecipitation protein;
in step S6, the pH is adjusted to 4.0-6.0.
2. The method for preparing tilapia mossambica-soybean meal coprecipitated protein according to claim 1, wherein in step S1, tilapia mossambica meat and defatted soybean meal powder are mixed in a mass ratio of 1:2-2:1 (dry basis), and ice distilled water is added according to material: water =1:7-10(g: mL).
3. The method for preparing tilapia mossambica-soybean meal coprecipitation protein according to claim 1, wherein the cosolvent comprises the following components in parts by weight: 3-7 parts of lecithin, 14-16 parts of mannose, 4-7 parts of silicon dioxide, 0.3-0.6 part of lysozyme, 13-17 parts of lysine, 15-20 parts of arginine and 9-14 parts of glutamic acid.
4. The method for preparing tilapia mossambica-soybean meal coprecipitation protein according to claim 3, wherein the cosolvent comprises the following components in parts by weight: lecithin 5, mannose 15, silicon dioxide 5.5, lysozyme 0.4, lysine 14, arginine 17 and glutamic acid 11.
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