CN111513263A - Method for improving animal cell source artificial meat morphology - Google Patents

Method for improving animal cell source artificial meat morphology Download PDF

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Publication number
CN111513263A
CN111513263A CN201910702856.6A CN201910702856A CN111513263A CN 111513263 A CN111513263 A CN 111513263A CN 201910702856 A CN201910702856 A CN 201910702856A CN 111513263 A CN111513263 A CN 111513263A
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meat
weight
animal
glutamine transaminase
added
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刘松
王兴隆
周景文
李江华
陈坚
堵国成
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Jiangnan University
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Jiangnan University
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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
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/10—Meat meal or powder; Granules, agglomerates or flakes
    • 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
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
    • A23L13/42—Additives other than enzymes or microorganisms in meat products or meat meals
    • A23L13/424—Addition of non-meat animal protein material, e.g. blood, egg, dairy products, fish; Proteins from microorganisms, yeasts or fungi
    • 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
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
    • A23L13/42—Additives other than enzymes or microorganisms in meat products or meat meals
    • A23L13/432—Addition of inorganic compounds, e.g. minerals; oligo-elements
    • 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
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
    • A23L13/48—Addition of, or treatment with, enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues

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  • Food Science & Technology (AREA)
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  • Meat, Egg Or Seafood Products (AREA)

Abstract

The invention discloses a method for improving animal cell source artificial meat morphology, and belongs to the technical field of food engineering. Mixing meat paste obtained by culturing animal stem cells in different parts by weight, salt, phosphate, water, casein powder and glutamine transaminase. Meanwhile, the invention provides a treatment method aiming at the animal cell culture finished product. The addition of glutamine transaminase promotes animal cells to be subjected to mass crosslinking, a mixture of phosphate, salt and water provides reaction conditions, and the addition of casein provides a bridge for the adhesion of the glutamine transaminase, so that the crosslinking degree is deepened. The meat analogue treated by the method has high compactness and good taste.

Description

Method for improving animal cell source artificial meat morphology
Technical Field
The invention relates to a method for improving the morphology of artificial meat derived from animal cells, belonging to the technical field of food engineering.
Background
Animal stem cell-derived meat analogue is a major strategy to solve food supply in the near future, and meat analogue cultured by animal stem cells has many advantages including controllable nutrients, no pollution, fast production speed, etc. However, because the meat analogue is cultured by in vitro cell culture, depending on the culture medium and culture dish, compared with the animal culture process, the meat quality produced by the training and growth of the animal body is relatively loose and is meat paste.
The glutamine transaminase is a natural adhesive which is commonly used in food, can catalyze protein in the food to generate covalent crosslinking, and can improve the shape and the phase of meat and improve the meat density and the lumpiness degree in the meat processing. At present, meat analogue cultured by animal stem cells has the problems of loose meat quality and poor taste, and glutamine transaminase has not been subjected to bonding attempt in the meat analogue, and the reaction time, the dosage and the bonding degree are unknown.
Therefore, it is an object of the present invention to provide a method for improving the morphological appearance of an artificial meat derived from animal cells, which is useful for industrial processing of artificial meat.
Disclosure of Invention
The invention aims to solve the problems of poor shape and taste of the artificial meat. The addition of glutamine transaminase and casein can promote deep crosslinking of animal cell surface glycoprotein, so as to increase compactness and improve sensory effect and taste.
The first purpose of the invention is to provide a method for improving the morphology of artificial meat derived from animal cells, which comprises the steps of mixing 74-88% of meat paste obtained by culturing animal stem cells, 0.5-2% of salt, 0.5-2% of phosphate, 10-20% of water, 0.5-1% of casein powder and 0.5-2% of glutamine transaminase of 100-120U/g substrate in parts by weight, and stirring after mixing.
In one embodiment of the invention, the meat paste obtained by culturing the animal stem cells is added in an amount of 80-85% by weight.
In one embodiment of the invention, the salt is added in an amount of 0.5-1% by weight.
In one embodiment of the present invention, the phosphate is added in an amount of 0.5 to 1% by weight.
In one embodiment of the present invention, the water is added in an amount of 15 to 20% by weight.
In one embodiment of the invention, the casein powder is added in an amount of 0.5-0.8% by weight.
In one embodiment of the invention, 100-120U/g substrate is added in an amount of 0.5-1% by weight.
In one embodiment of the invention, the stirring temperature is 23-27 ℃.
In one embodiment of the invention, the meat mixer is used for mixing for 10-25min at the rotation speed of 1000-.
Another object of the present invention is to provide an animal cell-derived meat analogue obtained by the above method.
The invention has the beneficial effects that:
(1) the animal stem cell meat analogue of the invention is added with casein and glutamine transaminase to catalyze the cross-linking reaction. Transglutaminase is an enzyme that catalyzes acyl group transfer between a gamma-carboxamide group (acyl donor) of a glutamine residue in a peptide chain and an acyl acceptor. When the acyl acceptor is primary amino group, the connection between protein molecule and small molecule primary amine is formed, when-amino group of lysine residue in polypeptide chain is acyl acceptor, the- (gamma-glutamic amino) lysine isopeptide bond (G-L bond) is formed in and between protein molecules, and the protein is cross-linked. Through the mode, the dispersed stem cells are catalytically aggregated into the blocks, the formed artificial meat has a more compact mouthfeel, and the problems that other recombined meat is not resistant to heating or meat blocks are scattered after heating and the like do not exist.
(2) The raw materials adopted by the invention are animal stem cells, and the meat blocks are formed by adhesion, so that the problem of shortage of meat resources is solved, and the meat-flavored fish is economical and convenient and has more vivid mouthfeel.
(3) Compared with the traditional pectin method, the artificial meat prepared by the invention has the advantages of good bonding effect, good thermal stability, no other peculiar smell, small taste difference between the prepared artificial meat and real meat, no obvious bonding trace and better appearance.
Drawings
FIG. 1: the instrument automatically gives a time-force value curve.
Detailed Description
Method for measuring enzyme activity of (mono) glutamine transaminase
1. Preparation of Standard Curve
Solution A: tris 9.688g, NH2OH-HCl 2.78G (100mM), GSH 1.229G, Z-Q-G4.048G (30mM) in 400mL water.
And B, liquid B: 3mol/L HCl, 12% trichloroacetic acid, 5% FeCl3Dissolved in 0.1mol/L HCl and the three solutions were mixed in equal amounts when used.
Weighing 64.8mg of standard L-glutamic acid-gamma-monohydroxamic acid (MW 162.1) and 10mL of 0.2mol/L Tris-HCl buffer (pH 6.0, same as solution A, but without hydroxylamine, GSH and substrate); sequentially diluting 5 gradients by a 2-fold dilution method with 0.2mol/L Tris-HCl buffer solution to obtain standard solutions with the concentrations of 6.48mg/mL, 3.24mg/mL, 1.62mg/mL, 0.81mg/mL and 0.405 mg/mL; respectively sucking up the standard solutions by a pipette, sucking up 200 mu L of each standard solution, placing the standard solution in a test tube, and preserving the temperature for 1min at 37 ℃; accurately adding 2mL of solution A preheated at 37 ℃ for 10min, and uniformly mixing by using an oscillating mixer; preserving the heat at 37 ℃ for 10min, then adding 2mL of liquid B, and uniformly mixing by using an oscillating mixer; separating insoluble substances at 3000rpm for 10 min; the absorbance of the supernatant was measured at 525nm 30min after addition of solution B.
Taking the amount of the absorbance to the hydroxamic acid as a straight line, obtaining a conversion coefficient K from the slope of the straight line, and calculating the production amount of the hydroxamic acid through K after the absorbance is obtained in the determination of the enzyme activity of the sample.
2. Enzyme activity assay
500mg of a sample to be tested was weighed into a 50mL beaker, and about 45mL of 0.2mol/L Tris-HCl buffer (pH 6.0) was added. Stir with a magnetic stirrer for 30min at room temperature. Transferring the sample to a 50mL volumetric flask, and diluting the volume to 50mL by using 0.2mol/L Tris-HCl buffer solution to be used as a sample to be detected. Accurately sucking 200 mu L of unknown sample to be detected into a test tube, and preserving the temperature for 1min at 37 ℃. 2mL of solution A preheated at 37 ℃ for 10min was added accurately, and mixed well by shaking the mixer. Then preserving the heat at 37 ℃ for 10min, then adding 2mL of liquid B, and uniformly mixing by using a shaking mixer. The insoluble matter was separated at 3000rpm for 10 min. The absorbance (At) of the supernatant was measured At 525nm 30min after addition of solution B.
Blank control group: pipette 200. mu.L of the sample to be tested into a test tube, and incubate at 37 ℃ for 1 min. Add 2mL of solution B and mix well with shaking the mixer. Preserving the heat at 37 ℃ for 10min, then adding 2mL of solution A which is preheated at 37 ℃ for 10min, and uniformly mixing by oscillating a mixer. Insoluble matter was separated at 3000rpm for 10 min. The absorbance (Ab) of the supernatant was measured at 525nm 30min after addition of solution B.
Definition of enzyme activity: one unit of enzyme activity is defined as the amount of enzyme that catalyzes the formation of a product from 1. mu. mol of substrate per minute.
1U-1 μmol product (or substrate)/min
TG enzyme activity (u/g) ═ K × (At-Ab) × (1/S) × (1/10)
K-conversion factor
S-weight of sample measured (g)
10-reaction time (min)
Method for measuring hardness and elasticity
Elasticity:
shear force assessment: the method comprises the steps of using a domestic C-LM2 type muscle tenderness meter, firstly carrying out pointer zero setting, taking a processed meat product sample, putting the processed meat product sample into a knife hole, pressing a switch to cut off the sample, recording kg data of a pointer on the tenderness meter, multiplying the kg data by a constant 0.2 according to the specification of the instrument, and obtaining the product of the constant and the shear force Fs.
Hardness:
1. test instrument and tool
Xlw (ec) intelligent electronic tensile tester, denland electromechanical technologies ltd. And a cylindrical probe with a diameter of 5 mm.
2. Procedure of the test
The meat product was prepared as a square sample having a height of 25mm and a bottom area of 40X 40 mm. The probe is mounted on the upper jaw of a tensile machine. The compression test by the tensile machine was started, and the sample was compressed at a rate of 1mm/s until the compression distance was 10mm, and then returned at the same rate. After a pause of 2s, the second compression was continued at the same speed, again for a compression distance of 10 mm. The instrument automatically gives a time-force value curve, as shown in fig. 1. And then calculating corresponding mass structure characteristic indexes in the following way:
hardness: the maximum peak in the first compression of the pressure curve is the internal binding force of the sample holding shape.
Example 1:
(1) pretreatment of raw materials
Cleaning the animal stem cell meat paste, and refrigerating at 0-4 ℃ for later use.
(2) Glutamine transaminase treatment of animal stem cells
Preparing raw material components of the impregnation liquid: 74-88% of minced meat, 0.5-2% of salt, 0.5-2% of phosphate, 10-20% of water, 0.5-1% of casein powder and 0.5-2% of glutamine transaminase.
If the concentration of salt or phosphate exceeds 2%, the taste of the meat is affected, and the meat loses too much water, while if the concentration is less than 0.5%, the catalysis by glutamine transaminase is not suitable. Because the main linking objects of the glutamine transaminase are glutamine and amino residues, and casein has a plurality of lysine and glutamine sites, economic cost is saved, and a linking bridge is provided for the glutamine transaminase, so that meat quality is linked more closely, and the taste is better. The use of glutamine transaminase in amounts below 0.5% requires a longer reaction time, while use of more than 2% leads to an excessively high product hardness.
The preparation process of the impregnation liquid comprises the following steps: adding weighed water into a stirrer, stirring at a high speed for 2 minutes, sequentially adding salt and phosphate, fully and uniformly mixing with casein, and then adding into the high-speed stirrer to stir for 10 minutes.
Dipping and rolling: putting the refrigerated animal stem cell minced meat into a rolling pot for dipping and rolling, then putting into a stirrer for stirring, and slowly adding glutamine transaminase during stirring to fully and uniformly mix the transaminase and the meat.
And stacking the stirred meat blocks in a special mold, and beating with fingers to remove air bubbles. The meat blocks with exhausted gas are put in a specific environment for enzyme catalytic reaction, the temperature is controlled at 23-27 ℃, and the time is set at 20 minutes. If the temperature is too high, the meat is easy to rot.
(3) Post-processing of
After the enzyme catalysis reaction is finished, quickly freezing at the temperature below minus 40 ℃, and freezing the finished product at the temperature below minus 20 ℃. This results in the formation of an animal stem cell meat analogue product. The hardness of the artificial meat is above 6500g, and the elasticity of the artificial meat is above 3000 g.
Example 2
The hardness of the meat analogue was 6025. + -. 112g, which was obtained by adjusting glutamine transaminase in the examples to 0.4% under the same conditions.
Example 3
The casein in the examples was adjusted to 0.3% and the conditions were otherwise unchanged, giving a meat analogue hardness of 6123. + -. 78 g.
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 in the appended claims.

Claims (10)

1. A method for improving the morphology of artificial meat derived from animal cells is characterized in that meat paste obtained by culturing animal dry cells is mixed with 0.5-2% of salt, 0.5-2% of phosphate, 10-20% of water, 0.5-1% of casein powder and 0.5-2% of glutamine transaminase with substrate of 100U/g and 120U/g in parts by weight, and the mixture is stirred after being mixed.
2. The method of claim 1, wherein the meat emulsion obtained by culturing the animal stem cells is 80-85% by weight.
3. The method of claim 1, wherein the salt is added in an amount of 0.5% to 1% by weight.
4. The method of claim 1, wherein the phosphate is added in an amount of 0.5% to 1% by weight.
5. The method of claim 1, wherein the moisture is added in an amount of 15 to 20% by weight.
6. The method of claim 1, wherein the casein powder is added in an amount of 0.5-0.8% by weight.
7. The process as claimed in claim 1, wherein 100-120U/g substrate is added in an amount of 0.5-1% by weight.
8. The method of claim 1, wherein the stirring temperature is 23-27 ℃.
9. The method as claimed in claim 1 or 8, wherein the stirring is carried out at a rotation speed of 1000-.
10. An animal cell-derived meat analogue obtained by the method according to any one of claims 1 to 9.
CN201910702856.6A 2019-07-31 2019-07-31 Method for improving animal cell source artificial meat morphology Pending CN111513263A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111518781A (en) * 2019-07-31 2020-08-11 江南大学 Glutamine transaminase complex enzyme and application thereof in artificial meat processing
WO2022261647A1 (en) * 2021-06-11 2022-12-15 Upside Foods, Inc. Comestible cell-based meat products comprising dry cell powder and methods of making such products

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103932223A (en) * 2014-04-30 2014-07-23 北京华都肉鸡公司 Chicken product prepared by restructuring ground meat and processing method of chicken product
CN106923216A (en) * 2017-02-28 2017-07-07 河南众品食业股份有限公司 A kind of meat product adhesive and its application in terms of meat products processing

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103932223A (en) * 2014-04-30 2014-07-23 北京华都肉鸡公司 Chicken product prepared by restructuring ground meat and processing method of chicken product
CN106923216A (en) * 2017-02-28 2017-07-07 河南众品食业股份有限公司 A kind of meat product adhesive and its application in terms of meat products processing

Non-Patent Citations (1)

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Title
赵鑫锐等: ""人造肉大规模生产的商品化技术"", 《食品与发酵工业》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111518781A (en) * 2019-07-31 2020-08-11 江南大学 Glutamine transaminase complex enzyme and application thereof in artificial meat processing
WO2022261647A1 (en) * 2021-06-11 2022-12-15 Upside Foods, Inc. Comestible cell-based meat products comprising dry cell powder and methods of making such products
CN117500385A (en) * 2021-06-11 2024-02-02 阿普赛德食品公司 Cell-based edible meat product comprising dried cell powder and method of making the same

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Application publication date: 20200811