阅读说明：本技术 一种提高动物细胞来源人造肉形态品相的方法 (Method for improving animal cell source artificial meat morphology ) 是由 刘松 王兴隆 周景文 李江华 陈坚 堵国成 于 2019-07-31 设计创作，主要内容包括：本发明公开了一种提高动物细胞来源人造肉形态品相的方法,属于食品工程技术领域。是将不同重量份数的动物干细胞培养所得肉糜、食盐、磷酸盐、水分、酪蛋白粉、谷氨酰胺转氨酶进行混合。同时,针对动物细胞培养成品,本发明提供了处理方法。通过添加谷氨酰胺转氨酶促使动物细胞进行大量交联,磷酸盐、食盐及水的混合物提供了反应条件,而酪蛋白的加入为谷氨酰胺转氨酶粘合提供了桥梁,以此加深了交联程度。通过该方法处理的人造肉具有高致密程度及口感。(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.)

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.

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.