Application of hemoglobin in plant-based / microorganism-based protein simulated meat food and biofortified drink
By preparing histidine-heme aggregate complexes, the problem of heme's insolubility in water was solved, improving the color and flavor of plant-based/microbial-based simulated meat products and reducing production costs.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-10-29
- Publication Date
- 2026-03-27
Smart Images
Figure HDA0004521541340000011 
Figure HDA0004521541340000012 
Figure HDA0004521541340000021
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of food technology, in particular to the application of hematin in plant-based / microorganism-based protein simulated meat food and bio-iron fortified liquid and solid drinks. BACKGROUND
[0002] Plant-based simulated meat mainly uses plant protein / microorganism-based as raw material, and processes plant tissue protein / microorganism-based into fiber structure similar to meat through electrospinning, extrusion technology and 3D printing technology, so as to simulate the structure, flavor and taste of real meat. At present, plant-based / microorganism-based simulated meat still has a large gap in texture, color, flavor and taste compared with real meat, and it is urgent to further improve the sensory properties of protein meat. The food pigments currently applied to protein meat include beet red, sorghum red, hemoglobin, etc. Among them, beet red, sorghum red, etc. are deep and unnatural in color, easy to oxidize and lack the luster of meat color, which seriously affects the color and flavor of protein meat; and the preparation cost and technical difficulty of hemoglobin are high, which increases the cost.
[0003] Hematin is known as "life pigment", and is the material basis for the color and fishy smell of real meat. Its preparation cost and technical difficulty are lower than those of hemoglobin, and it is the preferred substitute for hemoglobin. However, there are the following problems: (1) the structural characteristics of hematin make it insoluble in water and can only be dissolved in alkaline aqueous solution and organic reagents, which are not suitable for the preparation of plant-based / microorganism-based simulated protein food; (2) hematin is brown-green in production process, which cannot improve the color and taste of plant protein meat, and seriously hinders the application of hematin in plant-based / microorganism-based simulated protein food.
[0004] Therefore, how to solve the above technical problems is of great significance for reducing the cost of plant-based / microorganism-based simulated protein and effectively improving the color and taste of protein meat. SUMMARY
[0005] To solve the above technical problems, the present application provides an application of hematin in plant-based / microorganism-based protein simulated meat food and bio-iron fortified drinks, which is first prepared into a histidine-hematin aggregate complex under specific conditions, then freeze-dried and re-dissolved, and then used for preparing plant-based / microorganism-based protein simulated meat food or bio-iron fortified drinks.
[0006] The technical scheme of the present application is as follows: the application of hematin in plant-based / microorganism-based protein simulated meat food and bio-iron fortified drinks, comprising the following steps:
[0007] S1, preparation of histidine-heme aggregate complex: heme powder is added to PBS buffer solution with pH of 9.0±0.5, and stirred overnight to ensure heme is fully hydrated to obtain a heme alkaline solution;
[0008] Histidine powder is added to the above heme alkaline solution, and stirred to dissolve, and the obtained solution is reacted at 4±2℃ for 40-48h to obtain an amino acid-heme aggregate complex solution, which is further freeze-dried to obtain histidine-heme aggregate complex;
[0009] S2, the histidine-heme aggregate complex is re-dissolved in deionized water and then used to prepare plant-based protein / microorganism-based simulated meat food or bio-iron fortified beverage.
[0010] Further, the mass ratio of histidine powder to heme powder is (150-200):1.
[0011] Further, the concentration of heme in the heme alkaline solution is 0.1-0.3mg / mL.
[0012] Further, the heme includes but is not limited to chlorinated heme from animal blood, microbial heme, soybean heme or other heme derivative compounds.
[0013] Further, the histidine is L-histidine.
[0014] Further, the bio-iron fortified beverage includes but is not limited to solid beverage, liquid beverage.
[0015] The purpose of the present application is also to protect an amino acid-heme aggregate complex, which is the histidine-heme aggregate complex prepared in the above application.
[0016] The present application solves the problem that heme cannot be used in plant-based / microorganism-based simulated protein food because it is not soluble in water, and the problem that its color is brown-green, and the color and taste of plant / microorganism-based protein are poor, by preparing heme into histidine-heme aggregate complex solution under specific conditions, then freeze-drying it into powder, and further re-dissolving it and using it to prepare plant-based / microorganism-based protein simulated meat food or bio-iron fortified beverage, which is of great significance for reducing the production cost of plant-based protein simulated meat food and bio-iron fortified beverage, and improving its color and taste. BRIEF DESCRIPTION OF DRAWINGS
[0017] Figure 1 A is the ultraviolet absorption spectrum of heme dissolved in PBS solution and heme dissolved in dimethyl sulfoxide solution (DMSO), Figure 1B is the UV absorption spectrum of histidine-heme complex dissolved in PBS solution and heme dissolved in dimethyl sulfoxide solution (DMSO), Figure 1 C is the UV absorption spectrum of heme, histidine-heme complex and histidine dissolved in PBS solution, Figure 1 D is the reaction process prediction;
[0018] Figure 2 is the physical comparison chart of heme and histidine-heme complex;
[0019] Figure 3 is the color (a value) change of histidine-heme complex lyophilized powder of different concentrations after reconstitution in deionized water;
[0020] Figure 4 is the high performance liquid chromatogram of heme chloride and histidine-heme aggregate complex;
[0021] Figure 5 From left to right, respectively, are soybean protein isolate gels containing heme chloride and soybean protein isolate gels containing histidine-heme aggregate complex. DETAILED DESCRIPTION
[0022] The present application will be further described in conjunction with specific examples, so that those skilled in the art can more clearly understand the present application. The following examples are used to illustrate the present application, but are not intended to limit the scope of the present application. Based on the specific examples in the present application, all other examples obtained by those skilled in the art without making creative efforts are within the scope of protection of the present application.
[0023] In the embodiments of the present application, all raw material components are commercially available products well known to those skilled in the art unless otherwise specified; in the embodiments of the present application, the technical means used are conventional means well known to those skilled in the art unless otherwise specified.
[0024] The application of heme in plant-based / microorganism-based protein meat food simulation and bio-iron fortified drinks includes the following steps:
[0025] S1, preparation of histidine-heme aggregate complex: add heme powder to PBS buffer solution with pH of 9.0±0.5, stir overnight to ensure heme is fully hydrated, and obtain heme alkaline solution;
[0026] Add histidine powder to the above heme alkaline solution, stir to dissolve, and obtain the solution, which is further reacted at 4±2℃ for 40-48h to obtain an amino acid-heme aggregate complex solution, which is further lyophilized to obtain histidine-heme aggregate complex;
[0027] S2, the histidine-haem aggregate complex is dissolved in deionized water and then used to prepare plant-based protein simulated meat food or bio-iron fortified liquid and solid drinks.
[0028] The mass ratio of histidine powder to haem powder is (150-200): 1, preferably 200: 1. The concentration of haem in the haem alkaline solution is 0.1-0.3 mg / mL, preferably 0.2 mg / mL. The haem includes but is not limited to chlorinated haem from animal blood or other haem derivatives. The histidine is preferably L-histidine. The bio-iron fortified drink includes but is not limited to solid drinks, liquid drinks.
[0029] The application also provides an amino acid-haem aggregate complex, which is the histidine-haem aggregate complex prepared as described above. The amino acid-haem aggregate complex powder maintains good red color after being dissolved in deionized water and can be used to improve the color and provide a fishy smell of plant-based / microorganism-based simulated meat. It can also be widely used as an iron supplement in bio-iron fortified liquid and solid drinks and other types of food.
[0030] Example 1
[0031] This example provides an amino acid-haem aggregate complex, specifically a histidine-haem aggregate complex, which is prepared as follows:
[0032] (1) A 10 mM PBS buffer solution with a pH of 9.0 is prepared using deionized water, and haem powder is slowly added to the PBS and stirred overnight to ensure that the haem is fully hydrated, thereby obtaining a 0.2 mg / mL haem alkaline solution;
[0033] (2) L-histidine is added to the chlorinated haem solution at a mass ratio of L-histidine powder: haem powder = 200: 1, the solution is stirred, and the reaction is carried out at 4°C for 48 h, thereby obtaining an amino acid-haem aggregate complex solution;
[0034] (3) The amino acid-haem aggregate complex solution is freeze-dried at -40°C to obtain the product.
[0035] Comparative Example 1
[0036] This comparative example provides an amino acid-haem aggregate complex, specifically a histidine-haem aggregate complex, which is prepared as follows:
[0037] (1) hematin powder was slowly added into PBS buffer solution of 10 mM, pH 9.0 prepared with deionized water, and the hematin was fully hydrated by stirring overnight to obtain a 0.2 mg / mL hematin basic solution;
[0038] (2) L-histidine was added into the hematin chloride solution according to a mass ratio of L-histidine powder:hematin powder = 200:1, the solution was stirred and reacted at 4°C for 4 h to obtain an amino acid-hematin aggregate complex solution;
[0039] (3) The amino acid-hematin aggregate complex solution was freeze-dried at -40°C to obtain the product.
[0040] Comparative Example 2
[0041] The comparative example provides an amino acid-hematin aggregate complex, specifically a histidine-hematin aggregate complex, which is prepared as follows:
[0042] (1) hematin powder was slowly added into PBS buffer solution of 10 mM, pH 9.0 prepared with deionized water, and the hematin was fully hydrated by stirring overnight to obtain a 0.2 mg / mL hematin basic solution;
[0043] (2) L-histidine was added into the hematin chloride solution according to a mass ratio of L-histidine powder:hematin powder = 200:1, the solution was stirred and reacted at 25°C for 4 h to obtain an amino acid-hematin aggregate complex solution;
[0044] (3) The amino acid-hematin aggregate complex solution was freeze-dried at -40°C to obtain the product.
[0045] Comparative Example 3
[0046] The comparative example provides an amino acid-hematin aggregate complex, specifically a histidine-hematin aggregate complex, which is prepared as follows:
[0047] (1) hematin powder was slowly added into PBS buffer solution of 10 mM, pH 9.0 prepared with deionized water, and the hematin was fully hydrated by stirring overnight to obtain a 0.2 mg / mL hematin basic solution;
[0048] (2) L-histidine was added into the hematin chloride solution according to a mass ratio of L-histidine powder:hematin powder = 200:1, the solution was stirred and reacted at 25°C for 4 h to obtain an amino acid-hematin aggregate complex solution;
[0049] (3) The amino acid-hematin aggregate complex solution was freeze-dried at -40°C to obtain the product.
[0050] Comparative Example 4
[0051] This comparative example provides an amino acid-heme aggregate complex, specifically an arginine-heme aggregate complex, which is prepared as follows:
[0052] (1) A PBS buffer solution with pH 9.0 was prepared using deionized water, and heme powder was slowly added to the PBS, and stirred overnight to ensure that the heme was fully hydrated, to obtain a 0.2 mg / mL basic solution of heme;
[0053] (2) Arginine was added to the solution of chlorinated heme according to a mass ratio of L-histidine powder:heme powder = 200:1, the solution was stirred, and reacted at 25°C for 4 h, to obtain an amino acid-heme aggregate complex solution;
[0054] (3) The amino acid-heme aggregate complex solution was freeze-dried at -40°C to obtain the product.
[0055] Results analysis:
[0056] (1) The amino acid-heme aggregate complex solution obtained in step (2) in the examples and comparative examples was subjected to color determination using a color difference meter (UltraScan VIS, HunterLab, Virginia, USA), and the results are shown in Table 1:
[0057] Table 1 Color determination results in the examples and comparative examples
[0058] Item a* value Example 1 40.79±0.23 Comparative Example 1 36.85±0.26 Comparative Example 2 36.89±0.08 Comparative Example 3 35.49±0.21 Comparative Example 4 20.73±0.19
[0059] As can be seen from Table 1, the type of amino acid, temperature, and reaction time all affect the improvement effect on the color of heme.
[0060] (2) Determination of coordination compounds
[0061] UV-Vis spectrometer was used to determine the UV spectra of hemin in PBS solution, hemin in dimethyl sulfoxide solution, histidine-hemin complex in PBS solution and histidine in PBS solution, wherein the hemin-PBS solution (Hemin in PBS) was prepared according to the method in step (1) of Example 1; the histidine-hemin complex-PBS solution (Hemin-His in PBS) was the solution prepared in step (2) of Example 1; the hemin-dimethyl sulfoxide solution (Hemin in DMSO) was prepared by dissolving 2 mg of hemin powder in 10 mL of dimethyl sulfoxide; and the histidine-PBS solution (His in PBS) was prepared by dissolving 400 mg of histidine in 10 mL of PBS solution with pH = 9, and the results are shown in Figure 1 .
[0062] As can be seen from Figure 1 A, hemin in alkaline solution generates H-aggregated hemin aggregates; as can be seen from Figure 1 B and 1C, the UV spectrum of the histidine-hemin aggregate complex is obviously changed compared with that of hemin, indicating that histidine reacts with hemin to form a new coordination compound.
[0063] The obtained solution was subjected to color determination using a color difference meter, and the results are shown in Table 2 below:
[0064] Table 2: Test results of hemin-PBS solution and histidine-hemin complex-PBS solution
[0065] L a* b* Hemin 28.42±0.20 17.64±0.11 44.51±0.36 His-Hemin 24.85±0.18 40.79±0.23 39.24±0.24
[0066] As can be seen from Table 2, the a* value of the histidine-hemin complex obtained by reaction is significantly improved compared with hemin. At the same time, from the comparison of the actual objects in Figure 2 , it can be seen that the addition of histidine changes the color of the hemin solution from yellow-green to hemin.
[0067] The histidine-hemin aggregate complex powder obtained in step (3) of Example 1 was re-dissolved in deionized water to prepare solutions with different concentrations (5 mg / mL, 10 mg / mL, 20 mg / mL, 40 mg / mL), and the color was determined using a color difference meter, and the results are shown in Figure 3 . With the increasing concentration of the complex lyophilized powder, the a* value of the solution continuously rises, and when the concentration is 40 mg / mL, the color of the re-dissolved solution is basically the same as that of the solution before lyophilization.
[0068] (3) The solutions (40 mg / mL) of the heme chloride aqueous solution and the histidine-heme aggregate complex obtained in step (3) of Example 1, after redissolving in water, were determined using high performance liquid chromatography (HPLC). The heme chloride aqueous solution was prepared by adding 1 g of heme chloride to 100 mL of deionized water, stirring thoroughly, centrifuging, and collecting the supernatant to obtain the heme chloride aqueous solution. The HPLC conditions were as follows:
[0069] Chromatographic column: C18 column, 250 mm × 4.6 mm, particle size 5 μm; mobile phase: methanol: 0.6 acetic acid = 70:30 (volume ratio); flow rate: 1 mL / min; detection wavelength: 399 nm.
[0070] Heme solutions at concentrations of 0.25, 0.2, 0.15, 0.1, and 0.05 mg / mL were prepared using heme standards. The linear relationship between heme concentration and peak area was determined using high-performance liquid chromatography (HPLC), with the formula: y = 125754x + 669.41(R²). 2 =0.9993), from Figure 4 As can be seen, the peak area of heme content in heme chloride is 24, and the peak area of heme content in histidine-heme polymer complex solution is 14940.9. Substituting the peak areas into the above formula, we obtain that the solubility of heme in histidine-heme aggregate complex solution in step (2) of Example 1 is 0.1135 mg / mL.
[0071] The above results show that heme chloride is almost insoluble in deionized water. The histidine-heme aggregate complex obtained by the method of the present invention has a heme solubility of 0.11 mg / mL in deionized water. The method of the present invention can significantly improve the solubility of heme in neutral water.
[0072] (4) Application of the histidine-heme aggregate complex obtained in Example 1 in plant-based proteins.
[0073] The lyophilized histidine-heme aggregate complex obtained in step (3) of Example 1 was dissolved in 50 mL of water to prepare a reconstituted solution with a concentration of 40 mg / mL. Then, 6 g of soy protein isolate powder was added to 50 mL of distilled water and the above reconstituted solution, respectively, and stirred thoroughly. The mixture was then placed at 4°C overnight to ensure complete protein hydration. 20 U / g of glutamyl transaminase was added to the fully hydrated protein solution, and the mixture was stirred rapidly. The reaction was carried out at 50°C for 60 min to obtain soy protein isolate gel. After cooling at room temperature, the gel was placed in a 4°C refrigerator for later use. The soy protein isolate gel and the soy protein isolate gel containing the histidine-heme aggregate complex were photographed using a digital camera to observe their appearance changes. The results are as follows: Figure 5As shown, the addition of the histidine-heme aggregate complex significantly improved the color of the SPI gel.
[0074] It should be pointed out here that the above examples are only for further illustrating and describing the technical solutions of the present application, and are not for further limiting the technical solutions of the present application. The method of the present application is only a preferred embodiment, and is not used for limiting the protection scope of the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims
1. Use of hematin in plant-based / microorganism-based protein simulated meat food and bio-iron fortified beverage, characterized in that, The method comprises the following steps: S1, preparing a histidine-heme aggregate complex: adding heme powder into a PBS buffer solution with a pH of 9.0±0.5, stirring overnight to ensure that the heme is fully hydrated, and obtaining a heme alkaline solution; adding histidine powder into the heme alkaline solution, stirring to dissolve, and reacting the obtained solution at 4±2℃ for 40-48 h to obtain an amino acid-heme aggregate complex solution, which is further freeze-dried to obtain the histidine-heme aggregate complex; S2, re-dissolving the histidine-heme aggregate complex in deionized water and then using it to prepare plant-based / microorganism-based protein simulated meat food or bio-iron fortified liquid and solid drinks; The mass ratio of the histidine powder to the heme powder is (150-200):
1. The histidine is L-histidine.
2. Use of the hematin according to claim 1 in plant-based / microorganism-based protein simulated meat food and bio-iron fortified beverage, characterized in that, The concentration of the heme in the heme alkaline solution is 0.1-0.3 mg / mL.
3. Use of the heme according to claim 1 in plant-based / microorganism-based protein simulated meat food and bio-iron fortified beverage, characterized in that, The heme includes chlorinated heme from animal blood, microbial heme or soybean heme.
4. The use of the hematin according to claim 1 in plant-based / microorganism-based protein simulated meat food and bio-iron fortified beverage, characterized in that, The bio-iron fortified drink includes a solid drink and a liquid drink.
5. An amino acid-heme aggregate complex, characterized in that, The amino acid-heme aggregate complex is the histidine-heme aggregate complex prepared in any one of claims 1-4.
Citation Information
Patent Citations
Purification method of protoheme
CN1537860A
Hemin compound
GB8431398D0