Neurospora crassa HJDF, culture method thereof, product in 3D printing of artificial fish meat and application of product

Abstract

The invention provides neurospora crassa HJDF, a culture method thereof, a product in 3D printing of artificial fish meat and application of the product, and particularly belongs to the technical field of food processing. The Neurospora crassa HJDF disclosed by the invention is preserved in the General Microbiological Culture Collection Center of the China Committee for Culture Collection of Microorganisms, the preservation time is September 23, 2024, and the preservation number is CGMCC No.41518. The Neurospora crassa HJDF disclosed by the invention can be used for preparing the Neurospora crassa HJDF. The strain is high in protein content and complete and balanced in amino acid composition, and has natural'umami '. According to the method, the fiber arrangement of real fish can be effectively simulated by combining the micron-sized hypha protein raw material with a 3D printing method, and a new technical approach is provided for accurately copying the characteristics of the real fish in the future in flavor, nutrition and texture.

Description

Technical Field

[0001] The present invention belongs to the technical field of food processing, and in particular relates to Neurospora crassa HJDF and a culture method thereof, and products and applications thereof in 3D printing of artificial fish meat. Background Art

[0002] Animal meat is an important source of protein and other key nutrients, but with population growth and environmental pressure, traditional animal husbandry can no longer meet future protein needs, and it has an impact on the environment and animal welfare. Therefore, alternative proteins, such as plants, algae, microorganisms and insects, are gaining more and more attention due to their environmental protection and high efficiency. Although plant proteins are often considered to be nutritionally deficient due to incomplete amino acids, and the application of algae and insect proteins is limited by various factors, fungal protein shows great potential due to its relatively rich protein content, high production efficiency, environmental protection and no climate impact.

[0003] Among the many fungal proteins, Neurospora has become the focus because of its natural existence, long history and safety. It can not only decompose complex carbohydrates and proteins, but also grow in cheap carbon sources, with low production costs and high efficiency, making it very suitable for industrial production. In particular, Neurospora crassa, due to its rich protein content, shows great potential in the field of meat substitutes.

[0004] With the continuous advancement of 3D printing technology, this technology has been explored for the manufacture of foods with complex shapes and structures, especially in the preparation of meat substitutes. 3D printing technology can accurately replicate the external form of food, but the current challenge is to simulate the fiber texture and flavor of real fish. Summary of the invention

[0005] The purpose of the present invention is to provide Neurospora crassa HJDF and its cultivation method and products and applications in 3D printing artificial fish meat. The Neurospora crassa HJDF described in the present invention can prepare new 3D printing raw materials, simulate the morphology and texture of real fish meat, improve the quality of 3D printed fish meat substitutes, make them closer to real meat in taste and flavor, and provide new ideas for the application of meat substitutes with different needs.

[0006] The present invention provides a strain of Neurospora crassa ( Neurospora crassa ) HJDF, the crassa HJDF is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, the preservation date is September 23, 2024, and the preservation number is CGMCC No.41518.

[0007] The present invention also provides a method for culturing the Neurospora crassa HJDF described in the above technical solution, comprising the following steps: culturing the Neurospora crassa HJDF described in the above technical solution in a culture medium to obtain a Neurospora crassa HJDF culture.

[0008] The present invention also provides a Neurospora crassa HJDF mycelial protein biomass, and the preparation method of the Neurospora crassa HJDF mycelial protein biomass comprises the following steps: Inoculating the Neurospora crassa HJDF described in the above technical solution into a fermentation medium, and performing liquid submerged fermentation culture to obtain a liquid submerged fermentation product; treating the liquid submerged fermentation product at 65-70°C for 20-30 min, treating it at 90-95°C for 30-40 min, and cooling it to obtain a cooled fermentation product; The cooled fermented product is filtered, washed, and freeze-dried to obtain Neurospora crassa HJDF mycelial protein biomass.

[0009] The present invention also provides the use of the Neurospora crassa HJDF described in the above technical solution, the Neurospora crassa HJDF culture obtained by the cultivation method described in the above technical solution, or the Neurospora crassa HJDF mycelial protein biomass described in the above technical solution in the production of 3D printed artificial fish meat.

[0010] The present invention also provides a food printing ink based on Neurospora crassa HJDF, comprising the following raw materials for preparation: Neurospora crassa HJDF described in the above technical solution or Neurospora crassa HJDF culture obtained by the cultivation method described in the above technical solution or Neurospora crassa HJDF mycelial protein biomass described in the above technical solution and food acceptable auxiliary materials.

[0011] Preferably, the auxiliary materials include starch and water.

[0012] Preferably, the method for preparing the food printing ink comprises the following steps: preparing the Neurospora crassa HJDF mycelial protein biomass described in the above technical scheme based on the Neurospora crassa HJDF described in the above technical scheme, ultrafinely grinding the Neurospora crassa HJDF mycelial protein biomass, sieving, and obtaining mycelial protein powder; mixing the mycelial protein powder with a food-acceptable auxiliary material to obtain a food printing ink based on Neurospora crassa HJDF.

[0013] The present invention also provides a method for preparing 3D printed artificial fish meat based on Neurospora crassa HJDF described in the above technical solution, comprising the following steps: The food printing ink based on Neurospora crassa HJDF described in the above technical solution is introduced into a 3D printing device for 3D printing to obtain 3D printed artificial fish meat.

[0014] Preferably, the diameter of the 3D printing nozzle is 0.6-1.0 mm; the speed of the 3D printing is 10-40 mm / s; the barrel and nozzle temperatures of the 3D printing are 45-55° C. respectively; and the cooling platform temperature of the 3D printing is 15-25° C.

[0015] The present invention also provides 3D printed artificial fish meat prepared by the preparation method described in the above technical solution.

[0016] The present invention provides Neurospora crassa ( Neurospora crassa )HJDF. The present invention has excavated a strain of Neurospora crassa HJDF from traditional snacks in Fujian. The strain has the characteristics of safe source, high protein content, complete and balanced amino acid composition, natural "umami", etc., and has good application potential as a meat substitute. Traditional 3D printed fish meat cannot simulate the state of muscle fibers. The present invention uses the natural fiber state of mycelium to simulate, which can better form a texture similar to meat fibers. Because it is a natural fiber structure, the preparation method of the present invention is not costly. The present invention realizes large-scale cultivation of Neurospora crassa HJDF through deep fermentation, and then the obtained mycelial protein biomass is mixed with auxiliary materials after ultrafine grinding to prepare food printing ink. The mycelial protein biomass treated with ultrafine grinding can simulate the micron-level muscle fiber structure of fish meat, and combined with the 3D printing method, it can further simulate the millimeter-level fiber arrangement of fish meat, thereby accurately replicating the characteristics of real fish meat in structure and texture. The present invention not only improves the simulation degree of alternative protein products, but also provides a new direction for the development of food technology. In addition, compared with traditional dietary protein sources, the use of Neurospora crassa mycelial protein to produce 3D printed artificial fish meat is in line with the healthy, environmentally friendly and sustainable dietary concept of the current era. The present invention can achieve efficient and sustainable conversion and supply of dietary protein, providing a suitable solution for the growing population pressure and protein demand. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0018] Figure 1 A growth morphology diagram of Neurospora crassa HJDF provided by the present invention on a PDA culture medium; Figure 2 A phylogenetic tree diagram of Neurospora crassa HJDF provided by the present invention; Figure 3 A growth curve diagram of Neurospora crassa HJDF provided by the present invention; Figure 4 A diagram of the submerged fermentation products of Neurospora crassa HJDF provided by the present invention; Figure 5 A biomass diagram of mycelial protein of Neurospora crassa HJDF after freeze-drying provided by the present invention; Figure 6 This is a fluorescence microscopic structure diagram of Neurospora crassa HJDF after shearing provided by the present invention; Figure 7 A real-life picture of the Neurospora crassa HJDF-based 3D printed artificial fish meat provided by the present invention; Figure 8 This is a display of the finished product of the Neurospora crassa HJDF-based 3D printed artificial fish meat provided by the present invention.

[0019] Biological deposit information Neurospora crassa Neurospora crassa )HJDF, deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, referred to as CGMCC, on September 23, 2024. The address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the deposit number is CGMCC No.41518. DETAILED DESCRIPTION

[0020] The present invention provides a strain of Neurospora crassa ( Neurospora crassa) HJDF, the rough Neurospora HJDF is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration Committee, the preservation time is September 23, 2024, and the preservation number is CGMCC No.41518. The rough Neurospora HJDF described in the present invention is extracted and purified from the traditional fermented bean dregs snacks in Wuping, Fujian. The colony of the rough Neurospora HJDF described in the present invention is initially powdery and granular, and then develops into a fluffy state, with a color ranging from pink to light orange. The mycelium is loose, spreading around and has transverse septa. Conidia are spherical, with a smooth surface, spore septa, forming chains, and often gathering into clumps. The ascus is cylindrical, with a short stalk at the bottom and a ring at the top. Ascospores are elliptical, olive green at first, and then turn brown-black. The Neurospora crassa HJDF of the present invention grows best at a temperature of 25-30°C, with the highest and lowest initial growth pH being 8.0 and 3.0 respectively, and the optimal initial growth pH being 5.0; the mycelium germination stage is 12-24 hours. The present invention uses microbiological characteristics such as morphological characteristics, culture shape, physiological and biochemical characteristics, and ITS sequence (SEQ ID NO.1:) analysis to identify the strain as Ascomycota ( Ascomycetes ), Conchiales ( Sordariales )、Dung Shell Family( Sordariaceae ), Neurospora ( Neurospora Neurospora crassa Neurospora crassa The strain of the present invention has the characteristics of safe source, high bacterial protein content, complete and balanced amino acid composition, natural "umami taste", etc., and therefore shows good application potential as a meat substitute.

[0021] The present invention also provides a method for culturing Neurospora crassa HJDF described in the above technical solution, comprising the following steps: culturing the Neurospora crassa HJDF described in the above technical solution in a culture medium to obtain a culture of Neurospora crassa HJDF. In a specific embodiment, the culture medium may be a liquid fermentation medium. In a specific embodiment, the liquid fermentation medium may include 20-30 g / L of soluble starch; 10-15 g / L of yeast extract; 0.4-0.5 g / L of potassium chloride; 3-4 g / L of potassium dihydrogen phosphate and 0.4-0.5 g / L of magnesium sulfate heptahydrate. In a specific embodiment, the rotation speed of the culture may be 100-150 rpm. In a specific embodiment, the temperature of the culture may be 28-30° C. In a specific embodiment, the ventilation volume of the culture may be 0.08-0.5 vvm. In a specific embodiment, the culture time may be 3-5 d.

[0022] The Neurospora crassa HJDF culture obtained by the culture method described in the above technical solution is also within the protection scope of the present application. In the present invention, the types of the Neurospora crassa HJDF culture include Neurospora crassa HJDF fermentation products, Neurospora crassa HJDF metabolites or Neurospora crassa HJDF extracts.

[0023] The present invention also provides a Neurospora crassa HJDF mycelial protein biomass, and the preparation method of the Neurospora crassa HJDF mycelial protein biomass comprises the following steps: Inoculating the Neurospora crassa HJDF described in the above technical solution into a fermentation medium, and performing liquid submerged fermentation culture to obtain a liquid submerged fermentation product; treating the liquid submerged fermentation product at 65-70°C for 20-30 min, treating it at 90-95°C for 30-40 min, and cooling it to obtain a cooled fermentation product; The cooled fermented product is filtered, washed, and freeze-dried to obtain Neurospora crassa HJDF mycelial protein biomass.

[0024] The present invention inoculates the rough Neurospora HJDF described in the above technical solution into a fermentation medium, performs liquid deep fermentation culture, and obtains a liquid deep fermentation product. Before inoculation, the rough Neurospora HJDF can be cultured to obtain a seed solution. In a specific embodiment, the culture medium used in the preparation of the seed solution can be a PDA culture medium. In a specific embodiment, the inoculation amount of the inoculation can be 5-10%, specifically 5%, 7% or 10%. In a specific embodiment, the volume of the liquid deep fermentation culture can be 5-50L. In a specific embodiment, the culture medium used in the liquid deep fermentation culture can include soluble starch 20-30g / L; yeast extract 10-15g / L; potassium chloride 0.4-0.5g / L; potassium dihydrogen phosphate 3-4g / L and magnesium sulfate heptahydrate 0.4-0.5g / L. In a specific embodiment, the rotation speed of the liquid deep fermentation culture can be 100-150rpm. In a specific embodiment, the temperature of the liquid deep fermentation culture can be 28-30℃. In a specific embodiment, the aeration rate of the liquid submerged fermentation culture can be 0.08-0.5 vvm. In a specific embodiment, the liquid submerged fermentation culture time can be 3-5 days.

[0025] After obtaining the liquid submerged fermentation product, the present invention treats the liquid submerged fermentation product at 65-70° C. for 20-30 min, at 90-95° C. for 30-40 min, and cools to obtain a cooled fermentation product. The present invention provides the above two-step heat treatment, the first step of heat treatment can remove intracellular RNA; the second step of heat treatment of the biomass after RNA removal can inactivate fungal cells.

[0026] After obtaining the cooled fermented product, the present invention filters, washes, and freeze-dries the cooled fermented product to obtain the crassa HJDF mycelial protein biomass. In a specific embodiment, the filtration can be performed using a 100-mesh filter to separate the fermented product from the culture medium. Thereafter, washing is performed to completely remove the culture medium. In a specific embodiment, the washing can be performed using sterile water, and the number of washings can be 3 times. The present invention has no particular restrictions on the freeze-drying conditions, and conventional freeze-drying conditions can be used.

[0027] The present invention also provides the use of the Neurospora crassa HJDF described in the above technical solution, the Neurospora crassa HJDF culture obtained by the cultivation method described in the above technical solution, or the Neurospora crassa HJDF mycelial protein biomass described in the above technical solution in the production of 3D printed artificial fish meat.

[0028] The present invention also provides a food printing ink based on Neurospora crassa HJDF, comprising the following raw materials for preparation: the Neurospora crassa HJDF described in the above technical solution or the Neurospora crassa HJDF culture obtained by the culturing method described in the above technical solution or the Neurospora crassa HJDF mycelial protein biomass described in the above technical solution and an excipient acceptable to food. In a specific embodiment, the excipient includes starch and water. In a specific embodiment, the preparation method of the food printing ink comprises the following steps: preparing the Neurospora crassa HJDF mycelial protein biomass described in the above technical solution based on the Neurospora crassa HJDF described in the above technical solution, ultrafine grinding the Neurospora crassa HJDF mycelial protein biomass, sieving, and obtaining mycelial protein powder; mixing the mycelial protein powder with an excipient acceptable to food to obtain a food printing ink based on Neurospora crassa HJDF. The present invention prepares the Neurospora crassa HJDF mycelial protein biomass described in the above technical solution based on the Neurospora crassa HJDF described in the above technical solution, ultrafine grinding the Neurospora crassa HJDF mycelial protein biomass, sieving, and obtaining mycelial protein powder. In a specific embodiment, the sieving can be through a 60-100 mesh sieve. The mycelial protein powder is mixed with an excipient acceptable to food to obtain a food printing ink based on Neurospora crassa HJDF. In a specific embodiment, the mixing can be performed using a homogenizer. The homogenization conditions of the present invention can be 5000-7000 rpm and 25° C. In a specific embodiment, after the homogenization, the starch is heated at 80-90° C. for 20-30 minutes to achieve starch gelatinization. In a specific embodiment, when the excipients are starch and water, the raw materials for preparing the food printing ink include 84-86% water by mass, 3-5% mycelial protein powder by mass, and 10-13% starch by mass.

[0029] The present invention also provides a method for preparing 3D printed artificial fish meat based on Neurospora crassa HJDF described in the above technical solution, comprising the following steps: The food printing ink based on Neurospora crassa HJDF described in the above technical solution is introduced into a 3D printing device for 3D printing to obtain 3D printed artificial fish meat. In a specific embodiment, the device used for the 3D printing can be a 3D printer, specifically an extrusion 3D printer. The ink provided by the present invention can provide a moderate viscosity. If the colloid concentration is too high, it is not easy to extrude. If the colloid concentration is too low, the material is not easy to form after extrusion or it is easy to collapse after forming. The water content of the ink is moderate, so the finished product obtained by printing can better simulate the water-containing texture of real fish meat. In a specific embodiment, the diameter of the 3D printing nozzle is 0.6~1.0mm; the speed of the 3D printing is 10~40mm / s; the barrel and nozzle temperatures of the 3D printing are 45~55℃ respectively; the temperature of the cooling platform of the 3D printing is controlled between 15~25℃, so that the best molding effect of the ink on the cooling plate can be obtained. In a specific embodiment, the 3D printed fish meat printing model needs to be modeled using SolidWorks software and converted into STL format, and then the path slicing is performed using the software supporting the printer. The present invention uses micron-scale mycelium protein raw materials combined with a 3D printing method to effectively simulate the fiber arrangement of real fish meat, providing a new technical approach for accurately replicating the characteristics of real fish meat in terms of flavor, nutrition and texture in the future.

[0030] The present invention also provides 3D printed artificial fish meat prepared by the preparation method described in the above technical solution.

[0031] In order to further illustrate the present invention, the Neurospora crassa HJDF and its culture method, as well as the products and applications in 3D printing artificial fish meat provided by the present invention are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0032] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods; the materials and reagents used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0033] Example 1 Isolation and Identification of Neurospora crassa HJDF 1. Source of Neurospora crassa HJDF strain: isolated, purified and cultured from a traditional fermented bean dregs snack in Wuping County, Fujian Province, China.

[0034] 2. Colony characteristics: The colony morphology of Neurospora crassa HJDF on PDA plates is as follows Figure 1As shown. The colony of Neurospora crassa is initially powdery and granular, then develops into a velvety, pink to light orange color. The mycelium is loose, spreading and has transverse septa. The conidia are spherical, smooth, with septa, forming chains, often in clumps. The asci are cylindrical, with a short stalk at the bottom and a ring at the top. The ascospores are oval, olive green at first, and then turn brown-black.

[0035] 3. Growth characteristics: The best growth temperature is 25~30 ℃, the highest and lowest initial growth pH are 8.0 and 3.0 respectively, and the optimal initial growth pH is 5.0; the mycelium germination stage is 12~24h.

[0036] 4. Storage method of mycelium strains: Store in PDA slant tubes at 4°C.

[0037] 5. Molecular identification: Inoculate the liquid strain into the fermentation medium at a 10% inoculation rate, culture at 28-30°C for 3-5 days, collect the mycelium, and extract the total DNA of the mycelium using the CTAB method. Use the designed fungal ribosomal intergenic region (ITS) universal primers ITS1 / ITS4 to perform PCR amplification on the test DNA sequence.

[0038] Primer ITS1: 5′-TCC GTA GGT GAA CCT GCG G-3′ (SEQ ID NO. 2); Primer ITS4: 5′-TCC TCC GCT TAT TGA TAT GC-3′ (SEQ ID NO. 3).

[0039] After sequencing the PCR product, the ITS sequence is shown in SEQ ID NO. 1. The sequencing results were entered into the GeneBank database for BLAST comparison analysis, and then the neighbor-joining algorithm in the MEGA software was used to construct the NJ phylogenetic tree. The results are shown in Figure 2 As shown. Figure 2 It can be seen that Neurospora crassa, Neurospora intermedia and Neurospora tetrasperma The closest relationship is that of Neurospora crassa HJDF. Neurospora crassa The strain had the highest affinity, so it was named Neurospora crassa HJDF.

[0040] The strains were identified as Ascomycota ( Ascomycetes ), Conchiales ( Sordariales )、Dung Shell Family( Sordariaceae ), Neurospora ( Neurospora Neurospora crassa Neurospora crassa , and named it Neurospora crassa ( Neurospora crassa ) HJDF, the strain was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on September 23, 2024, and its deposit number is CGMCC NO: 41518. The viability of the culture was tested on September 23, 2023, and the result was viable.

[0041] Example 2 Submerged Fermentation Amplification and Nutritional Quality Evaluation of Neurospora crassa HJDF 1. Preparation of Neurospora crassa HJDF liquid seed solution After the mycelium fills the test tube or culture dish (90mm in diameter), inoculate it into the liquid culture medium. Burn the inoculation shovel over the flame to sterilize it, and after it cools down, gently scrape the mycelium (split the mycelium horizontally and vertically), trying to avoid the mother seed block. Inoculate the above mycelium into the PDB culture medium. One plate of mycelium can be inoculated with 2~3 bottles of seed liquid. Place it in a shaker for shake bottle fermentation. According to the growth curve of Neurospora crassa HJDF ( Figure 3 ), take the fermentation liquid in the logarithmic growth phase after culturing at 28℃ and 125rmp for 24h as the seed liquid.

[0042] 2. Submerged fermentation amplification of Neurospora crassa HJDF fermentation tank The composition of the culture medium used for HJDF fermentation of Neurospora crassa is: 30g / L soluble starch; 10g / L yeast extract; 0.4g / L potassium chloride; 4g / L potassium dihydrogen phosphate; 0.4g / L magnesium sulfate heptahydrate. Use deionized water to prepare a liquid culture medium with a volume of 60-70% of the volume of the fermenter, sterilize it at 121℃ for 20min in the tank, and then cool it for use. Inoculate the above seed liquid into the fermenter at 10% (v / v), control the temperature at 28-30℃, pH 4-6.5, speed 100-150rmp, ventilation 0.08-0.5vvm, and ferment for 4d.

[0043] After 4 days of cultivation, Neurospora crassa HJDF achieved rapid biomass accumulation, and its biomass after fermentation could reach 12-15 g / L. Figure 4 As shown, after the strain was submerged in a 50L fermenter, the biological density of the resulting fermentation broth was close to the volume limit.

[0044] 3. Protein content and quality detection of Neurospora crassa HJDF The HJDF cells of Neurospora crassa obtained through the fermentation process were separated from the culture medium, washed and freeze-dried, and their protein content and amino acid composition were accurately determined according to the national standards GB 5009.5-2016 and GB 5009.124-2016. The results are shown in Table 1. The protein content in the HJDF cells of Neurospora crassa accounted for 45-54% of the dry weight, which is higher than that of most plant proteins. HJDF of Neurospora crassa contains a wide range of amino acid spectrum, including 18 coded amino acids, 2 semi-essential amino acids and 8 essential amino acids, with a high content of essential amino acids. Among them, glutamic acid and alanine give the mycelium a special "umami taste". The high content of branched-chain amino acids (isoleucine, leucine and valine) makes the bacteria have the potential to promote muscle anabolism. In addition, lysine is the first limiting amino acid of plant protein (wheat / cereals) and is also abundant in HJDF of Neurospora crassa. These characteristics make HJDF of Neurospora crassa a meat substitute protein source with great potential, which can be applied to a variety of food applications in the future.

[0045] Table 1 Protein content and amino acid composition of Neurospora crassa HJDF

[0046] 4. Fat content and quality testing of Neurospora crassa HJDF The HJDF cells of Neurospora crassa obtained through the fermentation process were separated from the culture medium, washed and freeze-dried, and their fatty acid composition and content were accurately determined according to the national standard GB 5009.168-2016. The results are shown in Table 2. The composition ratio of these fatty acids indicates that compared with traditional meat products, the HJDF mycoprotein products of Neurospora crassa have potential health benefits in reducing the risk of cardiovascular disease.

[0047] Table 2 Fatty acid content and composition of Neurospora crassa HJDF

[0048] Example 3 Preparation of 3D printed artificial fish meat using Neurospora crassa mycelial protein HJDF 1. Production of Neurospora crassa HJDF mycelial protein The fermentation product was obtained by fermentation according to the method described in Example 2 to produce mycelial protein. The fermentation product was heat-treated, first placed at 65°C for 20 min to remove intracellular RNA. Then, the fermentation product was further heated to 95°C for 30 min to inactivate the fungal cells. The heat-treated fermentation product was filtered through a 100-mesh filter and washed three times with sterile water to completely remove the culture medium. After freeze-drying, the mycelial protein biomass of Neurospora crassa HJDF was obtained, such as Figure 5 shown.

[0049] 2. Preparation of Neurospora crassa HJDF 3D printed artificial fish meat The mycelial protein biomass of Neurospora crassa HJDF was crushed by an ultrafine grinder and passed through a 60-mesh sieve to obtain mycelial protein powder. Figure 6 The results showed that the mycelial protein biomass after pulverization can simulate the micron-scale muscle fiber structure of fish meat. Mycelial protein powder, wheat pregelatinized starch (Nantong Gaofeng Biotechnology Co., Ltd.) and water were mixed in a ratio of 4:11:85, mixed evenly in a homogenizer at 6000 rpm and 25°C, and then heated at 85°C for 20 minutes to achieve starch gelatinization to prepare food printing ink.

[0050] like Figure 7 As shown, the ink was introduced into the 3D printer, and a specific model was selected for printing. The parameters were set as follows: nozzle diameter 0.9 mm, printing speed 20 mm / s, barrel temperature 50 ° C, and the cooling platform temperature of 3D printing was controlled at 20 ° C. The printed product was left to stand at 4 ° C for 2 hours to set. Figure 8 The results showed that 3D printed mycelial protein fish meat can be prepared into an ideal shape on demand, with clear boundaries and smooth surface. 3D printing technology can effectively simulate the millimeter-level fiber arrangement of fish meat. Combined with micron-level mycelial protein raw materials, this technology can accurately simulate the characteristics of real fish meat in structure and texture.

[0051] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A strain of Neurospora crassa ( Neurospora crassa ) HJDF, characterized in that The Neurospora crassa HJDF is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on September 23, 2024, and the deposit number is CGMCC No.41518.

2. The method for culturing Neurospora crassa HJDF according to claim 1, characterized in that: The method comprises the following steps: culturing the Neurospora crassa HJDF according to claim 1 in a culture medium to obtain a Neurospora crassa HJDF culture.

3. A Neurospora crassa HJDF mycelial protein biomass, characterized in that: The method for preparing Neurospora crassa HJDF mycelial protein biomass comprises the following steps: inoculating the Neurospora crassa HJDF of claim 1 into a fermentation medium, and performing liquid submerged fermentation culture to obtain a liquid submerged fermentation product; treating the liquid submerged fermentation product at 65-70°C for 20-30 min, treating it at 90-95°C for 30-40 min, and cooling it to obtain a cooled fermentation product; The cooled fermented product is filtered, washed, and freeze-dried to obtain Neurospora crassa HJDF mycelial protein biomass.

4. Use of the Neurospora crassa HJDF according to claim 1, the Neurospora crassa HJDF culture obtained by the cultivation method according to claim 2, or the Neurospora crassa HJDF mycelial protein biomass according to claim 3 in the production of 3D printed artificial fish meat.

5. A food printing ink based on Neurospora crassa HJDF, characterized in that: The preparation method comprises the following raw materials: the Neurospora crassa HJDF according to claim 1 or the Neurospora crassa HJDF culture obtained by the cultivation method according to claim 2 or the Neurospora crassa HJDF mycelial protein biomass according to claim 3 and excipients acceptable to food.

6. The food printing ink according to claim 5, characterized in that The auxiliary materials include starch and water.

7. The food printing ink according to claim 5 or 6, characterized in that: The preparation method of the food printing ink comprises the following steps: preparing the Neurospora crassa HJDF mycelial protein biomass according to claim 3 based on the Neurospora crassa HJDF according to claim 1, ultrafinely grinding the Neurospora crassa HJDF mycelial protein biomass, sieving, and obtaining mycelial protein powder; mixing the mycelial protein powder with a food-acceptable auxiliary material to obtain a food printing ink based on Neurospora crassa HJDF.

8. A method for preparing 3D printed artificial fish meat based on Neurospora crassa HJDF according to claim 1, comprising the following steps: The food printing ink based on Neurospora crassa HJDF according to any one of claims 5 to 7 is introduced into a 3D printing device for 3D printing to obtain 3D printed artificial fish meat.

9. The preparation method according to claim 8, characterized in that: The diameter of the 3D printing nozzle is 0.6~1.0mm; the speed of the 3D printing is 10~40mm / s; the barrel and nozzle temperatures of the 3D printing are 45~55°C respectively; the cooling platform temperature of the 3D printing is 15~25°C.

10. 3D printed artificial fish meat prepared by the preparation method according to claim 8 or 9.

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