Trichosanthes kirilowii Maxim nutrition powder and preparation method and application thereof

Abstract

The invention belongs to the field of biological agriculture fermentation, and particularly relates to snakegourd fruit nutrition powder and a preparation method and application thereof.The snakegourd fruit nutrition powder is prepared by conducting primary subzero treatment on snakegourd fruit roots in liquid carbon dioxide, soaking the snakegourd fruit roots subjected to primary subzero treatment in egg yolk hydrolysate for treatment, soaking the snakegourd fruit roots subjected to hydrolysis egg yolk soaking in liquid nitrogen for secondary subzero treatment, and conducting secondary subzero treatment. Soaking the secondarily cryogenic trichosanthes kirilowii maxim roots in liquid carbon dioxide for three-time cryogenic treatment, and then inoculating a fermentation strain for fermentation, enzymolysis and drying. According to the fermentation method, the fermentation uniformity is good, the trichosanthes kirilowii maxim roots do not need to be crushed, and the problem that the trichosanthes kirilowii maxim roots are difficult to ferment due to the fact that raw materials are compact in structure, microbial inoculation and growth are limited, moisture and ventilation are uneven and the like can be solved by conducting subzero treatment on the trichosanthes kirilowii maxim roots three times in cooperation with egg yolk hydrolysate dipping treatment. The fermented feed liquid obtained by fermenting the trichosanthes kirilowii roots can be prepared into trichosanthes kirilowii nutrition powder, and the fermentation residues can be used as a pig feed additive.

Description

Technical Field

[0001] This invention belongs to the field of bio-agricultural fermentation technology, specifically relating to a Trichosanthes kirilowii nutrient powder, its preparation method, and its application. Background Technology

[0002] Nine-times steaming and nine-times sun-drying is one of the traditional processing techniques for Chinese medicinal herbs, also known as "nine-times steaming and nine-times sun-drying" or "nine-times processing." Its core steps involve steaming and sun-drying the medicinal materials nine times. Through repeated heat treatment and sun exposure, the properties, meridian tropism, toxicity, and efficacy of the medicinal materials are altered, achieving the goals of "reducing toxicity and increasing efficacy, facilitating preservation, and harmonizing yin and yang."

[0003] However, the nine-steaming and nine-drying process is inefficient, especially unsuitable for processing by-products such as Trichosanthes kirilowii roots. Therefore, it is necessary to develop a new processing method for Trichosanthes kirilowii roots, such as fermentation. However, fermentation of Trichosanthes kirilowii roots generally presents significant challenges, mainly including:

[0004] 1) The raw material structure of Trichosanthes kirilowii root is dense.

[0005] The high content of cellulose, hemicellulose, and lignin in root tissue makes it difficult for microorganisms to utilize them directly.

[0006] Pretreatment (such as crushing, enzymatic hydrolysis, or hot water soaking) is generally required to release fermentable sugars.

[0007] 2) Microbial inoculation and growth restriction

[0008] Oxygen deficiency, low pH, or the presence of antibacterial substances (such as phenols and saponins) inside the roots make fermentation difficult.

[0009] 3) Uneven moisture and ventilation

[0010] The large volume of the root tuber and the uneven distribution of internal moisture can lead to localized dryness or excessive moisture, which affects microbial activity.

[0011] Based on this, the present invention is proposed. Summary of the Invention

[0012] The purpose of this invention is to provide a Trichosanthes kirilowii nutrient powder, its preparation method, and its application, in order to solve the above-mentioned problems.

[0013] Firstly, a method for preparing Trichosanthes kirilowii nutritional powder includes the following steps:

[0014] S1. Pretreatment of Trichosanthes kirilowii root

[0015] After cutting the Trichosanthes root into sections, it is first soaked in liquid carbon dioxide for 75 seconds, then removed and brought back to room temperature. This is a cryogenic treatment, resulting in cryogenically treated Trichosanthes root.

[0016] After washing and sterilizing the eggs, separate the yolks, remove the yolk membrane, add water and stir to obtain egg yolk liquid; after preheating, add sodium ascorbate to dissolve and mix to obtain preheated egg yolk liquid; add Aspergillus yuzami acidic protease to the preheated egg yolk liquid, and enzymatically hydrolyze in an acidic environment. After the enzymatic hydrolysis is completed, cool to room temperature to obtain egg yolk hydrolysate.

[0017] The roots of Trichosanthes kirilowii, which have been subjected to deep-cold treatment, are dried in the sun, soaked in a hydrolysate of egg yolk, removed, and dried to obtain hydrolyzed egg yolk-infused Trichosanthes kirilowii roots.

[0018] The roots of Trichosanthes kirilowii, which were soaked in hydrolyzed egg yolk, were treated by immersing them in liquid nitrogen, then removed and brought back to room temperature. This was a second cryogenic treatment, resulting in the second cryogenic Trichosanthes kirilowii root.

[0019] The roots of Trichosanthes kirilowii were treated by immersing them in liquid carbon dioxide for two deep cryogenic treatments, then removed and allowed to return to room temperature. This was the third deep cryogenic treatment, resulting in the Trichosanthes kirilowii roots that had undergone three deep cryogenic treatments.

[0020] S2, vaccination

[0021] After activating and culturing Lactobacillus plantarum and yeast, seed liquid was prepared and inoculated into Trichosanthes kirilowii roots that had been subjected to three deep-cold treatments.

[0022] S3, Fermentation

[0023] Fermentation is carried out. After fermentation is complete, the mixture is crushed, filtered, and fermentation liquid and fermentation residue are obtained.

[0024] S4, Enzymatic hydrolysis and drying

[0025] Neutral protease was added to the fermentation liquid for enzymatic hydrolysis. After the enzymatic hydrolysis was completed, the mixture was filtered, vacuum concentrated, freeze-dried, pulverized and sieved to obtain the Trichosanthes kirilowii nutrient powder.

[0026] A further improvement involves immersing the Trichosanthes kirilowii root in liquid carbon dioxide for 70-80 seconds during a cryogenic treatment process.

[0027] In a further improvement, in S1, the solid content of the egg yolk liquid is 25±1%. After preheating at 50℃ for 10 minutes, sodium ascorbate is added to dissolve it. The amount of sodium ascorbate added is 0.1±0.01% of the mass of the egg yolk liquid.

[0028] In a further improvement, in S1, the amount of Aspergillus yusei acidic protease added is 0.9% of the mass of the preheated egg yolk liquid, and it is enzymatically hydrolyzed for 1 hour at 35±3℃ in an acidic environment with a pH of 4.0~4.5.

[0029] A further improvement is made in S1, where the Trichosanthes kirilowii root is soaked in egg yolk hydrolysate for 30±3 minutes.

[0030] Further improvements were made in S1, where, during the secondary cryogenic treatment, the hydrolyzed egg yolk-soaked Trichosanthes kirilowii root was immersed in liquid nitrogen for 22 seconds.

[0031] In S1, during the three cryogenic treatments, the Trichosanthes kirilowii root was immersed in liquid carbon dioxide for 120±3s during the second cryogenic treatment.

[0032] Further improvements were made in S3, with the following fermentation process: fermentation temperature of 30±2℃, stirring speed of 100±30r / min, aeration rate of 0.5±0.05VVM, and fermentation time of 24±1h; followed by static fermentation at 35℃ for 36±1h.

[0033] Further improvements were made in S4, where the amount of neutral protease added was 0.2% of the fermentation liquid, the hydrolysis temperature was 45±1℃, and the hydrolysis time was 1.5±0.1h.

[0034] Secondly, a Trichosanthes kirilowii nutrient powder is prepared using the above-mentioned preparation method.

[0035] Thirdly, the Trichosanthes kirilowii nutritional powder can be used as a raw material for manufacturing special beverages for diabetics.

[0036] According to traditional Chinese medicine records, Trichosanthes root "promotes the production of body fluids, quenches thirst, clears heat, and purges fire." The nutritional powder produced by this invention using a fermentation process can be taken with water or decocted for use in treating thirst and dry throat caused by fever. Especially for symptoms such as polydipsia and polyuria caused by diabetes, the beverage made from the Trichosanthes root nutritional powder not only helps lower blood sugar but also improves related symptoms such as thirst, irritability, frequent urination, and weight loss.

[0037] Fourthly, the fermentation residue obtained by the above preparation method can be used as an additive for pig feed.

[0038] This invention discovers that adding fermentation residue to pig feed can promote the digestion and absorption of dry matter, crude protein, crude fat, and crude fiber in pig feed, resulting in an apparent improvement in digestibility. This invention provides a specific application of fermentation residue produced from the fermentation of Trichosanthes kirilowii root in pig production, laying a favorable foundation for the efficient utilization of fermented Trichosanthes kirilowii root products in the pig farming industry and the development of feed resources.

[0039] Compared with the prior art, the beneficial effects of this invention are as follows:

[0040] 1. The fermentation method of the present invention has good fermentation uniformity and does not require crushing of Trichosanthes kirilowii root. By performing three deep cryogenic treatments on Trichosanthes kirilowii root and soaking it in egg yolk hydrolysate, the fermentation difficulties caused by the dense structure of the raw material, limited microbial inoculation and growth, and uneven moisture and aeration can be solved.

[0041] 2. Both the solid and liquid products from the fermentation of Trichosanthes kirilowii root can be effectively utilized. The fermentation liquid can be further refined to obtain Trichosanthes kirilowii nutrient powder with higher economic value, which can be used as a raw material for making special drinks for diabetics. As for the by-product fermentation residue, it can be used as a pig feed additive. Detailed Implementation

[0042] The present invention will be further described in detail below through specific embodiments.

[0043] Example 1

[0044] S1. Pretreatment of Trichosanthes kirilowii root

[0045] After cutting the Trichosanthes root into sections, soak it in liquid carbon dioxide for 75 seconds, then remove it and allow it to return to room temperature. This is a cryogenic treatment, resulting in cryogenically treated Trichosanthes root.

[0046] After washing and sterilizing eggs with ultraviolet light, the yolks were separated, the yolk membrane was removed, and deionized water was added to adjust the solids content to 25%. The mixture was stirred at 200 rpm for 5 minutes to obtain an egg yolk solution. The solution was preheated at 50°C for 10 minutes, and 0.1% sodium ascorbate (by weight of the egg yolk solution) was added to dissolve and mix to obtain a preheated egg yolk solution. Usami Aspergillus acidic protease was added to the preheated egg yolk solution at a concentration of 0.9% of the preheated egg yolk solution's mass. Enzymatic hydrolysis was carried out at 35°C for 1 hour in an acidic environment with a pH of 4.0–4.5. After hydrolysis, no inactivation was required; the solution was cooled to room temperature to obtain the egg yolk hydrolysate. The acidic environment of pH 4.0–4.5 was adjusted by adding citric acid solution or hydrochloric acid to the phosphate buffer. The Usami Aspergillus acidic protease was purchased from Beijing Bio-Tech Biotechnology Co., Ltd., which uses an acidic protease produced by Aspergillus Usami.

[0047] The roots of Trichosanthes kirilowii, once subjected to deep-cold treatment, are dried in the sun, soaked in egg yolk hydrolysate for 30 minutes, removed, and dried to obtain Trichosanthes kirilowii roots soaked in egg yolk hydrolysate.

[0048] The roots of Trichosanthes kirilowii, which are soaked in hydrolyzed egg yolk, are immersed in liquid nitrogen for 22 seconds, then removed and allowed to return to room temperature. This is a second cryogenic treatment, resulting in second-cryogenic Trichosanthes kirilowii roots.

[0049] The roots of Trichosanthes kirilowii were immersed in liquid carbon dioxide for 120 seconds after being subjected to two deep cryogenic treatments. After being removed and allowed to return to room temperature, this was the third deep cryogenic treatment, resulting in the Trichosanthes kirilowii roots subjected to three deep cryogenic treatments.

[0050] S2, vaccination

[0051] After activating and culturing Lactobacillus plantarum and yeast, a seed culture (with a viable count greater than or equal to 10⁻⁶) was prepared at a volume ratio of 2:1. 8 (CFU / mL), inoculated three times with Trichosanthes kirilowii roots at a 5% inoculation rate.

[0052] S3, Fermentation

[0053] Ferment at 30℃, 100r / min, and 0.5VVM for 24 hours, then let it stand at 35℃ for 36 hours. After fermentation, crush the solids to allow them to be further incorporated into the fermentation liquid. Filter to obtain fermentation liquid and fermentation residue.

[0054] S4, Enzymatic hydrolysis and drying

[0055] Neutral protease was added to the fermentation broth at a concentration of 0.2% of the broth. The mixture was then enzymatically hydrolyzed at 45°C for 1.5 hours, filtered, vacuum concentrated, freeze-dried, pulverized, and sieved to obtain the Trichosanthes kirilowii nutrient powder.

[0056] Tests showed that the trichosanthes fruit nutritional powder contained 0.51% quercetin (53.2% rutin conversion rate) and 0.28% cytidine, with a bitterness grade of I.

[0057] The methods for detecting quercetin and cytidine content were both high performance liquid chromatography-UV (HPLC-UV), with separation by a C18 reversed-phase column followed by detection at ultraviolet wavelength.

[0058] The higher the quercetin content after root fermentation, the better the fermentation process and microbial system, which can significantly increase the concentration of effective components and potential medicinal efficacy of Trichosanthes kirilowii root.

[0059] During the fermentation of traditional Chinese medicine, rutin is enzymatically converted by microorganisms into more easily absorbed flavonoids such as quercetin. The higher the rutin conversion rate detected after fermentation, the better the fermentation effect.

[0060] A higher cytidine content usually indicates a better nucleotide synthesis effect in the raw materials or fermentation system, resulting in a greater accumulation of nucleotides.

[0061] Bitterness levels were tested using a comparative sensory evaluation method, employing a 5-level bitterness scale (IV, I, almost no bitterness) for blind testing of the samples.

[0062] Example 2

[0063] S1. Pretreatment of Trichosanthes kirilowii root

[0064] After cutting the Trichosanthes root into sections, soak it in liquid carbon dioxide for 70 seconds, then remove it and allow it to return to room temperature. This is a cryogenic treatment, resulting in cryogenically treated Trichosanthes root.

[0065] After washing the eggs, sterilize them with ultraviolet light, separate the yolks, remove the yolk membrane, add deionized water to adjust the solids to 24%, and stir at 200 rpm for 5 minutes to obtain egg yolk liquid; preheat at 50℃ for 10 minutes, add 0.09% sodium ascorbate by weight of the egg yolk liquid to dissolve and mix, to obtain preheated egg yolk liquid; add Aspergillus yusei acidic protease to the preheated egg yolk liquid, the amount of Aspergillus yusei acidic protease added is 0.9% by weight of the preheated egg yolk liquid, and enzymatically hydrolyze at 35±3℃ for 1 hour in an acidic environment with pH 4.0~4.5. After enzymatic hydrolysis, no inactivation is required, cool to room temperature to obtain egg yolk hydrolysate.

[0066] The roots of Trichosanthes kirilowii, dried in the sun, were soaked in an egg yolk hydrolysate for 27 minutes, removed, and dried to obtain Trichosanthes kirilowii roots soaked in egg yolk hydrolysate.

[0067] The roots of Trichosanthes kirilowii, which are soaked in hydrolyzed egg yolk, are immersed in liquid nitrogen for 22 seconds, then removed and allowed to return to room temperature. This is a second cryogenic treatment, resulting in second-cryogenic Trichosanthes kirilowii roots.

[0068] The roots of Trichosanthes kirilowii were immersed in liquid carbon dioxide for 117 seconds after being subjected to two deep cryogenic treatments. After being removed and allowed to return to room temperature, the roots were obtained after three deep cryogenic treatments.

[0069] S2, vaccination

[0070] After activating and culturing Lactobacillus plantarum and yeast, a seed culture (with a viable count greater than or equal to 10⁻⁶) was prepared at a volume ratio of 2:1. 8 (CFU / mL), inoculated three times with Trichosanthes kirilowii roots at a 5% inoculation rate.

[0071] S3, Fermentation

[0072] Fermented at 28℃, 70r / min, and 0.55VVM for 25h, then allowed to stand at 35℃ for 37h. After fermentation, the mixture was crushed, filtered, and fermentation liquid and fermentation residue were obtained.

[0073] S4, Enzymatic hydrolysis and drying

[0074] Neutral protease was added to the fermentation broth at a concentration of 0.2% of the broth. The mixture was then enzymatically hydrolyzed at 44°C for 1.6 hours. After filtration, vacuum concentration, freeze-drying, pulverization, and sieving, the Trichosanthes kirilowii nutrient powder was obtained.

[0075] Tests showed that the trichosanthes fruit nutritional powder contained 0.47% quercetin (52.1% rutin conversion rate) and 0.23% cytidine, with a bitterness grade of I.

[0076] Example 3

[0077] S1. Pretreatment of Trichosanthes kirilowii root

[0078] After cutting the Trichosanthes root into sections, soak it in liquid carbon dioxide for 80 seconds, then remove it and allow it to return to room temperature. This is a cryogenic treatment, resulting in cryogenically treated Trichosanthes root.

[0079] After washing the eggs, sterilize them with ultraviolet light, separate the yolks, remove the yolk membrane, add deionized water to adjust the solids to 26%, and stir at 200 rpm for 5 minutes to obtain egg yolk liquid; preheat at 50℃ for 10 minutes, add 0.11% sodium ascorbate by weight of the egg yolk liquid to dissolve and mix, to obtain preheated egg yolk liquid; add Aspergillus yusei acidic protease to the preheated egg yolk liquid, the amount of Aspergillus yusei acidic protease added is 0.9% by weight of the preheated egg yolk liquid, and enzymatically hydrolyze at 35±3℃ for 1 hour in an acidic environment with pH 4.0~4.5. After enzymatic hydrolysis, no inactivation is required, cool to room temperature to obtain egg yolk hydrolysate.

[0080] The roots of Trichosanthes kirilowii, dried in the sun, were soaked in an egg yolk hydrolysate for 33 minutes, removed, and dried to obtain Trichosanthes kirilowii roots soaked in egg yolk hydrolysate.

[0081] The roots of Trichosanthes kirilowii, which are soaked in hydrolyzed egg yolk, are immersed in liquid nitrogen for 22 seconds, then removed and allowed to return to room temperature. This is a second cryogenic treatment, resulting in second-cryogenic Trichosanthes kirilowii roots.

[0082] The roots of Trichosanthes kirilowii were immersed in liquid carbon dioxide for 123 seconds after being subjected to a second cryogenic treatment. After being removed and allowed to return to room temperature, this was the third cryogenic treatment, resulting in Trichosanthes kirilowii roots subjected to a third cryogenic treatment.

[0083] S2, vaccination

[0084] After activating and culturing Lactobacillus plantarum and yeast, a seed culture (with a viable count greater than or equal to 10⁻⁶) was prepared at a volume ratio of 2:1. 8 (CFU / mL), inoculated three times with Trichosanthes kirilowii roots at a 5% inoculation rate.

[0085] S3, Fermentation

[0086] Fermented at 32℃, 130r / min, and 0.45VVM for 23 hours, then allowed to stand at 35℃ for 34 hours. After fermentation, the mixture was crushed, filtered, and fermentation liquid and fermentation residue were obtained.

[0087] S4, Enzymatic hydrolysis and drying

[0088] Neutral protease was added to the fermentation broth at a concentration of 0.2% of the broth. The mixture was then enzymatically hydrolyzed at 46°C for 1.4 hours. After filtration, vacuum concentration, freeze-drying, pulverization, and sieving, the Trichosanthes kirilowii nutrient powder was obtained.

[0089] Tests showed that the trichosanthes fruit nutritional powder contained 0.44% quercetin (51.8% rutin conversion rate) and 0.21% cytidine, with a bitterness grade of I.

[0090] Reference ratio 1

[0091] The only difference between this example and Example 1 is that in S1 of this example, the Trichosanthes kirilowii root is crushed, and after crushing, fine powder with a particle size <2mm is sieved out, retaining particles of 2~5mm. The remaining steps S2~S4 are the same. It is important that the Trichosanthes kirilowii root is not crushed too finely to avoid forming a slurry during fermentation, which would affect aeration.

[0092] Reference ratio 2

[0093] The only difference between this example and Example 1 is that in S1 of this example, the Trichosanthes kirilowii root is not subjected to three deep-cold treatments; the remaining steps S2 to S4 are the same, that is, the Trichosanthes kirilowii root is directly used for inoculation and fermentation.

[0094] Comparative Example 1

[0095] The only difference between this example and Example 1 is that this example uses a single deep-cold Trichosanthes kirilowii root inoculation and fermentation; all other aspects are the same.

[0096] Comparative Example 2

[0097] The only difference between this example and Example 1 is that this example uses a double-cold fermentation of Trichosanthes kirilowii root; all other aspects are the same.

[0098] Comparative Example 3

[0099] The only difference between this example and Example 1 is that frozen Trichosanthes kirilowii root was used for inoculation and fermentation in this example; all other aspects are the same. Frozen Trichosanthes kirilowii root refers to root that has been frozen in a freezer at -18°C for 1 hour.

[0100] In this experiment, a pungent, putrid odor was detected during random sampling during fermentation. However, no such odor was observed throughout the entire process in Example 1.

[0101] Comparative Example 4

[0102] The only difference between this example and Example 1 is that this example does not use egg yolk hydrolysate to treat the Trichosanthes kirilowii root. Instead, the first-cryogenic Trichosanthes kirilowii root is directly immersed in liquid nitrogen for 22 seconds, then removed and allowed to return to room temperature to obtain the second-cryogenic Trichosanthes kirilowii root; all other aspects are the same.

[0103] The fermentation residue from Example 1 was pale yellow or light brown. However, the fermentation residue in this example showed dark brown and black substances, indicating obvious abnormalities and suggesting fermentation failure and the production of unknown substances.

[0104] Comparative Example 5

[0105] The only difference between this example and Example 1 is that this example uses egg yolk liquid to treat the Trichosanthes kirilowii root soaking instead of egg yolk hydrolysate to treat the Trichosanthes kirilowii root, and finally obtains egg yolk-soaked Trichosanthes kirilowii root. The egg yolk-soaked Trichosanthes kirilowii root is immersed in liquid nitrogen for 22 seconds, taken out, and restored to room temperature to obtain secondary cryogenic Trichosanthes kirilowii root; the rest are the same.

[0106] In this case, the fermentation residue had a distinct rotten egg smell and contained at least one-third black matter, indicating that the fermentation failed and produced unknown substances.

[0107] Comparative Example 6

[0108] The only difference between this example and Example 1 is that the first, second, and third cryogenic treatments all use liquid carbon dioxide for 75 seconds; all other aspects are the same.

[0109] Comparative Example 7

[0110] The only difference between this example and Example 1 is that the first, second, and third cryogenic treatments all use liquid nitrogen for 22 seconds; all other aspects are the same.

[0111] The lactic acid content in the fermentation liquid of Example 1 was 0.92%, while the lactic acid content in the fermentation liquid of this example was 0.38%.

[0112] Comparative Example 8

[0113] The only difference between this example and Example 1 is that, in this example, Aspergillus oryzae acidic protease is used instead of Aspergillus umbellatus acidic protease; all other aspects are the same. Proteases produced by Aspergillus oryzae can be classified into three main categories based on their optimal pH for action: acidic proteases, neutral proteases, and alkaline proteases.

[0114] Comparative Example 9

[0115] The only difference between this example and Example 1 is that, in this example, Bacillus subtilis acidic protease is used instead of Aspergillus yusamycin acidic protease; all other aspects are the same.

[0116] Comparative Example 10

[0117] The only difference between this example and Example 1 is that, in this example, Aspergillus niger acidic protease is used instead of Aspergillus umbilicatedica acidic protease; all other aspects are the same. The Aspergillus niger acidic protease is refined from Aspergillus niger via fermentation and was purchased from Hebei Lihua Biotechnology Co., Ltd.

[0118] Characterization of fermentation uniformity

[0119] Gravimetric method: By sequentially removing impurities such as soluble sugars, fats, starches, and hemicellulose from the sample, retaining only cellulose, and calculating the mass fraction after drying to constant weight, the fermentation residue can be used to determine whether fermentation is complete. More complete fermentation indicates better fermentation uniformity.

[0120] 1) Take a sample from the fermentation residue, rinse it three times with distilled water, and dry it in a 60℃ oven until constant weight; crush the dried sample and pass it through an 80-mesh sieve to obtain the pretreated sample.

[0121] 2) Weigh 2.0000g of the pretreated sample (denoted as m0) and place it in an Erlenmeyer flask; add 50mL of 80% ethanol solution and reflux at 80℃ for 1h. Filter while hot and wash the filter residue 3 times (10mL each time) with hot distilled water.

[0122] 3) Transfer the filter residue back to the conical flask, add 50 mL of ether, soak at room temperature for 2 hours, filter by suction, and wash the filter residue twice with ether (5 mL each time).

[0123] 4) Add 50 mL of 10% hydrochloric acid to the filter residue, heat in an 80°C water bath for 2 hours, filter by suction, and wash the filter residue with hot distilled water until neutral.

[0124] 5) Dry the filter residue in an oven at 105℃ for 4 hours, then remove it and cool it in a desiccator for 30 minutes. Accurately weigh the mass of the filter residue and record it as m1.

[0125] Repeat step 5 until the difference between two weighings is ≤0.0002g (constant weight is achieved).

[0126] Residual unfermented cellulose content = m1 / m0, expressed as a percentage.

[0127] The test results of Examples 1, Reference Examples 1-2, Comparative Examples 1-2, and Comparative Examples 6-10 are shown in Table 1:

[0128] Table 1

[0129]

[0130] As demonstrated in Examples 1, 1-2, and 2, cryogenic treatment (first, second, and third cycles) deconstructs the cell structure of *Trichosanthes kirilowii* roots to varying degrees by freezing to specific levels. The cryogenically treated roots are then soaked in egg yolk hydrolysate. Certain components in the egg yolk hydrolysate, such as lecithin, may increase the release of soluble components from the root cells, thus benefiting subsequent fermentation. The combination of three cycles of cryogenic treatment and egg yolk hydrolysate filling works synergistically to significantly enhance the subsequent fermentation process and improve fermentation uniformity.

[0131] 2. After washing the fermentation residue with deionized water 2-3 times, dry it at a low temperature below 60℃, crush and sieve it to obtain the fermentation product. Mix the fermentation product with pig feed at a mass ratio of 1:12 and feed it to pigs for 40 days. The control group feed does not contain fermentation product, and all pig feed is from the same batch.

[0132] The test results of the fermentation product obtained in Example 1 are shown in Table 2:

[0133] Table 2

[0134]

[0135] As shown in Table 1, the digestibility of crude fat increased by 16.5%, and the digestibility of crude fiber increased by 29.9%.

[0136] In contrast, the digestibility of crude fat in reference ratio 1 decreased by 7.7%, and the digestibility of crude fiber decreased by 10.5%.

[0137] In contrast, the digestibility of crude fat in control group 8 decreased by 1.2%, while the digestibility of crude fiber increased by 2.0%.

[0138] In contrast, Comparative Example 9 showed a 5.6% increase in crude fat digestibility and a 6.9% increase in crude fiber digestibility.

[0139] In contrast, Comparative Example 8 showed a 1.5% increase in crude fat digestibility and a 3.8% increase in crude fiber digestibility.

[0140] 3. The fermented product from Example 1 was mixed with chicken feed at a mass ratio of 1:20 and fed to Sanhuang chickens for 7 days. The control group's feed did not contain the fermented product, and all chicken feeds were from the same batch. In the experimental group, more than 50% of the chickens showed abnormalities such as decreased appetite or refusal to eat (they were unwilling to eat even when provided with feed, and even became picky eaters), abdominal distension (the abdomen was obviously swollen and felt hard and bloated to the touch), and abnormal excretion.

[0141] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for preparing a Gualou nutritional powder, characterized in that, The method comprises the following steps: S1, pretreatment of the Trichosanthes root After the Trichosanthes root is cut into segments, it is first soaked in liquid carbon dioxide for 75 s, taken out and restored to room temperature, to obtain a first deep cold Trichosanthes root through a first deep cold treatment; After the egg is washed and sterilized, the yolk is separated, the yolk membrane is removed, and water is added to obtain a yolk liquid by stirring and mixing; After preheating, sodium ascorbate is added to dissolve and mix to obtain a preheated yolk liquid; the preheated yolk liquid is added with acid proteinase of Aspergillus usamii for enzymolysis in an acidic environment, and after the enzymolysis is completed, the yolk hydrolyzate is obtained by cooling to room temperature; The first deep cold Trichosanthes root is dried and soaked in the yolk hydrolyzate, taken out and dried to obtain a yolk hydrolyzate impregnated Trichosanthes root; The yolk hydrolyzate impregnated Trichosanthes root is soaked in liquid nitrogen, taken out and restored to room temperature, to obtain a second deep cold Trichosanthes root through a second deep cold treatment; The second deep cold Trichosanthes root is soaked in liquid carbon dioxide, taken out and restored to room temperature, to obtain a third deep cold Trichosanthes root through a third deep cold treatment; S2, inoculation After the Lactobacillus plantarum and the yeast are activated and cultured, a seed liquid is prepared and inoculated into the third deep cold Trichosanthes root; S3, fermentation Fermentation is carried out, and after the fermentation is completed, the fermented material is crushed, filtered to obtain a fermented liquid and a fermented residue; S4, enzymolysis and drying Neutral protease is added to the fermented liquid for enzymolysis, and after the enzymolysis is completed, the fermented liquid is filtered, vacuum concentrated, freeze-dried, crushed and sieved to obtain the Trichosanthes nutritional powder.

2. The method for preparing Trichosanthes kirilowii nutritional powder according to claim 1, characterized in that: In the first deep cold treatment process, the Trichosanthes root is soaked in liquid carbon dioxide for 70-80 s.

3. The method for preparing Trichosanthes kirilowii nutritional powder according to claim 1, characterized in that: In S1, the solid content of the yolk liquid is 25±1%, and after preheating at 50°C for 10 min, sodium ascorbate is added to dissolve, and the amount of sodium ascorbate added is 0.1±0.01% of the mass of the yolk liquid.

4. The method for preparing Trichosanthes kirilowii nutritional powder according to claim 1, characterized in that: In S1, the amount of acid proteinase of Aspergillus usamii added is 0.9% of the mass of the preheated yolk liquid, and the enzymolysis is carried out in an acidic environment with a pH of 4.0-4.5 at a temperature of 35±3°C for 1 h.

5. The method for preparing Trichosanthes kirilowii nutritional powder according to claim 1, characterized in that: In S1, the first deep cold Trichosanthes root is soaked in the yolk hydrolyzate for 30±3 min.

6. The method for preparing Trichosanthes kirilowii nutritional powder according to claim 1, characterized in that: In S1, in the second deep cold treatment process, the yolk hydrolyzate impregnated Trichosanthes root is soaked in liquid nitrogen for 22 s. And / or In S1, in the third deep cold treatment process, the second deep cold Trichosanthes root is soaked in liquid carbon dioxide for 120±3 s.

7. The method for preparing Trichosanthes kirilowii nutritional powder according to claim 1, characterized in that: In S3, the fermentation process is as follows: the fermentation temperature is 30±2°C, the stirring speed is 100±30 r / min, the aeration amount is 0.5±0.05 VVM, and the fermentation time is 24±1 h; then, the fermented material is placed at 35°C for 36±1 h of static fermentation; And / or In S4, the amount of neutral protease added is 0.2% of the mass of the fermented liquid, the enzymolysis temperature is 45±1°C, and the enzymolysis time is 1.5±0.1 h.

8. A Gualou nutritional powder, characterized in that, The Trichosanthes nutritional powder is prepared by the preparation method in any one of claims 1-7.

9. The application of the Trichosanthes nutritional powder in claim 8 in a special drink for diabetes.

10. A fermented residue, characterized in that, The fermented residue is used as an additive for pig feed, which is prepared by the preparation method in any one of claims 1-7.

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