Active probiotic soft sweets capable of improving intestinal health and enhancing immunity and high-temperature preparation method of active probiotic soft sweets

Abstract

The invention discloses active probiotic soft sweets capable of improving intestinal health and enhancing immunity and a high-temperature preparation method of the active probiotic soft sweets. The method comprises the following steps: preparing soluble carbohydrate into an adapter solution with the water activity of 0.7-0.9, the pH value of 5.0-7.0 and the temperature of 40-50 DEG C; adding a certain amount of bacillus coagulans powder into the adaptive solution, and keeping the temperature at 40-50 DEG C for 20-50 minutes to prepare a pre-activation solution; preparing a sugar glue solution; auxiliary materials are added into the carbohydrate gum solution to prepare a blended carbohydrate gum solution, the pH value of the blended carbohydrate gum solution is 3.0-6.0, and the temperature is 60-80 DEG C; adding the pre-activated solution into the blended carbohydrate glue solution, and preserving heat for 10-40 minutes at the temperature of 60-80 DEG C to passivate; and pouring the passivated sugar glue solution into a soft sweet mold at 65-85 DEG C for shaping, and drying until the moisture is 16-18% to obtain the active probiotic soft sweet. The active probiotic soft sweets prepared by the preparation method disclosed by the invention are high in survival rate of probiotics, can keep stable in the delivery period, and have the activity of improving intestinal health and enhancing immunity.

Description

Technical Field

[0001] The present invention relates to the field of food processing. More specifically, it relates to an active probiotic gummy candy for improving intestinal health and enhancing immunity and a high-temperature preparation method thereof. Background Art

[0002] Bacillus coagulans (Weizmannia coagulans, also known as Weizmannia coagulans), belonging to Gram-positive bacteria, has characteristics such as high temperature resistance, acid and alkali resistance, and strong stress resistance. As a beneficial probiotic strain, after oral ingestion, Bacillus coagulans can effectively maintain the immune homeostasis of the body, enhance the immune regulation ability, and at the same time exhibit the physiological activity of improving intestinal health.

[0003] In the production process of probiotic gummy candies, the main processes include material weighing, ingredient preparation, gelatin melting, sugar boiling, mixing, flavoring, color adjustment, shaping, etc. However, the high-temperature treatment of the syrup may cause lipid peroxidation of the cell membrane structure and conformational changes of functional proteins; moreover, the water migration in the shaping stage will also cause mechanical stress on the microenvironment for the survival of the bacteria. These factors jointly restrict the product R & D progress of probiotic gummy candies.

[0004] Although Bacillus coagulans is known to have characteristics such as high temperature resistance, acid and alkali resistance, and strong stress resistance, making it feasible to produce active probiotic gummy candies, most of these tolerance studies are conclusions drawn from single-factor tests. In the actual production of gummy candy products, multiple influencing factors are often faced with superimposed challenges. Especially sensitive conditions such as temperature, moisture, and acidity will cause serious harm to the activity of probiotics. It is very difficult to maintain a high survival rate of probiotics in gummy candies during the production process and the shelf life with the traditional gummy candy processing technology.

[0005] "The Possibility of the Application of Bacillus coagulans in Probiotic Candies" (Wang Zhen et al., The Possibility of the Application of Bacillus coagulans in Probiotic Candies, Food and Fermentation Industries, No. 6, 2012) conducted research on this. Without adding acid, when Bacillus coagulans was placed at 70 °C and 75 °C for 40 minutes, its activity was hardly affected. However, under acidic conditions, with the acidity calculated as citric acid being only 0.5%, the activity of Bacillus coagulans decreased by 15% - 20% after being placed for 40 minutes. In gummy candy products, acidity is an important part of the product's sensory properties. Usually, the appropriate acidity needs to be between 1% - 2%. And in the production process of gummy candies, the temperature and time of the liquid materials that probiotics will come into contact with are often more than the test conditions of 75 °C and 40 minutes. Therefore, after such processing conditions, the attenuation rate of the activity of probiotics in gummy candies is usually much higher than 20%. The produced probiotic gummy candies usually have a water content of more than 10% and are in an acidic condition. Under such conditions, the activity of probiotics will continue to decline. And as a relatively common circulating food, the shelf life of gummy candies usually needs to be more than 12 months. Therefore, it is very difficult to maintain the high viability of probiotics during the order period.

[0006] Gummy candies use water-soluble colloids for gel molding. Usually, it is necessary to pour the liquid material into the mold cavity by pouring, and then shape it by drying or cooling. Probiotics can usually only be added and mixed in the sugar solution during the blending process. If probiotics are added after shaping, they can only be coated on the surface of the candy body, which will cause problems such as uneven coating, poor stability, and affecting the appearance, and the feasibility is extremely poor. After blending, the temperature of the sugar solution is usually between 60 - 90 °C, and it contains 20 - 30% water. The solution is acidic, while the conditions suitable for the preservation of probiotics are low temperature, neutral pH, and low water activity. Therefore, these conditions of the blended sugar solution are very unfavorable for maintaining the activity of probiotics. The water content of the blended sugar solution has the most direct impact on the pouring fluidity. Reducing the water content will make the pouring fluidity poor, affect the molding effect, and even cause pre-gelling. The pH value of the blended sugar solution not only has a significant impact on the product flavor, but also colloids such as pectin need to gel under acidic conditions. Therefore, it is not very meaningful to increase the activity of probiotics by adjusting and reducing the water content or increasing the pH value of the blended sugar solution. Currently, only by reducing the temperature of the blended sugar solution and sacrificing the pouring fluidity to increase the survival rate of probiotics is relatively theoretically feasible.

[0007] Therefore, in current research on enhancing the activity of probiotics in probiotic gummies, most of it is achieved by encapsulating probiotics and reducing the processing temperature. Encapsulating probiotics requires the use of additional coating wall materials, increasing the operation process, while reducing the processing temperature will reduce the fluidity of the sugar solution, posing a great challenge to the dispersion uniformity, production smoothness, and molding integrity of the product. These methods for enhancing the activity of probiotics in gummies not only increase the time and difficulty of process operations, greatly limiting the selection of product components and processes, but also cannot solve the problem of a relatively high loss rate of viable probiotic counts during the shelf life.

[0008] In Chinese Patent Application CN113875870A, "A Probiotic Gummy and Its Preparation Method", after mixing sodium alginate solution, pectin solution, and Lactobacillus bulgaricus solution, it is solidified in a calcium ion environment, and then separated whey protein solution is added for further coating to form a probiotic gel. Then, the probiotic gel is added to the liquid material at 60 - 70°C and finally made into probiotic gummies. This invention not only has a cumbersome operation, requires specific materials such as sodium alginate and separated whey protein for pre - coating treatment, limiting the selection of formula components, but also the viable count continuously decays during storage, decreasing by 6 orders of magnitude after 3 months.

[0009] In Chinese Patent Application CN115812824A, "A Probiotic Pearl Gummy and Its Preparation", a method of preparing probiotic gummies with high viable counts is to first encapsulate probiotics into pearls and then put the pearls into the gummies. This method requires a pill - dropping machine to prepare probiotic pearls, not only needing multiple wall materials, but also having a cumbersome and complex operation process. Moreover, due to the temperature - sensitivity of the wall material of probiotic pearls itself, it can only be added to the liquid material at a temperature of 35 - 45°C, and it also cannot solve the process pain point of the need to add probiotics at a low temperature.

[0010] In Chinese Patent Application CN117598384A, "A Probiotic Hyaluronic Acid Gummy and a Method for Increasing the Viable Count of Probiotics in Gummies", first, a probiotic hyaluronic acid gel is prepared using sodium hyaluronate, and then the gel is put into the liquid material to prepare probiotic gummies. This not only has a cumbersome operation, but also the price of sodium hyaluronate is relatively high, without the economic benefits of practical application.

[0011] In the Chinese patent application CN118203062A, "Probiotic Gummy Candy and Preparation Process", a specific prebiotic combination is used and mixed with probiotics, and the probiotic gummy candy is made by adding the liquid material under the low-temperature condition of 45 - 55°C. The formula components and process temperature are also limited, and the promotion and application are quite limited. The method of low-temperature blending and pouring is indeed more operable than other means, but the cost of low temperature is that the viscosity of the blended sugar solution increases, the dispersion uniformity is poor, the bubbles brought in by stirring are difficult to eliminate, and due to limited fluidity after pouring, it may not be able to fill the mold cavity, resulting in molding defects, causing problems such as unsmooth production process and high defective product rate, and it is difficult to apply in actual production.

[0012] Therefore, finding a preparation method for high-activity probiotic gummy candy that does not require encapsulation of probiotics, does not require specific components, and can withstand high-temperature conditions during the processing process has high practical value and economic benefits. Summary of the Invention

[0013] The inventors' experiments found that the main reason for the attenuation of the probiotic activity of the probiotic gummy candy made from Bacillus coagulans is not that the spores are killed during the processing process, but that the environmental conditions during the processing process can cause some spores to be activated and transform into vegetative cells. Although vegetative cells have activity, compared with the spore form, their tolerance has decreased significantly, and they are easily inactivated when exposed to heat, acid, and high-moisture environments. Therefore, when using Bacillus coagulans to prepare active probiotic gummy candy, to maintain a high viable cell rate and the stability of the viable cell count during the shelf life, the key lies in avoiding the activation of spores and maintaining the complete form of spores, see Figure 1 .

[0014] The first object of the present invention is to provide a high-temperature preparation method for active probiotic gummy candy that improves intestinal health and enhances immunity. This method improves the traditional preparation process of gummy candy, can tolerate the high-temperature environment during the processing of gummy candy without encapsulating Bacillus coagulans, solves the defects caused by low-temperature pouring to maintain probiotic activity, and can also improve the survival rate of probiotics and maintain stability during the shelf life.

[0015] The second object of the present invention is to provide the active probiotic gummy candy that improves intestinal health and enhances immunity prepared by the above preparation method and its application in improving intestinal health and enhancing immunity.

[0016] To achieve the above object, the present invention adopts the following technical solutions:

[0017] In the first aspect, the present invention provides a high-temperature preparation method for active probiotic gummy candy that improves intestinal health and enhances immunity, including the following steps:

[0018] Prepare an adaptation solution with soluble carbohydrates having a water activity of 0.7 - 0.9, a pH value of 5.0 - 7.0, and a temperature of 40 - 50 °C;

[0019] Add a certain amount of Bacillus coagulans powder to the adaptation solution and incubate at 40 - 50 °C for 20 - 50 min to prepare a pre-activated solution;

[0020] Prepare a sugar gum solution;

[0021] Add excipients to the sugar gum solution to prepare a formulated sugar gum solution with a pH value of 3.0 - 6.0 and a temperature of 60 - 80 °C;

[0022] Add the pre-activated solution to the formulated sugar gum solution and incubate at 60 - 80 °C for 10 - 40 min for inactivation to prepare an inactivated sugar gum solution;

[0023] Pour the inactivated sugar gum solution into a gummy candy mold at 65 - 85 °C for shaping and dry to a moisture content of 16% - 18% to obtain active probiotic gummy candies.

[0024] In the high-temperature preparation method of the present invention, since Bacillus coagulans is pre-activated in a specific adaptation solution first, even when Bacillus coagulans contacts the high-temperature formulated sugar gum solution, for example, the temperature of the formulated sugar gum solution is 60 °C, 65 °C, 70 °C, 75 °C, 80 °C or any temperature range between these values, the attenuation of the viable bacteria rate of the prepared probiotic gummy candies does not exceed 10%, and its viable bacteria rate is close to or even exceeds that of probiotic gummy candies prepared by a low-temperature process. Moreover, this method enables Bacillus coagulans without encapsulation to tolerate a relatively high pouring temperature during the gummy candy processing, for example, the pouring temperature is 65 °C, 70 °C, 75 °C, 80 °C, 85 °C or any temperature range between these values, while maintaining a high survival rate, without deliberately reducing the pouring temperature, and the viable bacteria count of the prepared probiotic gummy candies can also remain stable during the shelf life.

[0025] Further, the soluble carbohydrates are selected from one or more of white granulated sugar, glucose syrup, maltitol, erythritol, and polydextrose.

[0026] Further, in the formula of the active probiotic gummy candies of the present invention, the amount of Bacillus coagulans powder generally accounts for 0.05% - 5% of the total raw material mass for preparing the active probiotic gummy candies; for example, if the total raw material mass for preparing the active probiotic gummy candies is 100 g, then the Bacillus coagulans powder in the raw materials accounts for 0.05 g - 5 g. The Bacillus coagulans powder used in the present invention is commercially purchased with a specification of 100 - 400 billion CFU / gram.

[0027] Further, in the present invention, the formulated amount of Bacillus coagulans powder needs to be added to the said adaptation liquid to prepare a solution with a mass concentration of 1%-20%, namely the pre-activation liquid.

[0028] Further, in order to fully dissolve the Bacillus coagulans powder in the adaptation liquid, when adding the Bacillus coagulans powder to the adaptation liquid, it is preferably added while stirring and mixing evenly; preferably, the stirring conditions are 200-500 rpm for 1-5 minutes.

[0029] Further, in the preparation method, for the step of adjusting the pH value, if the pH value of the prepared solution itself has reached the requirement, there is no need to additionally add an acidity regulator to adjust the pH value; if the pH value of the prepared solution itself does not meet the requirement, an acidity regulator needs to be used to adjust the pH value, and the acidity regulator used is selected from one or more of citric acid, malic acid, phosphoric acid, sodium citrate, and potassium citrate.

[0030] Further, the gum solution can be prepared by a method known in the art according to the type of the final gummy candy. The sugars used for preparing the gum solution are selected from one or more of granulated sugar, glucose syrup, maltitol, erythritol, polydextrose, and inulin; the gums used for preparing the gum solution are selected from one or more of gelatin, pectin, starch, and carrageenan.

[0031] Further, the auxiliary materials are selected from one or more of edible flavors, edible pigments, fruit juices, fruit powders, and acid solutions.

[0032] Exemplarily, the acids for preparing the acid solution are selected from one or more of citric acid, malic acid, phosphoric acid, sodium citrate, and potassium citrate.

[0033] Further, when preparing the blended gum solution, in order to meet the nutritional requirements of the gummy candy, nutrients can also be added;

[0034] Exemplarily, the nutrients are selected from one or more of vitamins, minerals, amino acids, unsaturated fatty acids, prebiotics, lipids, and plant extracts.

[0035] Further, since the blended gum solution has a certain viscosity, in order to fully dissolve the Bacillus coagulans in the blended gum solution, when adding the pre-activation liquid to the blended gum solution, it is preferably added while stirring and mixing evenly; preferably, the stirring conditions are 400-800 rpm for 1-3 minutes.

[0036] Further, according to the source of the selected gum, the type of the active probiotic gummy candy can be one of animal gum gummy candy, plant gum gummy candy, and animal-plant mixed gum gummy candy.

[0037] Furthermore, according to the classification of the selected gummy structure type, the type of the active probiotic gummy can be one of core gummies, coated gummies, aerated gummies, and double-layer gummies; generally, it can be poured using the passivated gum solution according to the conventional method. Exemplarily, when preparing aerated gummies, the passivated gum solution can be aerated and then poured and molded.

[0038] In a second aspect, the present invention provides an active probiotic gummy for improving intestinal health and enhancing immunity prepared by the above method.

[0039] The active Bacillus coagulans probiotic gummy prepared by the present invention can not only retain a high viable bacteria rate, but also maintain stable activity during the shelf life, and has been proven by experimental tests to have the effects of improving intestinal health and enhancing immunity.

[0040] The beneficial effects of the present invention are as follows:

[0041] Through a special preparation process, the present invention passivates Bacillus coagulans, enhances the stress resistance of the dormant bodies, makes the spore morphology more complete during the high-temperature processing, reduces the activation of the dormant bodies into vegetative bodies, and finally enables the viable bacteria count of the prepared gummy to continuously remain at a relatively high level. Although there are many studies on the characteristics of Bacillus coagulans itself at present, and there are also many reports and literatures on its culture and sporulation conditions, there are no relevant studies and reports on the stress ability and stress resistance performance of Bacillus coagulans in the high-temperature, acidic and high-moisture gum solution during the gummy processing. And due to the complexity of the gummy processing conditions, the conditions used for the research such as the screening, spore induction and culture of Bacillus coagulans are obviously difficult to be used as a reference and guidance. Description of the Drawings

[0042] Figure 1 It is a microscopic image of the spores and activated spores of Bacillus coagulans.

[0043] Figure 2 It is a graph of the changes in the levels of IgE and IgA antibodies in two groups before and after intervention.

[0044] Figure 3 It is a graph of the changes in the levels of C3 and C4 complements in two groups before and after intervention.

[0045] Figure 4 It is a graph of the changes in the levels of immune-related factors before and after intervention.

[0046] Figure 5 It is a graph of the effect of the intervention of Bacillus coagulans probiotic gummy on the composition of the intestinal microbiota. Note: A - chao1 index dilution curve; PD - whole - tree analysis; C - Chao1 index; D - observed species index; E - ace index; F - NMDS2 analysis.

[0047] Figure 6 Effect of the species distribution histogram of the gut microbiota at the phylum and genus levels of probiotic gummies of Bacillus coagulans on adult allergic rhinitis. Note: A - phylum level; B - genus level. Detailed implementation manner

[0048] To illustrate the present invention more clearly, the present invention will be further described below in conjunction with preferred embodiments. Those skilled in the art should understand that the content specifically described below is illustrative rather than restrictive, and should not be used to limit the protection scope of the present invention. Unless otherwise specified, the reagents involved in the embodiments of the present invention are all commercially available products and can be obtained through commercial channels. For the convenience of comparing the viable count of probiotic gummies prepared by different preparation methods, the Bacillus coagulans powder used in the examples and comparative examples is in a specification of 200 billion CFU / g, which does not mean that only raw materials of this specification can be used.

[0049] Example 1

[0050] This example provides an active probiotic pectin gummy and its high - temperature preparation process, including the following components in parts by weight: 86 parts of maltitol solution, 10 parts of erythritol, 1.8 parts of pectin, 0.6 parts of sodium citrate, 0.5 parts of orange essence, 0.1 part of β - carotene powder, 0.8 parts of citric acid, 0.2 part of Bacillus coagulans powder.

[0051] The active probiotic pectin gummy and its high - temperature preparation process of this example include the following steps:

[0052] (1) Prepare an adaptation solution with 2 parts of maltitol solution having a water activity of 0.78, a pH value of 6.6, and a temperature of 45°C for later use.

[0053] (2) Add 0.2 part of Bacillus coagulans powder to the adaptation solution in step (1) under the stirring conditions of 300 rpm for 3 minutes, and then keep it warm at 45°C for 40 minutes to prepare a pre - activation solution for later use.

[0054] (3) Dissolve 0.1 part of β - carotene powder with 2 times warm water to prepare a pigment solution for later use; dissolve 0.8 part of citric acid with 1 time hot water to prepare an acid solution for later use.

[0055] (4) Add 1.8 parts of pectin and 0.6 part of sodium citrate to 5 parts of erythritol and mix evenly. Disperse with 15 times the amount of warm water based on the amount of pectin and boil. After boiling, add the remaining 5 parts of erythritol and 84 parts of maltitol solution, and boil until the solids content is 75 ± 1 °Bx to prepare a sugar gum solution for later use.

[0056] (5) The prepared gum syrup in step (4) is added with the pigment solution, acid solution and 0.5 parts of orange essence prepared in step (3), and stirred evenly to prepare a blended gum syrup for standby. The pH value of the blended gum syrup is 4.6 and the temperature is 75 °C.

[0057] (6) The pre-activated solution prepared in step (2) is added to the blended gum syrup prepared in step (5) under the stirring condition of 600 rpm for 2 minutes, and passivation is completed by maintaining at 75 °C for 30 minutes.

[0058] (7) After passivation, the gum syrup is poured into a gummy candy mold for shaping under the condition of maintaining at 85 °C, dried until the moisture content is 16 - 18%, demolded, and polished to obtain active probiotic pectin gummy candies.

[0059] Example 2

[0060] This example provides an active probiotic prebiotic gelatin gummy candy and its high-temperature preparation process, including the following components in parts by weight: 70 parts of polydextrose solution, 22 parts of inulin, 5.8 parts of gelatin, 0.4 parts of strawberry essence, 0.1 parts of sodium citrate, 0.1 parts of purple carrot juice, 1.2 parts of citric acid, 0.4 parts of Bacillus coagulans powder.

[0061] The active probiotic prebiotic gelatin gummy candy and its high-temperature preparation process in this example include the following steps:

[0062] (1) 6 parts of polydextrose solution are made into an adapted solution with a water activity of 0.72, a pH value of 5.2, and a temperature of 50 °C.

[0063] (2) 0.4 parts of Bacillus coagulans powder are added to the adapted solution in step (1) under the stirring condition of 400 rpm for 4 minutes, and maintained at 50 °C for 30 minutes to prepare a pre-activated solution for standby.

[0064] (3) 0.1 parts of purple carrot juice are preheated for standby; 1.2 parts of citric acid and 0.1 parts of sodium citrate are dissolved with 1 times of hot water to prepare an acid solution for standby.

[0065] (4) 22 parts of inulin are added with 0.5 times of warm water and heated to boiling, and the remaining 64 parts of polydextrose solution are added and boiled until the solid content is 80 ± 1 °Bx to prepare a boiled sugar solution for standby; 5.8 parts of gelatin are completely dissolved with 1.5 times of hot water, and maintained at 60 °C for 30 minutes, then the surface bubbles are removed to prepare a gelatin solution for standby; the boiled sugar solution is cooled to below 90 °C, and the prepared gelatin solution is added and mixed evenly to prepare a gum syrup for standby.

[0066] (5) The prepared sugar gum solution in step (4) is added with the pigment solution, acid solution and 0.4 parts of strawberry essence prepared in step (3), and stirred evenly to prepare a blended sugar gum solution for standby. The pH value of the blended sugar gum solution is 3.9 and the temperature is 65°C.

[0067] (6) The pre-activated solution prepared in step (2) is added to the blended sugar gum solution prepared in step (5) under the stirring conditions of 800 rpm for 3 minutes, and passivation is completed by maintaining the temperature at 65°C for 40 minutes.

[0068] (7) After passivation, the sugar gum solution is poured into a soft candy mold for shaping under the condition of maintaining the temperature at 65°C, dried until the moisture content is 16-18%, demolded, and polished to obtain active probiotic prebiotic gelatin soft candy.

[0069] Example 3

[0070] This example provides an active probiotic aerated mixed soft candy and its high-temperature preparation process, including the following components in parts by weight: 61 parts of glucose syrup, 30 parts of granulated sugar, 6 parts of gelatin, 0.5 part of pectin, 0.5 part of peach essence, 0.2 part of sodium citrate, 0.2 part of purple carrot juice, 1.5 parts of citric acid, and 0.1 part of Bacillus coagulans powder.

[0071] The active probiotic aerated mixed soft candy and its high-temperature preparation process in this example include the following steps:

[0072] (1) 0.5 part of glucose syrup is made into an adapted solution with a water activity of 0.85, a pH value of 6.1, and a temperature of 40°C for standby.

[0073] (2) 0.1 part of Bacillus coagulans powder is added to the adapted solution in step (1) under the stirring conditions of 200 rpm for 1 minute, and incubated at 40°C for 20 minutes to prepare a pre-activated solution for standby.

[0074] (3) 0.2 part of purple carrot juice is preheated for standby; 1.5 parts of citric acid are dissolved in 1 times of hot water to prepare an acid solution for standby.

[0075] (4) 0.5 part of pectin and 0.2 part of sodium citrate are added to 3 parts of granulated sugar and mixed evenly, dispersed in warm water 30 times the amount of pectin and boiled; the remaining 27 parts of granulated sugar and 60.5 parts of glucose syrup are added and boiled until the solids content is 80±1°Bx to prepare a boiled sugar solution for standby; 6 parts of gelatin are dissolved thoroughly in 1.5 times of hot water, incubated at 60°C for 30 minutes, and the surface bubbles are removed to prepare a gelatin solution for standby; the boiled sugar solution is cooled to below 90°C, and the prepared gelatin solution is added and mixed evenly to prepare a sugar gum solution for standby.

[0076] (5) The prepared gum syrup in step (4) is added with the pigment solution, acid solution and 0.5 parts of peach essence prepared in step (3), and stirred evenly to prepare a blended gum syrup for standby. The pH value of the blended gum syrup is 3.5 and the temperature is 70°C.

[0077] (6) The pre-activated solution prepared in step (2) is added to the blended gum syrup prepared in step (5) under the stirring condition of 400 rpm for 1 minute, and passivated at 70°C for 20 minutes.

[0078] (7) After passivation, the gum syrup is whipped with an aeration device for 15 minutes to prepare a probiotic aerated gum syrup with a density of 1.05 - 1.1 g / cm 3 .

[0079] (8) The probiotic aerated gum syrup is poured into a soft candy mold under the condition of heat preservation at 75°C for shaping, dried until the moisture content is 16 - 18%, demolded, and polished to obtain active probiotic aerated mixed soft candies.

[0080] Example 4

[0081] This example provides a vitamin C active probiotic pectin soft candy and its high-temperature preparation process, including the following components in parts by weight: 60 parts of glucose syrup, 35 parts of granulated sugar, 1.6 parts of pectin, 0.4 parts of sodium citrate, 0.4 parts of lemon essence, 1.2 parts of vitamin C, 1.0 parts of citric acid, and 0.4 parts of Bacillus coagulans powder.

[0082] The vitamin C active probiotic pectin soft candy and its high-temperature preparation process in this example include the following steps:

[0083] (1) 4 parts of glucose syrup are made into an adapted solution with a water activity of 0.75, a pH value of 6.5, and a temperature of 45°C for standby.

[0084] (2) 0.4 parts of Bacillus coagulans powder are added to the adapted solution in step (1) under the stirring condition of 300 rpm for 3 minutes, and then incubated at 45°C for 40 minutes to prepare a pre-activated solution for standby.

[0085] (3) 1 part of citric acid is dissolved with 1 times of hot water to prepare an acid solution for standby; 1.2 parts of vitamin C are dissolved with 2 times of hot water to prepare a vitamin C solution for standby.

[0086] (4) 1.6 parts of pectin and 0.4 parts of sodium citrate are added to 5 parts of granulated sugar and mixed evenly, dispersed with 15 times of warm water according to the amount of pectin and boiled. After boiling, the remaining 30 parts of granulated sugar and 56 parts of glucose syrup are added, and boiled until the solid content is 75 ± 1°Bx to prepare a gum syrup for standby.

[0087] (5) The prepared sugar gum solution in step (4) is added with the chromogenic acid solution, vitamin C solution prepared in step (3), and 0.4 parts of lemon essence, and stirred evenly to prepare a blended sugar gum solution for standby. The pH value of the blended sugar gum solution is 3.5, and the temperature is 70 °C.

[0088] (6) The pre-activation solution prepared in step (2) is added to the blended sugar gum solution prepared in step (5) under the stirring conditions of 700 rpm for 2 minutes, and passivation is completed by maintaining at 70 °C for 30 minutes.

[0089] (7) After passivation, the sugar gum solution is poured into a soft candy mold for shaping under the heat preservation condition of 75 °C, dried until the moisture content is 16 - 18%, demolded, and polished to obtain vitamin C active probiotic pectin soft candies.

[0090] Comparative Example 1

[0091] The difference between this comparative example and Example 1 lies in step (1). In step (1) of this comparative example, 2 parts of maltitol solution are used to prepare an adapted solution with a water activity of 0.92, a pH value of 6.6, and a temperature of 45 °C for standby.

[0092] Comparative Example 2

[0093] The difference between this comparative example and Example 1 lies in step (1). In step (1) of this comparative example, 2 parts of maltitol solution are used to prepare an adapted solution with a water activity of 0.78, a pH value of 7.2, and a temperature of 45 °C for standby.

[0094] Comparative Example 3

[0095] The difference between this comparative example and Example 1 lies in step (2). In step (2) of this comparative example, 0.2 parts of Bacillus coagulans powder are added to the adapted solution in step (1) under the stirring conditions of 300 rpm for 3 minutes, and maintained at 30 °C for 40 minutes to prepare a pre-activation solution for standby.

[0096] Comparative Example 4

[0097] The difference between this comparative example and Example 1 lies in step (5). The pH value of the blended sugar gum solution in this comparative example is 6.5.

[0098] Comparative Example 5

[0099] The difference between this comparative example and Example 1 lies in step (6). Passivation is completed by maintaining at 50 °C for 30 minutes.

[0100] Test Example 1

[0101] The viable count of the active probiotic gummies prepared in Examples 1-4 and Comparative Examples 1-5 was detected respectively (according to the method for counting Weissella coagulans in QB / T 5949-2023), and the survival rate of probiotic bacteria during the processing was compared. The results are shown in Table 1.

[0102] Table 1 Detection results of viable count of active probiotic gummies in examples and comparative examples

[0103] Theoretical value (100 million CFU / g) Test result (100 million CFU / g) Survival rate Example 1 4.00 3.95 98.8％ Example 2 8.00 7.92 99.0％ Example 3 2.00 1.91 95.5％ Example 4 8.00 7.89 98.6％ Comparative example 1 4.00 3.64 91.0％ Comparative example 2 4.00 3.33 83.3％ Comparative example 3 4.00 3.67 91.8％ Comparative example 4 4.00 3.75 93.8％ Comparative example 5 4.00 2.91 72.8％

[0104] As can be seen from Table 1, the survival rate of the active probiotic gummies in the examples was significantly higher than that of most of the comparative examples, and the loss rate of viable bacteria was extremely low.

[0105] The accelerated stability and long-term stability tests were carried out on the active probiotic gummies prepared in Examples 1-4 and Comparative Examples 1-5 respectively. The detection results of viable count were used as the evaluation index. The conditions for the long-term stability test were temperature 25±2°C and humidity 25-65%RH, and the investigation time was 24 months; the conditions for the accelerated stability test were temperature 40±2°C and humidity 75±5%RH, and the investigation time was 3 months. The results are shown in Table 2 and Table 3.

[0106] Table 2 Detection results of viable count of active probiotic gummies in long-term stability in examples and comparative examples

[0107]

[0108] Table 3 Detection results of viable count of active probiotic gummies in accelerated stability in examples and comparative examples

[0109]

[0110] As can be seen from Table 2 and Table 3, the viable count of the active probiotic gummies in the examples was significantly higher than that of the comparative examples in terms of both shelf-life survival rate and accelerated survival rate. After the shelf-life or accelerated test, the survival rate of probiotics remained at a relatively high level.

[0111] Test Example 2

[0112] The active Bacillus coagulans probiotic gummies prepared in the present invention (the gummies prepared in Example 1) can not only retain a high viable bacteria rate, but also maintain stable activity during the shelf-life, and have been clinically tested and confirmed to have the effects of improving intestinal health and enhancing immunity.

[0113] 78 participants with allergic rhinitis aged 18-65 years old were selected and given placebo and Bacillus coagulans probiotic gummies respectively, 2 capsules were taken every day, and the intervention lasted for 8 weeks to study the immunomodulatory effect of Bacillus coagulans probiotic gummies and its influence on intestinal flora.

[0114] 1. Allergic rhinitis (AR) is a type I allergic reaction mainly mediated by immunoglobulin IgE after the body contacts allergens. Total serum immunoglobulin E (IgE) is a key factor triggering respiratory allergic inflammation, the main antibody involved in allergic reactions, and the only immunoglobulin that increases in the body of patients with allergic diseases. It is related to the severity of the disease and airway hyperresponsiveness. The total serum IgE level can be used as an important indicator of the immunopathological changes in allergic rhinitis. Serum IgA has immunoprecipitation and complement killing effects. An increase in IgA, accompanied by an increase in eosinophils and an increase in IgG, indicates that the body is in an allergic state. Figure 2 It was shown that there were no significant differences in the levels of IgA and IgE before and after the intervention in the placebo group. After 8 weeks of intervention in the group taking probiotic gummies containing Bacillus coagulans, the IgE level of the study participants decreased significantly from 13171.85 ± 2581.32 ng / mL to 10991.35 ± 1876.13 ng / mL (p < 0.01); the IgA level showed a downward trend (p < 0.05).

[0115] Figure 3 It was shown that when allergic rhinitis occurs, complement C3 binds to complement receptors and contracts airway smooth muscle. Complement C4 participates in the classical activation pathway. The serum complement C3 level increased in the placebo group, while the serum complement C3 level decreased significantly in the group taking probiotic gummies containing Bacillus coagulans (p < 0.05); there were no significant changes in the serum complement C4 levels in both groups after the intervention (p > 0.05).

[0116] Eosinophils (EOS) are downstream effector cells of type 2 cytokines (IL-4, IL-5, and IL-13). Their aggregation and activation in the respiratory mucosa can directly trigger respiratory inflammatory reactions, which is of great significance for judging the severity of allergic rhinitis and its diagnosis and treatment. From Figure 4 The results showed that after 8 weeks of intervention with probiotic gummies containing Bacillus coagulans, the serum EOS level showed a downward trend (p < 0.05), and the unqualified rate of (EOS) decreased from 20% before the intervention to 0% after the intervention.

[0117] 2. As the largest immune organ in the human body, the gut microbiota plays a crucial role in the human immune system, with approximately 60%-70% of immune cells located in the gastrointestinal tract. Dysregulation of the gut microbiota may lead to immune dysfunction, thereby increasing the risk of allergic rhinitis. Therefore, we conducted 16S rRNA gene sequencing (V3-V4 region) to investigate the effect of probiotic gummies containing Bacillus coagulans on the gut microbiota of patients with allergic rhinitis. Chao1 and observed_species mainly estimate the number of species (abundance) in the community, and the ace index (Abundance-based Coverage Estimator) is used to estimate species diversity and stability in an ecosystem. As Figure 5 shown, the Chao1, ace, and observed species indices were all significantly increased (p<0.01), indicating that the probiotic gummies containing Bacillus coagulans significantly increased the α-diversity of the gut microbiota in these patients. In addition, NMDS (non-metric multidimensional scaling) analysis shows that the distance between samples reflects their similarity in composition and abundance. As Figure 5 shown in F in

[0118] , after the intervention of probiotic gummies containing Bacillus coagulans, the bacterial communities were clearly clustered into two distinct groups, indicating a significant difference in the microbial composition between the two groups.

[0118] On this basis, we further analyzed the differences between the two groups at the phylum and genus levels. At the phylum level ( Figure 6 shown in A in Figure 6 ), compared with the placebo group, the relative abundances of Bacteroidetes, Deferribacterota, and Campilobacterota were significantly upregulated in the probiotic gummies containing Bacillus coagulans intervention group, while the relative abundances of Firmicutes and Proteobacteria were significantly downregulated. This regulation may reshape the anti-inflammatory and immunomodulatory functions of the gut microbiota, thereby inhibiting Th2-mediated inflammatory responses and alleviating the symptoms of allergic rhinitis (AR). At the genus level ( Figure 6 shown in B in

[0119] ), the intervention of probiotic gummies containing Bacillus coagulans upregulated beneficial short-chain fatty acid (SCFA)-producing genera, including Prevotella, Alistipes, Lachnoclostridium, and Dialister, while opportunistic pathogens and the Escherichia-Shigella genus were significantly downregulated.

[0119] The Bacillus coagulans probiotic gummies prepared in Example 2, Example 3, and Example 4 also exhibited the above activities. In summary, the Bacillus coagulans probiotic gummies can regulate the levels of immune factors in the body, enhance the body's immunity, regulate the intestinal flora, and have the potential to relieve the rhinitis symptoms of patients with allergic rhinitis.

[0120] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, rather than limiting the implementation manners of the present invention. For those of ordinary skill in the art, based on the above description, other different forms of changes or variations can be made. It is impossible to list all the implementation manners here. Any obvious changes or variations derived from the technical solutions of the present invention still fall within the protection scope of the present invention.

Claims

1. A high temperature preparation method of active probiotic soft candy for improving intestinal health and enhancing immunity, characterized in that: The following steps are involved: The soluble carbohydrate is made into an adaptation liquid having a water activity of 0.7-0.9, a pH of 5.0-7.0, and a temperature of 40-50°C; A certain amount of Bacillus coagulans powder is added to the adaptation solution, and the solution is kept warm at 40-50° C. for 20-50 minutes to prepare a pre-activated solution; preparing sugar gum solution; Adding auxiliary materials to the carbohydrate gum liquid to prepare a prepared carbohydrate gum liquid, wherein the pH value of the prepared carbohydrate gum liquid is 3.0-6.0 and the temperature is 60-80° C.; Adding the pre-activated liquid to the prepared sugar gum liquid, keeping it warm at 60-80° C. for 10-40 minutes for passivation, to prepare a passivated sugar gum liquid; The passivated sugar gum liquid is poured into a soft candy mold at 65-85° C. for shaping, and dried to a moisture content of 16-18%, thereby obtaining active probiotic soft candy.

2. The high temperature preparation method according to claim 1, characterized in that: The soluble carbohydrate is selected from one or more of white sugar, glucose syrup, maltitol, erythritol and polydextrose.

3. The high temperature preparation method according to claim 1, characterized in that: The amount of the Bacillus coagulans powder accounts for 0.05%-5% of the total raw material mass for preparing the active probiotic soft candy; Preferably, in the pre-activation solution, the mass concentration of Bacillus coagulans powder is 1%-20%; Preferably, when the Bacillus coagulans powder is added to the adaptation solution, stirring and mixing are performed while adding; Preferably, the stirring condition is 200-500 rpm, 1-5 min.

4. The high temperature preparation method according to claim 1, characterized in that: The acidity regulator used to adjust the pH value is selected from one or more of citric acid, malic acid, phosphoric acid, sodium citrate and potassium citrate.

5. The high temperature preparation method according to claim 1, characterized in that: The sugar used to prepare the sugar glue liquid is selected from one or more of white sugar, glucose syrup, maltitol, erythritol, polydextrose, and inulin; the gum used to prepare the sugar glue liquid is selected from one or more of gelatin, pectin, starch, and carrageenan.

6. The high temperature preparation method according to claim 1, characterized in that: The auxiliary material is selected from one or more of edible flavors, edible pigments, fruit juice, fruit powder, and acid solution; Preferably, the acid used to prepare the acid solution is selected from one or more of citric acid, malic acid, phosphoric acid, sodium citrate, and potassium citrate; Preferably, the preparation of the prepared carbohydrate gum solution also includes adding nutrients; Preferably, the nutrients are selected from one or more of vitamins, minerals, amino acids, unsaturated fatty acids, prebiotics, lipids and plant extracts.

7. The high temperature preparation method according to claim 1, characterized in that: When the pre-activation liquid is added to the prepared sugar gum liquid, stirring and mixing are performed while adding; Preferably, the stirring condition is 400-800 rpm, 1-3 min.

8. The high temperature preparation method according to claim 1, characterized in that: The active probiotic soft candy is one of animal gum soft candy, plant gum soft candy, and animal and plant mixed gum soft candy; Preferably, the active probiotic soft candy is a type of core-filled soft candy, coated soft candy, aerated soft candy, and double-layer soft candy.

9. An active probiotic soft candy for improving intestinal health and enhancing immunity, characterized in that: It is prepared by the high-temperature preparation method described in any one of claims 1 to 8.

10. Use of the active probiotic soft candy for improving intestinal health and enhancing immunity as claimed in claim 9 in the preparation of products for improving intestinal health and enhancing immunity.

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