CN105495553B - A kind of black fungus rich in selenium ferment slurries and its preparation method and application - Google Patents

Abstract

The invention discloses a selenium-enriched black fungus ferment liquid, which is prepared by the following method: (a) cultivating black fungus strains into black fungus mother species; (b) cultivating selenium-enriched black fungus mother species to obtain black fungus fermented mycelium ; (c) mixing the fermented mycelium of black fungus with water in proportion, grinding, and ultrasonication to obtain the fermented mycelia slurry of black fungus enriched in selenium; , dried to obtain dried fruiting body of black fungus; (e) crushing dry fruiting body of black fungus, and mixing with water in proportion, grinding, ultrasonication, to obtain black fungus fruiting body slurry enriched in selenium; (f) enriching selenium The black fungus fruit body slurry and the selenium-enriched black fungus fermented mycelium slurry are mixed in proportion to obtain the selenium-enriched black fungus fermented slurry. The method provided by the invention is simple, easy to operate, convenient and reliable, short in preparation time, low in cost, and the prepared product has stable and uniform properties, good taste, and strong blood lipid-lowering function. health food.

Description

A kind of selenium-enriched black fungus ferment liquid and its preparation method and application

technical field

The invention relates to a method for preparing a fungus health drink, in particular to a selenium-enriched black fungus fermented liquid and its preparation method and application.

Background technique

Black fungus, also known as black cabbage, mulberry fungus, this fungus, tree chicken, wood moth, wood antler, is named for its shape like an ear and its dark brown color. Black fungus is a plant of the family Auriculariaceae. Stomach and large intestine meridian, medical research proves that black fungus has natural nutritional and health effects, not only rich in nutrition but also can nourish qi, strengthen the body, nourish the kidney and stomach, etc. Black fungus also has the function of promoting blood circulation, anti-coagulation, anti-thrombosis, and lowering blood fat , Reduce blood viscosity, soften blood vessels and other functions, effectively reduce the occurrence of cardiovascular diseases and other functions. The development of deep-processing products of black fungus usually uses fungus as raw material, and the form of extract or fermented liquid is added to the product in proportion to reduce the weak fishy smell of the fungus in the product, or the fermented liquid is directly added to the product in proportion , and then supplemented with other nutritional raw materials to form a product with a certain taste and flavor.

Organic selenium can delay human aging and improve immunity, and its function of lowering blood fat and blood sugar is also recognized. After black fungus converts inorganic selenium into organic selenium, the nutritional and health effects of its corresponding products are significantly higher than those of ordinary black fungus products. Therefore, for the convenience of drinking, the research and development personnel in the industry have developed it into a nutrient solution by deep processing. At present, the most widely used method of deep processing is to soak, dry, grind, dissolve and pulp the selenium-enriched black fungus, and add it to other ingredients for seasoning and other processes. Processed into selenium-enriched black fungus drinks, but these products have the following defects: 1. the product has a heavy fishy smell and poor taste. It needs to be seasoned with a large amount of food flavoring additives during production. The composition of the product is complex and the safety is low. It is not suitable to be directly used as a health drink to promote the market; ②The nutrient content of the product is not suitable for control, the health care function is poor, and the properties are not stable enough, and the sensory quality is poor; Edible, the output is limited, and the fruiting bodies used for health care products are even less, the cultivation period from the fungus to the finished product of fungus is long, the production cost is high, the price in the promotion market is high, and the purchasing power of ordinary people is limited, which makes the promotion The application is blocked; etc. In addition, some experts focus their research on extracting the nutrients of selenium-enriched black fungus, such as extracting selenium polysaccharides. However, this kind of extraction method not only wastes a lot of raw materials, the extraction process is cumbersome and complicated, takes a long time, and the cost is high, but also the final product does not contain various nutrients, and its active ingredients are poorly absorbed and functional. Therefore, develop a kind of black fungus material that can fully utilize selenium-enriched black fungus raw material, reduce nutritional loss as far as possible, can reduce production cost again, guarantee the stability of product proterties, and retain suitable selenium content and abundant nutritional labeling, play good health care function Pure natural original ecological drinks or health care products have become an urgent need for the deep processing of black fungus.

Contents of the invention

The purpose of the present invention is to provide a selenium-enriched black fungus fermented liquid and its preparation method and application to solve the problems of unstable sensory properties, high additive content, high cost, low safety and poor health care function in existing black fungus deep-processed drinks The problem.

The present invention is achieved through the following technical solutions: a selenium-enriched black fungus fermented liquid prepared by the following method:

(a) Inoculate the activated black fungus strain on PDA slant medium and culture at 28°C for 8-12 days to obtain black fungus female species;

(b) inoculate the black fungus mother species in a selenium-enriched liquid medium, and culture it on a shaker at 23-28°C for 5-7 days to obtain a fermentation liquid; expand the culture, filter the expansion liquid to obtain selenium-enriched black fungus fermented mycelium;

(c) mixing the fermented mycelia of selenium-enriched black fungus with sterile water at a ratio of 1:5-15, grinding, and ultrasonicating to obtain a fermented mycelium of selenium-enriched black fungus;

(d) inoculating the black fungus strain on a selenium-enriched solid medium, and carrying out selenium-enriching cultivation on the black fungus strain to obtain selenium-enriched black fungus fruiting bodies, and drying to obtain a selenium-enriching black fungus fruiting body dry product;

(e) Soak the dried selenium-enriched black fungus fruit body in water at 40-50°C for 1-2h, remove impurities, dry at 90-100°C for 5-7h, and crush to 140-180 mesh to obtain selenium-enriched black Fungus fruiting body powder: mix the selenium-enriched black fungus fruiting body powder with water according to the material-to-water ratio of 1:10-30, soak for 1-1.5 h, grind, and ultrasonically obtain the selenium-enriching black fungus fruiting body slurry;

(f) Mixing the selenium-enriched black fungus fruit body slurry and the selenium-enriched black fungus fermented mycelium slurry at a volume ratio of 2:3 to obtain the selenium-enriched black fungus fermented slurry.

The preparation method of the selenium-enriched black fungus ferment liquid provided by the invention comprises the following steps:

(a) Inoculate the activated black fungus strain on PDA slant medium and culture at 28°C for 8-12 days to obtain black fungus female species;

(b) inoculate the black fungus mother species in a selenium-enriched liquid medium, and culture it on a shaker at 23-28°C for 5-7 days to obtain a fermentation liquid; expand the culture, filter the expansion liquid to obtain selenium-enriched black fungus fermented mycelium;

(c) mixing the fermented mycelia of selenium-enriched black fungus with sterile water at a ratio of 1:5-15, grinding, and ultrasonicating to obtain a fermented mycelium of selenium-enriched black fungus;

(d) inoculating the black fungus strain on a selenium-enriched solid medium, and carrying out selenium-enriching cultivation on the black fungus strain to obtain selenium-enriched black fungus fruiting bodies, and drying to obtain a selenium-enriching black fungus fruiting body dry product;

(e) Soak the dried selenium-enriched black fungus fruit body in water at 40-50°C for 1-2h, remove impurities, dry at 90-100°C for 5-7h, and crush to 140-180 mesh to obtain selenium-enriched black Fungus fruiting body powder: mix the selenium-enriched black fungus fruiting body powder with water according to the material-to-water ratio of 1:10-30, soak for 1-1.5 h, grind, and ultrasonically obtain the selenium-enriching black fungus fruiting body slurry;

(f) Mixing the selenium-enriched black fungus fruit body slurry and the selenium-enriched black fungus fermented mycelium slurry at a volume ratio of 2:3 to obtain the selenium-enriched black fungus fermented slurry.

The PDA slant culture medium in the step (a) of the present invention is prepared according to the following proportions: 20 g of potatoes, 20 g of glucose, 20 g of agar, and 1000 mL of water.

The selenium-enriched liquid medium in step (b) of the present invention is prepared according to the following proportions: 20 g of glucose, 2 g of yeast extract, 2 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate, 60 mg of sodium selenite, and 1000 mL of water.

The expanded cultivation in the step (b) of the present invention is to inoculate the fermented liquid with an inoculum size of 1:5-10 in the volume ratio in a fermenter equipped with the selenium-enriched liquid medium, at 23-28°C Expand the culture for 3-7 days.

The grinding process described in step (c) and step (e) of the present invention refers to the use of a colloid mill for grinding for 15-30 minutes; the ultrasound refers to the use of 20-25 kHz intermittent ultrasound for 30-40 minutes, with an interval of 1 minute every 2 minutes. Through moderate grinding and ultrasonic treatment, various nutrient solutes in the mycelium can be better dissolved.

The selenium-enriched solid medium in step (d) of the present invention is formed by mixing dry material and water at a mass ratio of 1:1.5-1.8; the formula of the dry material is: 40-50% of crushed corncobs, 40% of sawdust -46%, wheat bran 6-12%, sucrose 0.5-1.5%, gypsum 1-2%, urea 0.8%, potassium dihydrogen phosphate 0.5%; preferably, corncob powder 46%, sawdust 43%, wheat bran 8.2% %, sucrose 0.5%, gypsum 1%, urea 0.8%, potassium dihydrogen phosphate 0.5%.

The black fungus strain used in the present invention is preferably black fungus G6 strain, which is provided by Baoding Dongsheng Edible Fungus Station, and is a black fungus strain with strong selenium-enriching ability, high yield and stable performance.

The application of the selenium-enriched black fungus ferment liquid combined with food flavoring agent in the preparation of seasoning selenium-enriched black fungus ferment liquid.

The present invention also provides a seasoning selenium-enriched black fungus fermented liquid, honey is added to the selenium-enriched black fungus fermented liquid prepared in claim 1, and the volume ratio of adding honey to the selenium-enriched black fungus fermented liquid is 1:15 -20, stir well to obtain seasoned selenium-enriched black fungus fermented liquid.

The selenium-enriched black fungus fermented liquid provided by the present invention has been proved to have an excellent lipid-lowering effect through gavage experiments on experimental animals. Therefore, the product can be popularized and applied in the market as a low-cost blood lipid-lowering drink.

The selenium-enriched black fungus ferment liquid prepared by the present invention has a selenium content of 0.7-0.8 μg/mL, and it is suggested that the daily recommended drinking amount for adults is less than 60 mL, and the optimal recommended drinking amount is 50-60 mL.

The invention adopts the solid culture of selenium-enriched fruiting bodies of edible fungi and the deep layer liquid culture of mycelia, uses the mycelium slurry as a raw material, adds a certain proportion and concentration of selenium-enriched black fungus fruiting body slurry, and blends the preparation in a specific proportion A selenium-enriched black fungus fermented liquid health drink was developed. In this method, the mycelium of submerged liquid fermentation is mainly used as the main material, and submerged liquid fermentation is not affected by environmental conditions and sites. As long as the technology is mastered, equipment and production conditions are available, production can be carried out at any time; The selenium-enriched black fungus fruit body slurry prepared by a specific ratio and a specific process can be used as a thickener and stabilizer for the fermented liquid, thereby replacing chemical thickening and stabilizing ingredients, avoiding the problem of unstable product properties and easy precipitation. It makes the black fungus health food a real health food without any additives.

In addition, the present invention dissolves various nutrients contained in the mycelia through grinding combined with ultrasonic treatment under specific conditions for the submerged fermentation selenium-rich mycelium; the selenium-rich black fungus fruiting body crushing technology is applied to prepare selenium-rich black Fungus fruiting body slurry, adding selenium-enriched black fungus fruiting body slurry to mycelium slurry ensures the stability, viscosity and uniformity of the final fermentation liquid, and not only avoids the addition of additives such as thickeners and stabilizers , and make the nutritional components of the product itself more balanced and stable, reduce the content of insoluble matter, and make the deep processing and large-scale production of selenium-enriched black fungus more feasible; finally adding honey plays the role of masking and seasoning, making the final The prepared fermented drink is rich in the unique aroma of fungus, sweet and refreshing in the mouth, and more nutritious.

The method provided by the invention is simple, easy to operate, low in production conditions, short in preparation time, low in raw materials and production costs, and the prepared product has stable and uniform properties, no additives, high safety, good taste, and strong blood lipid-lowering functionality. A nutraceutical and health product that is easy to market, has low cost, high economic value, and is suitable for daily consumption by high-fat people.

Detailed ways

The following examples are used to further describe the present invention in detail, but do not limit the present invention in any form, without special instructions, each test is paralleled five times, and the obtained data is the average value of parallel tests.

The present invention screens the high-yield black fungus strains, that is, through experiments such as mycelial growth status, antagonism experiment and identification of cellulase activity, the growth performance of 60 black fungus strains cultivated in the collected Hebei area is studied, from which A high-yield black fungus strain G6 was selected (this strain can also be provided by Baoding Dongsheng Edible Fungus Station). The following examples all take the black fungus strain G6 as the test strain.

Example 1

(1) Cultivation of slant parent species: inoculate black fungus strain G6 on PDA slant medium, and prepare PDA slant medium according to: 20 grams of potato, 20 grams of glucose, 20 grams of agar, 1000 mL of water, and make PDA slant culture according to the routine base, inoculate black fungus strains on it, and cultivate at 28°C for 10 days to obtain black fungus female species;

(2) Erlenmeyer flask liquid fermentation: inoculate the female black fungus seed obtained in step (1) into a 500 ml triangular flask filled with 200 ml of selenium-enriched liquid medium, and inoculate 2 triangular flasks for each female seed, and cultivate it in selenium-enriched liquid medium The basic formula is: 20 grams of glucose, 2 grams of yeast extract, 2 grams of potassium dihydrogen phosphate, 1 gram of magnesium sulfate, 60 milligrams of sodium selenite, 1000 milliliters of water; shaker speed 160 r/min, fermented at 25 ℃ for 6 day, to get the fermentation liquid;

(3) Expanded cultivation: Inoculate the fermented liquid obtained in (2) into a 2-ton tank with an inoculum volume ratio of 1:10. The culture medium of the fermenter is the same as that of the Erlenmeyer flask, at 28°C, and the ventilation volume is 1:1.3 Fermented under the condition of 5 days, at this moment mycelia selenium content reaches 9.1 μ g/g, and the polysaccharide content of fermented liquid can reach 3.7 g/L, reaches the end point of fermentation, finishes fermentation; Selenium-enriched black fungus fermented mycelium, the mycelium was washed twice to remove impurities, and 13.2kg of relatively pure mycelium was obtained. After freeze-drying, 7.45kg of selenium-enriched black fungus fermented mycelium was obtained;

(4) Preparation of selenium-enriched black fungus fermented mycelia slurry: mix the dried selenium-enriched black fungus fermented mycelium obtained in step (3) with sterile water at a ratio of 1:15, grind for 15 minutes in a colloid mill, and then Intermittent treatment with 22 kHz ultrasonic waves for 30 minutes, with an interval of 1 minute every 2 minutes, so that the mycelium intracellular content is completely dissolved, and the selenium-enriched black fungus fermented mycelium slurry is made;

(5) Preparation of selenium-enriched black fungus fruiting bodies: the selenium-enriching black fungus fruiting bodies were obtained by cultivating the black fungus G6 strain on the selenium-enriched solid medium, wherein the selenium-enriched solid medium was mixed with dry material and water at a ratio of 1:1.5 , the formula of its dry material is crushed corn cob 46%, sawdust 43%, wheat bran 8.2%, sucrose 0.5%, gypsum 1%, urea 0.8%, potassium dihydrogen phosphate 0.5%, add water and mix well, then pack 17cm×38cm After packing, autoclave for 2 hours with a sterilization pressure of 0.14-0.15MP. After the sterilization, the bag is cooled to below 30°C, and the fermented liquid prepared in step (2) is inoculated on the selenium-enriched solid culture On the base, inoculate 30mL per bag, fasten the mouth and culture, culture at 25°C, keep ventilated, 40 days, when black primordia appear on the surface of the bacteria bag, 120 holes are opened on the surface of each bag, and placed vertically in an open space In the area, spray water every day to keep it moisturized until the fungus grows and harvest, and the harvested fungus is dried or dried to obtain the selenium-enriched black fungus fruiting body dry product;

(6) Preparation of selenium-enriched black fungus fruiting body slurry: select dry selenium-enriching black fungus fruiting bodies that are shiny, fleshy, large, free of impurities, mildew-free, loose, and smooth, and soak them in warm water at 45°C for 1 h, then remove the root impurities, bake at 95°C for 6 h until completely dry, and grind the obtained black fungus to more than 150 mesh to obtain the raw material of selenium-enriched black fungus fruit body powder that meets the technological conditions. Add water to black fungus fruit body powder and water in a ratio of 1:20, soak for 1 hour at room temperature, grind for 15 minutes in a colloid mill, and then treat it intermittently with 22 kHz ultrasonic wave for 30 minutes, with an interval of 1 minute every 2 minutes, to obtain selenium-enriched black fungus Fruiting body slurry;

(7) Preparation of selenium-enriched black fungus fermentation liquid: mix 300mL of selenium-enriched black fungus fermented mycelia slurry and 200mL of selenium-enriched black fungus fruiting body slurry, add 30mL of honey, and mix the three to obtain selenium-enriched black fungus Yeast liquid. After testing, its selenium content was 0.8 μg/mL.

Example 2

(1) Cultivation of slant parent species: inoculate black fungus strain G6 on PDA slant medium, and prepare PDA slant medium according to: 20 grams of potato, 20 grams of glucose, 20 grams of agar, 1000 mL of water, and make PDA slant culture according to the routine base, inoculate black fungus strains on it, and cultivate at 28°C for 8 days to obtain black fungus female species;

(2) Erlenmeyer flask liquid fermentation: inoculate the female black fungus seed obtained in step (1) into a 500 ml triangular flask filled with 200 ml of selenium-enriched liquid medium, and inoculate 2 triangular flasks for each female seed, and cultivate it in selenium-enriched liquid medium Basic formula: 20 grams of glucose, 2 grams of yeast extract, 2 grams of potassium dihydrogen phosphate, 1 gram of magnesium sulfate, 60 mg of sodium selenite, 1000 ml of water; shaker speed 160 r/min, fermented at 23 ° C for 7 days , to obtain fermentation broth;

(3) Expanded cultivation: Inoculate the fermented liquid obtained in (2) into a 2-ton tank at an inoculum amount of 1:8 by weight. The culture medium of the fermenter is the same as that of the Erlenmeyer flask, at 23°C, and the ventilation volume is 1:1.3 Under the conditions of fermenting for 8 days, the fermentation was completed; the fermented liquid was filtered through a plate and frame filter to obtain the fermented mycelium of selenium-enriched black fungus, and the mycelium was washed twice to remove impurities and obtain a relatively pure mycelium. After freeze-drying, the dried product of fermented mycelium of selenium-enriched black fungus is obtained;

(4) Preparation of selenium-enriched black fungus mycelium slurry: mix the dried mycelium obtained in step (3) with sterile water at a ratio of 1:15, grind for 20 minutes in a colloid mill, and then intermittently treat with 22 kHz ultrasonic waves 35 minutes, every 2 minutes intermittently for 1 minute, so that the mycelium intracellular content is completely dissolved, and the mycelium slurry is made;

(5) Preparation of selenium-enriched black fungus fruiting bodies: the selenium-enriching black fungus fruiting bodies were obtained by cultivating the black fungus G6 strain on the selenium-enriched solid medium, wherein the selenium-enriched solid medium was mixed with dry material and water at a ratio of 1:1.6 , the formula of the dry material is 50% of crushed corncobs, 40.2% of sawdust, 6% of wheat bran, 1.5% of sucrose, 1% of gypsum, 0.8% of urea, 0.5% of potassium dihydrogen phosphate, mix well with water, and then pack it into a 17cm×38cm container Polypropylene plastic bag, autoclaved for 2 hours after filling, the sterilization pressure is 0.14-0.15MP, after the bag is cooled to below 30°C after sterilization, inoculate the fermented liquid prepared in step (2) on the selenium-enriched solid medium First, inoculate 30 ml per bag, tie the mouth and then culture, culture at 25°C, keep ventilated, 120 holes are opened on the surface of each bag when black primordia appear on the surface of the bacteria bag for 40 days, and placed vertically in an open space In the area, spray water every day to keep it moisturized until the fungus grows and harvest, and the harvested fungus is dried or dried to obtain the selenium-enriched black fungus fruiting body dry product;

(6) Preparation of selenium-enriched black fungus fruiting body slurry: select dry selenium-enriching G6 black fungus fruiting bodies that are shiny, fleshy, large, free of impurities, mildew-free, loose, and smooth, and soaked at 40°C in warm water for 2 h, then remove the root impurities, bake at 90°C for 7 h until completely dry, and grind the obtained black fungus to more than 180 mesh to obtain the raw material of black fungus fruit body powder that meets the technological conditions. Add water to the fruiting body powder and water at a ratio of 1:10, soak at room temperature for 1 hour, grind with a colloid mill for 20 minutes, and then treat it intermittently with 22 kHz ultrasonic waves for 35 minutes, with an interval of 1 minute every 2 minutes, to obtain the selenium-enriched black fungus fruiting body slurry ;

(7) Preparation of selenium-enriched black fungus fermented liquid: mix 300mL of fermented mycelium and 200mL of black fungus fruiting body, add 25mL of honey, and mix the three to obtain selenium-enriched black fungus fermented liquid. After testing, its selenium content was 0.76 μg/mL.

Example 3

(1) Cultivation of slant parent species: inoculate black fungus strain G6 on PDA slant medium, and prepare PDA slant medium according to: 20 grams of potato, 20 grams of glucose, 20 grams of agar, 1000 mL of water, and make PDA slant culture according to the routine base, inoculate black fungus strains on it, and cultivate at 28°C for 12 days to obtain black fungus female species;

(2) Erlenmeyer flask liquid fermentation: inoculate the female black fungus seed obtained in step (1) into a 500 ml triangular flask filled with 200 ml of selenium-enriched liquid medium, and inoculate 2 triangular flasks for each female seed, and cultivate it in selenium-enriched liquid medium Basic formula: 20 grams of glucose, 2 grams of yeast extract, 2 grams of potassium dihydrogen phosphate, 1 gram of magnesium sulfate, 60 mg of sodium selenite, 1000 ml of water; shaker speed 160 r/min, fermented at 28 ℃ for 5 days , to obtain fermentation broth;

(3) Expanded cultivation: Inoculate the fermented liquid obtained in (2) into a 2-ton tank with an inoculum volume ratio of 1:5. The culture medium of the fermenter is the same as that of the Erlenmeyer flask. At 25°C, the ventilation volume is 1:1.3 Ferment for 3 days under certain conditions, and finish the fermentation; filter the fermented liquid through a plate and frame filter to obtain the fermented mycelium of selenium-enriched black fungus, wash the mycelium twice to remove impurities, and obtain relatively pure selenium-enriched black fungus fermented mycelium;

(4) Preparation of selenium-enriched black fungus mycelium slurry: use the selenium-enriched black fungus fermented mycelium obtained in step (3), mix it with sterile water at a ratio of 1:5, grind it with a colloid mill for 30 minutes, and then use 22 KHz ultrasonic waves are intermittently treated for 40 minutes, with an interval of 1 minute every 2 minutes, so that the mycelium intracellular content is completely dissolved, and the mycelium slurry is made;

(5) Preparation of selenium-enriched black fungus fruiting bodies: the selenium-enriching black fungus fruiting bodies were obtained by cultivating the black fungus G6 strain on the selenium-rich solid medium, wherein the selenium-rich solid medium was mixed with dry material and water at a ratio of 1:1.8 , the formula of the dry material is 45.2% of crushed corncobs, 40% of wood chips, 12% of wheat bran, 0.5% of sucrose, 1% of gypsum, 0.8% of urea, 0.5% of potassium dihydrogen phosphate, mix well with water, and then pack it into a 17cm×38cm container Polypropylene plastic bag, autoclaved for 2 hours after filling, the sterilization pressure is 0.14-0.15MP, after the bag is cooled to below 30°C after sterilization, inoculate the fermented liquid prepared in step (2) on the selenium-enriched solid medium First, inoculate 30 ml per bag, tie the mouth and then culture, culture at 25°C, keep ventilated, 120 holes are opened on the surface of each bag when black primordia appear on the surface of the bacteria bag for 40 days, and placed vertically in an open space In the area, spray water every day to keep it moisturized until the fungus grows and harvest, and the harvested fungus is dried or dried to obtain the selenium-enriched black fungus fruiting body dry product;

(6) Preparation of selenium-enriched black fungus fruiting body slurry: select dry selenium-enriching G6 black fungus fruiting bodies that are shiny, fleshy, large, free of impurities, mildew-free, loose, and smooth, and soaked at 50°C in warm water for 1.5 h, then remove the root impurities, bake at 100°C for 5 h until completely dry, and grind the obtained black fungus to more than 140 mesh to obtain the raw material of black fungus fruit body powder that meets the technological conditions. Add water to fruiting body powder and water at a ratio of 1:30, soak at room temperature for 1.5 h, grind with a colloid mill for 30 minutes, and then treat it intermittently with 22 kHz ultrasonic waves for 40 minutes, with an interval of 1 minute every 2 minutes, to obtain the fruiting body of black fungus rich in selenium slurry;

(7) Preparation of selenium-enriched black fungus fermented liquid: mix 300 mL of fermented mycelium slurry and 200 mL of black fungus fruiting body liquid to obtain selenium-enriched black fungus fermented liquid. After testing, its selenium content was 0.74 μg/mL.

Comparative example 1

(1) Cultivation of slant parent species: inoculate black fungus strain G6 on PDA slant medium, and prepare PDA slant medium according to: 20 grams of potato, 20 grams of glucose, 20 grams of agar, 1000 mL of water, and make PDA slant culture according to the routine base, inoculate black fungus strains on it, and cultivate at 28°C for 10 days to obtain black fungus female species;

(2) Erlenmeyer flask liquid fermentation: inoculate the female black fungus seed obtained in step (1) into a 500 ml triangular flask filled with 200 ml of selenium-enriched liquid medium, and inoculate 2 triangular flasks for each female seed, and cultivate it in selenium-enriched liquid medium The basic formula is: 20 grams of glucose, 2 grams of yeast extract, 2 grams of potassium dihydrogen phosphate, 1 gram of magnesium sulfate, 60 milligrams of sodium selenite, 1000 milliliters of water; shaker speed 160 r/min, fermented at 25 ℃ for 6 day, to get the fermentation liquid;

(3) Expanded cultivation: Inoculate the fermented liquid obtained in (2) into a 2-ton tank with an inoculum volume ratio of 1:10. The culture medium of the fermenter is the same as that of the Erlenmeyer flask, at 28°C, and the ventilation volume is 1:1.3 Fermented for 5 days under certain conditions, and the fermentation was completed; the expansion medium was filtered through a plate and frame filter to obtain the fermented mycelium of selenium-enriched black fungus, and the mycelium was washed twice to remove impurities and obtain a relatively pure mycelium , after freeze-drying, the dry product of fermented mycelium of selenium-enriched black fungus is obtained;

(4) Preparation of selenium-enriched black fungus fermented mycelia slurry: mix the dried selenium-enriched black fungus fermented mycelium obtained in step (3) with sterile water at a ratio of 1:15, grind for 15 minutes in a colloid mill, and then Intermittent treatment with 22 kHz ultrasonic waves for 30 min, with an interval of 1 min every 2 min, so that the mycelium intracellular substance was completely dissolved, and the selenium-enriched black fungus fermentation mycelium slurry was prepared; the volume ratio of the selenium-enriched black fungus fermentation mycelium slurry was Add honey to taste at 1:15 to get black fungus nutrient solution.

Embodiment 4 Sensory quality and health care function detection of the product prepared by the present invention

(1) Sensory and physical and chemical testing

(1) Sensory indicators: the sensory products prepared in Examples 1-3 of the present invention are light brown homogeneous viscous liquids, with stable and uniform properties and no precipitation; there is a compound fragrance of Inonotus obliquus and black fungus, and no peculiar smell. Sweet and sour taste after adding honey; the sensory perception of the product prepared in Comparative Example 1 is light brown, viscous, unstable in character, easy to layer, with a small amount of precipitation; slightly peculiar smell.

(2) Physical and chemical indicators: It is tested that the pH value of the prepared product is within the range of 6.6-6.8, and the selenium content reaches 0.7-0.8 μg/mL.

(3) Hygienic indicators: in line with GB19297-2003 hygienic standards for fruit and vegetable juice beverages.

From the sensory comparison, it can be seen that if the fermented liquid is prepared with mycelium alone, since the biomass of the fermented mycelium is relatively small, its viscosity and stability are poor, and the sensory organs are easy to separate and precipitate. The present invention overcomes the defects of the prior art. By adding a certain proportion of black fungus fruiting body powder, it can be used as a thickener for the fermented liquid, and supplements the lack of mycelium biomass, ensuring that the fermented liquid drink has a certain Viscosity, make the feed liquid uniform. The drink combines the stickiness of fungus powder and the taste-masking effect of honey to ensure the stability and palatability of the fermented liquid, making the drink rich in the unique aroma of fungus and sweet in the mouth. The invention not only avoids adding thickeners and stabilizers, but also has more balanced and stable nutritional components in the product, reduces the content of insoluble matter, and makes the deep processing and large-scale production of the selenium-enriched black fungus more feasible.

(2) Health function testing

(1) Sample

Dosage setting of the black fungus fermented liquid drink prepared in Example 1: the product is prepared with physiological saline so that the volume ratio concentration is 30%, 60%, and 100%, which correspond to three dosage groups of low, medium, and high respectively.

(2) Experimental animals

Male and female BAL B/c mice and basic feed (purchased from the Department of Experimental Animals, Hebei Medical University), weighing 20-25g.

High-fat feed formula: basic feed 78.8%, egg yolk powder 10%, lard 10%, cholesterol 1%, bile salt 0.2%.

(3) Reagents

Serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) assay kits, etc.

(4) Test method

① Feeding experiment: BAL B/c mice were fed with basal diet for 7 days, and then the tail blood was collected to measure coagulation time, serum total cholesterol (TC), serum triglyceride (triglyceride, TG), and serum high-density lipoprotein Cholesterol (high density lipoprotein cholesterol, HDL-C) and serum low density lipoprotein cholesterol (lowdensity lipoprotein cholesterol, LDL-C) value. According to the TC value, they were randomly divided into 5 groups, 10 rats/group, which were blank group, high-fat control group, low, medium and high dose groups of black fungus fermented liquid drink and control group 1. The blank group freely ingested the basic feed and gavage with normal saline (2 mL/cause) every day; the high-fat control group freely ingested high-fat feed and gavaged normal saline every day (2 mL/cause); each feeding group freely ingested high-fat feed , gavage with selenium-enriched G6 black fungus fermented liquid (3 mL/cause) every day, and fed continuously for 45 days. After fasting for 16 hours before the end of the experiment, blood was collected from the orbital venous plexus to measure the coagulation time, TC, TG, HDL-C and LDL-C values. The experiment was repeated 3 times, the average value was taken, and the new multiple range test was performed on the data using DPS data analysis software.

② Determination of various blood lipid indexes: Determination of coagulation time by capillary glass tube method; TG, TC, HDL-C, LDL-C were determined by automatic biochemical analyzer according to kit instructions. The above indicators were entrusted to Baoding Hospital of Traditional Chinese Medicine for testing. The experimental data is processed by DPS7.05 statistical software.

(5) Experimental results

The indicators of each group of mice before and after the experiment were obtained by measurement, and the results are shown in Table 1. The coagulation time is different among the groups, among which the high dose group prolongs the coagulation time of the experimental animals compared with the blank control group and the high-fat control group, and the middle and low dose group also has a longer coagulation time than the blank control group. It shows that a certain dose of fermented liquid product has some anticoagulant effects on the blood of normal and high fat BAL B/c mice. After intragastric administration, the TC values of the blank group, the middle dose group and the high dose group were all lower than the high fat control group, and the low dose group had no significant difference compared with the high fat control group and the blank group, indicating that the fermented liquid product has a certain effect on high fat animals. Reduce the function of TC, but the effect on normal experimental animals is not obvious. The TG value of the experimental animals in the middle dose group and the high dose group was significantly lower than that of the high fat control group and the blank control group, and the difference between the middle dose group and the high fat control group and the blank control group was significant, indicating that long-term use of selenium-enriched black fungus yeast The serum product has the function of significantly reducing the TG of normal and high-fat experimental animals within a certain concentration range, but the lipid-lowering function of the high-dose fermented serum product is somewhat reduced. The HDL-C value of the blank group, high-dose group, middle-dose group and low-dose group was higher than that of the high-fat control group. Except for the high-fat control group, there was no obvious difference among the other experimental groups, but the HDL-C value increased with the increase of dose. The increasing trend shows that high-dose selenium-enriched black fungus fermented liquid products can significantly increase the HDL-C level of high-fat experimental animals, and has no obvious effect on normal experimental animals. The LDL-C values of the experimental animals in the blank group and the low, middle and high dose groups were significantly lower than those in the high fat control group. The difference between the high dose group and the blank control group was not obvious, but compared with the middle and low dose group, the difference was obvious, indicating that the yeast Serum products have little effect on reducing LDL-C in normal experimental animals, but can significantly reduce LDL-C in high-fat experimental animals.

Table 1 Blood lipids of mice in each group after feeding black fungus fermented liquid drink

Note: Different lowercase letters in the same column indicate significant differences at the 0.05 level, and different uppercase letters indicate significant differences at the 0.01 level.

(6 Conclusion

Using BAL B/c mice as experimental animals, the blood lipid-lowering function test of selenium-enriched black fungus fermented liquid was carried out by the lipid metabolism disorder model-preventive test sample method. /c mice have obvious anticoagulant effect, can reduce the total cholesterol, triglyceride and low-density lipoprotein cholesterol in the serum of high-fat BAL B/c mice, and can increase the high-density lipoprotein cholesterol in the serum of high-fat BAL B/c mice Lipoprotein cholesterol content; for normal BAL B/c mice, the effect of this product on reducing total cholesterol and low-density lipoprotein cholesterol and increasing high-density lipoprotein cholesterol is not obvious, but it can significantly reduce the content of triglyceride in serum .

The invention utilizes the self-adhesiveness of the black fungus after homogenization, and uses an appropriate amount of black fungus fruit bodies as a food thickener. Firstly, there is no need to add chemical thickening ingredients, and secondly, the slurry avoids precipitation, making the functional food truly healthy and non-toxic. Harmful green food. It not only enhances the safety of products, but also provides a reference method for the healthy development of the food industry, which is the first case in China. The fermented liquid prepared by the invention is used as a green food, and after eating the black fungus fermented liquid, it can not only enhance people's physique, but also can be used as a health care product to prevent the occurrence of various diseases such as blood coagulation. With the continuous improvement of people's living standards and the increasing number of patients with cardiovascular and cerebrovascular diseases and diabetes, people will have more and more demands for this selenium-enriched fermented liquid drink similar to black fungus. Therefore, this product has a wide range of applications prospect.

Claims (8)

1. a selenium-enriched black fungus fermented liquid, is characterized in that, is prepared by the following method: (a) Inoculate the activated black fungus strain on PDA slant medium and culture at 28°C for 8-12 days to obtain black fungus female species; (b) inoculate the black fungus mother species in a selenium-enriched liquid medium, and culture it in a shaking table at 23-28°C for 5-7 days to obtain a fermentation liquid; expand the culture, filter the expansion liquid to obtain selenium-enriched black fungus fermented mycelium; (c) mixing the fermented mycelia of selenium-enriched black fungus with sterile water at a ratio of 1:5-15, grinding, and ultrasonicating to obtain a fermented mycelium of selenium-enriched black fungus; (d) inoculating the black fungus strain on a selenium-enriched solid medium, and carrying out selenium-enriching cultivation on the black fungus strain to obtain selenium-enriched black fungus fruiting bodies, and drying to obtain a selenium-enriching black fungus fruiting body dry product; (e) Soak the dried selenium-enriched black fungus fruit body in water at 40-50°C for 1-2h, remove impurities, dry at 90-100°C for 5-7h, and crush to 140-180 mesh to obtain selenium-enriched black Fungus fruiting body powder: mix the selenium-enriched black fungus fruiting body powder with water according to the ratio of material to water of 1:10-30, soak for 1-1.5 h, grind, and ultrasonically obtain the selenium-enriching black fungus fruiting body slurry; (f) Mixing the selenium-enriched black fungus fruit body slurry and the selenium-enriched black fungus fermented mycelium slurry at a volume ratio of 2:3 to obtain the selenium-enriched black fungus fermented slurry. 2. The selenium-enriched black fungus ferment liquid according to claim 1, characterized in that the selenium-enriched liquid medium in step (b) is prepared according to the following proportions: 20 g of glucose, 2 g of yeast extract, 2 g of potassium dihydrogen phosphate, Magnesium sulfate 1 g, sodium selenite 60 mg, water 1000 mL. 3. The selenium-enriched black fungus ferment liquid according to claim 1, characterized in that, the ultrasonic in step (c) and step (e) both refers to the use of 20-25kHz intermittent ultrasonic for 30-40min, with an interval of 1min every 2min. 4. The selenium-enriched black fungus fermented liquid according to claim 1, characterized in that the selenium-enriched solid medium in step (d) is formed by mixing dry material and water at a mass ratio of 1:1.5-1.8; The formula of said dry material is: crushed corn cob 40-50%, sawdust 40-46%, wheat bran 6-12%, sucrose 0.5-1.5%, gypsum 1-2%, urea 0.8%, potassium dihydrogen phosphate 0.5%. 5. a preparation method of selenium-enriched black fungus fermented liquid, is characterized in that, comprises the following steps: (a) Inoculate the activated black fungus strain on PDA slant medium and culture at 28°C for 8-12 days to obtain black fungus female species; (b) inoculate the black fungus mother species in a selenium-enriched liquid medium, and culture it in a shaking table at 23-28°C for 5-7 days to obtain a fermentation liquid; expand the culture, filter the expansion liquid to obtain selenium-enriched black fungus fermented mycelium; (c) mixing the fermented mycelia of selenium-enriched black fungus with sterile water at a ratio of 1:5-15, grinding, and ultrasonicating to obtain a fermented mycelium of selenium-enriched black fungus; (d) inoculating the black fungus strain on a selenium-enriched solid medium, and carrying out selenium-enriching cultivation on the black fungus strain to obtain selenium-enriched black fungus fruiting bodies, and drying to obtain a selenium-enriching black fungus fruiting body dry product; (e) Soak the dried selenium-enriched black fungus fruit body in water at 40-50°C for 1-2h, remove impurities, dry at 90-100°C for 5-7h, and crush to 140-180 mesh to obtain selenium-enriched black Fungus fruiting body powder: mix the selenium-enriched black fungus fruiting body powder with water according to the material-to-water ratio of 1:10-30, soak for 1-1.5 h, grind, and ultrasonically obtain the selenium-enriching black fungus fruiting body slurry; (f) Mixing the selenium-enriched black fungus fruit body slurry and the selenium-enriched black fungus fermented mycelium slurry at a volume ratio of 2:3 to obtain the selenium-enriched black fungus fermented slurry. 6. an application of the selenium-enriched black fungus fermented liquid as claimed in claim 1 in the preparation of a blood fat-lowering health drink. 7. The application of the selenium-enriched black fungus fermented liquid as claimed in claim 1 together with a food flavoring agent in the preparation of seasoning selenium-enriched black fungus fermented liquid drink. 8. a seasoning selenium-enriched black fungus fermented liquid, it is characterized in that, in the prepared selenium-enriched black fungus fermented liquid of described claim 1, honey is added, and the volume ratio of adding honey and selenium-enriched black fungus fermented liquid is 1: 15-20, stir well to obtain seasoned selenium-enriched black fungus fermented liquid.