CN114424815B - Fermented black bean sprout powder with effects of dispelling effects of alcohol and protecting liver and preparation method thereof - Google Patents
Fermented black bean sprout powder with effects of dispelling effects of alcohol and protecting liver and preparation method thereof Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/70—Germinated pulse products, e.g. from soy bean sprouts
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- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- C12N1/14—Fungi; Culture media therefor
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- C12N1/18—Baker's yeast; Brewer's yeast
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention discloses fermented black bean sprout powder with the effects of dispelling effects of alcohol and protecting liver and a preparation method thereof, and relates to the technical field of food fermentation. The results show that the fermented black bean sprout powder can reduce the blood alcohol concentration of a alcoholism rat, reduce the level of serum AST, ALT, ALP, inhibit the release of TNF-alpha, IL-6 and IL-1 beta to reduce inflammatory reaction, improve the activity of antioxidant enzymes such as ADH, ALDH, CAT in the liver and further inhibit oxidative stress reaction. The research shows that the medicine and food homologous fermented black bean sprout powder can improve the liver injury degree of rats caused by acute alcoholism, shows that the fermented black bean sprout powder has a certain alcohol dispelling and liver protecting effect, and can be developed as a health care product for dispelling alcohol and protecting liver.
Description
Technical Field
The invention relates to the technical field of food fermentation, in particular to fermented black bean sprout powder with the effects of dispelling the effects of alcohol and protecting liver and a preparation method thereof.
Background
Alcoholic liver disease caused by drinking is one of the most common liver diseases in the world, and in recent years, as the intake of alcohol increases, the incidence of alcoholic liver disease is also increasing, which is attracting attention worldwide. Research shows that oxidative stress is a main mechanism of the pathogenesis of alcoholic liver diseases, and alcohol generates a large amount of active oxygen in the metabolic process, so that the oxidative stress reaction and abnormal lipid metabolism of an organism are caused, and the health of a human body is seriously influenced. The enhancement of the antioxidant capacity of organisms is considered as an effective way for preventing and treating alcoholic liver diseases, but most of the current medicines for dispelling effects of alcohol and protecting liver are combined medicines, have certain side effects, and are to be developed into a healthy product with the effects of dispelling effects of alcohol and protecting liver.
With the improvement of the living standard of people, the development of functional foods is also more rapid. At present, the processing mode of the black beans is single, so that the nutrition value of the black beans is not effectively utilized. The composite probiotics fermentation is favored because of the important functions of enhancing the flavor, enhancing the fermentation performance, improving the fermentation efficiency and the like, and DU-106 is an aerobic lactic acid bacillus with extremely high proliferation capability, the genome of the composite probiotics fermentation lacks virulence genes, and the DU-106 has good functions in the aspect of collaborative fermentation. At present, the bean products with the effect of dispelling the effects of alcohol are not more, and researches show that the bean products such as black beans, natto and mung beans have good effects of dispelling the effects of alcohol and protecting liver. The fermented black bean sprout powder is a medicinal and edible product prepared by taking black beans as raw materials, adding various auxiliary materials and fermenting various strains. The fermented black bean sprout powder developed by the invention is rich in various active ingredients, such as soybean isoflavone and procyanidine, which have the functions of resisting oxidization, protecting liver and the like, and can inhibit liver injury by reducing the level of intracellular ROS.
In conclusion, the invention explores the effect of the self-developed fermented black bean sprout powder on the aspects of dispelling the effects of alcohol and protecting the liver, and provides application basis for the high added value utilization of black beans.
Disclosure of Invention
In order to solve the problems, the invention takes black beans as raw materials, and is supplemented with kudzuvine root, kudzuvine flower and hovenia dulcis thunb, and the fermented black bean sprout powder is produced by fermentation of DU-106 composite strains, so that the invention is used for preventing alcoholic liver diseases and improving the industrial value of the black beans.
A preparation method of fermented black bean sprout powder with effects of dispelling effects of alcohol and protecting liver comprises the following steps:
s1, selecting: selecting black beans produced in the current year, and removing wrinkled, moldy, worm-eroded and deteriorated black beans;
s2, germination: cleaning the selected black beans with running water, draining, adding clear water to submerge the surface of the black beans for 1-2cm, soaking for 12-24h, taking out the black beans, slightly draining water, spreading on 1-2 layers of bamboo plaiting plates, covering with wet towels, sprouting in a incubator in a dark place, replacing the towels every 4-8h, and obtaining sprouted black beans after 36-72 h;
s3, steaming and sterilizing: placing black beans with good germination conditions into a sterilizing pot for sterilizing treatment, and cooling for later use;
s4, smashing: placing the cooled black bean sprouts after sterilization treatment into a pulverizer to be crushed into coarse sand-shaped broken black bean sprouts for standby;
s5, auxiliary material treatment: boiling radix Puerariae, flos Puerariae Lobatae, and semen Hoveniae in water for 1-1.5 hr, and filtering to obtain adjuvant filtrate;
s6, uniformly mixing, namely scattering the auxiliary material filtrate obtained in the step S5 into the crushed black bean sprouts obtained in the step S4, and stirring the crushed black bean sprouts while scattering until the crushed black bean sprouts are fully and uniformly mixed;
s7, strain activation and expansion culture:
inoculating Rhizopus chinensis 12 into a slant activating (PDA) culture medium, culturing in a 28-32deg.C incubator for 48-72 hr, picking up the activated Rhizopus chinensis 12, and shake culturing in a liquid culture medium at 28-32deg.C for 48-72 hr to obtain Rhizopus chinensis 12 seed solution;
with reference to the method, after activating the beer yeast, performing expansion culture to obtain beer yeast seed liquid;
weighing 0.2g of lactobacillus DU-106, adding into 100mL of lactobacillus proliferation liquid culture medium, and culturing at 35-37 ℃ for 12-24 hours to obtain lactobacillus DU-106 seed liquid;
s8, primary fermentation: inoculating 10% -15% (v/w) of rhizopus chinensis 12 into the crushed black bean sprouts treated in the step S6, fermenting for 3-7 days at 20-35 ℃, and until the crushed black bean sprouts show full white hyphae;
s9, secondary fermentation: fully mixing crushed black bean sprouts homogenate of overgrown white hypha with 1% -2% (v/w) beer yeast and 0.25% -0.5% (v/w) DU-106, and continuing fermenting for 7-15 days to obtain fermented black bean sprouts;
s10, drying and sieving powder: drying the fermented black bean sprouts for 24-48h by adopting a vacuum freeze drying method, crushing the dried fermented black bean sprouts, and sieving the crushed fermented black bean sprouts with a 80-mesh sieve to obtain the fermented black bean sprout powder.
Preferably, the preparation method of the liquid medium for the expansion culture in step S7 is as follows:
200g of fresh potato, 20g of glucose, 1000mL of distilled water and autoclaving at 121 ℃ for 15min.
Preferably, the preparation method of the lactobacillus proliferation liquid medium in step S7 is as follows:
glucose 5g, protein 5g, yeast extract 1g, calcium chloride 0.1g, agar 15g, distilled water 1000mL, pH6.5, and sterilization at 115℃for 20min.
Preferably, in step S2, the incubator temperature is maintained at 25-30 ℃.
Preferably, in the step S3, the temperature of the sterilizing pot is 121 ℃ and the sterilizing process is carried out for 15min.
The invention has the beneficial effects that:
the study uses an acute alcoholic liver injury rat as a model to evaluate the anti-alcohol effect of the fermented black bean sprout powder. The results show that the fermented black bean sprout powder can reduce the blood alcohol concentration of a alcoholism rat, reduce the level of serum AST, ALT, ALP, inhibit the release of TNF-alpha, IL-6 and IL-1 beta to reduce inflammatory reaction, improve the activity of antioxidant enzymes such as ADH, ALDH, CAT in the liver and further inhibit oxidative stress reaction. The research shows that the medicine and food homologous fermented black bean sprout powder can improve the liver injury degree of rats caused by acute alcoholism, shows that the fermented black bean sprout powder has a certain alcohol dispelling and liver protecting effect, and can be developed as a health care product for dispelling alcohol and protecting liver.
Drawings
FIG. 1 is a bar graph of liver injury associated enzyme levels in rat serum;
FIG. 2 is a bar graph of liver injury associated enzyme levels in rat liver tissue;
FIG. 3 is a bar graph of serum inflammatory factor levels in rats;
FIG. 4 is a bar graph of blood lipid levels in rat serum;
FIG. 5 is a bar graph of the levels of rat serum alcohol metabolism related enzymes;
FIG. 6 is a bar graph of ethanol metabolism related enzyme levels in rat liver tissue;
FIG. 7 is a bar graph of rat liver antioxidant-associated enzyme levels.
Detailed Description
The technical scheme of the present invention will be described in further detail with reference to the accompanying drawings in the embodiments of the present invention, but the present invention is not limited to the following embodiments. All other embodiments, which are derived from the embodiments of the invention without creative efforts of a person skilled in the art, belong to the protection scope of the present invention.
A preparation method of fermented black bean sprout powder with effects of dispelling effects of alcohol and protecting liver comprises the following steps:
s1, selecting: selecting black beans produced in the current year, and removing wrinkled, moldy, worm-eroded and deteriorated black beans;
s2, germination: cleaning the selected black beans with running water, draining, adding clear water to submerge the surface of the black beans for 1-2cm, soaking for 12-24h, taking out the black beans, slightly draining water, spreading on 1-2 layers of bamboo plaiting plates, covering with wet towels, germinating in a incubator at 25-30 ℃ in a dark place, replacing the towels every 4-8h, and obtaining germinated black beans after 36-72 h;
s3, steaming and sterilizing: placing black beans with good germination condition in a sterilizing pot at 121 ℃ for 15min for sterilization treatment, and cooling for later use;
s4, smashing: placing the cooled black bean sprouts after sterilization treatment into a pulverizer to be crushed into coarse sand-shaped broken black bean sprouts for standby;
s5, auxiliary material treatment: boiling radix Puerariae, flos Puerariae Lobatae, and semen Hoveniae in water for 1-1.5 hr, and filtering to obtain adjuvant filtrate;
s6, uniformly mixing, namely scattering the auxiliary material filtrate obtained in the step S5 into the crushed black bean sprouts obtained in the step S4, and stirring the crushed black bean sprouts while scattering until the crushed black bean sprouts are fully and uniformly mixed;
s7, strain activation and expansion culture:
inoculating Rhizopus chinensis 12 into a slant activating (PDA) culture medium, culturing in a 28-32deg.C incubator for 48-72 hr, picking up the activated Rhizopus chinensis 12, and shake culturing in a liquid culture medium at 28-32deg.C for 48-72 hr to obtain Rhizopus chinensis 12 seed solution;
with reference to the method, after activating the beer yeast, performing expansion culture to obtain beer yeast seed liquid;
weighing 0.2g of lactobacillus DU-106, adding into 100mL of lactobacillus proliferation liquid culture medium, and culturing at 35-37 ℃ for 12-24 hours to obtain lactobacillus DU-106 seed liquid;
s8, primary fermentation: inoculating 10% -15% (v/w) of rhizopus chinensis 12 into the crushed black bean sprouts treated in the step S6, fermenting for 3-7 days at 20-35 ℃, and until the crushed black bean sprouts show full white hyphae;
s9, secondary fermentation: fully mixing crushed black bean sprouts homogenate of overgrown white hypha with 1% -2% (v/w) beer yeast and 0.25% -0.5% (v/w) DU-106, and continuing fermenting for 7-15 days to obtain fermented black bean sprouts;
s10, drying and sieving powder: drying the fermented black bean sprouts for 24-48h by adopting a vacuum freeze drying method, crushing the dried fermented black bean sprouts, and sieving the crushed fermented black bean sprouts with a 80-mesh sieve to obtain the fermented black bean sprout powder.
Specifically, the preparation method of the liquid medium for the expansion culture in the step S7 is as follows:
200g of fresh potato, 20g of glucose, 1000mL of distilled water and autoclaving at 121 ℃ for 15min.
Specifically, the preparation method of the lactobacillus proliferation liquid culture medium in the step S7 is as follows:
glucose 5g, protein 5g, yeast extract 1g, calcium chloride 0.1g, agar 15g, distilled water 1000mL, pH6.5, and sterilization at 115℃for 20min.
In order to explore the efficacy of dispelling the effects of alcohol and protecting liver of the fermented black bean sprout powder, 2 experimental verification is carried out respectively:
comparative example 1:
and (3) pouring normal saline into the stomach of the experimental rat, and finally pouring the normal saline into the stomach for molding.
Comparative example 2:
the experimental rats were perfused with physiological saline, and finally the model was perfused with 12 mL/(kg.bw) 52℃Erguotou.
Comparative example 3:
the experimental rats were filled with 2 g/(kg.bw) of infinite Jin Zhao capsules and finally the molds were filled with 12 mL/(kg.bw) of 52℃red star Erguotou.
Example 1:
the experimental rats were filled with fermented black bean sprout powder at a gastric dose of 0.5 g/(kg.bw), and finally were subjected to gastric filling with 12 mL/(kg.bw) of 52℃Erguotou white spirit.
Example 2:
the experimental rat is filled with fermented black bean sprout powder with the gastric dose of 1 g/(kg.bw), and finally 12 mL/(kg.bw) of 52-degree red star Erguotou white spirit is used for filling the stomach.
Experimental example 1: evaluation of anti-hangover Effect
2.1 laboratory animals
SPF-class male SD rats, 40, weighing 180 g-220 g, were offered by the medical laboratory animal center in Guangdong province.
2.2 Experimental methods
After experimental animals were adaptively fed for one week, SD rats were randomly divided into control 1, control 2, control 3 and example 1, 10 rats per group, and each group was filled with corresponding samples according to 10 mL/(kg. Bw) of gastric juice, 0.9% physiological saline was filled in control 1 and control 2, 2 g/(kg. Bw) of infinite Jin Zhao capsules were filled in control 3, 0.5 g/(kg. Bw) of fermented black bean sprout powder was filled in example 1, 2 times per day, and 4 hours intervals. After the last administration for 30min, 12 mL/(kg.bw) of 52-degree red star Erguotou white spirit was administered by gastric lavage to cause acute alcoholism in SD rats in each group except for comparative example 1, and an equal amount of 0.9% physiological saline was administered in comparative example 1. The orbital vein of the rat was sampled 1h, 2h, and 4h after the gastric lavage of alcohol, respectively.
2.3 evaluation of anti-hangover Effect
As can be seen from Table 1, the ethanol concentration of comparative example 1 was zero, and the ethanol concentration of comparative example 2 was higher than that of the other groups, indicating that the modeling of acute alcoholism was successful. The blood alcohol concentration of the rats in the comparison example 2 rises sharply after drinking for 2 hours, which is significantly higher than that of the rats in the comparison example 3 and the experimental example 1 (P < 0.01), the blood alcohol concentration of the rats in each group is reduced after drinking for 4 hours, the blood alcohol level of the rats in the experimental example is the lowest, and the difference reaches an extremely significant level (P < 0.01) compared with the rats in the comparison example 2. The result shows that the fermented black bean sprout powder can effectively inhibit the rise of blood alcohol concentration and has the function of reducing blood alcohol, and the purpose of dispelling the effects of alcohol can be achieved by reducing the blood alcohol concentration.
TABLE 1 Effect of fermented Black Bean sprout powder on acute alcoholism rat blood alcohol concentration
Table 1 notes: # p represents a significant difference (P < 0.05) from comparative example 1; * p represents a significant difference (P < 0.05) from comparative example 2; ## p represents a very significant difference (P < 0.01) from comparative example 1; ** p represents the extremely significant difference (P < 0.01) from comparative example 2, as follows.
Experimental example 2: evaluation of anti-hangover and liver-protecting efficacy
3.1 laboratory animals
SPF-class male SD rats, 40, weighing 180 g-220 g, were offered by the medical laboratory animal center in Guangdong province.
3.2 Experimental methods
After the experimental animals were fed adaptively for one week, the rats were randomly divided into control 1 (0.9% physiological saline), control 2 (0.9% physiological saline), control 3 (2 g/(kg·bw) Infinite Jin Zhao capsule), example 1 (0.5 g/(kg·bw)), and example 2 (1 g/(kg·bw)). Each group of rats was subjected to gastric lavage with 10 mL/(kg·bw) of each group of corresponding samples once per 1 day for 7 days, and after the last administration for 30min, 12 mL/(kg·bw) of 52 ° red star Erguotou white spirit was added to each group of rats except for comparative example 1, and after 12h of no water forbidden, rats were anesthetized with 10% chloral hydrate, abdominal artery blood collection was performed on the rats, and the livers of the rats were stored in a refrigerator at-80 ℃ for later use.
3.3 efficacy evaluation
3.3.1 liver function index
As shown in FIG. 1, compared with comparative example 1, the AST, ALT, ALP enzyme activities of the rat serum of comparative example 2 are significantly higher than those of comparative example 1 (P < 0.01); in fig. 2, the liver AST, ALT, ALP enzyme activity of the rat in the comparative example 1 is also obviously higher than that in the comparative example 2, which shows that acute alcoholism can lead to the rapid increase of the liver injury related enzyme activity of the rat, and the modeling is successful. As shown in fig. 1A, both experimental examples 1 and 2 were able to reduce serum AST enzyme activity, and the difference of experimental example 2 reached a significant level (P < 0.05), and experimental example 1 did not have a significant difference; FIGS. 1B and 2B illustrate that fermented black bean sprout powder and Infinite Jin Zhao capsule can reduce serum and liver ALT enzyme activities; as shown in FIGS. 1C and 2C, each treatment group was able to reduce ALP enzyme activity in rat serum and liver, with the difference reaching a very significant level (P < 0.01). Experimental results show that acute alcoholism can cause acute liver injury of rats, so that related enzyme activity is increased, and the fermented black bean sprout powder reduces the liver injury degree caused by alcohol by reducing the enzyme activity of AST, ALT, ALP.
Fig. 1 notes: # p represents a significant difference (P < 0.05) from comparative example 1; * p represents a significant difference (P < 0.05) from comparative example 2; ## p represents a very significant difference (P < 0.01) from comparative example 1; ** p represents the extremely significant difference (P < 0.01) from comparative example 2, as follows.
3.3.2 inflammatory factor index
Alcohol intake stimulates liver macrophages, which in turn promotes the release of inflammatory factors TNF-alpha, IL-1 beta, IL-6. As shown in fig. 3, the levels of TNF- α, IL-1β, IL-6 in comparative example 2 were significantly increased (P < 0.01) compared to comparative example 1, and both the group of comparative example 3 and the experimental example were able to suppress the increase in the level of inflammatory factors induced by alcohol, except that the effect of example 2 on the inhibition of TNF- α levels was not significant, and the inhibition effect was extremely significant (P < 0.01), indicating that fermented black bean sprout powder could reduce the inflammatory reaction caused by alcohol by inhibiting the release of inflammatory factors.
3.3.3 evaluation of blood lipid
As shown in fig. 4A, acute alcoholic liver injury resulted in a significant increase in TC (P < 0.01) compared to comparative example 1, while both comparative example 3 and example had a significant inhibition of TC increase (P < 0.01), and example 2 had a better inhibition effect; similarly, each of the administration examples also significantly reduced the TG level compared to comparative example 2, and comparative example 2 had a TG level higher than that of comparative example 1, but the difference was not significant (fig. 4B). As shown in FIGS. 4C and 4D, the acute alcoholic liver injury had no significant effect on the levels of LDL-C and HDL-C, and control 2 significantly reduced the level of LDL-C. The results show that the fermented black bean sprout powder can improve lipid metabolism disorder induced by alcohol and can obviously reduce the levels of serum TC and TG.
3.3.4 ethanol Metabolic enzyme index
As shown in FIGS. 5 and 6, the enzyme levels involved in alcohol metabolism in the body were reduced in the serum and liver of comparative example 2 as compared with comparative example 1, and the ADH in the serum of comparative example 2 was extremely different from that of comparative example 1 (P < 0.01). And the ADH enzyme activity of the rats subjected to the treatment intervention of the fermented black bean sprout powder is higher than that of the control example 2; correlation of CYP2E1 with another pathway of alcohol metabolism As shown in FIGS. 5C and 6C, the CYP2E1 content of control 2 was significantly increased (P < 0.01) compared with control 1, the CYP2E1 level in serum of each administration group was significantly lower than that of control 2 (P < 0.01), and the CYP2E1 level in liver was also lower than that of control 2, but the difference was insignificant (P > 0.05). Experimental results show that the alcohol stimulation can reduce the enzyme activities of ADH and ALDH in serum and liver tissues of rats, and simultaneously improve the enzyme activities of CYP2E1, and the intervention of the fermented black soybean sprout powder can improve the enzyme activities of ADH and ALDH and reduce the level of CYP2E 1.
3.3.5 antioxidant enzyme index
CAT, SOD, GSH-Px in the liver plays an important role in the antioxidant capacity of rat liver, the measurement results of enzyme activity related to the antioxidant effect in the rat liver are shown in figure 7, the enzyme activity of the liver CAT, SOD, GSH-Px can be obviously reduced by alcohol treatment, and the reduction of the antioxidant enzyme activity of the liver can be prevented to different degrees by pretreatment of fermented black bean sprout powder of the rat for 7 days. As shown in fig. 7A and 7C, both example 1 and example 2 can significantly improve the activity of CAT and SOD (P < 0.01), and comparative example 3 can improve the activity of CAT without significant difference (P < 0.05) compared to comparative example 2. Example 2 in FIG. 7B is able to significantly increase GSH-Px enzyme activity (P < 0.01) compared to comparative example 2, whereas example 1 has no significant difference in the increase effect (P < 0.01). Experimental results show that the fermented black bean sprout powder can improve the antioxidant capacity of rats with acute alcoholic liver injury, and clear active oxygen generated by ethanol metabolism, so that the fermented black bean sprout powder has the effects of dispelling the effects of alcohol and protecting the liver.
In conclusion, the study uses an acute alcoholic liver injury rat as a model to evaluate the anti-alcohol effect of the fermented black bean sprout powder. The results show that the fermented black bean sprout powder can reduce the blood alcohol concentration of a alcoholism rat, reduce the level of serum AST, ALT, ALP, inhibit the release of TNF-alpha, IL-6 and IL-1 beta to reduce inflammatory reaction, improve the activity of antioxidant enzymes such as ADH, ALDH, CAT in the liver and further inhibit oxidative stress reaction. The research shows that the medicine and food homologous fermented black bean sprout powder can improve the liver injury degree of rats caused by acute alcoholism, shows that the fermented black bean sprout powder has a certain alcohol dispelling and liver protecting effect, and can be developed as a health care product for dispelling alcohol and protecting liver.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present disclosure describes embodiments, not every embodiment is provided with a separate embodiment, and that this description is provided for clarity only, and that the disclosure is not limited to the embodiments described in detail below, and that the embodiments described in the examples may be combined as appropriate to form other embodiments that will be apparent to those skilled in the art.
Claims (1)
1. The preparation method of the fermented black bean sprout powder with the effects of dispelling the effects of alcohol and protecting the liver is characterized by comprising the following steps of:
s1, selecting: selecting black beans produced in the current year, and removing wrinkled, moldy, worm-eroded and deteriorated black beans;
s2, germination: cleaning the selected black beans with running water, draining, adding clear water to submerge the surface of the black beans for 1-2cm, soaking for 12-24h, taking out the black beans, slightly draining water, spreading on 1-2 layers of bamboo plaiting plates, covering with wet towels, sprouting in a incubator in a dark place, keeping the temperature of the incubator at 25-30 ℃, replacing the towels every 4-8h, and obtaining sprouted black beans after 36-72 h;
s3, steaming and sterilizing: placing black beans with good germination condition into a sterilizing pot for sterilizing at 121 ℃ for 15min, and cooling for later use;
s4, smashing: placing the cooled black bean sprouts after sterilization treatment into a pulverizer to be crushed into coarse sand-shaped broken black bean sprouts for standby;
s5, auxiliary material treatment: boiling radix Puerariae, flos Puerariae Lobatae, and semen Hoveniae in water for 1-1.5 hr, and filtering to obtain adjuvant filtrate;
s6, uniformly mixing, namely scattering the auxiliary material filtrate obtained in the step S5 into the crushed black bean sprouts obtained in the step S4, and stirring the crushed black bean sprouts while scattering until the crushed black bean sprouts are fully and uniformly mixed;
s7, strain activation and expansion culture:
inoculating Rhizopus chinensis 12 into a slant activating (PDA) culture medium, culturing in a 28-32deg.C incubator for 48-72 hr, picking up the activated Rhizopus chinensis 12, and shake culturing in a liquid culture medium at 28-32deg.C for 48-72 hr to obtain Rhizopus chinensis 12 seed solution;
with reference to the method, after activating the beer yeast, performing expansion culture to obtain beer yeast seed liquid;
weighing 0.2g of lactobacillus DU-106, adding into 100mL of lactobacillus proliferation liquid culture medium, and culturing at 35-37 ℃ for 12-24 hours to obtain lactobacillus DU-106 seed liquid;
the preparation method of the liquid culture medium for the expansion culture comprises the following steps:
200g of fresh potatoes, 20g of glucose, 1000mL of distilled water and autoclaving at 121 ℃ for 15min;
the preparation method of the lactobacillus proliferation liquid culture medium comprises the following steps:
glucose 5g, protein 5g, yeast extract 1g, calcium chloride 0.1g, agar 15g, distilled water 1000mL, pH6.5, and sterilization at 115 ℃ for 20min;
s8, primary fermentation: inoculating 10% -15% (v/w) of rhizopus chinensis 12 into the crushed black bean sprouts treated in the step S6, fermenting for 3-7 days at 20-35 ℃, and until the crushed black bean sprouts show full white hyphae;
s9, secondary fermentation: fully mixing crushed black bean sprouts homogenate of overgrown white hypha with 1% -2% (v/w) beer yeast and 0.25% -0.5% (v/w) DU-106, and continuing fermenting for 7-15 days to obtain fermented black bean sprouts;
s10, drying and sieving powder: drying the fermented black bean sprouts for 24-48h by adopting a vacuum freeze drying method, crushing the dried fermented black bean sprouts, and sieving the crushed fermented black bean sprouts with a 80-mesh sieve to obtain the fermented black bean sprout powder.
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| CN108094794A (en) * | 2017-12-31 | 2018-06-01 | 佛山市艳晖生物科技有限公司 | A kind of complex microorganism drink of relieving alcoholism and protecting liver and preparation method thereof |
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