Preparation method of mulberry fruit wine with probiotic effect

Abstract

The preparation method comprises the following steps: squeezing mulberry fruits to obtain mulberry fruit juice and mulberry fruit residues, fermenting the mulberry fruit juice with saccharomyces cerevisiae to prepare the mulberry fruit wine, carrying out micro-electrolysis on the mulberry fruit residues, filtering, carrying out reverse osmosis desalination treatment on the filtrate, and concentrating and sterilizing to obtain a concentrated solution; adding the concentrated solution into mulberry juice which is fermented for 2 days, inoculating lactobacillus acidophilus for anaerobic fermentation, and recording the fermentation liquor after fermentation as fermentation liquor A; uniformly mixing the fermentation liquor A, lactose and mulberry fruit juice fermented for 4 days, continuing to ferment, and filtering and aging after fermentation to obtain the mulberry fruit wine. The technology is extremely simple, the production cost is low, the mulberry fruit wine residues are recycled while the probiotic effect of the mulberry fruit wine is remarkably improved, and the method has important significance for improving the quality of the mulberry fruit wine and recycling the mulberry fruit wine residues.

Description

Technical Field

[0001] The invention relates to the field of fruit wine processing, in particular to a method for preparing mulberry wine. Background Art

[0002] Mulberry, also known as mulberry fruit, mulberry date, etc., is the mature fruit of the perennial woody plant of the Moraceae family, the mulberry tree. Studies have shown that mulberry contains a full range of amino acids, as many as 18 kinds, and the total content is 1 times higher than that of fruits such as bananas, grapes, pears and citrus. In addition, mulberry also contains a variety of vitamins and minerals, and is a nutritious seasonal fruit. Mulberry is very easy to spoil. In the season when mulberries are abundant, processing mulberry fruits can effectively solve the problem of seasonal overcapacity of mulberry fruits and reduce the market entry of fresh mulberries. Deep processing of mulberry fruits can effectively promote the development of mulberry planting and processing industries and improve the commercial value of mulberry fruits. Mulberry wine is an alcoholic beverage brewed by fermentation using mulberry fruits as raw materials. Mulberry wine is rich in polyphenols, flavonoids, anthocyanins and various ester substances, which give mulberry wine a variety of nutrients and special flavors. Therefore, mulberry wine is deeply loved by consumers.

[0003] At present, mulberry wine is mainly brewed by fermenting mulberry juice after squeezing with brewer's yeast as the main fermentation strain. Mulberry juice itself also contains a variety of organic acids. During the fermentation process, brewer's yeast will also produce organic acids including acetic acid, butyric acid and heptanoic acid. Therefore, mulberry wine brewed by conventional methods has the disadvantage of a "sour" taste. In addition, because there are fewer substances with a "mulberry flavor" remaining in the juice, mulberry wine also has the disadvantage of not having a strong "mulberry flavor". The above shortcomings significantly affect the quality of mulberry wine and consumers' acceptance of mulberry wine.

[0004] Improving the "sour and astringent" taste of mulberry wine and giving it a stronger "mulberry flavor" have always been research hotspots in the mulberry wine industry. For example, Li Ruiyi et al. found that the filtration method has a significant impact on the quality of mulberry wine. Both cardboard coarse filtration-membrane fine filtration composite filtration and cardboard coarse filtration-centrifugal fine filtration treatments can significantly reduce the aroma substance content of mulberry wine, among which the aroma substance loss rate of mulberry wine after cardboard coarse filtration-centrifugal fine filtration treatment is lower. Wang Yu et al. found that pre-treatment of mulberry slurry with different pectinases before fermentation can significantly improve the quality of mulberry wine, and different pectinases have significantly different effects on the total acid and flavor substance content in mulberry wine. Luo Wenshan et al. studied that composite fermentation (non-brewing yeast and brewing yeast are used together for the fermentation of mulberry wine) can significantly improve the quality of mulberry wine; the mulberry wine that meets the fermentation requirements has a lower total acid content and a stronger wine and fruit aroma.

[0005] Although there have been reports on improving the quality of fruit wine, there have been no reports on imparting prebiotic effects to mulberry wine. Therefore, the present invention introduces pectin oligosaccharides with prebiotic effects into mulberry wine, hoping to increase the prebiotic effects of mulberry wine while also solving the problems of mulberry wine quality such as "sour" taste and "mulberry flavor" not being obvious. Summary of the invention

[0006] The purpose of the present invention is to solve the above technical problems and provide a method for preparing mulberry wine which can impart probiotic effects to mulberry wine while solving the problems of "sour and astringent" taste and lack of "mulberry flavor" of mulberry wine and significantly improving the quality of mulberry wine.

[0007] The method for preparing mulberry wine of the present invention comprises the steps of squeezing mulberry fruits to obtain mulberry juice and mulberry pomace, fermenting the mulberry juice with brewer's yeast to prepare mulberry wine,

[0008] The mulberry pomace is subjected to micro-electrolysis and then filtered, and the filtrate is subjected to reverse osmosis desalination treatment and then concentrated and sterilized to obtain a concentrated solution;

[0009] The concentrate was added to mulberry juice that had been fermented for 2 days and inoculated with Lactobacillus acidophilus for anaerobic fermentation. The fermentation liquid at the end of the fermentation was recorded as fermentation liquid A.

[0010] The fermentation liquid A, lactose and mulberry juice that has been fermented for 4 days are mixed evenly and then continued to ferment. After the fermentation is completed, the mixture is filtered and aged to obtain mulberry wine.

[0011] The conditions of the micro-electrolysis are as follows: distilled water is added to the mulberry pomace at a mass ratio of 1:30, graphite is used as the negative electrode, platinum is used as the positive electrode, the distance between the electrodes is 40 cm, sodium nitrate electrolyte is added to the mulberry pomace treated with water at a mass ratio of 0.5:100, and electrolysis is performed at a voltage of 25 V for 12 hours under stirring conditions.

[0012] The filtrate is subjected to reverse osmosis desalination treatment and concentrated to 1 / 10 of the original solution, and then sterilized to obtain a concentrated solution;

[0013] The mixing mass ratio of the concentrated liquid to the mulberry juice fermented for 2 days is 1:50-70, and the fermentation conditions are: anaerobic fermentation at 37° C. for 24-36 hours.

[0014] The inoculation amount of Lactobacillus acidophilus in the mulberry juice fermented for 2 days is 0.8-1.6 ml / L, and the concentration of lactic acid bacteria used in the Lactobacillus acidophilus is (2-7)×10 6 cfu / mL.

[0015] The strain number of the Lactobacillus acidophilus is CICC 6083.

[0016] The mixing mass ratio of the fermentation liquid A, lactose and mulberry juice fermented for 4 days is 1:(0.2-0.4):(10000-50000).

[0017] The fermentation liquid A, lactose and mulberry juice fermented for 4 days are uniformly mixed, and then vacuum oxygen extraction treatment is performed, and then fermentation is continued for 3 days and then aged to obtain mulberry wine.

[0018] In order to solve the problems existing in the background technology, the inventors have made the following improvements:

[0019] The inventor has found that Lactobacillus acidophilus can improve the "sour and astringent" taste of fruit wine to a certain extent, but due to the presence of brewer's yeast, Lactobacillus acidophilus cannot directly grow and breed in partially fermented mulberry juice, and cultivating Lactobacillus acidophilus with nutrients of other ingredients and then adding them to mulberry wine will affect the "mulberry taste" of mulberry wine. For this reason, the inventor considers making full use of mulberry wine dregs, and extracting pectin oligosaccharides with probiotic effects therefrom by micro-electrolysis method, which can be used as nutrients to promote the growth and breeding of Lactobacillus acidophilus, and promote Lactobacillus acidophilus to grow and breed in partially fermented mulberry juice, thereby solving the problem that Lactobacillus acidophilus cannot grow and breed in partially fermented mulberry juice. And the pectin oligosaccharides for cultivating Lactobacillus acidophilus are extracted from mulberry wine dregs, which will increase the mulberry taste unique to mulberry wine, and further improve the quality and mouthfeel of wine.

[0020] The micro-electrolysis method is a prior art. As an example, the micro-electrolysis method described in the patent (ZL201910646941.5) can be used. For mulberry pomace, the conditions of the micro-electrolysis are: distilled water is added to the mulberry pomace at a mass ratio of 1:30, graphite is used as the negative electrode, platinum is used as the positive electrode, and the distance between the electrodes is 40cm. Sodium nitrate electrolyte is added to the mulberry pomace treated with water at a mass ratio of 0.5:100, and electrolysis is performed under a voltage of 25V for 12 hours under stirring conditions. Then, the fermentation liquid A of Lactobacillus acidophilus cultured with partially fermented mulberry juice is added to the mulberry juice that has been fermented for 4 days, and lactose is added to further ferment the mulberry juice with brewer's yeast, and then conventional subsequent filtration and aging are performed to obtain mulberry wine that not only has probiotic effects, but also has rich "sour taste" and strong "mulberry flavor", and the quality is significantly improved.

[0021] Furthermore, the addition ratio of the concentrate, the strain number, concentration, addition amount, culture time, mixing ratio, and lactose dosage and ratio of the Lactobacillus acidophilus have a certain influence on the quality of the final mulberry wine. For example, when the mixing ratio of the concentrate and the mulberry wine that has been fermented for 2 days is higher than 1:50, the Lactobacillus acidophilus will multiply rapidly in the fermentation liquid A and produce a large amount of acid; after the fermentation liquid A is mixed with the mulberry wine that has been fermented for 4 days, the Lactobacillus acidophilus will not adapt to the alcoholic fermentation juice, and then die quickly, and will not achieve the purpose of assisting the brewing yeast in the co-fermentation. When the mixing ratio of the concentrate and the mulberry wine that has been fermented for 2 days is lower than 1:70, the content of Lactobacillus acidophilus in the fermentation liquid A is low after the fermentation is completed. After mixing with the mulberry wine that has been fermented for 4 days, the number of Lactobacillus acidophilus is low in the subsequent fermentation with the brewing yeast, and the flavor and sourness of the mulberry wine cannot be produced due to the low content. Similarly, the selection of the concentration of lactobacillus within the above range, the addition amount, and the mixing ratio of the fermentation liquid and lactose to the mulberry juice fermented for 4 days is also based on the above considerations.

[0022] Further, in the method of the present invention, the pectin oligosaccharide concentrate is added to the mulberry juice that has been fermented for 2 days and accessed to an appropriate amount of Lactobacillus acidophilus, rather than the unfermented original mulberry juice, considering that the mulberry juice fermented for 2 days contains a certain amount of saccharomyces cerevisiae and the alcohol content is not high. At this time, the Lactobacillus acidophilus is accessed to allow the Lactobacillus acidophilus to adapt to the living environment containing trace alcohol and yeast and to reproduce in large quantities under this environment, increase the number of Lactobacillus acidophilus and metabolize to produce some flavor substances. The mixing time of the fermented liquid A containing Lactobacillus acidophilus is in the mulberry juice fermented for 4 days, and the mulberry juice at this time contains a large amount of saccharomyces cerevisiae and a relatively high alcohol content. Under this condition, Lactobacillus acidophilus has a certain effect on the growth and metabolism of saccharomyces cerevisiae, but it is impossible to replace saccharomyces cerevisiae as the main fermentation strain; with the subsequent further fermentation (alcohol content is further improved), the growth and metabolism of Lactobacillus acidophilus will be inhibited until death; and in this process, the substances produced by the metabolism of Lactobacillus acidophilus and the introduced pectin oligosaccharides will significantly change the flavor of mulberry wine.

[0023] Furthermore, there are many types of Lactobacillus acidophilus reported so far, and studies have found that not all Lactobacillus acidophilus are suitable for application in the method of the present invention. If Lactobacillus acidophilus numbered CICC 6074 is selected to replace the Lactobacillus acidophilus used in the present invention, the "sour and astringent" taste of mulberry wine cannot be significantly improved, and an abnormal sour smell will be produced, further reducing the wine taste of mulberry wine.

[0024] Further oxygen extraction treatment after the fermentation liquid A is mixed with the mulberry wine fermented for 4 days is also essential; if oxygen extraction treatment is not performed, the oxygen introduced during the mixing process will cause the produced ethanol to be oxidized into acetic acid during the aging process, thereby reducing the quality of the mulberry wine.

[0025] Beneficial effects:

[0026] The present invention appropriately adjusts and improves the existing mulberry wine brewing process, combines micro-electrolysis technology, and fully utilizes the nutrients in the mulberry pomace, a by-product of mulberry wine brewing, without significantly increasing the production cost. The present invention solves the problems of "sour and astringent" taste and lack of "mulberry flavor" of the mulberry wine, while making the mulberry wine have a prebiotic effect, significantly improving the nutritional value of the mulberry wine, and making the mulberry wine have higher commodity value and market competitiveness. DETAILED DESCRIPTION

[0027] In the following embodiments, the subsequent treatment refers to the filtration, aging, canning, packaging and other treatment measures commonly used in the mulberry wine brewing process. Those skilled in the art can select the steps and conditions for subsequent treatment according to specific needs. This is the prior art and will not be described in detail.

[0028] Among them, the strain number of Lactobacillus acidophilus in the following examples is CICC 6083,

[0029] Embodiment 1:

[0030] The squeezed mulberry juice is added to the activated brewer's yeast seed solution, and fermented at 28-30° C. for 7 days. After the fermentation is completed, the mulberry wine is obtained after filtering, clarifying and aging.

[0031] At the same time, distilled water was added to the mulberry pomace after juicing at a ratio of 1:30, with graphite as the negative electrode, platinum as the positive electrode, and the distance between the electrodes was 40 cm. Sodium nitrate electrolyte was added to the mulberry pomace treated with water at a ratio of 0.5:100, and electrolysis was carried out at 25V voltage under stirring conditions for 10 hours. . The electrolyzed mulberry pomace was filtered and the filtrate was collected. After the sodium chloride in the filtrate was removed by reverse osmosis treatment, the filtrate was concentrated to 10% of the mass of the original solution by a rotary vacuum concentrator and sterilized. After the concentrate was added to the mulberry juice that had been fermented for 2 days at a mass ratio of 1:50, 2×10 colonies were inoculated into the mulberry juice at a ratio of 1.6 mL / L. 6 cfu / mL of Lactobacillus acidophilus, and anaerobically fermented at 37°C for 24 hours to obtain fermentation liquid A. Fermentation liquid A containing Lactobacillus acidophilus and lactose were mixed evenly with mulberry juice that had been fermented for 4 days in a mass ratio of 1:0.2:10000 and vacuumed, and fermented for 3 days at the temperature and humidity required for mulberry wine fermentation. After the fermentation was completed, the insoluble matter in the fermentation liquid was filtered out and aged for 1 year according to the aging method of mulberry wine.

[0032] The lactic acid content in mulberry wine was determined to be 0.543 mg / L, the acetic acid content was 15.77 mg / L, the butyric acid content was 0.68 mg / L, the ethyl acetate content was 60.13 mg / L, the ethyl lactate content was 42.11 mg / L, the diethyl amber content was 18.21 mg / L, the ethyl benzoate content was 0.56 mg / L, the lauric aldehyde content was 0.27 mg / L, and the 2-phenylethanol content was 78.19 mg / L. According to sensory evaluation, the mulberry wine has a layered sour taste and astringency; it has a strong frankincense smell but does not cover the wine taste, and has a strong "mulberry flavor".

[0033] The mulberry wine was heated at 100°C for 25 minutes by heating and evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium in an amount of 3wt% and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way of culturing bifidobacteria (37°C, anaerobic culture for 24h); after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid was increased by 0.549 (the OD600 value of bifidobacteria cultured in the MRS liquid culture medium without adding mulberry wine was used as the control group). The results show that the pectin treated above can significantly promote the growth of bifidobacteria and has a probiotic effect (the same as in the subsequent embodiments).

[0034] Embodiment 2:

[0035] In this embodiment, the inoculation amount of Lactobacillus acidophilus is 0.8 mL / L, the fermentation time after inoculation is 36 h, the mass ratio of the fermentation liquid A and lactose to the mulberry juice fermented for 4 days is 1:0.4:30000, and the other parameters are the same as those in Example 1.

[0036] The lactic acid content in mulberry wine was determined to be 0.423 mg / L, the acetic acid content was 15.97 mg / L, the butyric acid content was 0.61 mg / L, the ethyl acetate content was 60.88 mg / L, the ethyl lactate content was 48.37 mg / L, the diethyl amber content was 20.16 mg / L, the ethyl benzoate content was 0.61 mg / L, the lauric aldehyde content was 0.33 mg / L, and the 2-phenylethanol content was 80.91 mg / L. According to sensory evaluation, the mulberry wine has a layered sour taste and astringency; it has a frankincense smell and does not cover the wine taste, and has a strong "mulberry flavor".

[0037] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium in an amount of 3wt% and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24h) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.413.

[0038] Embodiment 3:

[0039] In this example, the concentration of Lactobacillus acidophilus during inoculation was 5×10 6 cfu / mL, the inoculation amount was 1.2 mL / L, the fermentation time after inoculation was 30 h, the mixing ratio of fermentation liquid A and lactose to mulberry juice that had been fermented for 4 days was 1:0.3:50000, and the other parameters were the same as in Example 1.

[0040] The lactic acid content in mulberry wine was determined to be 0.397 mg / L, the acetic acid content was 15.91 mg / L, the butyric acid content was 0.88 mg / L, the ethyl acetate content was 68.14 mg / L, the ethyl lactate content was 47.68 mg / L, the diethyl amber content was 22.27 mg / L, the ethyl benzoate content was 0.90 mg / L, the lauric aldehyde content was 0.36 mg / L, and the 2-phenylethanol content was 60.11 mg / L. According to sensory evaluation, the mulberry wine has a rich sour taste and astringency, with a slight frankincense smell and a strong "mulberry flavor".

[0041] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium in an amount of 3wt% and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24h) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.484.

[0042] Embodiment 4:

[0043] In this embodiment, the inoculation amount of Lactobacillus acidophilus is 1.6 mL / L, the fermentation time after inoculation is 24 h, the mass ratio of the fermentation liquid A and lactose to the mulberry juice fermented for 4 days is 1:0.2:10000, and the other parameters are the same as those in Example 3.

[0044] The lactic acid content in mulberry wine was determined to be 0.566 mg / L, the acetic acid content was 15.79 mg / L, the butyric acid content was 0.77 mg / L, the ethyl acetate content was 73.45 mg / L, the ethyl lactate content was 48.53 mg / L, the diethyl amber content was 25.13 mg / L, the ethyl benzoate content was 0.79 mg / L, the lauric aldehyde content was 0.49 mg / L, and the 2-phenylethanol content was 87.14 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste and astringency; it had a strong frankincense smell but did not cover the wine taste, and had a strong "mulberry flavor".

[0045] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium in an amount of 3wt% and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24h) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.551.

[0046] Embodiment 5:

[0047] In this example, the colony count of Lactobacillus acidophilus during inoculation was 7×10 6 cfu / m, the mass ratio of fermentation liquid A and lactose to mulberry juice fermented for 4 days was 1:0.4:30000, and other parameters were the same as those in Example 3.

[0048] The lactic acid content in mulberry wine was determined to be 0.564 mg / L, the acetic acid content was 15.22 mg / L, the butyric acid content was 0.75 mg / L, the ethyl acetate content was 66.77 mg / L, the ethyl lactate content was 61.18 mg / L, the diethyl amber content was 25.23 mg / L, the ethyl benzoate content was 0.70 mg / L, the lauric aldehyde content was 0.41 mg / L, and the 2-phenylethanol content was 82.19 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste and astringency; it had a strong frankincense smell but did not cover the wine taste, and had a strong "mulberry flavor".

[0049] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.517.

[0050] Embodiment 6:

[0051] In this example, the colony count of Lactobacillus acidophilus during inoculation was 7×10 6 cfu / m, the mass ratio of fermentation liquid A and lactose to mulberry juice fermented for 4 days was 1:0.3:50000, and other parameters were the same as those in Example 2.

[0052] The lactic acid content in mulberry wine was determined to be 0.358 mg / L, the acetic acid content was 15.34 mg / L, the butyric acid content was 0.93 mg / L, the ethyl acetate content was 72.05 mg / L, the ethyl lactate content was 49.59 mg / L, the diethyl amberate content was 23.34 mg / L, the ethyl benzoate content was 0.92 mg / L, the lauric aldehyde content was 0.37 mg / L, and the 2-phenylethanol content was 63.02 mg / L. According to sensory evaluation, the mulberry wine has a layered sour taste and a mild astringency; it has a slight frankincense smell and a strong "mulberry flavor".

[0053] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.474.

[0054] Embodiment 7:

[0055] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:60, and other parameters are the same as those in Example 1.

[0056] The lactic acid content in mulberry wine was determined to be 0.523 mg / L, the acetic acid content was 17.19 mg / L, the butyric acid content was 0.61 mg / L, the ethyl acetate content was 58.87 mg / L, the ethyl lactate content was 40.16 mg / L, the diethyl amber content was 16.19 mg / L, the ethyl benzoate content was 0.48 mg / L, the lauric aldehyde content was 0.25 mg / L, and the 2-phenylethanol content was 74.55 mg / L. According to sensory evaluation, the mulberry wine has a layered sour taste and astringency; it has a strong frankincense smell but does not cover the wine taste, and has a strong "mulberry flavor".

[0057] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.418.

[0058] Embodiment 8:

[0059] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:60, and other parameters are the same as those in Example 2.

[0060] The lactic acid content in mulberry wine was determined to be 0.405 mg / L, the acetic acid content was 17.15 mg / L, the butyric acid content was 0.57 mg / L, the ethyl acetate content was 58.66 mg / L, the ethyl lactate content was 45.98 mg / L, the diethyl amber content was 17.23 mg / L, the ethyl benzoate content was 0.57 mg / L, the lauric aldehyde content was 0.27 mg / L, and the 2-phenylethanol content was 77.99 mg / L. Sensory evaluation shows that mulberry wine has a layered sour taste and astringency; it has a frankincense smell and does not cover the wine taste, and has a strong "mulberry flavor".

[0061] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.323.

[0062] Embodiment 9:

[0063] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:60, and other parameters are the same as those in Example 3.

[0064] The lactic acid content in mulberry wine was determined to be 0.368 mg / L, the acetic acid content was 17.22 mg / L, the butyric acid content was 0.74 mg / L, the ethyl acetate content was 65.89 mg / L, the ethyl lactate content was 45.39 mg / L, the diethyl amber content was 19.98 mg / L, the ethyl benzoate content was 0.82 mg / L, the lauric aldehyde content was 0.33 mg / L, and the 2-phenylethanol content was 58.46 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste and a mild astringency; it had a slight frankincense smell and a strong "mulberry flavor".

[0065] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.363.

[0066] Embodiment 10:

[0067] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:60, and other parameters are the same as those in Example 4.

[0068] The lactic acid content in mulberry wine was determined to be 0.549 mg / L, the acetic acid content was 17.18 mg / L, the butyric acid content was 0.72 mg / L, the ethyl acetate content was 65.71 mg / L, the ethyl lactate content was 43.15 mg / L, the diethyl amberate content was 22.86 mg / L, the ethyl benzoate content was 0.76 mg / L, the lauric aldehyde content was 0.41 mg / L, and the 2-phenylethanol content was 85.60 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste and astringency; it had a strong frankincense smell but did not cover the wine taste, and had a strong "mulberry flavor".

[0069] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.314.

[0070] Embodiment 11:

[0071] In this embodiment, the ratio of the concentrate to the mulberry wine that has been fermented for 2 days is 1:60, and other parameters are the same as those in Example 5.

[0072] The lactic acid content in mulberry wine was determined to be 0.533 mg / L, the acetic acid content was 16.54 mg / L, the butyric acid content was 0.65 mg / L, the ethyl acetate content was 64.99 mg / L, the ethyl lactate content was 59.18 mg / L, the diethyl amber content was 23.69 mg / L, the ethyl benzoate content was 0.58 mg / L, the lauric aldehyde content was 0.35 mg / L, and the 2-phenylethanol content was 78.94 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste and astringency; it had a strong frankincense smell but did not cover the wine taste, and had a strong "mulberry flavor".

[0073] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium in an amount of 3wt% and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24h) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.389.

[0074] Embodiment 12:

[0075] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:60, and other parameters are the same as those in Example 6.

[0076] The lactic acid content in mulberry wine was determined to be 0.366 mg / L, the acetic acid content was 16.88 mg / L, the butyric acid content was 0.79 mg / L, the ethyl acetate content was 70.19 mg / L, the ethyl lactate content was 47.11 mg / L, the diethyl amber content was 20.94 mg / L, the ethyl benzoate content was 0.78 mg / L, the lauric aldehyde content was 0.31 mg / L, and the 2-phenylethanol content was 58.19 mg / L. According to sensory evaluation, the mulberry wine has a layered sour taste and a mild astringency; it has a slight frankincense smell and a strong "mulberry flavor".

[0077] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.344.

[0078] Embodiment 13:

[0079] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:70, and other parameters are the same as those in Example 1.

[0080] The lactic acid content in mulberry wine was determined to be 0.501 mg / L, the acetic acid content was 18.41 mg / L, the butyric acid content was 0.54 mg / L, the ethyl acetate content was 55.17 mg / L, the ethyl lactate content was 37.82 mg / L, the diethyl amberate content was 14.15 mg / L, the ethyl benzoate content was 0.33 mg / L, the lauric aldehyde content was 0.17 mg / L, and the 2-phenylethanol content was 70.17 mg / L. According to sensory evaluation, mulberry wine has a layered sour taste and astringency; it has a frankincense smell and is used to cover the wine taste, and has a strong "mulberry flavor".

[0081] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.254.

[0082] Embodiment 14:

[0083] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:70, and other parameters are the same as those in Example 2.

[0084] The lactic acid content in mulberry wine was determined to be 0.383 mg / L, the acetic acid content was 18.35 mg / L, the butyric acid content was 0.43 mg / L, the ethyl acetate content was 55.88 mg / L, the ethyl lactate content was 43.15 mg / L, the diethyl amber content was 14.79 mg / L, the ethyl benzoate content was 0.39 mg / L, the lauric aldehyde content was 0.21 mg / L, and the 2-phenylethanol content was 74.15 mg / L. According to sensory evaluation, the mulberry wine has a layered sour taste and a mild astringency; it has a slight frankincense smell and a strong "mulberry flavor".

[0085] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.264.

[0086] Embodiment 15:

[0087] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:70, and other parameters are the same as those in Example 3.

[0088] The lactic acid content in mulberry wine was determined to be 0.333 mg / L, the acetic acid content was 18.81 mg / L, the butyric acid content was 0.66 mg / L, the ethyl acetate content was 63.19 mg / L, the ethyl lactate content was 42.07 mg / L, the diethyl amber content was 17.55 mg / L, the ethyl benzoate content was 0.69 mg / L, the lauric aldehyde content was 0.21 mg / L, and the 2-phenylethanol content was 55.81 mg / L. According to sensory evaluation, the mulberry wine has a layered sour taste and a mild astringency; it has a slight frankincense smell and a strong "mulberry flavor".

[0089] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.233.

[0090] Embodiment 16:

[0091] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:70, and other parameters are the same as those in Example 4.

[0092] The lactic acid content in mulberry wine was determined to be 0.517 mg / L, the acetic acid content was 18.56 mg / L, the butyric acid content was 0.68 mg / L, the ethyl acetate content was 63.44 mg / L, the ethyl lactate content was 41.01 mg / L, the diethyl amber content was 19.94 mg / L, the ethyl benzoate content was 0.66 mg / L, the lauric aldehyde content was 0.29 mg / L, and the 2-phenylethanol content was 82.13 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste and astringency; it had a strong frankincense smell and did not cover the wine taste, and had a strong "mulberry flavor".

[0093] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.203.

[0094] Embodiment 17:

[0095] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:70, and other parameters are the same as those in Example 5.

[0096] The lactic acid content in mulberry wine was determined to be 0.503 mg / L, the acetic acid content was 16.96 mg / L, the butyric acid content was 0.49 mg / L, the ethyl acetate content was 61.03 mg / L, the ethyl lactate content was 56.82 mg / L, the diethyl amberate content was 20.49 mg / L, the ethyl benzoate content was 0.31 mg / L, the lauric aldehyde content was 0.28 mg / L, and the 2-phenylethanol content was 73.16 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste and astringency; it had a strong frankincense smell and did not cover the wine taste, and had a strong "mulberry flavor".

[0097] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.247.

[0098] Embodiment 18:

[0099] In this embodiment, the mass ratio of the concentrated liquid to the mulberry juice that has been fermented for 2 days is 1:70, and other parameters are the same as those in Example 6.

[0100] The lactic acid content in mulberry wine was determined to be 0.348 mg / L, the acetic acid content was 18.13 mg / L, the butyric acid content was 0.64 mg / L, the ethyl acetate content was 65.44 mg / L, the ethyl lactate content was 43.77 mg / L, the diethyl amber content was 18.14 mg / L, the ethyl benzoate content was 0.55 mg / L, the lauric aldehyde content was 0.23 mg / L, and the 2-phenylethanol content was 55.14 mg / L. According to sensory evaluation, the mulberry wine has a layered sour taste and a mild astringency; it has a slight frankincense smell and a strong "mulberry flavor".

[0101] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.205.

[0102] Comparative Example 1:

[0103] In this example, the mass ratio of the concentrate to the mulberry juice that had been fermented for 2 days was 1:45, and the colony count of Lactobacillus acidophilus during inoculation was 7×10 6 The cfu / mL inoculation amount was 1.6 mL / L, the fermentation time after inoculation was 24 h, the mixing ratio of fermentation liquid A and lactose to mulberry wine that had been fermented for 4 days was 1:0.4:10000, and the other parameters were the same as in Example 1.

[0104] The lactic acid content in mulberry wine was determined to be 0.967 mg / L, the acetic acid content was 15.11 mg / L, the butyric acid content was 0.99 mg / L, the ethyl acetate content was 69.55 mg / L, the ethyl lactate content was 70.61 mg / L, the diethyl amber content was 18.93 mg / L, the ethyl benzoate content was 0.98 mg / L, the lauric aldehyde content was 0.37 mg / L, and the 2-phenylethanol content was 64.33 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste but a distinct lactic acid taste, and the astringency was not significant; the strong frankincense smell covered the wine taste, and the "mulberry flavor" was strong.

[0105] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium in an amount of 3wt% and the MRS culture medium was sterilized, and then bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24h) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.562.

[0106] Comparative Example 2:

[0107] In this example, the mass ratio of the concentrate to the mulberry juice that has been fermented for 2 days is 1:74, the fermentation time after inoculation is 36 hours, the mixing ratio of fermentation liquid A and lactose to mulberry wine that has been fermented for 4 days is 1:0.2:50000, and the other parameters are the same as those in Comparative Example 1.

[0108] The lactic acid content in mulberry wine was determined to be 0.108 mg / L, the acetic acid content was 17.49 mg / L, the butyric acid content was 0.36 mg / L, the ethyl acetate content was 50.18 mg / L, the ethyl lactate content was 37.75 mg / L, the diethyl amber content was 11.54 mg / L, the ethyl benzoate content was 0.27 mg / L, the lauric aldehyde content was 0.11 mg / L, and the 2-phenylethanol content was 48.19 mg / L. According to sensory evaluation, mulberry wine tastes sour and astringent, with obvious astringency; there is no frankincense smell, and there is a "mulberry flavor" but it is not significant.

[0109] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.110.

[0110] Comparative Example 3:

[0111] In this example, the mass ratio of the concentrate to the mulberry juice that has been fermented for 2 days is 1:45, and other parameters are the same as those in Comparative Example 2.

[0112] The lactic acid content in mulberry wine was determined to be 0.779 mg / L, the acetic acid content was 15.39 mg / L, the butyric acid content was 0.78 mg / L, the ethyl acetate content was 65.88 mg / L, the ethyl lactate content was 67.34 mg / L, the diethyl amberate content was 17.46 mg / L, the ethyl benzoate content was 0.82 mg / L, the lauric aldehyde content was 0.26 mg / L, and the 2-phenylethanol content was 60.17 mg / L. Sensory evaluation showed that the mulberry wine had a layered sour taste but a distinct lactic acid taste, and the astringency was not significant; the strong frankincense smell covered the wine taste, and the "mulberry flavor" was strong.

[0113] The mulberry wine was heated at 100°C for 25 minutes by heating evaporation to remove most of the alcohol in the wine. The mulberry wine treated above was added to the MRS liquid culture medium at an addition amount of 3%, and the MRS culture medium was sterilized, and then the bifidobacteria were cultured in the conventional way (37°C, anaerobic culture for 24 hours) of bifidobacteria; after the fermentation liquid was diluted by 10, the OD600 value of the fermentation liquid increased by 0.427.

[0114] The above embodiments are only examples, and do not mean that the present invention can only obtain mulberry wine of the quality in the above embodiments. Those skilled in the art, based on actual needs, according to the principles of the present invention and under the control conditions of the present invention, adjust the parameters such as the ratio of the concentrate to the mulberry wine that has been fermented for 2 days, the inoculation concentration and amount of Lactobacillus acidophilus and the fermentation time after inoculation, the mixing ratio of the fermentation liquid A and lactose to the mulberry wine that has been fermented for 4 days, so as to obtain mulberry wine with different sour taste, different types of "mulberry flavor" aroma and different probiotic abilities.

Claims

1. A method for preparing mulberry wine with probiotic effects, comprising squeezing mulberry fruits to obtain mulberry juice and mulberry pomace, and fermenting the mulberry juice with brewer's yeast to prepare mulberry wine, characterized in that: The mulberry pomace is subjected to micro-electrolysis and then filtered, and the filtrate is subjected to reverse osmosis desalination treatment and then concentrated and sterilized to obtain a concentrated solution; The concentrate was added to mulberry juice that had been fermented for 2 days and inoculated with Lactobacillus acidophilus for anaerobic fermentation. The fermentation liquid at the end of the fermentation was recorded as fermentation liquid A. The fermentation liquid A, lactose and mulberry juice that has been fermented for 4 days are mixed evenly and then continued to ferment. After the fermentation is completed, the mixture is filtered and aged to obtain mulberry wine.

2. The method for preparing the mulberry wine with probiotic effects as claimed in claim 1, wherein: The conditions of the micro-electrolysis are as follows: distilled water is added to the mulberry pomace at a mass ratio of 1:30, graphite is used as the negative electrode, platinum is used as the positive electrode, the distance between the electrodes is 40 cm, sodium nitrate electrolyte is added to the mulberry pomace treated with water at a mass ratio of 0.5:100, and electrolysis is performed at a voltage of 25 V for 10 hours under stirring conditions.

3. The method for preparing mulberry wine with probiotic effects as claimed in claim 1, wherein the filtrate is concentrated to 1 / 10 of the mass of the original solution after reverse osmosis desalination treatment and then sterilized to obtain a concentrated solution.

4. The method for preparing the mulberry wine with probiotic effects as claimed in claim 1, wherein: The mixing mass ratio of the concentrated liquid to the mulberry juice fermented for 2 days is 1:50-70, and the fermentation conditions are: anaerobic fermentation at 37° C. for 24-36 hours.

5. The method for preparing the mulberry wine with probiotic effects as claimed in any one of claims 1 to 4, wherein: The inoculation amount of Lactobacillus acidophilus in the mulberry juice fermented for 2 days is 0.8-1.6 ml / L, and the concentration of lactic acid bacteria used in the Lactobacillus acidophilus is (2-7)×10 6 cfu / mL.

6. The method for preparing the mulberry wine with probiotic effects as claimed in claim 5, wherein: The strain number of the Lactobacillus acidophilus is CICC 6083, and the fermentation conditions are: anaerobic fermentation time at 37° C. is 24 to 36 hours.

7. The method for preparing the mulberry wine with probiotic effects as claimed in claim 1, wherein: The mixing mass ratio of the fermentation liquid A, lactose and mulberry juice fermented for 4 days is 1:(0.2-0.4):(10000-50000).

8. The method for preparing the mulberry wine with probiotic effects as claimed in claim 1, wherein: The fermentation liquid A, lactose and mulberry juice fermented for 4 days are uniformly mixed, and then oxygen is extracted by vacuum treatment, and then fermented for 3 days and aged to obtain the product.