TW202100177A - Panax lycium fermented liquid and making method thereof - Google Patents
Panax lycium fermented liquid and making method thereof Download PDFInfo
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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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
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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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
- A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
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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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
Description
本發明關於一種發酵液飲品,尤其是指一種人參枸杞發酵液飲品及其製法。The invention relates to a fermented liquid drink, in particular to a ginseng and wolfberry fermented liquid drink and a preparation method thereof.
由微生物發酵的食品具有更易於消化、香味增加、儲藏時間提升等優勢。例如:泡菜、納豆、水果醋等等都是廣受歡迎的發酵類食品。Foods fermented by microorganisms have advantages such as easier digestion, increased flavor, and improved storage time. For example: kimchi, natto, fruit vinegar, etc. are all popular fermented foods.
在飲品界中,發酵飲品也愈來愈受到消費者的喜愛。發酵飲品常以乳品、蔬菜、水果、豆類等為原料基底,再經由不同種類的微生物作用而製得。In the beverage industry, fermented beverages are becoming more and more popular among consumers. Fermented beverages are often prepared with dairy, vegetables, fruits, beans, etc. as the raw material base, and then through the action of different types of microorganisms.
有鑑於此,本發明提供一種人參枸杞發酵液飲品及其製法,其能進一步提升發酵飲品的機能。於此,以中藥材為基底,精選高效能的菌種,並且經由磁場發酵過程以製得人參枸杞發酵液。In view of this, the present invention provides a ginseng wolfberry fermented liquid beverage and a preparation method thereof, which can further improve the function of the fermented beverage. Here, based on Chinese medicinal materials, high-efficiency strains are selected, and the fermentation liquid of ginseng and wolfberry is prepared through a magnetic field fermentation process.
在一實施例中,人參枸杞發酵液的製備方法包括:提供培養液;將培養液及複數菌種在磁場內進行發酵4~15.5日以得到發酵原液;調整發酵原液的糖度以形成人參枸杞發酵液。其中,培養液包括由1重量份的人參、1重量份的枸杞及50重量份的水所形成的人參枸杞溶液及人參枸杞溶液總重的10%的葡萄糖,複數菌種包括相對於培養液為0.01%~0.5%的酵母菌、相對於培養液為0.01%~0.25%的乳酸菌及相對於培養液為3~10%的醋酸菌。In one embodiment, the preparation method of ginseng wolfberry fermentation broth includes: providing a culture broth; fermenting the culture broth and plural strains in a magnetic field for 4 to 15.5 days to obtain a fermentation stock solution; adjusting the sugar content of the fermentation stock solution to form a ginseng wolfberry fermentation liquid. Among them, the culture solution includes a ginseng wolfberry solution formed by 1 part by weight of ginseng, 1 part by weight of wolfberry and 50 parts by weight of water, and 10% glucose of the total weight of the ginseng wolfberry solution. The plural bacterial species include 0.01%~0.5% yeast, 0.01%~0.25% lactic acid bacteria relative to the culture solution, and 3~10% acetic acid bacteria relative to the culture solution.
在一實施例中,將培養液及菌種在磁場內進行發酵的步驟包括:將酵母菌在磁場內進行發酵1~2.5日後形成第一初發酵液;添加乳酸菌於第一初發酵液內並在磁場內發酵1~3日後形成第二初發酵液;添加醋酸菌於第二初發酵液內並於磁場內發酵3~10日後形成初發酵液;過濾初發酵液以得到發酵原液。In one embodiment, the step of fermenting the culture broth and strains in a magnetic field includes: fermenting yeasts in a magnetic field for 1 to 2.5 days to form a first initial fermentation broth; adding lactic acid bacteria in the first initial fermentation broth and Ferment in a magnetic field for 1 to 3 days to form a second primary fermentation broth; add acetic acid bacteria to the second primary fermentation broth and ferment in a magnetic field for 3 to 10 days to form a primary fermentation broth; filter the primary fermentation broth to obtain the fermentation stock.
在一實施例中,過濾初發酵液的步驟包括在60℃下減壓濃縮及以200mesh過濾初發酵液。In one embodiment, the step of filtering the initial fermentation broth includes concentrating under reduced pressure at 60° C. and filtering the initial fermentation broth with 200 mesh.
在一實施例中,發酵原液的糖度小於等於4°Bx。In one embodiment, the sugar content of the fermentation stock solution is less than or equal to 4°Bx.
在一實施例中,提供培養液的步驟包括:混合人參枸杞溶液及葡萄糖形成混合液;將混合液於50-100℃下萃取0.5-1.5小時以得到培養液。In one embodiment, the step of providing the culture solution includes: mixing the ginseng wolfberry solution and glucose to form a mixed solution; extracting the mixed solution at 50-100° C. for 0.5-1.5 hours to obtain the culture solution.
在一實施例中,酵母菌為Saccharomyces cerevisiae ,乳酸菌為Lactobacillus plantarum ,醋酸菌為Acetobacter aceti 。In one embodiment, the yeast is Saccharomyces cerevisiae , the lactic acid bacteria is Lactobacillus plantarum , and the acetic acid bacteria is Acetobacter aceti .
在一實施例中,調整發酵原液的糖度的步驟包括:添加寡糖至發酵原液以使發酵原液的糖度達到40°Bx以形成人參枸杞發酵液。In one embodiment, the step of adjusting the sugar content of the fermentation stock solution includes: adding oligosaccharides to the fermentation stock solution so that the sugar content of the fermentation stock solution reaches 40°Bx to form a ginseng wolfberry fermentation broth.
在一實施例中,本發明提供一種人參枸杞發酵液,係經由上述的人參枸杞發酵液的製備方法所製得。In one embodiment, the present invention provides a ginseng wolfberry fermentation broth, which is prepared by the above-mentioned preparation method of ginseng wolfberry fermentation broth.
在一實施例中,人參枸杞發酵液的總多酚含量為436µg/ml以上。In one embodiment, the total polyphenol content of the fermentation broth of ginseng and wolfberry is more than 436 μg/ml.
在一實施例中,人參枸杞發酵液的黃酮含量為12703µg/ml以上。In one embodiment, the flavonoid content of the fermentation broth of ginseng wolfberry is more than 12703 µg/ml.
綜上,根據任一實施例的人參枸杞發酵液的製備方法,其能增添人參枸杞發酵液的機能性,縮短發酵時間,並提升人參枸杞發酵液內的多酚及黃酮含量。在一實施例中,在發酵過程中分次依序加入複數菌種並且控制使用各菌種的發酵時間,以使得菌種能有最佳的生長,並且進一步提升人參枸杞發酵液的有效成分濃度。在一實施例中,藉由濃縮初發酵液以利後續進行人參枸杞發酵液的風味及口感的調整。在一實施例中,藉由監控發酵過程中的PH值及糖度來確保發酵過程順利完成。在一實施例中,藉由將葡萄糖與人參枸杞溶液一起加熱有助於溶解葡萄糖,並且避免培養液受到污染。在一實施例中,利用寡糖調整甜度口感,以降低熱量。In summary, the preparation method of ginseng wolfberry fermentation broth according to any embodiment can increase the functionality of the ginseng wolfberry fermentation broth, shorten the fermentation time, and increase the content of polyphenols and flavonoids in the ginseng wolfberry fermentation broth. In one embodiment, during the fermentation process, multiple strains are added sequentially and the fermentation time of each strain is controlled, so that the strains can grow optimally, and the effective ingredient concentration of the ginseng wolfberry fermentation broth is further increased . In one embodiment, the initial fermentation broth is concentrated to facilitate subsequent adjustment of the flavor and mouthfeel of the ginseng and wolfberry fermentation broth. In one embodiment, the pH value and sugar content during the fermentation process are monitored to ensure the smooth completion of the fermentation process. In one embodiment, heating the glucose and ginseng wolfberry solution together helps to dissolve the glucose and avoid contamination of the culture solution. In one embodiment, oligosaccharides are used to adjust the sweetness and mouthfeel to reduce calories.
於下列實施方式的說明中,除非另有相關說明,則「%」符號是指重量百分比。In the description of the following embodiments, unless otherwise specified, the symbol "%" refers to percentage by weight.
參考圖1。在一實施例中,人參枸杞發酵液的製備方法包括下列步驟:提供培養液(步驟S01)、將培養液及複數菌種在磁場內進行發酵4~15.5日以得到發酵原液(步驟S02)、以及調整發酵原液的糖度(Degrees Brix,°Bx)以完成人參枸杞發酵液(步驟S03)。Refer to Figure 1. In one embodiment, the preparation method of ginseng wolfberry fermentation broth includes the following steps: providing a culture broth (step S01), fermenting the broth and plural strains in a magnetic field for 4 to 15.5 days to obtain a fermentation stock (step S02), And adjust the degree of brix (Degrees Brix, °Bx) of the fermentation stock solution to complete the ginseng wolfberry fermentation broth (step S03).
在步驟S01中,培養液包括由1重量份的人參、1重量份的枸杞及50重量份的水所形成的人參枸杞溶液及人參枸杞溶液總重的10%的葡萄糖。在一些實施例中,人參(Panax ginseng )係採用無炮製過的乾燥白蔘根部。枸杞(Lycium chinense )係採用乾燥枸杞果實。In step S01, the culture solution includes a ginseng wolfberry solution formed by 1 part by weight of ginseng, 1 part by weight of wolfberry and 50 parts by weight of water, and 10% glucose of the total weight of the ginseng and wolfberry solution. In some embodiments, ginseng ( Panax ginseng ) uses unprocessed dry white ginseng roots. Lycium chinense ( Lycium chinense ) uses dried medlar fruits.
在一實施例中,步驟S01可包括:依上述比例混合人參、枸杞及葡萄糖形成混合液;以及將混合液於50-100℃下萃取0.5-1.5小時以得到培養液。於此,將葡萄糖與人參枸杞溶液一併進行萃取程序,藉以助於葡萄糖溶解並且還可以避免污染。In one embodiment, step S01 may include: mixing ginseng, wolfberry, and glucose in the above-mentioned ratio to form a mixed solution; and extracting the mixed solution at 50-100° C. for 0.5-1.5 hours to obtain a culture solution. Here, the glucose and ginseng wolfberry solution are combined to perform an extraction process to help dissolve the glucose and avoid contamination.
在另一個實施例中,步驟S01包括:將人參、枸杞及水於50-100℃下萃取0.5-1.5小時形成水萃液;以及將葡萄糖加入水萃液以得到培養液。於此,將人參和枸杞單獨進行萃取程序,以利於人參及/或枸杞內有效成分的確實地釋出。In another embodiment, step S01 includes: extracting ginseng, wolfberry and water at 50-100° C. for 0.5-1.5 hours to form an aqueous extract; and adding glucose to the aqueous extract to obtain a culture solution. Here, the ginseng and wolfberry are extracted separately to facilitate the reliable release of the effective ingredients in the ginseng and/or wolfberry.
在一些實施例中,萃取係指將混合液維持在50-100℃並靜置0.5-1.5小時。在一實施例中,萃取係指將混合液維持在95℃並靜置1小時。In some embodiments, extraction refers to maintaining the mixed solution at 50-100°C and standing for 0.5-1.5 hours. In one embodiment, extraction refers to maintaining the mixed solution at 95° C. and letting it stand for 1 hour.
在一實施例中,培養液的糖度為9°Bx~10°Bx。於此,足夠的糖度可以確保後續發酵的順利進行,菌種可以有足夠的養份。In one embodiment, the sugar content of the culture solution is 9°Bx~10°Bx. In this regard, sufficient sugar content can ensure the smooth progress of subsequent fermentation, and the bacteria can have sufficient nutrients.
後續,將培養液及複數菌種在磁場內進行發酵4~15.5日以得到發酵原液(步驟S02)。其中,複數菌種包括相對於培養液為0.01%~0.5%的酵母菌、相對於培養液為0.01%~0.25%的乳酸菌及相對於培養液為3~10%的醋酸菌(Acetobacter aceti )。在一實施例中,培養液不另濾除其內部的固形物(萃取後的人參和/或枸杞)直接加入菌種進行發酵,藉以利用菌種進一步提取固形物中的活性成分。Subsequently, the culture broth and the plurality of bacterial species are fermented in a magnetic field for 4 to 15.5 days to obtain a fermentation stock solution (step S02). Among them, the plural bacterial species include 0.01% to 0.5% yeast relative to the culture solution, 0.01% to 0.25% lactic acid bacteria relative to the culture solution, and 3-10% Acetobacter aceti relative to the culture solution. In one embodiment, the culture solution is directly added to the strain for fermentation without filtering out the internal solids (extracted ginseng and/or wolfberry), so that the strains are used to further extract the active ingredients in the solids.
在一實施例中,酵母菌可以是市售的啤酒酵母(Saccharomyces cerevisiae )。舉例而言,向財團法人食品工作發展研究所所採購寄存編號BCRC20271(國際寄存ATCC26602)菌株的啤酒酵母。In one embodiment, the yeast may be commercially available Saccharomyces cerevisiae ( Saccharomyces cerevisiae ). For example, purchase beer yeast with the strain number BCRC20271 (International Deposit ATCC26602) from the Food Work Development Research Institute.
在一實施例中,乳酸菌可以是為市售的胚芽乳酸桿菌(Lactobacillus plantarum)、市售的嗜熱鏈球菌(Streptococcus thermophiles )或植物乳桿菌。舉例而言,採用寄存編號BCRC910805(國際寄存DSM33108)菌株的胚芽乳酸桿菌TCI028或寄存編號BCRT910760(國際寄存DSM32451)菌株的胚芽乳酸桿菌TCI378。再舉例而言,採用寄存編號BCRC910636(國際寄存DSM28121)菌株的嗜熱鏈球菌TCI633。In one embodiment, the lactic acid bacteria may be commercially available Lactobacillus plantarum, commercially available Streptococcus thermophiles or Lactobacillus plantarum. For example, use the Lactobacillus embryonicum TCI028 with the deposit number BCRC910805 (International Deposit DSM33108) strain or the Lactobacillus embryonicum TCI378 with the deposit number BCRT910760 (International Deposit DSM32451) strain. For another example, the strain of Streptococcus thermophilus TCI633 with the deposit number BCRC910636 (International Deposit DSM28121) is used.
在一實施例中,向美國菌種中心(American Type Culture Collection)採購寄存編號BCRC11688(國際寄存ATCC15973)菌株的醋酸菌。In one embodiment, the acetic acid bacteria with the deposit number BCRC11688 (International Deposit ATCC15973) strain is purchased from the American Type Culture Collection.
參考圖2。在一實施例中,步驟02可包括:將酵母菌在磁場內進行發酵1~2.5日後形成第一初發酵液(步驟S21);添加乳酸菌於第一初發酵液內並在磁場內發酵1~3日後形成第二初發酵液(步驟S22);添加醋酸菌於第二初發酵液內並於磁場內發酵3~10日後形成初發酵液(步驟S23);以及過濾初發酵液以得到發酵原液(步驟S24)。Refer to Figure 2. In an embodiment, step 02 may include: fermenting yeasts in a magnetic field for 1 to 2.5 days to form a first initial fermentation broth (step S21); adding lactic acid bacteria to the first initial fermentation broth and fermenting in the magnetic field for 1~ A second primary fermentation broth is formed after 3 days (step S22); acetic acid bacteria are added to the second primary fermentation broth and fermented in a magnetic field to form a primary fermentation broth after 3-10 days (step S23); and the primary fermentation broth is filtered to obtain a fermentation stock (Step S24).
在步驟S21中,透過先添加酵母菌可以使培養液發酵以產生酒精;於此,酒精有利提取出人參與枸杞內不同的有效成份。在一些實施例中,第一初發酵液的PH值小於4,且其糖度約為9°Bx。In step S21, by first adding yeast, the culture solution can be fermented to produce alcohol; here, the alcohol is advantageous in extracting different effective components in the wolfberry. In some embodiments, the PH value of the first initial fermentation broth is less than 4, and its sugar content is about 9°Bx.
在步驟S22中,透過添加乳酸菌可以使得第一初發酵液內的葡萄糖被進一步消耗而降低糖度並且產生乳酸,以降低第一初發酵液的PH值;於此,降低第一初發酵液的PH值有利於進一步提取人參與枸杞內其他不同有效成分。在一些實施例中,第二初發酵液的PH值小於3.5,且其糖度約為6°Bx。In step S22, by adding lactic acid bacteria, the glucose in the first initial fermentation broth can be further consumed to reduce the sugar content and produce lactic acid, so as to reduce the PH value of the first initial fermentation broth; here, reduce the pH of the first initial fermentation broth. The value is conducive to the further extraction of other different effective ingredients in the Chinese wolfberry. In some embodiments, the PH value of the second primary fermentation broth is less than 3.5, and its sugar content is about 6°Bx.
在步驟S23中,透過添加醋酸菌可以使得第二初發酵液內的酒精被消耗,而進一步再降低葡萄糖的含量。在一些實施例中,初發酵液的PH值小於3.5,且其糖度為約為3°Bx。In step S23, the alcohol in the second primary fermentation broth can be consumed by adding acetic acid bacteria, and the glucose content can be further reduced. In some embodiments, the PH value of the initial fermentation broth is less than 3.5, and its sugar content is about 3°Bx.
在步驟S24的一實施例中,可以是過濾並濃縮初發酵液以得到發酵原液。在一示範例中,濃縮方式可以採取在60℃下的減壓濃縮,以及過濾方式可以是以200mesh過濾。在一些實施例中,發酵原液的糖度約為2°Bx。在一些實施例中,發酵原液的PH值小於3.5。In an embodiment of step S24, the primary fermentation liquid may be filtered and concentrated to obtain the fermentation raw liquid. In an exemplary embodiment, the concentration method may be concentration under reduced pressure at 60° C., and the filtration method may be 200 mesh filtration. In some embodiments, the sugar content of the fermentation stock is about 2°Bx. In some embodiments, the pH value of the fermentation stock is less than 3.5.
在步驟S03的一實施例中,可以是添加寡糖至發酵原液以使發酵原液的糖度達到40°Bx以形成人參枸杞發酵液。於此,寡糖係指由3~10個單醣分子聚合而成的低聚糖。其中,寡糖可為果寡糖、半乳寡糖、木寡糖、異麥芽寡糖等等。在一實施例中,所添加的寡糖可為含40%~70%異麥芽寡糖的寡糖溶液。In an embodiment of step S03, oligosaccharides may be added to the fermentation stock solution so that the sugar content of the fermentation stock solution reaches 40°Bx to form a ginseng wolfberry fermentation broth. Here, oligosaccharides refer to oligosaccharides formed by the polymerization of 3-10 monosaccharide molecules. Among them, the oligosaccharides can be fructooligosaccharides, galactooligosaccharides, xylo-oligosaccharides, isomalto-oligosaccharides and the like. In an embodiment, the added oligosaccharide may be an oligosaccharide solution containing 40% to 70% isomalt oligosaccharide.
在一實施例中,步驟S03所得的人參枸杞發酵液的總多酚含量為436.33µg/ml。在一實施例中,人參枸杞發酵液的黃酮含量為12703µg/ml。In one embodiment, the total polyphenol content of the ginseng wolfberry fermentation broth obtained in step S03 is 436.33 µg/ml. In one embodiment, the flavonoid content of the ginseng wolfberry fermentation broth is 12703 µg/ml.
示範例一:人參枸杞發酵液的製備Demonstration example 1: Preparation of ginseng and wolfberry fermentation broth
將人參、枸杞、水與葡萄糖混合成為混合液。在混合液中,人參為1重量份、枸杞為1重量份以及水為50重量份,並且葡萄糖為人參、枸杞與水所形成的人參枸杞溶液總重的10%。接著,將混合液於95℃下萃取1小時以得到培養液。待培養液冷卻之後置入其外圍設置有磁鐵的發酵桶內。添加相對於培養液為0.01%的BCRC20271菌株之啤酒酵母於發酵桶中的培養液內,然後於30℃下進行發酵1天以形成第一初發酵液。再添加相對於培養液為0.05%的BCRC910805菌株之胚芽乳酸桿菌於發酵桶中的第一初發酵液內,然後進行發酵1天以形成第二初發酵液。最後添加相對於培養液為5%的BCRC11688菌株之醋酸菌於發酵桶中的第二初發酵液內,然後進行發酵5天以得到發酵原液。於得到發酵原液後,在60℃下減壓濃縮及以200mesh過濾初發酵液以得到發酵原液。然後,以異麥芽寡糖60%及發酵原液40%之比例進行混合以使發酵原液的糖度達到40°Bx而形成A組人參枸杞發酵液。於此,發酵桶外圍設置的磁鐵能在發酵桶內形成磁場,以致前述於發酵桶內的溶液在磁場內進行各階段的發酵。Mix ginseng, wolfberry, water and glucose into a mixed solution. In the mixed solution, ginseng is 1 part by weight, wolfberry is 1 part by weight, and water is 50 parts by weight, and glucose is 10% of the total weight of the ginseng and wolfberry solution formed by ginseng, wolfberry and water. Next, the mixed solution was extracted at 95°C for 1 hour to obtain a culture solution. After the culture solution is cooled, it is placed in a fermenting barrel equipped with a magnet on its periphery. The brewer's yeast of BCRC20271 strain at 0.01% relative to the culture broth was added to the culture broth in the fermenter, and then fermented at 30°C for 1 day to form the first initial fermentation broth. Then add 0.05% of the BCRC910805 strain Lactobacillus embryo to the first primary fermentation broth in the fermenter, and then ferment for 1 day to form the second primary fermentation broth. Finally, 5% of the acetic acid bacteria of the BCRC11688 strain was added to the second primary fermentation broth in the fermentation tank, and then the fermentation was carried out for 5 days to obtain the fermentation stock solution. After obtaining the fermentation stock, concentrate under reduced pressure at 60° C. and filter the initial fermentation broth with 200 mesh to obtain the fermentation stock. Then, the ratio of 60% isomalt-oligosaccharide and 40% of the fermentation stock is mixed so that the sugar content of the fermentation stock reaches 40°Bx to form a group A ginseng wolfberry fermentation broth. Here, the magnet provided on the periphery of the fermenting barrel can form a magnetic field in the fermenting barrel, so that the aforementioned solution in the fermenting barrel undergoes various stages of fermentation in the magnetic field.
另外,採用與A組人參枸杞發酵液相同的原料比例與發酵程序,但其發酵桶外圍並未設置磁鐵,來製得B組人參枸杞發酵液。In addition, the same raw material ratio and fermentation procedure as the fermentation broth of group A ginseng and wolfberry are used, but the magnet is not set on the periphery of the fermentation barrel to prepare the fermentation broth of group B ginseng and wolfberry.
實驗一:發酵階段PH值及糖度數值的測定結果Experiment 1: Measurement results of PH value and sugar content in the fermentation stage
在製備過程中各階段形成的溶液的PH值及糖度數值測定如下。The pH value and sugar content of the solution formed at each stage of the preparation process are determined as follows.
表一
由上述表一可見,A組人參枸杞發酵液在各階段的PH值及糖度都比B組人參枸杞發酵液更低;其代表相較於B組人參枸杞發酵液,A組人參枸杞發酵液的發酵作用的進行更為順利,其菌種的活性較佳,代謝產物較多,且葡萄糖的被利用率較佳。換句話說,在磁場內發酵可以明顯提升發酵的效率。It can be seen from the above table that the PH value and sugar content of the ginseng and wolfberry fermentation broth of group A are lower than those of the ginseng wolfberry fermentation broth of group B at all stages; its representative is compared with the ginseng wolfberry fermentation broth of group B. The fermentation proceeded more smoothly, the activity of the bacteria was better, the metabolites were more, and the utilization rate of glucose was better. In other words, fermentation in a magnetic field can significantly improve the efficiency of fermentation.
實驗二:總多酚含量測試Experiment 2: Total polyphenol content test
分別以前述示範例一所得的A組人參枸杞發酵液及B組人參枸杞發酵液為樣本。將各樣本以水稀釋10倍後取100mL到離心管中。接著,加入500μL之Folin-Ciocalteu酚試劑至離心管中與稀釋後的樣本混合並靜置3分鐘後,再加入400μL之7.5%碳酸鈉混勻靜置30分鐘後以得到待測反應溶液。於二次靜置後,取200μL之待測反應溶液至96孔板中,並測量待測反應溶液於750nm下之吸光值。Take the group A ginseng wolfberry fermentation broth and the B group ginseng wolfberry fermentation broth obtained in Example 1 above as samples. Dilute each sample 10 times with water and take 100 mL into a centrifuge tube. Then, add 500 μL of Folin-Ciocalteu phenol reagent to the centrifuge tube and mix with the diluted sample and let it stand for 3 minutes, then add 400 μL of 7.5% sodium carbonate and mix and let it stand for 30 minutes to obtain the reaction solution to be tested. After standing for the second time, take 200 μL of the reaction solution to be tested into a 96-well plate, and measure the absorbance of the reaction solution to be tested at 750 nm.
並且,以沒食子酸(Gallic acid)作為標準品製作標準曲線。於此,配置0μL/mL、20μL/mL、40μL/mL、60μL/mL、80μL/mL、及100μL/mL之沒食子酸的標準溶液,並分別取100μL之各濃度的標準溶液至10mL離心管中。加入500μL之Folin-Ciocalteu酚試劑至離心管內與標準溶液混合並靜置3分鐘後,再加入400μL之7.5%碳酸鈉混勻靜置30分鐘後以得到標準反應溶液。取200μL之標準反應溶液96孔板中,並測量其在750nm下之吸光值,以獲得標準曲線。In addition, a standard curve was prepared using gallic acid as a standard. Here, prepare 0μL/mL, 20μL/mL, 40μL/mL, 60μL/mL, 80μL/mL, and 100μL/mL gallic acid standard solutions, and take 100μL of each concentration standard solution to 10mL centrifugation In the tube. Add 500μL of Folin-Ciocalteu phenol reagent to the centrifuge tube and mix with the standard solution and let it stand for 3 minutes, then add 400μL of 7.5% sodium carbonate and mix and let it stand for 30 minutes to obtain the standard reaction solution. Take 200μL of the standard reaction solution in a 96-well plate, and measure its absorbance at 750nm to obtain a standard curve.
接著,利用標準曲線將待測反應溶液的吸光值換算成總多酚含量。於此,可得到A組人參枸杞發酵液總多酚含量為436.33µg/ml及B組人參枸杞發酵液中總多酚含量202µg/ml,如圖3所示。Then, use the standard curve to convert the absorbance value of the reaction solution to be tested into the total polyphenol content. From this, the total polyphenol content of the fermentation broth of group A ginseng and wolfberry is 436.33μg/ml and the total polyphenol content of the fermentation broth of group B ginseng and wolfberry is 202μg/ml, as shown in Figure 3.
由實驗結果可知,A組人參枸杞發酵液總多酚含量較B組人參枸杞發酵液總多酚含量提升21%;基此可知在磁場內進行發酵更有利於有效成份的提取。It can be seen from the experimental results that the total polyphenol content of the fermentation broth of ginseng and wolfberry in group A is increased by 21% compared with the total polyphenol content of the fermentation broth of ginseng and wolfberry in group B; this shows that fermentation in a magnetic field is more conducive to the extraction of effective ingredients.
實驗三:總黃酮含量測試Experiment 3: Total flavonoid content test
分別以前述示範例一所得的A組人參枸杞發酵液及B組人參枸杞發酵液為樣本。將各樣品以水稀釋20倍至1200ul,並分別加入200μL的5%亞硝酸鈉並混合後靜置6分鐘後,再加入200μL的10%硝酸鋁並混合後靜置6分鐘,接著加入2ml的4%氧氧化鈉並混合,最後再加入1.4ml的水並混合以得到待測反應溶液。將待測反應溶液至96孔板中,並以分光光度計測量待測反應溶液在500nm下的吸光值。Take the group A ginseng wolfberry fermentation broth and the B group ginseng wolfberry fermentation broth obtained in Example 1 above as samples. Dilute each sample 20 times with water to 1200ul, and add 200μL of 5% sodium nitrite and mix and let it stand for 6 minutes, then add 200μL of 10% aluminum nitrate and mix and let it stand for 6 minutes, then add 2ml of 4% oxygen sodium oxide and mix, and finally add 1.4ml of water and mix to obtain the reaction solution to be tested. Put the test reaction solution into a 96-well plate, and measure the absorbance value of the test reaction solution at 500 nm with a spectrophotometer.
並且,以芸香苷(rutin)作為標準品以製作標準曲線。於此,配置0μg/mL、400μg/mL、600μg/mL、1000μg/mL及1200μg/mL之沒芸香苷的標準溶液。將各標準溶液分別加入200μL的5%亞硝酸鈉並混合後靜置6分鐘後,再加入200μL的10%硝酸鋁並混合後靜置6分鐘,接著加入2ml的4%氧氧化鈉並混合,最後再加入1.4ml的水並混合以得到待測標準品溶液。將200μL的待測標準品溶液移至96孔板中,並以分光光度計測量待測反應溶液在500nm下的吸光值分別為0μg/mL的吸光值為0.035、400μg/mL的吸光值為0.13、600μg/mL的吸光值為0.183、1000μg/mL的吸光值為0.273及1200μg/mL的吸光值為0.335,並以上述結果依線性回歸計算以獲得標準曲線。In addition, rutin was used as a standard to prepare a standard curve. Here, prepare standard solutions of Brutin at 0μg/mL, 400μg/mL, 600μg/mL, 1000μg/mL and 1200μg/mL. Add 200μL of 5% sodium nitrite to each standard solution and mix and let it stand for 6 minutes, then add 200μL of 10% aluminum nitrate and mix and let it stand for 6 minutes, then add 2ml of 4% sodium oxynitride and mix. Finally, add 1.4ml of water and mix to obtain the standard solution to be tested. Transfer 200μL of the standard solution to be tested to a 96-well plate, and use a spectrophotometer to measure the absorbance of the reaction solution to be tested at 500nm. The absorbance at 500nm is 0.035 and the absorbance at 400μg/mL is 0.13. The absorbance value of 600μg/mL is 0.183, the absorbance value of 1000μg/mL is 0.273, and the absorbance value of 1200μg/mL is 0.335, and the above results are calculated by linear regression to obtain the standard curve.
接著,利用標準曲線將待測反應溶液的吸光值換算成總黃酮含量。於此,可得到A組人參枸杞發酵液的總黃酮含量為12703µg/ml及B組人參枸杞發酵液的總黃酮含量8315µg/ml,如圖4所示。Then, use the standard curve to convert the absorbance value of the reaction solution to be tested into the total flavonoid content. From this, the total flavonoid content of the ginseng wolfberry fermentation broth of group A was 12703 µg/ml and the total flavonoid content of ginseng wolfberry fermentation broth of group B was 8315 µg/ml, as shown in Figure 4.
由實驗結果可知,A組人參枸杞發酵液的總黃酮含量較B人參枸杞發酵液的總黃酮含量提升52%;基此可知在磁場內進行發酵能大幅提升有效成份的提取量。According to the experimental results, the total flavonoid content of the fermentation broth of ginseng and wolfberry of group A was increased by 52% compared with the total flavonoid content of the fermentation broth of ginseng and wolfberry of B; based on this, it can be seen that fermentation in a magnetic field can greatly increase the extraction of effective ingredients.
實驗四:粒線體活性實驗Experiment 4: Mitochondrial activity experiment
此實驗採用小鼠骨骼肌肉細胞(C2C12,ATCC® CRL-1772)。於此,培養基包括DMEM細胞培養液(Dulbecco’s modified Eagle’s medium)、1%的青霉素-鏈黴素(Penicillin-streptomycin, Gibco)以及10%胎牛血清(FBS, Gibco)。This experiment uses mouse skeletal muscle cells (C2C12, ATCC® CRL-1772). Here, the medium includes DMEM cell culture medium (Dulbecco’s modified Eagle’s medium), 1% penicillin-streptomycin (Gibco) and 10% fetal bovine serum (FBS, Gibco).
將細胞分為四組進行,第一組到第三組包括2ml上述培養基並接種1×105 的細胞及樣本,第一組樣本為前述示範例一所得的A組人參枸杞發酵液,第二組樣本為B組人參枸杞發酵液、第三組樣本為示範例一的製備過程中所形成的培養液,第四組細胞未作施加任何溶液處理(即空白對照組)。上述各組在37℃下培養24小時。於此,第一組到第三組的培養基內樣本濃度為0.03125%。The cells were divided into four groups. The first to third groups included 2ml of the above-mentioned medium and inoculated with 1×10 5 cells and samples. The first group of samples was the fermentation broth of group A ginseng and wolfberry obtained in Example 1 above, and the second The samples in group B are the fermentation broth of ginseng and wolfberry, the samples in the third group are the culture broth formed in the preparation process of Example 1, and the cells in the fourth group are not treated with any solution (ie blank control group). The above groups were cultured at 37°C for 24 hours. Here, the sample concentration in the culture medium of the first to third groups is 0.03125%.
於24小時後,採用BD™,MitoScreen,(JC-1)染劑對空白及處理後的各組細胞進行染色15min,再以JC-1分析緩衝液清洗二次,然後採用BD Accuri C6 Plus流式細胞儀量測PBMC粒線體的膜電位,以得到粒線體的膜電位的活性數據。為避免實驗操作誤差,各組以相同步驟操作三次,實驗數據如下表二所示:After 24 hours, the blank and treated cells were stained with BD™, MitoScreen, (JC-1) dye for 15 minutes, and then washed twice with JC-1 analysis buffer, and then BD Accuri C6 Plus was used. The cytometer measures the membrane potential of PBMC mitochondria to obtain mitochondrial membrane potential activity data. In order to avoid experimental operation errors, each group performed the same steps three times. The experimental data is shown in Table 2 below:
表二
依上述三組實驗數據之平均值繪製圖5。由圖5可見,分析換算之後得到第一組相對活性為47.2%,第二組相對活性為40.1%,第三組相對活性為38.0%,第四組相對活性為38.92;基此可知,A組人參枸杞發酵液及B組人參枸杞發酵液皆可以提升骨骼肌細胞中粒線體的活性,其中A組人參枸杞發酵液提升幅度最明顯。Figure 5 is drawn based on the average of the above three sets of experimental data. It can be seen from Figure 5 that after analysis and conversion, the relative activity of the first group is 47.2%, the relative activity of the second group is 40.1%, the relative activity of the third group is 38.0%, and the relative activity of the fourth group is 38.92; Both the ginseng and wolfberry fermentation broth and the B group ginseng and wolfberry fermentation broth can increase the activity of mitochondria in skeletal muscle cells, and the A group ginseng and wolfberry fermentation broth has the most significant increase.
實驗五:基礎代謝率提升實驗Experiment 5: Basal metabolic rate improvement experiment
此實驗採用小鼠骨骼肌肉細胞(C2C12,ATCC CRL-1772)。於此,培養基包括DMEM細胞培養液(Dulbecco’s modified Eagle’s medium)、3.7g/L碳酸氫鈉、100I.U.青霉素、100μg/mL鏈黴素(Penicillin-streptomycin, Gibco)以及10%胎牛血清(FBS, Gibco)。This experiment uses mouse skeletal muscle cells (C2C12, ATCC CRL-1772). Here, the culture medium includes DMEM cell culture medium (Dulbecco's modified Eagle's medium), 3.7g/L sodium bicarbonate, 100I.U. penicillin, 100μg/mL streptomycin (Penicillin-streptomycin, Gibco) and 10% fetal bovine serum ( FBS, Gibco).
將細胞分為四組進行,第一組到第三組包括2ml上述培養基並接種1×105 的細胞及樣本,第一組樣本為前述示範例一所得的A組人參枸杞發酵液,第二組樣本為B組人參枸杞發酵液、第三組樣本為示範例一的製備過程中所形成的培養液,第四組細胞未作施加任何溶液處理(即空白對照組)。於此,第一組到第三組的培養基內樣本濃度為0.03125%。The cells were divided into four groups. The first to third groups included 2ml of the above-mentioned medium and inoculated with 1×10 5 cells and samples. The first group of samples was the fermentation broth of group A ginseng and wolfberry obtained in Example 1 above, and the second The samples in group B are the fermentation broth of ginseng and wolfberry, the samples in the third group are the culture broth formed in the preparation process of Example 1, and the cells in the fourth group are not treated with any solution (ie blank control group). Here, the sample concentration in the culture medium of the first to third groups is 0.03125%.
接下來,依下述程序處理各組細胞。將細胞以條件培養基(含有1%FBS及1%馬血清的DMEM)進行培育以分化出粒線體,添加上述樣本處理細胞48小時,以100μL丙酮酸緩衝液裂解細胞,將細胞在4℃下以10000g離心10分鐘並收集上清液,在上清液內添加20μL溶液液(Lysates)並添加丙酮酸測定緩衝液至總體積50μL,混入反應混合物(丙酮酸測定緩衝液46μL、丙酮酸探針2μL及酶混合液2μL)並培育30分鐘,並以分光光度計測量待測反應溶液在570nm下的吸光值。Next, process each group of cells according to the following procedure. Incubate the cells with conditioned medium (DMEM containing 1% FBS and 1% horse serum) to differentiate into mitochondria, add the above sample to treat the cells for 48 hours, lyse the cells with 100 μL pyruvate buffer, and place the cells at 4°C Centrifuge at 10000g for 10 minutes and collect the supernatant. Add 20μL of solution (Lysates) to the supernatant and add pyruvate assay buffer to a total volume of 50μL. Mix into the reaction mixture (pyruvate assay buffer 46μL, pyruvate probe 2μL and enzyme mixture 2μL) and incubate for 30 minutes, and measure the absorbance of the reaction solution to be tested at 570nm with a spectrophotometer.
並且,以丙酮酸作為標準品以製作標準曲線。於此,配置0、2、4、6、8及10nmol/μL之丙酮酸的標準溶液。依上述結果與標準曲線依線性回歸計算以獲得丙酮酸鹽的含量,以來確認細胞的代謝狀態。於此,所測得丙酮酸鹽相對含量如圖6所示。In addition, pyruvic acid was used as a standard to prepare a standard curve. Here, prepare 0, 2, 4, 6, 8 and 10 nmol/μL standard solutions of pyruvate. According to the above results and the standard curve, the pyruvate content can be obtained by linear regression calculation to confirm the metabolic state of the cells. Here, the measured relative content of pyruvate is shown in Figure 6.
由圖6可見,相較於空白對照組(即以第四組細胞的相對表現率為1的情況),第一組細胞的相對表現率為3.23,第二組細胞的相對表現率為2.39,第三組細胞的相對表現率為2.34。可知,B組人參枸杞發酵液可以提升細胞的基礎代謝率達239%,而A組人參枸杞發酵液更能提升細胞的基礎代謝率達320%。It can be seen from Figure 6 that compared to the blank control group (that is, the relative expression rate of the fourth group of cells is 1), the relative expression rate of the first group of cells is 3.23, and the relative expression rate of the second group of cells is 2.39. The relative expression rate of the third group of cells was 2.34. It can be seen that the ginseng and wolfberry fermentation broth of group B can increase the basal metabolic rate of cells by 239%, and the ginseng and wolfberry fermentation broth of group A can even increase the basal metabolic rate of cells by 320%.
實驗六:提升免疫力基因表現實驗Experiment 6: Gene performance experiment to enhance immunity
此實驗採用人類單核球細胞 (THP-1細胞株, ATCC TIB202)。於此,培養基包括培養液(RPMI-1640, Gibco)為基底,添加有10%胎牛血清(Gibco)、0.05Mm二硫基乙醇、100units/ml青霉素和100μg/mL鏈黴素。This experiment uses human monocytes (THP-1 cell line, ATCC TIB202). Here, the medium includes a culture medium (RPMI-1640, Gibco) as a base, supplemented with 10% fetal bovine serum (Gibco), 0.05 Mm disulfide ethanol, 100 units/ml penicillin and 100 μg/mL streptomycin.
實驗共分四組進行,第一組到第三組為實驗組,第一組樣本為前述示範例一所得的A組人參枸杞發酵液,第二組樣本為B組人參枸杞發酵液、第三組樣本為示範例一的製備過程中所形成的培養液,第四組細胞未作施加任何溶液處理(即空白對照組)。The experiment was divided into four groups. The first to third groups were experimental groups. The first group of samples was the fermentation broth of ginseng and wolfberry from the first example, the second group was the fermentation broth of ginseng and wolfberry from the third group. Group samples are the culture solution formed during the preparation process of Exemplary Example 1, and the fourth group of cells are not treated with any solution (ie, blank control group).
接下來,依下述程序處理細胞。提取2ml上述培養基並接種1×105 的細胞,添加上述樣本以處理細胞持續24小時,利用RNA提取試劑(Geneaid)提取RNA,用逆轉錄酶(SuperScript®III)將RNA反轉錄為cDNA,並藉由ABI Step One Plus Real-Time PCR system來執行使用qPCR來對目標基因進行定量。PCR反應的熔化曲線(melting curve)是在定量即時PCR反應期間進行確認。採用SCORE方法來確認基因表達的相對定量。Next, process the cells according to the following procedure. Extract 2ml of the above medium and inoculate 1×10 5 cells, add the above sample to treat the cells for 24 hours, use RNA extraction reagent (Geneaid) to extract RNA, use reverse transcriptase (SuperScript®III) to reverse transcription of RNA into cDNA, and The ABI Step One Plus Real-Time PCR system is used to perform qPCR to quantify the target gene. The melting curve of the PCR reaction is confirmed during the quantitative real-time PCR reaction. The SCORE method was used to confirm the relative quantification of gene expression.
採用下表三所式的引子(Primer)以聚合酶連鎖反應(PCR)儀觀察THP-1細胞株的IL-1β及IL-10的基因表現率,其結果如圖7及圖8。The gene expression rate of IL-1β and IL-10 of the THP-1 cell line was observed with a polymerase chain reaction (PCR) instrument using the primers shown in the following table three (Primer). The results are shown in Figure 7 and Figure 8.
表三
由圖7可見,相較於空白對照組(相對表現率為1的情況),第一組細胞的IL-1β的相對表現率為1.53,第二組細胞的IL-1β的相對表現率為0.41,而第三組細胞的IL-1β的相對表現率為0.36。在由圖8可見,相較於空白對照組(相對表現率為1的情況),第一組細胞的IL-10的相對表現率為1.14,第二組細胞的IL-10的相對表現率為0.87,而第三組細胞的IL-10的相對表現率為0.6。可知,人參枸杞發酵液可以提升免疫力。It can be seen from Figure 7 that compared to the blank control group (the relative expression rate is 1), the relative expression rate of IL-1β of the first group of cells is 1.53, and the relative expression rate of IL-1β of the second group of cells is 0.41 , And the relative expression rate of IL-1β in the third group of cells was 0.36. It can be seen from Figure 8 that compared to the blank control group (the relative expression rate is 1), the relative expression rate of IL-10 in the first group of cells is 1.14, and the relative expression rate of IL-10 in the second group of cells is 1.14 0.87, while the relative expression rate of IL-10 in the third group of cells was 0.6. It can be seen that the fermentation liquid of ginseng and wolfberry can enhance immunity.
在一些實施例中,所得的人參枸杞發酵液可進一步作為食品添加物(food additive),以致得含有人參枸杞發酵液的食品組合物。於此,能藉由習知方法於原料製備時添加任一實施例的人參枸杞發酵液,或是於食品的製作過程中添加任一實施例的人參枸杞發酵液,而與任一種可食性材料配製成供人類與非人類動物攝食的食品產品(即食品組合物)。In some embodiments, the obtained ginseng and wolfberry fermentation liquid can be further used as a food additive, so that a food composition containing ginseng and wolfberry fermentation liquid is obtained. Herein, the ginseng wolfberry fermentation liquid of any embodiment can be added during the preparation of raw materials by a conventional method, or the ginseng wolfberry fermentation liquid of any embodiment can be added during the food production process, and it can be combined with any edible material Formulated into food products (ie food compositions) for human and non-human animals to eat.
在一些實施例中,食品產品種類包括但不限於:飲料(beverages)、發酵食品(fermented foods)、烘培產品(bakery products)、健康食品(health foods)以及膳食補充品(dietary supplements)。In some embodiments, the types of food products include, but are not limited to: beverages, fermented foods, bakery products, health foods, and dietary supplements.
雖然本發明的技術內容已經以較佳實施例揭露如上,然其並非用以限定本發明,任何熟習此技藝者,在不脫離本發明之精神所作些許之更動與潤飾,皆應涵蓋於本發明的範疇內,因此本發明之保護範圍當視後附之申請專利範圍所界定者為準。Although the technical content of the present invention has been disclosed in the preferred embodiments as above, it is not intended to limit the present invention. Anyone who is familiar with this technique and makes some changes and modifications without departing from the spirit of the present invention should be covered by the present invention Therefore, the scope of protection of the present invention shall be subject to the scope of the attached patent application.
S01~S24:步驟S01~S24: steps
[圖1] 係本發明一實施例之發酵液的製法的流程圖。 [圖2] 係圖1步驟S02的細部的流程圖。 [圖3] 係總多酚含量測試結果數據圖。 [圖4] 係總黃酮含量測試結果數據圖。 [圖5] 係相對粒線體活性檢測結果數據圖。 [圖6] 係相對丙酮酸鹽含量結果數據圖。 [圖7] 係IL-1B引子的基因的相對表現率數據圖。 [圖8] 係IL-10引子的基因的相對表現率數據圖。[Figure 1] is a flowchart of a method for preparing fermentation broth according to an embodiment of the present invention. [Fig. 2] It is a detailed flowchart of step S02 in Fig. 1. [Figure 3] Department of total polyphenol content test results data diagram. [Figure 4] A diagram showing the test results of total flavonoids. [Figure 5] The data graph of the relative mitochondrial activity detection results. [Figure 6] The relative pyruvate content result data graph. [Figure 7] The relative expression rate data graph of the gene of the IL-1B primer. [Figure 8] The relative expression rate data graph of the gene of the IL-10 primer.
S01~S03:步驟 S01~S03: steps
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CN112868963A (en) * | 2021-02-02 | 2021-06-01 | 台健生物科技(福建)有限公司 | Preparation method and application of ginseng enzyme |
CN115969033A (en) * | 2021-10-15 | 2023-04-18 | 百岳特生物技术(上海)有限公司 | Preparation method and application of elderberry magnetic field fermentation product |
TWI817632B (en) * | 2022-07-29 | 2023-10-01 | 統欣生物科技股份有限公司 | Composition of probiotics plus Chinese herbal medicine and its use |
CN116172192B (en) * | 2023-03-30 | 2024-07-26 | 广州五科生物科技有限公司 | High-activity composite ferment powder and preparation method thereof |
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JP2006199591A (en) * | 2005-01-17 | 2006-08-03 | Toyo Shinyaku:Kk | Lever function-improving agent |
US20080182309A1 (en) * | 2006-08-21 | 2008-07-31 | Emtech, Llc | Method and apparatus for magnetic fermentation |
CN102178311B (en) * | 2011-03-29 | 2012-12-19 | 丁政然 | Ginseng-vinegar beverage and preparation method of the ginseng-vinegar beverage |
CN102260714B (en) * | 2011-06-24 | 2013-04-03 | 河南科技大学 | Method for producing ethanol by magnetic-field-assisted gas stripping fermentation |
CN102940288B (en) * | 2012-11-19 | 2013-10-30 | 商丘市饮之健食品有限公司 | Composite enzyme beverage and preparation process method thereof |
CN102994362B (en) * | 2012-12-06 | 2014-03-12 | 陕西科技大学 | Method for fermenting apple vinegar |
US9877494B2 (en) * | 2014-08-21 | 2018-01-30 | Shantung HSU | Active fermentation process and fermented liquid and drinks made by using the same |
CN104479987B (en) * | 2014-12-30 | 2018-11-27 | 丁政然 | The preparation method and its product of pine needle fermented ginseng vinegar |
CN104450488B (en) * | 2015-01-16 | 2017-07-07 | 王体军 | A kind of ginseng medlar vinegar and preparation method thereof |
CN204625600U (en) * | 2015-05-13 | 2015-09-09 | 新疆恒安食品有限公司 | A kind of cider vinegar fermentor tank and deeply filter mixing device |
CN106085788B (en) * | 2016-08-11 | 2020-01-10 | 山西双锦生物科技有限公司 | Ginseng and medlar vinegar and preparation method and application thereof |
CN107343584A (en) * | 2017-05-17 | 2017-11-14 | 天津岂均生物科技有限公司 | Fermented beverage and preparation method with anti-fatigue effect |
CN107048390A (en) * | 2017-05-19 | 2017-08-18 | 程君 | Make the composition and its method of ferment |
CN107164178B (en) * | 2017-05-31 | 2020-11-13 | 兰考侨兴酒业有限公司 | Preparation method of medlar and ginseng wine |
CN107647232A (en) * | 2017-09-21 | 2018-02-02 | 广西南宁桂尔创环保科技有限公司 | A kind of preparation method of tremella beverage |
CN109527146A (en) * | 2017-09-22 | 2019-03-29 | 勐海茶业有限责任公司 | A kind of fruits and vegetables Pu'er tea mixed fermentation beverage and preparation method thereof |
CN107686806A (en) * | 2017-10-10 | 2018-02-13 | 中国科学院西北高原生物研究所 | A kind of brewing method of wolfberry fruit vinegar |
CN108850924A (en) * | 2018-07-20 | 2018-11-23 | 扬生(南召)生物科技有限公司 | A kind of fructus lycii probiotics ferment and preparation method thereof |
CN108991507A (en) * | 2018-07-20 | 2018-12-14 | 扬生(南召)生物科技有限公司 | A kind of fig probiotics ferment and preparation method thereof |
CN108949491A (en) * | 2018-08-28 | 2018-12-07 | 北京方旭食品有限公司 | A kind of acid pears vinegar and its brewing method |
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