TWI802136B - Fermentation of plants and their products by pure bacteria inoculation and their application in anti-oxidation - Google Patents
Fermentation of plants and their products by pure bacteria inoculation and their application in anti-oxidation Download PDFInfo
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Abstract
本發明揭示一種用於製備一發酵產物的方法及其所製得的發酵產物。本發明亦揭示由上述方法所製得的發酵產物可被用來抗氧化。The invention discloses a method for preparing a fermentation product and the prepared fermentation product. The present invention also discloses that the fermented product prepared by the above method can be used for anti-oxidation.
Description
本發明是有關於一種用於製備一發酵產物的方法及其所製得的發酵產物。本發明亦有關於使用該發酵產物來抗氧化。The present invention relates to a method for preparing a fermentation product and the obtained fermentation product. The present invention also relates to the use of the fermentation product for anti-oxidation.
已知在好氧生物體(aerobic organisms)利用氧氣進行呼吸反應(respiration)的過程中會生成活性氧族(reactive oxygen species, ROS)以及自由基(free radicals)。當活性氧族以及自由基的數量超過細胞或組織本身的抗氧化能力時，即會形成氧化性壓力(oxidative stress)。已有報導指出，氧化性壓力在老化(aging)以及發炎(inflammation)的過程中扮演一個重要的角色。It is known that reactive oxygen species (reactive oxygen species, ROS) and free radicals (free radicals) are generated during the respiration process of aerobic organisms using oxygen. When the amount of reactive oxygen species and free radicals exceeds the antioxidant capacity of cells or tissues, oxidative stress will be formed. It has been reported that oxidative stress plays an important role in the process of aging and inflammation.
目前所使用的抗氧化藥物在臨床應用上存在有療效不佳以及容易產生副作用(side effect)的問題。因此，本領域的相關研究人員開始嘗試從天然植物來源中尋找有用的活性組分(active components)來發展出具有抗氧化活性(anti-oxidative activity)並且不會產生非所欲的副作用的藥物。例如，CN 107348272 A揭示一種益生菌蔬果酵素飲料，其是藉由將蔬果拿來以副乾酪乳酸桿菌( Lactobacillus paracasei)、植物乳酸桿菌( Lactobacillus plantarum)、瑞士乳酸桿菌( Lactobacillus helveticus)以及鼠李糖乳酸桿菌( Lactobacillus rhamnosus)所構成的乳酸菌組合進行發酵培養而被製得。該益生菌蔬果酵素飲料經實驗而被證實具有抗氧化的效用。 The currently used antioxidant drugs have the problems of poor curative effect and easy side effects in clinical application. Therefore, relevant researchers in this field began to try to find useful active components from natural plant sources to develop drugs with anti-oxidative activity and without unwanted side effects. For example, CN 107348272 A discloses a probiotic vegetable and fruit enzyme drink, which is obtained by adding fruits and vegetables to Lactobacillus paracasei ( Lactobacillus paracasei ), plant Lactobacillus ( Lactobacillus plantarum ), Swiss Lactobacillus ( Lactobacillus helveticus ) and rhamnose The lactic acid bacteria combination composed of Lactobacillus rhamnosus is prepared by fermenting and cultivating. The probiotic vegetable and fruit enzyme drink has been proved to have anti-oxidative effects through experiments.
雖然已有上述的專利前案，本領域的相關研究人員仍致力於開發出具有抗氧化活性的植物材料的發酵產物的方法。Although the above-mentioned prior patents exist, relevant researchers in this field are still working on methods for developing fermentation products of plant materials with antioxidant activity.
發明概要Summary of the invention
於本發明中，申請人嘗試將73種植物材料(plant material)拿來以酵母菌、醋酸菌、乳酸菌、米麴菌以及紅麴菌進行發酵，由此所得到之複合植物材料的發酵產物(fermentation products)經實驗而被證實具有優異的抗氧化活性(anti-oxidative activity)。In the present invention, the applicant tried to ferment 73 kinds of plant materials with yeast, acetic acid bacteria, lactic acid bacteria, Koji oryzae and Rhododendron, and the resulting composite plant material fermentation product ( Fermentation products) have been proved by experiments to have excellent anti-oxidative activity (anti-oxidative activity).
於是，在第一個方面，本發明提供一種用於製備一發酵產物的方法，其包括： (a) 將一植物材料與一酵母菌進行發酵培養，藉此而得到一階段發酵培養物； (b)將該一階段發酵培養物與一醋酸菌進行發酵培養，藉此而得到二階段發酵培養物； (c) 將該二階段發酵培養物與一乳酸菌進行發酵培養，藉此而得到三階段發酵培養物；以及 (d)將該三階段發酵培養物與一米麴菌以及一紅麴菌進行發酵培養，藉此而得到四階段發酵培養物， 其中，該植物材料包含下列四群： i. 蓮藕、蘆筍、萵苣、西紅柿、蕎麥、番薯葉、茼蒿、菠菜、銀耳、甜椒、油菜、茄子、西芹、空心菜、青花椰菜、青椒、黃豆芽、黃秋葵、甘藍、芥藍、苦瓜、番石榴、橄欖、柿子、枇杷、竹筍、蓮子、四季豆、香菇、黃瓜、南瓜、芥菜、黑豆、豌豆、莧菜、蕪菁、李子以及木瓜； ii. 紅蘿蔔、芋頭、山藥、甜菜根、白蘿蔔以及番薯； iii. 青梅、鳳梨、葡萄、藍莓、檸檬、櫻桃、楊梅、水蜜桃、沙梨、葡萄柚、柳橙、柑橘、菠蘿蜜、草莓、火龍果、奇異果、蘋果以及楊桃；以及 iv. 紫蘇、蒲公英、烏梅、紅棗、黑芝麻、白芝麻、薑、甘草、枸杞、桑葉以及肉桂。 Thus, in a first aspect, the present invention provides a method for preparing a fermentation product comprising: (a) fermenting a plant material with a yeast to obtain a one-stage fermentation culture; (b) performing fermentation culture on the first-stage fermentation culture and an acetic acid bacterium, thereby obtaining a second-stage fermentation culture; (c) fermenting the two-stage fermentation culture with a lactic acid bacterium to obtain a three-stage fermentation culture; and (d) fermenting the three-stage fermentation culture with a koji bacterium and a red yeast rice bacterium, thereby obtaining a four-stage fermentation culture, Among them, the plant material includes the following four groups: i. Lotus root, asparagus, lettuce, tomato, buckwheat, sweet potato leaves, chrysanthemum, spinach, white fungus, sweet pepper, rape, eggplant, celery, water spinach, broccoli, green pepper, soybean sprouts, okra, cabbage, kale, bitter gourd, Guava, olives, persimmons, loquats, bamboo shoots, lotus seeds, green beans, shiitake mushrooms, cucumbers, pumpkins, mustard greens, black beans, peas, amaranths, turnips, plums, and papayas; ii. Carrots, taro, yams, beetroots, white radishes and sweet potatoes; iii. Greengage, pineapple, grape, blueberry, lemon, cherry, bayberry, peach, pear, grapefruit, orange, mandarin, jackfruit, strawberry, dragon fruit, kiwi, apple and star fruit; and iv. Basil, dandelion, dark plum, red dates, black sesame, white sesame, ginger, licorice, wolfberry, mulberry leaves and cinnamon.
在第二個方面，本發明提供一種發酵產物，它是藉由一如上所述的方法而被製得。In a second aspect, the present invention provides a fermentation product produced by a method as described above.
在第三個方面，本發明提供一種如上所述的發酵產物供應用於製備一用來抗氧化之組成物的用途。In a third aspect, the present invention provides a use of the above-mentioned fermentation product for preparing a composition for anti-oxidation.
在第四個方面，本發明提供一種用於抗氧化的方法，其包括對一有此需要的個體投予一如上所述的組成物。In a fourth aspect, the present invention provides a method for anti-oxidation comprising administering to a subject in need thereof a composition as described above.
本發明的上述以及其它目的、特徵與優點，在參照以下的詳細說明與較佳實施例後，將變得明顯。The above and other objects, features and advantages of the present invention will become apparent with reference to the following detailed description and preferred embodiments.
發明的詳細說明Detailed Description of the Invention
要被瞭解的是：若有任何一件前案刊物在此被引述，該前案刊物不構成一個下述承認：在台灣或任何其他國家之中，該前案刊物形成本技藝中的常見一般知識之一部分。It is to be understood that if any prior publication is cited herein, that prior publication does not constitute an acknowledgment that, in Taiwan or any other country, that prior publication forms a common practice in the art part of knowledge.
為了這本說明書之目的，將被清楚地瞭解的是：文字“包含有(comprising)”意指“包含但不限於”，以及文字“包括(comprises)”具有一對應的意義。For the purposes of this specification, it will be clearly understood that the word "comprising" means "including but not limited to", and that the word "comprises" has a corresponding meaning.
除非另外有所定義，在本文中所使用的所有技術性與科學術語具有熟悉本發明所屬技藝的人士所共同瞭解的意義。一熟悉本技藝者會認知到許多與那些被描述於本文中者相似或等效的方法和材料，它們可被用於實施本發明。當然，本發明決不受到所描述的方法和材料之限制。Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. Of course, the invention is in no way limited by the methods and materials described.
本發明提供一種用於製備一發酵產物的方法，其包括： (a) 將一植物材料(plant material)與一酵母菌進行發酵培養，藉此而得到一階段發酵培養物； (b)將該一階段發酵培養物與一醋酸菌進行發酵培養，藉此而得到二階段發酵培養物； (c) 將該二階段發酵培養物與一乳酸菌進行發酵培養，藉此而得到三階段發酵培養物；以及 (d)將該三階段發酵培養物與一米麴菌以及一紅麴菌進行發酵培養，藉此而得到四階段發酵培養物， 其中，該植物材料包含下列四群： i. 蓮藕、蘆筍、萵苣、西紅柿、蕎麥、番薯葉、茼蒿、菠菜、銀耳、甜椒、油菜、茄子、西芹、空心菜、青花椰菜、青椒、黃豆芽、黃秋葵、甘藍、芥藍、苦瓜、番石榴、橄欖、柿子、枇杷、竹筍、蓮子、四季豆、香菇、黃瓜、南瓜、芥菜、黑豆、豌豆、莧菜、蕪菁、李子以及木瓜； ii. 紅蘿蔔、芋頭、山藥、甜菜根、白蘿蔔以及番薯； iii. 青梅、鳳梨、葡萄、藍莓、檸檬、櫻桃、楊梅、水蜜桃、沙梨、葡萄柚、柳橙、柑橘、菠蘿蜜、草莓、火龍果、奇異果、蘋果以及楊桃；以及 iv. 紫蘇、蒲公英、烏梅、紅棗、黑芝麻、白芝麻、薑、甘草、枸杞、桑葉以及肉桂。 The invention provides a method for preparing a fermentation product, comprising: (a) fermenting a plant material (plant material) with a yeast to obtain a one-stage fermentation culture; (b) performing fermentation culture on the first-stage fermentation culture and an acetic acid bacterium, thereby obtaining a second-stage fermentation culture; (c) fermenting the two-stage fermentation culture with a lactic acid bacterium to obtain a three-stage fermentation culture; and (d) fermenting the three-stage fermentation culture with a Koji oryzae bacterium and a Rhododendron bacterium, thereby obtaining a four-stage fermentation culture, Among them, the plant material includes the following four groups: i. Lotus root, asparagus, lettuce, tomato, buckwheat, sweet potato leaves, chrysanthemum, spinach, white fungus, sweet pepper, rape, eggplant, celery, water spinach, broccoli, green pepper, soybean sprouts, okra, cabbage, kale, bitter gourd, Guava, olives, persimmons, loquats, bamboo shoots, lotus seeds, green beans, shiitake mushrooms, cucumbers, pumpkins, mustard greens, black beans, peas, amaranths, turnips, plums, and papayas; ii. Carrots, taro, yams, beetroots, white radishes and sweet potatoes; iii. Greengage, pineapple, grape, blueberry, lemon, cherry, bayberry, peach, pear, grapefruit, orange, mandarin, jackfruit, strawberry, dragon fruit, kiwi, apple and star fruit; and iv. Perilla, dandelion, dark plum, red dates, black sesame, white sesame, ginger, licorice, wolfberry, mulberry leaves and cinnamon.
較佳地，該酵母菌是啤酒酵母菌( Saccharomyces cerevisiae)。 Preferably, the yeast is Saccharomyces cerevisiae .
較佳地，該醋酸菌是選自於由下列所構成的群組：醋酸菌( Acetobacter aceti)、木質醋酸菌( Gluconacetobacter xylinus)，以及它們的組合。 Preferably, the acetic acid bacteria is selected from the group consisting of: Acetobacter aceti , Gluconacetobacter xylinus , and combinations thereof.
較佳地，該乳酸菌是選自於由下列所構成的群組：嗜酸乳桿菌( Lactobacillus acidophilus)、保加利亞乳桿菌( Lactobacillus Bulgaricus)、胚芽乳桿菌( Lactobacillus plantarum)、副乾酪乳桿菌( Lactobacillus paracasei)、嗜熱鏈球菌( Streptococcus thermophilus)、菊糖芽孢乳桿菌( Sporolactobacillus inulinus)、凝結芽孢桿菌( Bacillus coagulans)、雙叉型雙叉桿菌( Bifidobacterium bifidum)，以及它們的組合。 Preferably, the lactic acid bacteria are selected from the group consisting of Lactobacillus acidophilus , Lactobacillus Bulgaricus , Lactobacillus plantarum , Lactobacillus paracasei ), Streptococcus thermophilus, Sporolactobacillus inulinus , Bacillus coagulans , Bifidobacterium bifidum , and combinations thereof.
依據本發明，適用於製備酵母菌、醋酸菌、乳酸菌、米麴菌與紅麴菌之接種源的培養基包括，但不限於：YM肉湯培養基(YM broth)、AC肉湯培養基(AC broth)以及MRS肉湯培養基(MRS broth)。According to the present invention, the medium suitable for preparing the inoculum source of yeast, acetic acid bacteria, lactic acid bacteria, Koji oryzae and Rhododendron include, but not limited to: YM broth, AC broth And MRS broth (MRS broth).
此外，該培養基亦可包含有一選自於下列群組中的天然物質：白米(milled rice)[例如，稉米(japonica rice)]、糙米(brown rice)、紫米(black rice)、糯米(waxy rice)，以及它們的組合。較佳地，該培養基進一步包含有稉米。In addition, the culture medium may also contain a natural substance selected from the group consisting of milled rice [for example, japonica rice], brown rice, black rice, waxy rice rice), and their combinations. Preferably, the medium further contains japonica rice.
依據本發明，該植物材料可以是未經加工處理的(unprocessed)(亦即新鮮摘採的)，或者是經過一選自於由下列所構成之群組中的加工處理所製得的：乾燥處理(drying treatment)、研磨處理(grinding treatment)、切碎處理(chopping treatment)、粉碎處理(comminuting treatment)，以及它們的組合。在本發明的一個較佳具體例中，該植物材料是經過切碎處理而製得。According to the present invention, the plant material may be unprocessed (i.e. freshly picked) or obtained through a process selected from the group consisting of: drying (drying treatment), grinding treatment (grinding treatment), chopping treatment (chopping treatment), comminuting treatment (comminuting treatment), and combinations thereof. In a preferred embodiment of the present invention, the plant material is obtained by chopping.
依據本發明，在進行該方法的步驟(a)中的發酵培養的期間，可藉由添加一醣類來調整糖度，較佳地，該糖度被維持在15至20 Brix。According to the present invention, during the fermentation culture in the step (a) of the method, the sugar content can be adjusted by adding a sugar, preferably, the sugar content is maintained at 15 to 20 Brix.
依據本發明，該醣類可選自於由下列所構成的群組：葡萄糖、果糖、蔗糖、以蔗糖為主要成分的食用糖，以及它們的組合。According to the present invention, the sugar can be selected from the group consisting of glucose, fructose, sucrose, table sugar with sucrose as the main component, and combinations thereof.
較佳地，該醣類是以蔗糖為主要成分的食用糖。適用於本發明的以蔗糖為主要成分的食用糖包括，但不限於：冰糖、白砂糖以及赤砂糖(它亦被稱為紅糖或黑糖)。在本發明的一個較佳具體例中，該以蔗糖為主要成分的食用糖是赤砂糖。Preferably, the sugar is edible sugar with sucrose as the main component. Table sugars suitable for the present invention with sucrose as the main component include, but are not limited to: rock sugar, white granulated sugar and brown granulated sugar (it is also called brown sugar or black sugar). In a preferred embodiment of the present invention, the edible sugar mainly composed of sucrose is brown sugar.
依據本發明，在該方法的步驟(b)中，第i群與第ii群植物材料的一階段發酵培養物可先被混合，再進行該使用醋酸菌的發酵培養，而得到一個第i-ii群植物材料的二階段發酵培養物。According to the present invention, in step (b) of the method, the one-stage fermentation cultures of group i and group ii plant materials can be mixed first, and then the fermentation culture using acetic acid bacteria is carried out to obtain an i- A two-stage fermentation culture of group ii plant material.
依據本發明，在該方法的步驟(c)中，第i-ii群植物材料的二階段發酵培養物與第iii群植物材料的二階段發酵培養物可先被混合，再進行該使用乳酸菌的發酵培養，而得到一個第i-iii群植物材料的三階段發酵培養物。According to the present invention, in step (c) of the method, the second-stage fermentation culture of group i-ii plant material and the second-stage fermentation culture of group iii plant material can be mixed first, and then the fermentation using lactic acid bacteria is carried out. fermentation to obtain a three-stage fermentation culture of group i-iii plant material.
依據本發明，在該方法的步驟(d)中，第i-iii群植物材料的三階段發酵培養物與第iv群植物材料的三階段發酵培養物可先被混合，再進行該使用米麴菌與紅麴菌的發酵培養，而得到一個第i-iv群植物材料的四階段發酵培養物。According to the present invention, in the step (d) of the method, the three-stage fermentation culture of plant material of group i-iii and the three-stage fermentation culture of plant material of group iv may be mixed first, and then the use of rice koji Bacteria and red yeast rice fermented to obtain a four-stage fermentation culture of the i-iv group plant material.
依據本發明，該四階段發酵培養物可進一步進行一固-液分離處理(solid-liquid separation treatment)，俾以移除固體部分。According to the present invention, the four-stage fermentation culture can be further subjected to a solid-liquid separation treatment to remove the solid portion.
依據本發明，該固-液分離處理可選自於由下列所構成的群組：離心處理(centrifugation treatment)、過濾處理(filtration treatment)、濃縮處理(concentration treatment)，以及它們的組合。在本發明的一個較佳具體例中，該固-液分離處理是離心處理。According to the present invention, the solid-liquid separation treatment can be selected from the group consisting of: centrifugation treatment, filtration treatment, concentration treatment, and combinations thereof. In a preferred embodiment of the present invention, the solid-liquid separation treatment is centrifugation treatment.
如本文中所使用的，術語“發酵(fermentation)”、“培養(culturing)”以及“培育(cultivation)”可被交換地使用。有關發酵的操作程序與參數條件等是落在熟習此項技術之人士的專業素養與例行技術範疇內。As used herein, the terms "fermentation", "culturing" and "cultivation" are used interchangeably. The operating procedures and parameter conditions related to fermentation fall within the professional quality and routine technical scope of those who are familiar with this technology.
可瞭解到的是，有關發酵的操作條件會進一步隨著所使用的菌株以及該菌株與該植物材料的用量比例等因素而被變動，以便達致最佳的發酵效果。而這些操作條件的選擇是熟習此項技藝者能例行性地自行決定的。It can be understood that the operating conditions related to fermentation will be further changed with factors such as the strain used and the ratio of the strain to the plant material in order to achieve the best fermentation effect. The selection of these operating conditions is within the routine discretion of those skilled in the art.
較佳地，該發酵可在一範圍落在25℃至30℃內的溫度下被進行。在本發明的一個較佳具體例中，該發酵是在28℃下被進行。Preferably, the fermentation is carried out at a temperature ranging from 25°C to 30°C. In a preferred embodiment of the present invention, the fermentation is carried out at 28°C.
本發明亦提供一種發酵產物，它是藉由使用一如上所述的方法而被製得。The present invention also provides a fermented product prepared by using a method as described above.
此外，本發明亦提供一種如上所述的發酵產物供應用於製備一用來抗氧化之組成物的用途。In addition, the present invention also provides a use of the above-mentioned fermentation product for preparing a composition for anti-oxidation.
依據本發明，該組成物可以是藥學組成物(pharmaceutical composition)。According to the present invention, the composition may be a pharmaceutical composition.
依據本發明，該藥學組成物可進一步包含有一被廣泛地使用於藥物製造技術之藥學上可接受的載劑(pharmaceutically acceptable carrier)。例如，該藥學上可接受的載劑可包含一或多種選自於下列的試劑：溶劑(solvent)、緩衝液(buffer)、分解劑(decomposer)、賦形劑(excipient)、安定劑(stabilizing agent)、稀釋劑(diluent)、潤濕劑(wetting agent)、潤滑劑(lubricant)以及類似之物。有關這些試劑的選用與數量是落在熟習此項技術之人士的專業素養與例行技術範疇內。According to the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier (pharmaceutically acceptable carrier) which is widely used in pharmaceutical manufacturing technology. For example, the pharmaceutically acceptable carrier may contain one or more reagents selected from the group consisting of: solvent, buffer, decomposer, excipient, stabilizing agent), diluent, wetting agent, lubricant, and the like. The selection and quantities of these reagents are within the professionalism and routine skill of those skilled in the art.
依據本發明，該組成物可以是食品組成物(food composition)，例如，呈一食品添加物(food additive)的形式，其可以被添加至一可食性材料(edible material)中以製備一供人類或動物食用的食品產品。依據本發明，該食物產品的種類可包括，但不限於：奶粉(milk powder)、發酵乳(fermented milk)、優格(yogurt)、乳酪(butter)、飲料(beverages)(例如，茶、咖啡)、機能性飲料(functional beverages)、麵製品(flour product)、烘焙食品(baked foods)、甜點(confectionery)、糖果(candies)、發酵食品(fermented foods)、動物飼料(animal feeds)、保健食品(health foods)、嬰兒食品(infant food)，以及膳食補充品(dietary supplements)。According to the invention, the composition may be a food composition, for example, in the form of a food additive which may be added to an edible material to prepare a food for human consumption. or food products for animal consumption. According to the present invention, the kind of this food product can include, but not limited to: milk powder (milk powder), fermented milk (fermented milk), yogurt (yogurt), cheese (butter), beverage (beverages) (for example, tea, coffee ), functional beverages, flour products, baked foods, confectionery, candies, fermented foods, animal feeds, health food (health foods), baby food (infant food), and dietary supplements (dietary supplements).
本發明亦提供一種用於抗氧化的方法，其包括對一有此需要的個體投予一如上所述的組成物。The present invention also provides a method for anti-oxidation, which comprises administering a composition as described above to an individual in need thereof.
如本文中所使用的，術語“投予(administering)”以及“投藥(administration)”可被交換地使用，並且意指藉由任何合適的途徑來對一個體導入(introducing)、提供(providing)或遞送(delivering)一預定的活性成分以執行其預期的效用。As used herein, the terms "administering" and "administration" are used interchangeably and mean introducing, providing, Or delivering (delivering) a predetermined active ingredient to perform its intended effect.
如本文中所使用的，術語“個體(subject)”意指任何感興趣的哺乳類動物，諸如人(humans)、猴子(monkeys)、牛(cows)、綿羊(sheep)、馬(horses)、豬(pigs)、山羊(goats)、狗(dogs)、貓(cats)、小鼠(mice)以及大鼠(rats)。 較佳實施例之詳細說明 As used herein, the term "subject" means any mammal of interest, such as humans, monkeys, cows, sheep, horses, pigs (pigs), goats (goats), dogs (dogs), cats (cats), mice (mice) and rats (rats). Detailed Description of the Preferred Embodiment
本發明將就下面的實施例來做進一步說明，但應瞭解的是，該等實施例僅是供例示說明用，而不應被解釋為本發明的實施上的限制。
實施例 一般實驗材料： 1. 在下面的實施例中所使用的植物材料包括：青梅(plum)(
Prunus mumeSieb. et Zucc.)、鳳梨(pineapple)(
Ananas comosus)、葡萄(grape)(
Vitis vinifera)、藍莓(blueberry)(
Cyanococcus)、檸檬(lemon)(
Citrus limon)、櫻桃(cherry)(
Prunus avium)、楊梅(Chinese bayberry)(
Myrica rubra)、水蜜桃(peach)(
Prunus persica)、沙梨(Chinese pear)(
Pyrus pyrifolia)、葡萄柚(grapefruit)(
Citrus paradisi)、柳橙(orange)(
Citrus sinensis)、柑橘(citrus)(
Citrus reticulata)、菠蘿蜜(Jackfruit)(
Artocarpus heterophyllus)、草莓(strawberry)(
Fragaria ananassa)、火龍果(dragon fruit)[
Hylocereus undatus(Haw.) Britt. et Rose]、奇異果(Kiwifruit)(
Actinidia chinensisPlanch.)、蘋果(apple)(
Malus pumila)、楊桃(star fruit)(
Averrhoa carambola)、木瓜(papaya)(
Carica papaya)、蓮藕(lotus root)(
Nelumbo nucifera Gaertn)、蘆筍(asparagus)(
Asparagus officinalis)、萵苣(lettuce)(
Lactuca sativa)、西紅柿(tomato)(
Solanum lycopersicum)、蕎麥(buckwheat)(
Fagopyrum esculentum)、番薯葉(sweet potato leaf)(
Ipomoea batatas)、茼蒿(garland chrysanthemum)(
Glebionis coronaria)、菠菜(spinach)(
Spinacia oleracea)、銀耳(white fungus)(
Tremella fuciformis)、甜椒(sweet pepper)(
Capsicum annuum var. grossum)、油菜(rapeseed)(
Brassica campestris)、茄子(aubergine)(
Solanum melongena)、西芹(celery)(
Apium graveolens)、空心菜(water spinach)(
Ipomoea aquatica)、青花椰菜(broccoli)(
Brassica oleracea var. italica)、黃豆芽(bean sprout)[
Glycine max (L.) Merr.]、秋葵(okra)(
Abelmoschus esculentus)、甘藍(cabbage)(
Brassica oleracea)、芥蘭(Chinese kale)(
Brassica oleracea Alboglabra Group)、苦瓜(bitter melon)(
Momordica charantia)、番石榴(guava)(
Psidium guajava)、橄欖(olive)(
Olea europaea)、柿子(persimmon)(
Diospyros kaki)、枇杷(loquat)(
Eriobotrya japonica)、竹筍(bamboo shoot)(
Leleba oldhami)、蓮子(lotus seed)(
Nelumbo nucifera Gaertn.)、四季豆(green bean)(Phaseolus vulgaris)、香菇(shiitake fungus)(
Lentinula edodes)、黃瓜(cucumber)(
Cucumis sativus)、南瓜(pumpkin)(
Cucurbita moschata)、芥菜(mustard)(
Brassica juncea)、黑豆(black bean)(
Phaseolus vulgaris)、豌豆(garden pea)(
Pisum sativum)、莧菜(edible amaranth)(Amaranthus gangeticus)、蕪菁(turnip)(
Brassica rapa subsp. rapa)、李子(plum)(
Prunus domestica)、紅蘿蔔(carrot)(
Daucus carotasubsp.
sativus)、芋頭(taro)(
Colocasia esculenta)、山藥(Chinese yam)(
Dioscorea batatas)、甜菜根(beetroot)(
Beta vulgaris)、白蘿蔔(white radish)(
Raphanus sativus var. Longipinnatus)、番薯(sweet potato)(
Ipomoea batatas)、紫蘇(perilla)(
Perilla frutescens var. crispa)、蒲公英(Mongolian Dandelion)(
Taraxacum)、烏梅(mume fruit)(
Prunus mume)、紅棗(Chinese date)(
Zizyphus jujuba)、黑芝麻(black sesame seeds)(
Semen Sesami Nigrum)、白芝麻(white sesame seeds)(
Sesamum indicum L.)、薑(ginger)(
Zingiber officinale)、甘草(liquorice)(
Glycyrrhiza uralensis)、枸杞(wolfberry)(
Lycium chinense)、桑葉(mulberry leave)(
Morus alba)、肉桂(cinnamon)(
Cinnamomum cassia)，分別購自於台中果菜批發市場以及新世代生藥行。
2. 在下面的實施例中所使用的酵母菌(yeast)、醋酸菌(acetic acid bacteria)、乳酸菌(lactic acid bacteria)以及米麴菌(
Aspergillus oryzae)與紅麴菌(
Monascus purpureus)皆是得自於台灣的食品工業發展研究所(Food Industry Research and Development Institute, FIRDI)的生物資源保存及研究中心(Bioresource Collection and Research Center, BCRC)(300新竹市食品路331號，台灣)，並且已分別被整合於下面的表1中。
表1. 所使用的各個菌株
將上面“一般實驗材料”的第2項當中所述的1種酵母菌菌株接種至如上面表2中所示的YM肉湯培養基中，繼而於26℃下以120 rpm予以震盪培養歷時60小時，所形成的培養物被使用作為酵母菌菌株的接種源。 2. 製備醋酸菌菌株的接種源： One of the yeast strains described in item 2 of "General Experimental Materials" above was inoculated into the YM broth medium shown in Table 2 above, followed by shaking culture at 120 rpm at 26°C for 60 hours , the resulting culture was used as an inoculum source for the yeast strain. 2. Prepare the inoculum source of the acetic acid bacteria strain:
將上面“一般實驗材料”的第2項當中所述的2種醋酸菌菌株分別接種至AC肉湯培養基(AC broth)(廠牌為HIMEDIA，貨號為M875)中，繼而於26℃下以120 rpm予以震盪培養歷時60小時，所形成的培養物被使用作為醋酸菌菌株的接種源。 3. 製備乳酸菌菌株的接種源： Inoculate the two acetic acid bacteria strains mentioned in item 2 of the above "General Experimental Materials" into AC broth (HIMEDIA, product number: M875) respectively, and then inoculate at 26°C at 120 rpm was shaken for 60 hours, and the resulting culture was used as the inoculum source of the acetic acid bacteria strain. 3. Prepare the inoculum source of the lactic acid bacteria strain:
將上面“一般實驗材料”的第2項當中所述的8種乳酸菌菌株分別接種至MRS肉湯培養基(MRS broth)(廠牌為HIMEDIA，貨號為M369)中，繼而於35℃下以120 rpm予以震盪培養歷時60小時，所形成的培養物被使用作為乳酸菌菌株的接種源。 4. 製備米麴菌與紅麴菌的接種源： Inoculate the 8 kinds of lactic acid bacteria strains mentioned in item 2 of the above "General Experimental Materials" into MRS broth medium (MRS broth) (the brand is HIMEDIA, the article number is M369), and then incubate at 35°C at 120 rpm Shaking culture was performed for 60 hours, and the resulting culture was used as an inoculation source of the lactic acid bacteria strain. 4. Prepare the inoculum source of rice yeast and red yeast rice:
將上面“一般實驗材料”的第2項當中所述的米麴菌與紅麴菌菌株分別接種至經蒸煮的稉米(japonica rice)，繼而於35℃至40℃下靜置培養歷時50小時，所形成的培養物分別被使用作為米麴菌與紅麴菌的接種源。 實驗方法： The strains of Koji oryzae and Koji Koji described in item 2 of the above "General Experimental Materials" were respectively inoculated into boiled japonica rice, followed by static culture at 35°C to 40°C for 50 hours, The resulting cultures were used as inoculum sources for Koji oryzae and Rhododendron, respectively. experimental method:
首先，將在上面“一般實驗材料”的第1項當中所述的73種植物材料以水清洗乾淨且予以風乾(air-dried)，接著依照下面表3將該等植物材料分成4群(亦即第i群至第iv群)，然後將每一種植物材料切碎並將這4群的植物材料各別置於4個0.5噸的發酵槽中，繼而添加適量的砂糖(購自於台灣糖業股份有限公司)來完全覆蓋該等發酵槽內經切碎的植物材料。
表3. 各群所含有的植物材料種類
之後，將上面“實驗材料”的第1項當中所得到的1種酵母菌菌株的接種源以一為0.1% (v/v)的接種量分別接種至該4個發酵槽中並予以混合均勻，接著藉由使用糖度計(saccharometer)來測量所形成的混合物的糖度(Brix)，然後依據測得的數值來將該混合物的糖度維持在15至20 Brix，繼而令該等發酵槽在一為28℃的溫度下靜置發酵歷時30天，藉此而分別得到4個一階段發酵培養物(亦即，第i群至第iv群植物材料的一階段發酵培養物)。之後，將第i群與第ii群植物材料的一階段發酵培養物混合，而得到第i-ii群植物材料的一階段發酵培養物。Afterwards, the inoculum source of 1 yeast strain obtained in item 1 of the above "experimental material" was inoculated into the 4 fermentation tanks with an inoculum amount of 0.1% (v/v) and mixed evenly , then by using a saccharometer (saccharometer) to measure the Brix of the mixture formed, and then maintain the Brix of the mixture at 15 to 20 Brix according to the measured value, and then make the fermentation tanks in a The static fermentation lasted 30 days at a temperature of 28° C., thereby obtaining four first-stage fermentation cultures (ie, the first-stage fermentation cultures of group i to group iv plant materials). Thereafter, group i is mixed with the primary fermentation culture of group ii plant material to obtain a primary fermentation culture of group i-ii plant material.
接著，將上面“實驗材料”的第2項當中所得到的2種醋酸菌菌株的接種源分別以一為0.2% (v/v)的接種量同時接種至第i-ii群、第iii群與第iv群植物材料的一階段發酵培養物中並予以混合均勻，繼而在一為28℃的溫度、一為55 L/min的通氣量以及一為1,700 rpm的攪拌速率下進行發酵歷時30天，藉此而分別得到3個二階段發酵培養物(亦即，第i-ii群、第iii群與第iv群植物材料的二階段發酵培養物)。之後，將第i-ii群植物材料的二階段發酵培養物與第iii群植物材料的二階段發酵培養物混合，而得到第i-iii群植物材料的二階段發酵培養物。Then, the inoculum sources of the 2 kinds of acetic acid bacteria strains obtained in item 2 of the above "experimental materials" were simultaneously inoculated into groups i-ii and group iii with an inoculation amount of 0.2% (v/v) respectively. Into a one-stage fermentation culture of group iv plant material and mixed uniformly, followed by fermentation for 30 days at a temperature of 28°C, an aeration rate of 55 L/min and a stirring rate of 1,700 rpm , thereby obtaining three second-stage fermentation cultures (that is, the second-stage fermentation cultures of group i-ii, group iii, and group iv plant materials) respectively. Thereafter, the second-stage fermentation culture of group i-ii plant material is mixed with the second-stage fermentation culture of group iii plant material to obtain a second-stage fermentation culture of group i-iii plant material.
之後，將上面“實驗材料”的第3項當中所得到的8種乳酸菌菌株的接種源分別以一為0.002% (v/v)的接種量同時接種至第i-iii群與第iv群植物材料的二階段發酵培養物中並予以混合均勻，繼而在一為28℃的溫度下靜置發酵歷時30天，藉此而分別得到2個三階段發酵培養物(亦即，第i-iii群與第iv群植物材料的三階段發酵培養物)。之後，將第i-iii群植物材料的三階段發酵培養物與第iv群植物材料的三階段發酵培養物混合，而得到第i-iv群植物材料的三階段發酵培養物。Afterwards, the inoculum sources of 8 kinds of lactic acid bacteria strains obtained in item 3 of the above "experimental materials" were respectively inoculated to the i-iii group and the iv group plants at the same time with an inoculum amount of 0.002% (v/v) The two-stage fermentation cultures of the materials were mixed uniformly, and then left to ferment at a temperature of 28°C for 30 days, thereby obtaining two three-stage fermentation cultures (that is, groups i-iii Three-stage fermentation culture with group iv plant material). Thereafter, the three-stage fermentation culture of group i-iii plant material is mixed with the three-stage fermentation culture of group iv plant material to obtain a three-stage fermentation culture of group i-iv plant material.
之後，將上面“實驗材料”的第4項當中所得到的米麴菌與紅麴菌的接種源分別以一為0.1% (v/v)的接種量接種至第i-iv植物材料的三階段發酵培養物中並予以混合均勻，繼而在一為28℃的溫度下靜置發酵歷時至少365天，藉此而得到第i-iv群植物材料的四階段發酵培養物。After that, inoculate the inoculum sources of Koji oryzae and Rhododendron obtained in Item 4 of the above "Experimental Materials" to the three inoculums of the i-iv plant materials with an inoculum amount of 0.1% (v/v) respectively. The four-stage fermentation culture of group i-iv plant materials is obtained by mixing them uniformly in the stage fermentation culture, and then standing and fermenting at a temperature of 28° C. for at least 365 days.
之後，所得到的四階段發酵培養物以1,300 rpm來進行離心歷時10分鐘，繼而收取上澄液，藉此而得到複合植物材料的發酵產物。 實施例 2. 本發明的發酵產物中的游離胺基酸 (free amino acid) 含量之測定 實驗方法： Afterwards, the obtained four-stage fermentation culture was centrifuged at 1,300 rpm for 10 minutes, and then the supernatant liquid was collected, thereby obtaining the fermentation product of the composite plant material. Embodiment 2. The assay method of the free amino acid (free amino acid) content in the fermented product of the present invention :
有關本發明的發酵產物的游離胺基酸含量之測定是委託台灣的食品工業發展研究所並使用離子交換層析法(ion exchange chromatography)來代為進行。 結果： The determination of the free amino acid content of the fermented product of the present invention was entrusted to Taiwan's Institute of Food Industry Development and performed by using ion exchange chromatography (ion exchange chromatography). result:
本實驗所測得的結果被顯示於下面的表4中。從表4可見，本發明的發酵產物含有數種游離胺基酸且總游離胺基酸含量高達164.1 mg/100 g。
表4. 本發明的發酵產物中的游離胺基酸種類及其含量
有關本發明的發酵產物的抗氧化酵素活性之測定是委託台灣檢驗科技股份有限公司(SGS TAIWAN LTD)的超微量工業安全實驗室(Ultra Trace Industrial Safety Hygiene)並使用化學發光光譜儀(Chemiluminescence Spectrometer)來進行似超氧化物歧化酶(superoxide dismutase-like, SOD-like)活性的檢測。 結果： The determination of the antioxidant enzyme activity of the fermentation product of the present invention is entrusted to the Ultra Trace Industrial Safety Hygiene of Taiwan Inspection Technology Co., Ltd. (SGS TAIWAN LTD) and uses a chemiluminescence spectrometer (Chemiluminescence Spectrometer) to The detection of superoxide dismutase-like (SOD-like) activity was carried out. result:
實驗結果顯示：本發明的發酵產物的似超氧化物歧化酶活性為24,300 U/g，這表示本發明的發酵產物能夠展現出優異的抗氧化效用。The experimental results show that the superoxide dismutase-like activity of the fermented product of the present invention is 24,300 U/g, which means that the fermented product of the present invention can exhibit excellent antioxidant effect.
綜合以上的實驗結果可知：本發明的發酵產物富含有多種游離胺基酸以及具有優異的抗氧化活性，而具有發展成為一用來抗氧化的食品產品的高潛力。Based on the above experimental results, it can be seen that the fermentation product of the present invention is rich in various free amino acids and has excellent antioxidant activity, and has high potential to be developed into a food product for antioxidant.
於本說明書中被引述之所有專利和文獻以其整體被併入本案作為參考資料。若有所衝突時，本案詳細說明(包含界定在內)將佔上風。All patents and literature cited in this specification are hereby incorporated by reference in their entirety. In case of conflict, the detailed description of the case (including definitions) will prevail.
雖然本發明已參考上述特定的具體例被描述，明顯地在不背離本發明之範圍和精神之下可作出很多的修改和變化。因此意欲的是，本發明僅受如隨文檢附之申請專利範圍所示者之限制。While the invention has been described with reference to specific examples thereof, obviously many modifications and variations can be made without departing from the scope and spirit of the invention. It is therefore intended that the present invention be limited only as indicated by the claims attached hereto.
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| TW110145897A TWI802136B (en) | 2021-12-08 | 2021-12-08 | Fermentation of plants and their products by pure bacteria inoculation and their application in anti-oxidation |
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| TW (1) | TWI802136B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102940288A (en) * | 2012-11-19 | 2013-02-27 | 商丘市饮之健食品有限公司 | Composite enzyme beverage and preparation process method thereof |
| CN109567161A (en) * | 2018-12-03 | 2019-04-05 | 苏剑锋 | A kind of selenium-rich blueberry enzyme beverage and preparation method thereof |
| TWM611003U (en) * | 2021-01-06 | 2021-04-21 | 邱聖斌 | Vegetable and fruit herbal fermentation device capable of enhancing enzyme activity of superoxide dismutase |
-
2021
- 2021-12-08 TW TW110145897A patent/TWI802136B/en active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102940288A (en) * | 2012-11-19 | 2013-02-27 | 商丘市饮之健食品有限公司 | Composite enzyme beverage and preparation process method thereof |
| CN109567161A (en) * | 2018-12-03 | 2019-04-05 | 苏剑锋 | A kind of selenium-rich blueberry enzyme beverage and preparation method thereof |
| TWM611003U (en) * | 2021-01-06 | 2021-04-21 | 邱聖斌 | Vegetable and fruit herbal fermentation device capable of enhancing enzyme activity of superoxide dismutase |
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| TW202322830A (en) | 2023-06-16 |
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