TW202216986A - Culture medium and culturing method of polypore mycelial pellets - Google Patents
Culture medium and culturing method of polypore mycelial pellets Download PDFInfo
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本發明是有關於一種培養真菌的方法,特別是一種培養多孔菌菌絲球的培養液及方法。The present invention relates to a method for culturing fungi, in particular to a culture solution and method for culturing polyporous fungus mycelial balls.
多孔菌(polypores)中許多菌種具有多種有效物質(如:三萜類化合物、酚類化合物、聚酮化合物、多醣體及固醇類等)而具有藥用價值(例如:靈芝、雲芝、牛樟芝、茯苓等)。上述有效物質不僅可存在於子實體,也可存在於菌絲體中,因此菌絲體也具有藥用價值。Many species of polypores have a variety of effective substances (such as: triterpenoids, phenolic compounds, polyketides, polysaccharides and sterols, etc.) and have medicinal value (such as: Ganoderma lucidum, Yunzhi, Antrodia camphorata, Poria, etc.). The above-mentioned effective substances can exist not only in the fruit body, but also in the mycelium, so the mycelium also has medicinal value.
依據培養基的類型,培養藥用真菌的方法可大致分為兩類:固態醱酵及液態醱酵,其中固態醱酵多用以生產子實體,且液態培養多用以生產菌絲體。相較於固態醱酵,液態醱酵之條件較容易操控,且相較於子實體,菌絲體的生產週期較短,因此液態培養具有大規模生產的潛力。According to the type of medium, the methods of culturing medicinal fungi can be roughly divided into two categories: solid-state fermentation and liquid fermentation. Among them, solid-state fermentation is mostly used to produce fruiting bodies, and liquid culture is mostly used to produce mycelium. Compared with solid fermentation, the conditions of liquid fermentation are easier to control, and the production cycle of mycelium is shorter than that of fruiting bodies, so liquid culture has the potential for large-scale production.
根據培養液的成分及培養方式,菌絲體在培養液中可形成絲狀或球狀等形態,其中菌絲體的形態可影響總產量。具體而言,絲狀菌絲體會增加培養液的黏度及非牛頓流變性,引發不良的質傳(如氧氣或溫度的傳遞),導致產量下降。相反地,在低密度的狀況下,球狀菌絲體(菌絲球,pellets)無上述問題,因此藉由調整培養方式可改變菌絲球的型態,從而提高菌絲體之總產量。According to the composition of the culture medium and the culture method, the mycelium can form a filamentous or spherical shape in the culture medium, and the shape of the mycelium can affect the total yield. Specifically, filamentous mycelium increases the viscosity and non-Newtonian rheology of the culture medium, causing poor mass transfer (such as oxygen or temperature transfer), resulting in decreased yield. On the contrary, under the condition of low density, spherical mycelium (pellets) does not have the above problems, so by adjusting the culture method, the shape of the mycelium can be changed, thereby increasing the total yield of mycelium.
因此,亟需一種培養菌絲球的方法,以提高菌絲體的總產量。Therefore, there is an urgent need for a method for culturing mycelial balls to improve the total yield of mycelium.
因此,本發明之一樣態是提供一種培養菌絲球的方法,其係在培養液中添加非水解動物性蛋白來提高菌絲球的數量及生物量。Therefore, one aspect of the present invention is to provide a method for culturing mycelial balls, which is to add non-hydrolyzed animal protein to the culture solution to increase the number and biomass of the mycelial balls.
根據本發明之上述之態樣,提出一種培養菌絲球的方法。上述方法可例如利用培養液對多孔菌菌絲體進行液態醱酵步驟,以獲得菌絲球,其特徵在於培養液可包含但不限於馬鈴薯右旋糖培養液(potato dextrose broth,PDB)及非水解動物性蛋白(unhydrolyzed animal protein),且非水解動物性蛋白於PDB的含量可例如小於或等於1重量百分比(wt%)。According to the above aspect of the present invention, a method for culturing mycelial balls is provided. The above-mentioned method can, for example, use a culture solution to perform a liquid fermentation step on the mycelium of the polyporous fungus to obtain mycelial balls, and it is characterized in that the culture solution can include but is not limited to potato dextrose broth (PDB) and non- Hydrolyzed animal protein (unhydrolyzed animal protein), and the content of the unhydrolyzed animal protein in the PDB may be, for example, less than or equal to 1 weight percent (wt%).
依據本發明上述之實施例,非水解動物性蛋白可例如來源自蛋黃。According to the above-mentioned embodiment of the present invention, the non-hydrolyzed animal protein can be derived from egg yolk, for example.
依據本發明上述之實施例,培養液可選擇性包含0.25 wt%至0.75 wt%之酵母萃取物。According to the above-mentioned embodiment of the present invention, the culture medium may optionally contain 0.25 wt % to 0.75 wt % of yeast extract.
依據本發明上述之實施例,在進行液態醱酵步驟前,上述方法可選擇性包含對培養液進行熱不活化(heat-inactivated)處理。According to the above-mentioned embodiment of the present invention, before the liquid fermentation step, the above-mentioned method can optionally include heat-inactivating the culture medium.
依據本發明上述之實施例,多孔菌可包含但不限於擬層孔菌屬(Fomitopsis spp.)及靈芝屬(Ganoderma spp.)。According to the above-mentioned embodiments of the present invention, the polypore bacteria may include, but are not limited to, Fomitopsis spp. and Ganoderma spp.
依據本發明上述之實施例,液態醱酵步驟的溫度可例如為23°C至29°C,且液態醱酵步驟之轉速可例如為100 rpm至150 rpm。According to the above-mentioned embodiment of the present invention, the temperature of the liquid fermentation step can be, for example, 23°C to 29°C, and the rotational speed of the liquid fermentation step can be, for example, 100 rpm to 150 rpm.
本發明之另一樣態是提供一種培養菌絲球的培養液,以應用於上述培養菌絲球的方法。Another aspect of the present invention is to provide a culture solution for culturing mycelial balls, which can be applied to the above-mentioned method for culturing mycelial balls.
根據本發明之上述之態樣,提出一種培養菌絲球的培養液,其特質在於馬鈴薯右旋糖培養液(PDB)含有小於或等於1 wt%之非水解動物性蛋白。According to the above aspect of the present invention, a culture solution for culturing mycelial balls is provided, which is characterized in that the potato dextrose culture solution (PDB) contains less than or equal to 1 wt% of non-hydrolyzed animal protein.
依據本發明上述之實施例,培養液可選擇性包含0.25 wt%至0.75 wt%之酵母萃取物。According to the above-mentioned embodiment of the present invention, the culture medium may optionally contain 0.25 wt % to 0.75 wt % of yeast extract.
依據本發明上述之實施例,非水解動物性蛋白可例如來源自蛋黃。According to the above-mentioned embodiment of the present invention, the non-hydrolyzed animal protein can be derived from egg yolk, for example.
依據本發明上述之實施例,培養液可例如經熱不活化處理。According to the above-described embodiments of the present invention, the culture medium may be, for example, heat inactivated.
應用本發明之培養菌絲球的方法,可顯著提升菌絲球數量及生物量,從而提高菌絲體的總產量。By applying the method for culturing mycelial balls of the present invention, the number and biomass of the mycelial balls can be significantly increased, thereby increasing the total yield of the mycelium.
本發明所提到的單數形式「一」、「一個」和「所述」包含複數引用,除非上下文另有明確規定。數值範圍(如10%至11%的A)若無特定說明皆包含上、下限值(即10%≤A≤11%);數值範圍若未界定下限值(如低於0.2%的B,或0.2%以下的B),則皆指其下限值可能為0(即0%≤B≤0.2%)。上述用語是用以說明及理解本發明,而非用以限制本發明。References herein to the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. The numerical range (such as 10% to 11% of A) includes the upper and lower limit values unless otherwise specified (
本發明提供的培養菌絲球(pellets)的方法,其利用含非水解動物性蛋白(unhydrolyzed animal protein)之馬鈴薯右旋糖培養液(potato dextrose broth,PDB)對多孔菌之菌絲體(mycelium)進行液態醱酵步驟,以提高菌絲球數量及生物量。The method for culturing mycelium balls (pellets) provided by the present invention utilizes the potato dextrose broth (PDB) containing unhydrolyzed animal protein (unhydrolyzed animal protein) to treat the mycelium (mycelium) of polyporous fungus. ) to carry out a liquid fermentation step to increase the number and biomass of mycelial balls.
所述「菌絲體」是多細胞真菌經液態醱酵步驟培養所獲得的菌絲之集合。菌絲體的形態多變,可為菌絲自由分散的絲狀菌絲體,或為菌絲高度糾纏的球狀菌絲體(即菌絲球),其中菌絲體的形態會因為基因、細胞壁組成、菌種大小、生長速度、培養基之碳/氮比、pH值、溫度、離子濃度及所受剪力等因素而改變。The "mycelium" is a collection of hyphae obtained by culturing multicellular fungi through a liquid fermentation step. The shape of the mycelium is variable, it can be a filamentous mycelium with freely dispersed mycelium, or a spherical mycelium with a highly entangled mycelium (ie, mycelium ball). Cell wall composition, strain size, growth rate, carbon/nitrogen ratio of the medium, pH, temperature, ion concentration, and shear force.
所述「多孔菌」是指一群擔子柄生長在孔狀子實層的擔子菌。一般而言,多孔菌的子實體質地較硬且較不易腐敗,因此又有硬菇的俗名。依據現代的真菌分類系統,多孔菌包含多孔菌目(Polyporales)及鏽革孔菌目(Hymenochaetales),其中多孔菌目包含靈芝科(Ganoderma)、擬層孔菌科(Fomitopsidaceae)及多孔菌科(Polyporaceae)等。在一實施例中,多孔菌可包含擬層孔菌屬(Fomitopsis)及靈芝屬(Ganoderma)。在一實施例中,多孔菌包含熱帶靈芝(Ganoderma tropicum)、有柄樹舌靈芝(Ganoderma gibbosum)、松生擬層孔菌(Fomitopsis pinicola)或同為靈芝屬或擬層孔菌屬之其他菌株。The "polypore" refers to a group of basidiomycetes with basidiomycetes growing in the pore-shaped fruit layer. Generally speaking, the fruiting body of Polyporus is harder and less prone to spoilage, hence the common name of Hard Mushroom. According to the modern fungal classification system, polyporous fungi include Polyporales and Hymenochaetales, among which Polyporales include Ganoderma, Fomitopsidaceae and Polyporaceae (Fomitopsidaceae). Polyporaceae) and so on. In one embodiment, the polyporous bacteria may include Fomitopsis and Ganoderma. In one embodiment, the polypore comprises Ganoderma tropicum, Ganoderma gibbosum, Fomitopsis pinicola, or other strains of the same genus Ganoderma or Pseudomonas.
所述PDB可例如以習知配方(新鮮馬鈴薯及葡萄糖)配製而成,或是利用市售的PDB粉末加水配製而成。The PDB can be formulated, for example, with a conventional formula (fresh potato and glucose), or by using commercially available PDB powder plus water.
所述「非水解動物性蛋白」是使用動物性蛋白但排除使用酵素水解處理。在一實施例中,非水解動物性蛋白可例如源自於任何鳥類或爬蟲類的蛋黃,如:雞蛋、鴨蛋、鵪鶉蛋或蜥蜴蛋。在一實施例中,上述蛋黃之劑型可例如將蛋經噴霧乾燥處理後之粉末,如蛋黃粉。The "non-hydrolyzed animal protein" uses animal protein but excludes the use of enzymatic hydrolysis. In one embodiment, the non-hydrolyzed animal protein can be derived, for example, from the egg yolk of any bird or reptile, such as egg, duck, quail or lizard eggs. In one embodiment, the above-mentioned dosage form of egg yolk can be, for example, powder after spray-drying the egg, such as egg yolk powder.
經實驗證實,非水解動物性蛋白可有效提高菌絲球數量及生物量,從而提高菌絲體的總產量,其中菌絲球數量及生物量隨非水解動物性蛋白之含量的增加而提高。然而,如果非水解動物性蛋白之含量大於1重量百分比(wt%),則不僅無法提高菌絲球數量及生物量,還可能造成污染。在一實施例中,非水解動物性蛋白於PDB的含量是0.25 wt%。Experiments have confirmed that non-hydrolyzed animal protein can effectively increase the number and biomass of mycelial balls, thereby increasing the total yield of mycelium. The number and biomass of mycelial balls increase with the increase of the content of non-hydrolyzed animal protein. However, if the content of non-hydrolyzed animal protein is greater than 1 weight percent (wt%), not only the number and biomass of mycelial balls cannot be increased, but also pollution may be caused. In one embodiment, the content of non-hydrolyzed animal protein in PDB is 0.25 wt%.
在一實施例中,上述培養液可選擇性包含酵母萃取物。酵母萃取物的使用量不限,可例如0.25 wt%至0.75 wt%。In one embodiment, the above-mentioned culture medium may optionally contain yeast extract. The amount of yeast extract used is not limited, and can be, for example, 0.25 wt% to 0.75 wt%.
在一實施例中,在進行液態醱酵步驟前,可選擇性包含對培養液進行熱不活化(heat-inactivated)處理。熱不活化處理可以任何習知熱不活化之方法進行。在一實施例中,熱不活化處理可例如以100°C至150°C之溫度、1.0 kg/cm 2至1.5 kg/cm 2之壓力進行達10至30分鐘。 In one embodiment, before the liquid fermentation step, it may optionally include heat-inactivating the culture medium. The heat inactivation treatment can be carried out by any known heat inactivation method. In one embodiment, the heat inactivation treatment may be performed, for example, at a temperature of 100°C to 150°C and a pressure of 1.0 kg/cm 2 to 1.5 kg/cm 2 for 10 to 30 minutes.
在一實施例中,液態醱酵步驟的溫度可以習知多孔菌較偏好生長的溫度進行,如:23°C至29°C。在一實施例中,液態醱酵步驟之轉速為100 rpm至150 rpm,以使菌絲體形成菌絲球。In one embodiment, the temperature of the liquid fermentation step can be performed at a temperature at which polypore bacteria prefer to grow, such as 23°C to 29°C. In one embodiment, the rotational speed of the liquid fermentation step is 100 rpm to 150 rpm, so that the mycelium can form a mycelial ball.
以下利用數個實施例以說明本發明之應用,然其並非用以限定本發明,本發明技術領域中具有通常知識者,在不脫離本發明之精神和範圍內,當可作各種之更動與潤飾。Several embodiments are used below to illustrate the application of the present invention, but they are not intended to limit the present invention. Those with ordinary knowledge in the technical field of the present invention can make various changes and modifications without departing from the spirit and scope of the present invention. retouch.
實施例一、分析不同成分對菌絲球數量的影響
利用田口分析法分析馬鈴薯粉[製造商:美國百信(Basic American Foods);品名:美味快熟馬鈴薯粉]、葡萄糖、酵母萃取物及蛋黃粉分別對菌絲球數量的影響效果,在此以相對貢獻百分比(relative percentage contribution)評估。首先,利用表1之配方分別加水並進行熱不活化步驟(溫度:121°C、壓力:1.2kg/cm
2、時間:25分鐘),以配製組別1至9的培養液。接著,在25°C下以130 rpm之轉速對熱帶靈芝(Ganoderma tropicum)之菌絲體(直徑為0.8 cm之菌塊,其中菌塊是利用PDA培養瓊脂於25°C下培養後獲得)進行液態醱酵步驟達7天,並計算菌絲球之數量。上述熱帶靈芝可為採集自臺灣屏東大武山[基因銀行(GeneBank)編號為MK007291.1]或同為熱帶靈芝之其他菌株,例如寄存於生物資源保存及研究中心(Bioresourse Collection and Research Center,BCRC)中寄存編號BCRC 35346、BCRC 36209、BCRC 37032、BCRC 37041、BCRC 37065、BCRC 37067或BCRC 37122之菌株。將結果列於表1中。上述蛋黃粉是新鮮的雞蛋蛋黃經噴霧乾燥處理形成。
Taguchi analysis method was used to analyze the effect of potato flour [manufacturer: Basic American Foods; product name: delicious quick-cooking potato flour], glucose, yeast extract and egg yolk powder on the number of mycelial balls respectively. Relative percentage contribution assessment. First, using the formula in Table 1, water was added and a heat inactivation step (temperature: 121°C, pressure: 1.2 kg/cm 2 , time: 25 minutes) was respectively added to prepare the culture solutions of
表1 Table 1
然後,計算不同成分之貢獻度,並將結果列於圖1。圖1係顯示根據本發明一實施例之不同成分對菌絲球數量變化之相對貢獻度的長條圖,其中橫軸表示成分,縱軸表示以所有成分之貢獻度之總和為100%,評估各成分之相對貢獻度。如圖1所示,蛋黃粉的相對貢獻度最高,表示蛋黃粉是上述四個成分中,對菌絲球數量多寡影響效果較佳的成分。Then, the contribution of different components is calculated, and the results are shown in Figure 1. 1 is a bar graph showing the relative contribution of different components to the change in the number of mycelial balls according to an embodiment of the present invention, wherein the horizontal axis represents the components, and the vertical axis represents the sum of the contributions of all components as 100%. The relative contribution of each component. As shown in Figure 1, the relative contribution of egg yolk powder is the highest, indicating that egg yolk powder is the component that has a better effect on the number of mycelial balls among the above four components.
實施例二、評估蛋黃粉含量對靈芝菌絲球數量的影響Embodiment 2. Evaluate the influence of egg yolk powder content on the number of Ganoderma lucidum mycelial balls
利用市售之馬鈴薯右旋糖培養液(potato dextrose broth,PDB)粉末加水配製1倍PDB。接著,在PDB中加入蛋黃粉,以分別配製含有0.25 wt%、0.5 wt%、0.75 wt%及1 wt%蛋黃粉之培養液,並進行熱不活化步驟,其中蛋黃粉經熱不活化步驟後會懸浮於培養液。利用100 mL培養液在250 mL錐形瓶對上述熱帶靈芝之菌絲體進行上述液態醱酵步驟,以產生菌絲球。將液態醱酵步驟後培養液倒入培養皿中,如圖2A至圖2E所示。如圖2A至圖2E係顯示根據本發明一實施例之以含有0 wt%(圖2A)、0.25 wt%(圖2B)、0.5 wt%(圖2C)、0.75 wt%(圖2D)及1 wt%(圖2E)蛋黃粉之培養液進行液態醱酵步驟之結果的照片。如圖2A至圖2E所示,菌絲球數量隨著蛋黃粉含量的增加而增加。Use commercially available potato dextrose broth (potato dextrose broth, PDB) powder to add water to prepare 1-fold PDB. Next, egg yolk powder was added to the PDB to prepare a culture solution containing 0.25 wt %, 0.5 wt %, 0.75 wt % and 1 wt % egg yolk powder respectively, and a heat inactivation step was performed, wherein the egg yolk powder was subjected to the heat inactivation step. suspended in the culture medium. The above-mentioned liquid fermentation step was performed on the mycelium of Ganoderma lucidum in a 250 mL conical flask using 100 mL of culture medium to produce mycelial balls. Pour the culture solution after the liquid fermentation step into a petri dish, as shown in Figure 2A to Figure 2E. FIG. 2A to FIG. 2E show that according to an embodiment of the present invention, it contains 0 wt % ( FIG. 2A ), 0.25 wt % ( FIG. 2B ), 0.5 wt % ( FIG. 2C ), 0.75 wt % ( FIG. 2D ) and 1 wt % Photographs of the results of the liquid fermentation step with wt% (FIG. 2E) egg yolk powder broth. As shown in Fig. 2A to Fig. 2E, the number of mycelial balls increased with the increase of egg yolk powder content.
接著,計算每單位體積之菌絲球數量,並將結果列於圖3中。圖3係顯示根據本發明一實施例之蛋黃粉含量對菌絲球數的長條圖,其中橫軸是蛋黃粉含量,縱軸是每單位體積之菌絲球數量,且不同字母(a、b及c)表示利用最小顯著差異法(least-significant difference,LSD)進行統計分析後各組間具有顯著差異(P<0.05)。如圖3所示,相較於未添加蛋黃粉之PDB,含有蛋黃粉之培養液所獲得之每單位體積之菌絲球數量顯著較高。此外,每單位體積之菌絲球數量隨著蛋白粉含量的增加而上升,顯示蛋白粉對菌絲球數具有劑量依賴效果(dose dependent effect)。Next, the number of mycelial balls per unit volume was calculated and the results are shown in FIG. 3 . 3 is a bar graph showing the content of egg yolk powder versus the number of mycelial balls according to an embodiment of the present invention, wherein the horizontal axis is the content of egg yolk powder, the vertical axis is the number of mycelial balls per unit volume, and different letters (a, b and c) indicate that there are significant differences between groups (P<0.05) after statistical analysis using the least-significant difference (LSD) method. As shown in FIG. 3 , the number of mycelial balls per unit volume obtained from the broth containing egg yolk powder was significantly higher than that of PDB without the addition of egg yolk powder. In addition, the number of mycelial balls per unit volume increased with the increase of protein powder content, indicating that protein powder had a dose dependent effect on the number of mycelial balls.
然後,利用市售軟體IMAGE J分析菌絲球之球徑,並將結果列於圖4中。圖4係顯示根據本發明一實施例之蛋黃粉含量對菌絲球球徑的長條圖,其中橫軸表示蛋黃粉含量,縱軸表示球徑,且相同字母(a)表示利用最小顯著差異法進行統計分析後不具有統計上的顯著差異(P<0.05)。如圖4所示,不同含量之蛋黃粉所培養出的菌絲球在球徑上不具有統計上的顯著差異,其中球徑為0.33 cm至0.36 cm。詳細而言,菌絲球數量隨著蛋黃粉含量增加而上升,而當菌絲球數量少的時候,菌絲球球徑的差異大(變異數大),但當菌絲球數量多的時候,菌絲球球徑差異變小(變異數小)。Then, the spherical diameter of the mycelial balls was analyzed using the commercially available software IMAGE J, and the results are shown in FIG. 4 . 4 is a bar graph showing the content of egg yolk powder versus the diameter of mycelial balls according to an embodiment of the present invention, wherein the horizontal axis represents the content of egg yolk powder, the vertical axis represents the diameter of the ball, and the same letter (a) represents the use of the least significant difference There was no statistically significant difference (P<0.05) after statistical analysis. As shown in Figure 4, the mycelial balls cultured with different contents of egg yolk powder have no statistically significant difference in ball diameter, wherein the ball diameter is 0.33 cm to 0.36 cm. In detail, the number of mycelial balls increased with the increase of egg yolk powder content, and when the number of mycelial balls was small, the difference in the diameter of the mycelial balls was large (large number of variation), but when the number of mycelial balls was large , the difference in the diameter of the mycelial ball becomes smaller (the number of variation is small).
將菌絲球烘乾並測量其乾重,以評估蛋黃粉含量對生物量之影響,並將結果列於圖5中。圖5係顯示根據本發明一實施例之蛋黃粉含量對生物量的長條圖,其中橫軸是蛋黃粉含量,縱軸是每單位體積之生物量,且不同英文字母表示利用最小顯著差異法進行統計分析後各組間具有顯著差異(P<0.05)。如圖5所示,相較於未添加蛋黃粉之PDB,添加0.25 wt%蛋黃粉之培養液所培養之菌絲球的生物量顯著較高,顯示蛋黃粉可有效增加生物量。其次,生物量隨著蛋黃粉含量的增加而上升,顯示生物量與蛋黃粉具有劑量依賴效果。Mycelium balls were dried and their dry weight was measured to evaluate the effect of egg yolk powder content on biomass, and the results are shown in Figure 5. 5 is a bar graph showing the content of egg yolk powder versus biomass according to an embodiment of the present invention, wherein the horizontal axis is the egg yolk powder content, the vertical axis is the biomass per unit volume, and different English letters indicate the use of the least significant difference method After statistical analysis, there were significant differences among the groups (P<0.05). As shown in Figure 5, compared with the PDB without egg yolk powder, the biomass of mycelial balls cultured in the culture solution with 0.25 wt% egg yolk powder was significantly higher, indicating that egg yolk powder can effectively increase the biomass. Secondly, the biomass increased with the increase of egg yolk powder content, showing a dose-dependent effect of biomass and egg yolk powder.
以下利用具有特定含量之蛋黃粉的培養液,評估其對於提升其他多孔菌菌絲球數量的影響。The following uses a culture medium with a specific content of egg yolk powder to evaluate its effect on increasing the number of other polyporous fungus mycelial balls.
實施例三、評估實施例二之培養液對多孔菌菌絲球數量的影響Example 3. Evaluation of the influence of the culture solution of Example 2 on the number of polyporous fungus mycelial balls
分別配製含或不含0.25 wt%蛋黃粉之培養液,以對表2所列之多孔菌之菌絲體進行上述液態醱酵步驟。表2顯示菌種的學名、18S rRNA序列對應的基因銀行(GeneBank)編號及培養天數,其中菌種1至菌種3係採自於臺灣屏東大武山,分別為有柄樹舌靈芝(Ganoderma gibbosum)、靈芝屬菌種(Ganoderma sp.)及松生擬層孔菌(Fomitopsis pinicola),但菌種1至菌種3也可以是相同菌種的其他菌株,舉例而言,菌種1可為寄存編號為BCRC 36441之有柄樹舌靈芝、菌種2可為寄存編號為BCRC 37046或BCRC 37047之菌株,且菌種3可為BCRC 32527、BCRC 35303或公告於2007年11月28日的韓國專利KR 100778941 B1所述之松生擬層孔菌,其寄存機構為韓國農業遺傳資源保藏中心(Korean Agricultural Culture Collection,KACC),且寄存編號為KACC 93042 P。A culture solution containing or not containing 0.25 wt% egg yolk powder was prepared, respectively, to carry out the above-mentioned liquid fermentation step on the mycelia of the polypore fungi listed in Table 2. Table 2 shows the scientific name of the bacterial species, the corresponding GeneBank number of the 18S rRNA sequence and the number of days of cultivation, wherein the
表2 Table 2
進行液態醱酵步驟,再計算菌絲球數量,並將結果列於圖6中。圖6係根據本發明之一實施例之不同菌種以含或不含蛋黃粉之培養液培養之菌絲球數量的直條圖,其中橫軸表示菌種,縱軸表示每單位體積菌絲球數量,且直條601及603分別表示不添加及添加蛋黃粉。如圖6所示,相較於不添加蛋黃粉之PDB,添加蛋黃粉之培養液所培養之菌絲球數量較多,顯示蛋黃粉可增加有孔菌之菌絲球數量。The liquid fermentation step was carried out, and the number of mycelial balls was counted, and the results were listed in Figure 6. 6 is a bar graph of the number of mycelial balls cultured with different bacterial species in a culture solution with or without egg yolk powder according to an embodiment of the present invention, wherein the horizontal axis represents bacterial species, and the vertical axis represents mycelium per unit volume number of balls, and bars 601 and 603 indicate no and added egg yolk powder, respectively. As shown in FIG. 6 , compared with the PDB without egg yolk powder, the number of mycelial balls cultured in the culture solution with egg yolk powder was higher, indicating that egg yolk powder can increase the number of mycelial balls of foramen.
由上述實施例可知,應用本發明之培養菌絲球的方法,可藉由增加有孔菌之菌絲球數量及生物量來提高菌絲體之產量。It can be seen from the above examples that the method for culturing mycelial balls of the present invention can increase the yield of mycelium by increasing the number and biomass of mycelial balls of foraminifera.
需補充的是,本發明雖以特定的菌種、特定的菌株、特定的濃度及/或特定的分析方法做為例示,說明本發明之培養菌絲球的方法在提升菌絲球及生物量之功效,惟本發明所屬技術領域中任何具有通常知識者可知,本發明並不限於此,在不脫離本發明之精神和範圍內,本發明之培養菌絲球的方法及培養液亦可使用其他的菌種、其他的菌株、其他的濃度及/或其他的分析方法說明功效。It should be added that although the present invention takes a specific bacterial species, a specific strain, a specific concentration and/or a specific analysis method as an example, it is explained that the method for culturing the mycelial ball of the present invention can improve the mycelial ball and the biomass. However, anyone with ordinary knowledge in the technical field to which the present invention belongs will know that the present invention is not limited to this. Other species, other strains, other concentrations, and/or other analytical methods illustrate efficacy.
雖然本發明已以數個實施例揭露如上,然其並非用以限定本發明,在本發明所屬技術領域中任何具有通常知識者,在不脫離本發明之精神和範圍內,當可作各種之更動與潤飾,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 present invention has been disclosed above with several embodiments, it is not intended to limit the present invention. Anyone with ordinary knowledge in the technical field to which the present invention belongs, without departing from the spirit and scope of the present invention, can make various Therefore, the scope of protection of the present invention should be determined by the scope of the appended patent application.
601,603:直條601,603: Straight
為讓本發明之上述和其他目的、特徵、優點與實施例能更明顯易懂,所附圖式之詳細說明如下: [圖1]係顯示根據本發明一實施例之不同成分對菌絲球數量變化之相對貢獻度的長條圖。 [圖2A]至[圖2E]係顯示根據本發明一實施例之以含有0 wt%、0.25 wt%、0.5 wt%、0.75 wt%及1 wt%蛋黃粉之培養液進行液態醱酵步驟之結果的照片。 [圖3]係顯示根據本發明一實施例之蛋黃粉含量對菌絲球數的長條圖。 [圖4]係顯示根據本發明一實施例之蛋黃粉含量對菌絲球球徑的長條圖。 [圖5]係顯示根據本發明一實施例之蛋黃粉含量對生物量的長條圖。 [圖6]係根據本發明之一實施例之不同菌種以含或不含蛋黃粉之培養液培養之菌絲球數量的直條圖。 In order to make the above and other objects, features, advantages and embodiments of the present invention more clearly understood, the detailed description of the accompanying drawings is as follows: [ Fig. 1 ] is a bar graph showing the relative contribution of different components to the change in the number of mycelial balls according to an embodiment of the present invention. [FIG. 2A] to [FIG. 2E] show the process of liquid fermenting step with the culture solution containing 0 wt%, 0.25 wt%, 0.5 wt%, 0.75 wt% and 1 wt% egg yolk powder according to an embodiment of the present invention Photo of the result. [ Fig. 3 ] is a bar graph showing the content of egg yolk powder versus the number of mycelial balls according to an embodiment of the present invention. [ Fig. 4 ] is a bar graph showing the content of egg yolk powder versus the diameter of mycelial balls according to an embodiment of the present invention. [ Fig. 5 ] is a bar graph showing the content of egg yolk powder versus biomass according to an embodiment of the present invention. [ Fig. 6 ] is a bar graph of the number of mycelial balls cultured by different strains in a culture solution with or without egg yolk powder according to an embodiment of the present invention.
國內寄存資訊(請依寄存機構、日期、號碼順序註記) 無 國外寄存資訊(請依寄存國家、機構、日期、號碼順序註記) 無 Domestic storage information (please note in the order of storage institution, date and number) none Foreign deposit information (please note in the order of deposit country, institution, date and number) none
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