TW201439317A - Taiwanofungus camphoratus fungus culturing method and culturing substrate ingredient - Google Patents

Taiwanofungus camphoratus fungus culturing method and culturing substrate ingredient Download PDF

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TW201439317A
TW201439317A TW102112867A TW102112867A TW201439317A TW 201439317 A TW201439317 A TW 201439317A TW 102112867 A TW102112867 A TW 102112867A TW 102112867 A TW102112867 A TW 102112867A TW 201439317 A TW201439317 A TW 201439317A
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TWI577797B (en
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Shi-Min Chen
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Cheng Lin Agricultural Biotechnology Co Ltd
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Abstract

This invention relates to a Taiwanofungus camphoratus fungus culturing method and a culturing substrate ingredient, characterized by comprising the following steps: 1) a primary fungus culturing step for transferring wild Taiwanofungus camphoratus fungus to a culture plate containing culturing substrate; 2) a transfer-culturing step for transferring the wild Taiwanofungus camphoratus fungus to a culturing container greater than the culture plate for culturing, wherein the culturing container contains culturing solution at this stage; 3) a transform-culturing step for adding aquilaria chips to the culturing container when the mycelia of the Taiwanofungus camphoratus fungus grow richer in the culturing container so as to enable the mycelia to fully absorb triterpene compound ingredients in the aquilaria during growth.

Description

牛樟芝菌體培養方法及培養基底成份 Bacillus subtilis cell culture method and medium bottom composition

本發明係為一種牛樟芝菌體培養方法及培養基底成份,尤指一種使用培養容器培養牛樟芝菌體之培養方法與培養基底成份。 The invention relates to a method for cultivating the cells of the genus Antrodia camphorata and a bottom component of the medium, in particular to a culture method for cultivating A. sinensis cells and a bottom component of the culture medium.

牛樟樹是台灣特有植物,受到極度的重視,並列為保育類植物,而牛樟芝菌(Antrodia comphorata)又只長於牛樟樹,其他樹木無法長出牛樟芝,牛樟芝對抑制癌症、預防癌症的轉移、B型肝炎、糖尿病、高血壓等疾病具有功效,並在抗氧化、抗血液中凝血、及腸胃疾病上也有相當的功效,牛樟芝的成份複雜,其中重要成份包括有:三萜類化合物(Triterpenoids)、超氧歧化酵素(SOD:Superoxide Dismutase)、腺甘(Adenosine)、β-D-葡聚醣(β-D-glucan)等營養成份;其中三萜類化合物是牛樟芝最重要的化學成份之一,也是牛樟芝之萃取物中苦味成份的來源。以目前的研究結果得知,在每種靈芝內的三萜類含量約1~3%,而單牛樟芝一種就擁有15~45%的三萜類含量,三萜類的主要功能有抑制癌細胞的生長、抑制組織胺的釋放,防止過敏、促進肝功能、促進血小板凝集、以及降血脂等作用,因此三萜類的含量及種類愈多,則愈有醫療價值,實驗證實:三萜類化合物能有效的抑制血管收縮素轉化脢酵素(ACE:Angiotensin Converting Enzyme)的活性,進而降低血壓,另外三萜類也具有抗發炎之功用。 Burdock is a unique plant in Taiwan, which is highly valued and listed as a conservation plant. Antrodia comphorata is only longer than burdock. Other trees cannot grow Antrodia camphorata. The inhibition of cancer and prevention of cancer by Antrodia camphorata, type B Hepatitis, diabetes, high blood pressure and other diseases have efficacy, and also have considerable effects in anti-oxidation, anti-blood coagulation, and gastrointestinal diseases. The composition of Antrodia camphorata is complex, among which important ingredients include: Triterpenoids, Super Nutrient components such as SOD (Superoxide Dismutase), Adenosine (Adenosine), and β-D-Glucan (β-D-glucan); among them, triterpenoids are one of the most important chemical constituents of Antrodia camphorata. The source of bitter ingredients in the extract of Antrodia camphorata. According to the current research results, the content of triterpenoids in each ganoderma lucidum is about 1 to 3%, while that of monoterpenoids has 15 to 45% triterpenoids. The main function of triterpenoids is to inhibit cancer cells. Growth, inhibition of histamine release, prevention of allergies, promotion of liver function, promotion of platelet aggregation, and blood lipid lowering, so the more the content and variety of triterpenoids, the more medical value, experimental confirmation: triterpenoids It can effectively inhibit the activity of angiotensin-converting enzyme (ACE: Angiotensin Converting Enzyme), thereby lowering blood pressure, and the other triterpenoids also have anti-inflammatory effects.

但是牛樟芝菌在生長上為實體屬多年生,具有強烈樟樹香味,外型呈板狀或鐘狀,新鮮者表面呈鮮紅橘色,成熟後顏色呈桔色或黃色,菌蓋半圓形,牛樟芝只寄生在野生牛樟樹洞中,牛樟樹為台灣特有的木,生長於海拔450~2000公尺的山區,近年來因為大幅採發後牛樟木已顯得非常稀有,列為生態保護的樹種;過去以人工培養牛樟芝菌的方式為在加工切塊後的牛樟樹段木上植菌,並透過環境控制使其長成後再取出成熟的牛樟芝菌進行加工製作成營養食品;然而在市場大量需求狀況下,業者必須大量培養牛樟芝菌,但是在牛樟樹段木取得困難且成本過高的狀況下,遂有業者開發出固態培養法及液態培養法兩種主要技術;其中在固態培養法為將牛樟芝菌種以包裝容器進行菌絲體培養,內含有纖維物、醣類、五穀雜糧類等培養基底,促使該牛樟芝菌能獲得與野生牛樟芝相近的成份,培養時間約三個月以上,其成份中之多糖體與三帖類會因培養基的內容物而改變;另一種液態培養法,其技術手段為在液體發酵容器中進行菌種液體發酵以收取菌絲體,培養時間較短、成本較低,但同時液態培養法製造出來的牛樟芝菌絲體之營養成份相對上較低。 However, the growth of A. faecalis is a perennial growth of the genus. It has a strong eucalyptus scent, and its appearance is plate-shaped or bell-shaped. The surface of the fresh person is bright red orange. The color is orange or yellow after ripening. The cover is semi-circular. Parasitic in the wild burdock tree hole, burdock tree is a unique wood in Taiwan, growing in the mountainous area at 450~2000 meters above sea level. In recent years, because of the large-scale harvesting, the burdock tree has become very rare and classified as an ecologically protected tree species; The method of cultivating Antrodia camphorata is to plant bacteria on the burdock tree after processing and dicing, and to grow into a nutritious food by processing the mature burdock bacillus through environmental control; however, under the market demand, The industry must cultivate a large number of burdock bacillus, but under the condition that the burdock tree is difficult and the cost is too high, the cockroach has developed two main technologies: solid culture method and liquid culture method; The mycelial culture is carried out in a packaging container, which contains a medium such as a fibrous substance, a saccharide or a whole grain, and the bacillus is obtained. The similar composition of wild Antrodia camphorata, the culture time is about three months, the polysaccharide and the three types of the ingredients will change due to the content of the medium; the other liquid culture method, the technical means is to carry on the bacteria in the liquid fermentation container The liquid fermentation is to take the mycelium, the cultivation time is shorter, and the cost is lower, but at the same time, the nutrient composition of the mycelium of Antrodia camphorata produced by the liquid culture method is relatively low.

前述之牛樟芝菌取代培養方式,其雖然可以減低對牛樟樹段木的依賴及需要,但是轉植後卻因為牛樟芝菌後續成長中缺乏牛樟木所含有三帖類成份的吸收,反致使轉植後的牛樟芝菌所含的三萜類化合物等成份比率降低或者根本不存在。 The above-mentioned method of substituting the culture of Antrodia camphorata can reduce the dependence on the burdock tree and the need, but after the transplant, the lack of absorption of the three kinds of ingredients in the subsequent growth of the burdock bacillus is reversed. The ratio of components such as triterpenoids contained in Antrodia camphorata is reduced or does not exist at all.

本發明係為一種牛樟芝菌體培養方法及培養基底成份,其培養方法主要特徵在於包括有以下步驟:1)初菌培養步驟:將野生牛樟芝菌 移植到培養皿中,其中該培養皿置放有培養基底;2)轉植培養步驟:至將野生牛樟芝菌轉植到容積較培養皿更大的培養容器中培養,此一階段培養容器內附有培養液;3)轉化培養步驟:當牛樟芝菌體之菌絲於培養容器中至更飽滿時,於培養容器中加入沉香木屑驅使菌絲在成長過程中,充分吸收沉香之三萜類化合物成份。 The invention relates to a method for cultivating the cells of the burdock and the bottom of the culture medium, and the main method of the cultivating method comprises the following steps: 1) the initial culture step: the wild bacterium Transplanted into a culture dish, wherein the culture dish is placed with a medium bottom; 2) a planting culture step: to transplant the wild A. sinensis to a larger culture vessel having a larger volume than the culture dish, and the first stage culture vessel is attached There is a culture solution; 3) transformation culture step: when the mycelium of A. sinensis is more fully filled in the culture container, the agarwood chips are added to the culture container to drive the hyphae during the growth process, and the agaric compound is fully absorbed. .

本發明之培養基底成份特徵在於:一種牛樟芝菌體培養基底成份,其使用於牛樟芝菌轉植培養容器內,其成份包含培養液及增長基底,其特徵在於該增長基底包含有三萜類化合物成份;其中該增長基底為沉香木屑。 The bottom component of the present invention is characterized in that: a bottom component of the Astragalus membranaceus cell culture medium, which is used in a B. gingivalis culture container, the composition comprising a culture liquid and a growth substrate, wherein the growth substrate comprises a triterpenoid component; Wherein the growth substrate is agarwood chips.

圖1:係本發明之實施步驟流程圖。 Figure 1 is a flow chart showing the steps of the implementation of the present invention.

本發明係為一種牛樟芝菌體培養方法及培養基底成份,尤可藉由特殊培養基底的選用,使牛樟芝菌體更富含有更多的三萜類化合物(Triterpenoids),主要包括有以下步驟: The invention relates to a method for cultivating the cells of the Antrodia camphorata and the bottom of the culture medium, and particularly, the selection of the special medium bottom makes the Antrodia camphorata more abundant in Triterpenoids, mainly including the following steps:

1.初菌培養步驟:在進行培養過程前,先採集野生牛樟芝菌,再將野生牛樟芝菌移植到培養皿中,其中該培養皿置放有培養基底,該些培養基底包括有:酵母萃取物、葡萄糖、蒟蒻粉、糖蜜、馬鈴薯萃取物,其中該酵母萃取物性質上為提供野生牛樟芝菌繁衍的營養主劑使用,而蒟蒻粉提供凝聚結合而提供增加野生牛樟芝菌附著生長的功效,搭配葡萄糖、糖蜜、馬鈴薯萃取物等物質,提供野生牛樟芝菌初期生長時吸收的養分,並增加富含的養分。 1. Initial bacterial culture step: Before carrying out the culture process, the wild A. faecalis is collected, and then the wild A. faecalis is transplanted into a culture dish, wherein the culture dish is placed with a culture bottom, and the bottom of the medium includes: yeast extract , glucose, glutinous powder, molasses, potato extract, wherein the yeast extract is used to provide a nutrient main agent for the propagation of wild A. angustifolia, and the glutinous rice provides agglomeration and provides the effect of increasing the growth and growth of wild A. sinensis, with glucose , molasses, potato extract and other substances, provide nutrients absorbed by the wild growth of A. sinensis, and increase the nutrient rich.

2.轉植培養步驟:牛樟芝菌植入培養皿後成長面積至培養皿70%~80%面積時,將牛樟芝菌轉植到容積較培養皿更大的培養容器中培養,此一階段培養容器內附有培養液,該培養液之成份包括有:酵母萃取物、葡萄糖、糖蜜、馬鈴薯萃取物及牛樟樹木屑,該牛樟木屑可使牛樟芝菌體之菌絲成長更佳。 2. Replanting culture step: When the growth area of the burdock bacillus is 70%~80% in the culture dish, the A. faecalis is transplanted into a larger culture container than the culture dish. The first stage culture container A culture solution is provided, and the composition of the culture solution includes: yeast extract, glucose, molasses, potato extract and burdock wood chips, and the burdock wood can make the hyphae of the burdock bacillus fungus grow better.

3.轉化培養步驟:當牛樟芝菌體之菌絲於培養容器中至更飽滿時,於培養容器中加入一增長基底成份及五穀類為本發明重要的技術特徵,在本實施例中該增長基底為沉香木屑,主要在於沉香木屑含有三萜類化合物(Hydroxyhopanone).透過此一培養技術可驅使菌絲在成長成牛樟芝成菌過程中,充分吸收沉香之三萜類化合物成份,促使其更具更高比例的三萜類化合物成份,使之經濟效益更高,而沉香木屑中更富含有沉香醇(Linalool)之香料成份,更可促使牛樟芝菌在此一程序培養後散發出獨特的香氣,使其在採集後製作成營養食品時添加氣味,增進使用者在食用時的味覺效果;在此步驟的加入三萜類化合物成份的增長基底不以沉香木屑為限,其他含有三萜類化合物成份之植物成份皆屬可應用之範疇。 3. Transformation culture step: when the hyphae of the burdock bacillus is more fully filled in the culture vessel, adding a growth base component and grains to the culture vessel is an important technical feature of the invention, and in the present embodiment, the growth substrate For agarwood sawdust, mainly because the agarwood sawdust contains a trioxon compound (Hydroxyhopanone). Through this culture technique, the hyphae can be driven into the process of growing into a burdock, and fully absorb the astringent compound of the agarwood, thereby promoting a higher proportion of the triterpenoid compound, so that the economic benefit is higher, and The agarwood chips are richer in the fragrance ingredients of Linalool, which can also promote the unique aroma of the burdock bacillus in this program, so that it can add odor when it is made into nutritious food after collection, and enhance the user's The taste effect when eating; the growth substrate of the triterpenoids added in this step is not limited to agarwood chips, and other plant ingredients containing triterpenoids are applicable.

自前述步驟中可知,本發明主要的技術重點在於本發明透過特殊培養步驟及培養基底的組成,使原本轉植培養導致牛樟芝菌內含三萜類化合物等有效成份偏低的狀況獲得較佳的改善,在不大量使用牛樟樹段木又可獲得更高營養成份比率的功效下,對於環境保護改善及促進人體健康兩個需求上獲得相輔相成的效果。 It can be seen from the foregoing steps that the main technical focus of the present invention is that the present invention achieves better conditions such as the low active ingredient content of the triterpenoids in the burdock bacteria by the special culture step and the composition of the bottom of the culture medium. Improvement, in the absence of a large number of use of burdock section wood and the ability to obtain higher nutrient ratios, the two requirements for environmental protection and human health have achieved complementary effects.

Claims (9)

一種牛樟芝菌體培養方法,其包括有以下步驟:初菌培養步驟:將野生牛樟芝菌移植到培養皿中,其中該培養皿置放有培養基底;轉植培養步驟:野生牛樟芝菌移植到培養皿中培養至預設面積比率後,將牛樟芝菌轉植到容積較培養皿更大的培養容器中培養,此一階段培養容器內附有培養液;轉化培養步驟:當牛樟芝菌體之菌絲於培養容器中至更飽滿時,於培養容器中加入包含有三萜類化合物成份之增長基底,驅使菌絲在成長過程中吸收增長基底之三萜類化合物成份。 The invention relates to a method for cultivating a strain of Antrodia camphorata, which comprises the following steps: a step of cultivating the initial bacteria: transplanting the wild A. faecalis into a culture dish, wherein the culture dish is placed with a medium bottom; and the step of cultivating the culture: transplanting the wild A. faecalis to the culture dish After culturing to a preset area ratio, the A. faecalis is transplanted into a culture vessel having a larger volume than the culture dish, and the culture medium is attached to the culture vessel in the first stage; the transformation culture step: when the mycelium of the burdock bacillus When the culture vessel is fuller, a growth substrate containing a triterpenoid component is added to the culture vessel to drive the hyphae to absorb the growth of the triterpenoid component of the substrate during growth. 如申請專利範圍第1項所述之牛樟芝菌體培養方法,其中該野生牛樟芝菌移植到培養皿中培養之預設面積比率為培養面積至培養皿70%~80%面積。 The method for cultivating the Astragalus sinensis cell as described in claim 1, wherein the ratio of the predetermined area of the wild A. faecalis strain cultured to the culture dish is from 70% to 80% of the culture area to the culture dish. 如申請專利範圍第1項所述之牛樟芝菌體培養方法,其中該初菌培養步驟之培養基底包括有:酵母萃取物、葡萄糖、蒟蒻粉、糖蜜、馬鈴薯萃取物,該蒟蒻粉提供凝聚結合而提供增加野生牛樟芝菌附著生長的功效。 The method for cultivating an Antrodia camphorata cell as described in claim 1, wherein the bottom of the culture step of the initial bacterial culture step comprises: a yeast extract, glucose, glutinous rice flour, molasses, and potato extract, wherein the glutinous rice powder provides agglomeration and binding. Provides the effect of increasing the growth and growth of wild A. serrata. 如申請專利範圍第1項所述之牛樟芝菌體培養方法,其中該增長基底為沉香木屑。 The method for cultivating an Antrodia camphorata cell as described in claim 1, wherein the growth substrate is agarwood sawdust. 如申請專利範圍第1項所述之牛樟芝菌體培養方法,其中該轉植培養步驟之培養液成份包括有:酵母萃取物、葡萄糖、糖蜜、馬鈴薯萃取物及牛樟樹木屑。 The method for cultivating an Antrodia camphorata cell as described in claim 1, wherein the composition of the culture solution of the transplanting culture step comprises: yeast extract, glucose, molasses, potato extract, and burdock shavings. 如申請專利範圍第1項所述之牛樟芝菌體培養方法及培養基底成份,其中 該轉化培養步驟於培養容器中加入五穀類。 For example, the method for cultivating the Astragalus membranaceus cells and the composition of the culture medium according to Item 1 of the patent application scope, wherein The transformation culture step adds grains to the culture vessel. 一種牛樟芝菌體培養基底成份,其使用於牛樟芝菌轉值培養容器內,其成份包含培養液及增長基底,其特徵在於該增長基底包含有三萜類化合物成份。 The base component of the Astragalus membranaceus cell culture medium is used in a transformation culture container of Antrodia camphorata, and the composition thereof comprises a culture liquid and a growth substrate, wherein the growth substrate comprises a triterpenoid compound component. 如申請專利範圍第7項所述之牛樟芝菌體培養基底成份,其中該增長基底為沉香木屑。 The bottom material of the Astragalus membranaceus cell culture medium according to claim 7, wherein the growth substrate is agarwood wood chips. 如申請專利範圍第7項所述之牛樟芝菌體培養基底成份,其中該培養液成份包括有:酵母萃取物、葡萄糖、糖蜜、馬鈴薯萃取物及牛樟樹木屑。 The bottom material of the Astragalus bacillus cell culture medium according to claim 7, wherein the culture liquid component comprises: yeast extract, glucose, molasses, potato extract and burdock wood chips.
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CN108377839A (en) * 2018-04-09 2018-08-10 张国清 A kind of breeding method of Antrodia camphorata

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TWI422680B (en) * 2010-12-31 2014-01-11 Oasis Bio Tech Co Ltd Culture medium for culturing fruiting bodies of antrodia cinnamomea and method for culturing the same
CN102488719A (en) * 2011-12-25 2012-06-13 南京农业大学 Method for improving triterpene output of Ganoderma lucidum liquid fermented mycelia

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CN104371931A (en) * 2014-10-20 2015-02-25 中山安荞生物科技有限公司 Taiwanofungus camphorates culture medium and culturing method
CN108377839A (en) * 2018-04-09 2018-08-10 张国清 A kind of breeding method of Antrodia camphorata

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