200825177 九、發明說明： 【發明所屬之技術領域】 本發明係有關於一種桑黃與中草藥共發酵製成方法， 桑黃利用液態培養基及配合最佳產量控制條件下與中草藥 單方或複方共發酵，比桑黃單獨發酵產生較佳菌絲及多醣 體產量及提升抗癌之效果，實為一充分符合產業利用性之 製造方法。 Φ 【先前技術】 \桑黃爲裂蹄木層孔菌，學名Phllinus linteus，是寄生 於活桑樹上之真菌類，呈黑色瘤狀，質地堅硬，内铡成鮮 艷之黃色，所以稱之爲桑黃，於明代李時珍的「本草綱目 」即有S己載，桑黃性寒、味微苦、能利五臟、排毒氣、止 血等’在韓國及日本民間使用上則有治腹痛、利尿、抗腫 j、健胃、止瀉等功能，根據韓日的研究結果指出，從桑 黃分離出的多醣體，能明顯刺激T細胞的免疫反應，分析 •桑黃多醣體抵抗腫瘤生長及轉移的活性，是透過活化身體 之免疫機制系統，雖然有自日本引進桑黃萃取物，但價格 叩貴’且功效及純度多不明確，故欲利用桑黃於液態培養 基亚配合最佳之碳源、氮源、無機鹽類、p H值、轉速條 件下發酵’讀升桑黃之_絲及多體產量，並於桑黃最 佳條件下與中草藥單方或複方共發酵，以增加菌絲及多醣 體產量，並提升抗癌之效果。 有鑑於上述之缺失弊端，本發明人認為其有急待改進 之必要，乃思及改良創作之意念，及其一貫秉持具有之優 5 200825177 良。又,十理心，針對既有之缺失加以改良，經多方設計探討 亚夕-人修正改良，終乃有本發明桑黃與中草藥共發酵製成 方法’係為-充分符合實用進步性與產業利用性之設計者 〇 【發明内容】 本發明之桑黃與中草藥共發酵製成方法〔請參閱第一 圖〕，於液態培養基（工）加入桑黃菌株（2)配合最佳 產f控制條件（3)與中草藥（4)單方或複方共發酵， 其中： ㈣培養基（！），由馬鈴#萃取出之成份與葡萄醣 此合，最佳比例爲馬鈐薯萃取物（p〇tat〇 2⑽g几 、葡萄醣（Glucose)20g/L組成者； 桑黃菌株（2 )，爲裂蹄木層孔菌，學名phUinus linteus，一種珍貴的藥用菌，具有抗腫瘤及提高免疫力之 生理活性，加入液態培養基（i )内並配合最佳產量控制 _ 條件（3)培養者； 最佳產量控制條件（3)，爲葡萄醣（giucose)濃度 4 %、玉米粉（corn steep powder)濃度工％、氯化鈣 (Cacl2)濃度〇·1%、環境酸鹼值p H 4、轉速每分鐘丄2 5 轉（rpm)，可控制最佳菌絲及多醣體產量者； 中草藥（4)，由中草藥（4)内萃取出之成份，中 草樂（4)單獨一種稱爲單方或中草藥（4)混合二種以 上稱爲複方，桑黃菌株（2)於最佳產量控制條件（3 ) 與中草藥（4)單方或複方共發酵，比桑黃菌株（2)單 6 200825177 獨發酵時產生較佳菌絲及多酿體產量以及提升抗癌之效果 者。 【實施方式】 實施目的： 卜先找出最佳產量控制條件（3)，如最適當之碳源、 氣，、無機鹽類、PΗ值、轉速，礙源可被菌類吸收代謝 以提供忐1，氮源及無機鹽類影響菌絲及多醣體產量，以 %使桑Η產生最佳多醣體及菌絲產量，在多醣體產量上需達 到6 g/1 L以上，在菌絲總重達到i 〇 g/1 L以上，以利 ^技產業上量產’接著，取十四種中草藥（4)半枝連、 玄參、五味子、枸杞、仙逢、黃考、蒲公英、白花蛇舌草 三=真子、玫瑰、絞股藍、紅景天、刺五加、人參，在桑 黃最佳產量控制條件（3 )下與單方（中草藥.（4 )單獨 一種）或複方（中草藥（4)混合二種以上）共發酵，找 出比桑黃菌株（2)單獨發酵時產生較佳菌絲及多醣體產 _量，並選取人類大腸癌細胞（sw 細胞（A431)、人類前列腺癌細胞（PC_3S 類肝癌細胞（HepaG2 )四種癌細胞做腫瘤細胞存活測定（ MTT assay)，以比較桑黃菌株（2)單獨發酵及桑普 菌株（2)與中草藥（4)共發酵抗癌之效果。 ’、 實施步驟： 取桑黃菌株（2 )於固態培養基，培養至長到八分滿 ，先將250ml三角瓶内裝液態培養基（1)〔請參閱第 一圖〕，再將每個三角瓶内接種桑黃菌株（2)，將最佳 7 200825177 產量控制條件（3 )使用碳源爲葡萄醣（giucose)濃度4 %、使用氮源爲玉米粉（corn steep p〇wder)濃度1 %、使 用無機鹽類爲氯化鈣（Cac〗2)濃度〇·ι%加入液態培養基（1 )内，並於環境酸鹼值ρ Η 4、轉速每分鐘i 2 5轉（rpm )進行發酵，觀察桑黃菌株（2)在第六、八、十、十二 、十四天菌絲總重之變化〔請參閱第二圖〕，菌絲總重第 六天約爲7 g/1 L、第八天約爲丄2 g/1 L、第十天約爲 1 Θ g/1 L·第十一天約爲1 7 g/i l、第十四天約爲2 0 g/1 L，自第八天開始可達產生菌絲總重丄〇忌八L以 上標準，觀察桑黃菌株（2 )在第六、八、十、十二、十 四天多醣體產量之變化〔請參閱第三圖〕，多醣體產量第 六天約爲9 g/1 L、第八天約爲7 g/1 L、第十天約爲 7· 5g/l L、第十二天約a 7 σ/ι τ 铱丄 十 八、]爲（g/1 L·、弟十四天約爲5· 5 L，多膽體產量在第六至十二天可達6g/lL以上的桿準 馨 •，然後，先將十四種中草藥（4)(半枝連、黨參、五 子、枸杞、仙渔、黃#、蒲公英、白花蛇舌草°、 玫瑰、絞股藍、紅景天、刺五加、人參）^水者，、接 浪縮並於6 0。C乾燥，取得中草藥萃取物 桑黃菌株（2 )於最佳產量控制條件（3 )添加雜： 4)單方或複方共發酵實驗〔請參閱第—圖〕 :（ 株（2)添加單方中草藥（4)蒲公英共發酵時〔=菌 第四、五圖〕’以桑黃菌株（2)於最佳產量控制：：閱 3)單獨發酵做爲照對組，將桑黃菌株（2) ^士件（ 草藥（4)蒲公英於最佳產量控制條件（ 早方中 ^卜共發酵， 8 200825177 使用三種不同蒲公英濃度爲〇._、0.05%、0.1%做爲實 =組’觀察土：、八、十、十二、十四天菌絲及多醣體產 置變化，觀察菌絲產量之變化發現濃度〇 〇1%、〇 〇5%、200825177 IX. Description of the invention: [Technical field of invention] The present invention relates to a method for co-fermenting mulberry yellow and Chinese herbal medicine, and mulberry yellow is co-fermented with Chinese herbal medicine unilaterally or compoundly under the condition of liquid medium and optimal yield control. Compared with the production of better hyphae and polysaccharides and the effect of improving anti-cancer effect, it is a manufacturing method that fully meets the industrial applicability. Φ [Previous technique] \Sanghuang is a bacterium of the genus Phlinatus linteus, a fungus that is parasitic on live mulberry trees. It is black-like, with a hard texture and a bright yellow inside. It is called mulberry yellow. In the Ming Dynasty, Li Shizhen's "Compendium of Materia Medica" has its own load, mulberry yellow cold, slightly bitter taste, able to benefit the five internal organs, detoxification gas, hemostasis, etc. In the Korean and Japanese folk use, it has abdominal pain, diuresis, anti-tumor, According to the results of Korea and Japan, the polysaccharides isolated from mulberry can significantly stimulate the immune response of T cells, and analyze the activity of mulberry polysaccharides against tumor growth and metastasis. The immune system that activates the body, although it has introduced mulberry extract from Japan, but the price is expensive, and the efficacy and purity are not clear. Therefore, it is necessary to use mulberry in the liquid medium to match the best carbon source, nitrogen source and inorganic. Salt, p H value, rotation speed conditions, fermentation, read the mulberry yellow silk and multi-body yield, and co-ferment with Chinese herbal medicine unilateral or compound under the optimal conditions of mulberry yellow to increase the production of hyphae and polysaccharide And to enhance the anti-cancer effect. In view of the above-mentioned shortcomings, the inventor believes that it has the urgent need to improve, but also thinks about the idea of improving creation, and it has always maintained its superiority. In addition, the ten principles, the improvement of the existing defects, through the multi-party design to explore the Asian-human correction and improvement, the end of the invention is the method of co-fermentation of mulberry yellow and Chinese herbal medicines - is fully in line with practical progress and industry UTILITY DESIGNER 发明 【Abstract 】 The method for co-fermentation of mulberry yellow and Chinese herbal medicine of the present invention (please refer to the first figure), adding mulberry yellow strain (2) in liquid medium (work) with the optimal production f control condition (3) Co-fermentation with Chinese herbal medicine (4) unilateral or compound, wherein: (4) Medium (!), the ingredients extracted by Ma Ling # and glucose are combined, the optimal ratio is horse yam extract (p〇tat〇2(10)g Glucose 20g / L composition; Mulberry strain (2), is a bacterium of the genus phUinus linteus, a precious medicinal fungus, has anti-tumor and immune activity to enhance immunity, added to the liquid Medium (i) with optimal yield control _ condition (3) culture; optimal yield control condition (3), giucose concentration 4%, corn steep powder concentration, chlorination Calcium (Cacl2) Degree 〇·1%, environmental pH value p H 4, rotation speed 丄25 rpm (rpm), can control the best hyphae and polysaccharide yield; Chinese herbal medicine (4), extracted from Chinese herbal medicine (4) Ingredients, Zhongcao (4) alone is called unilateral or Chinese herbal medicine (4) mixed more than two kinds called compound, mulberry strain (2) in optimal yield control conditions (3) and Chinese herbal medicine (4) unilateral or compound Co-fermentation, B. sinensis strain (2) Single 6 200825177 Produces better hyphae and multi-breast yield and enhances anti-cancer effect when fermentation alone. [Embodiment] Objective: To find out the optimal yield control conditions (3) If the most suitable carbon source, gas, inorganic salt, PΗ value, and rotational speed, the source of the disorder can be absorbed and metabolized by the fungus to provide 忐1, and the nitrogen source and inorganic salt affect the production of hyphae and polysaccharide. % makes mulberry produce the best polysaccharide and hyphae production, and needs to reach 6 g/1 L or more in the yield of polysaccharides, and the total weight of mycelium reaches i 〇g/1 L or more, in order to facilitate mass production in the technology industry. 'Next, take fourteen kinds of Chinese herbal medicines (4) Half-branched, Scrophulariaceae, Schisandra, Poria, Xianfeng, Huang Kao, Pugong , Hedyotis diffusa three = true son, rose, Gynostemma pentaphyllum, Rhodiola, Acanthopanax senticosus, ginseng, under the optimal yield control conditions of mulberry (3) and unilateral (Chinese herbal medicine. (4) alone) or compound (Chinese herbal medicine (4) Mixing two or more kinds of co-fermentation to find out the best hyphae and polysaccharide production when fermenting alone (2), and selecting human colorectal cancer cells (sw cells (A431), human prostate) Cancer cells (PC_3S hepatocarcinoma cells (HepaG2), four cancer cells were used for tumor cell survival assay (MTT assay) to compare the fermentation of mulberry yellow strain (2) and Sample strain (2) and Chinese herbal medicine (4) to co-fermentate anticancer The effect. ', implementation steps: Take the mulberry strain (2) in solid medium, culture until the end of eight minutes, first 250ml flask containing liquid medium (1) [please refer to the first picture], then each triangle bottle Inoculation of mulberry strain (2), the best 7 200825177 yield control conditions (3) using carbon source for glucose (giucose concentration 4%), using nitrogen source for corn flour (corn steep p〇wder) concentration of 1%, use The inorganic salt is calcium chloride (Cac 2) concentration 〇·ι% is added to the liquid medium (1), and the fermentation is carried out at an environmental pH value of ρ Η 4 and a rotation speed of i 2 5 rpm (rpm). The total weight of mycelium in the sixth, eighth, tenth, twelfth and fourteenth day of the yellow strain (2) (see the second figure), the total weight of the mycelium is about 7 g/1 L on the sixth day, the eighth The day is about 2 g/1 L, the tenth day is about 1 Θ g/1 L. The eleventh day is about 17 g/il, and the fourteenth day is about 20 g/1 L. At the beginning of the day, the total weight of mycelium can be raised to the standard of more than eight L. The changes of polysaccharide yield in the sixth, eighth, tenth, twelfth and fourteenth days of the mulberry strain (2) were observed (please refer to the third figure). Polysaccharide The yield is about 9 g/1 L on the sixth day, about 7 g/1 L on the eighth day, about 7·5 g/l L on the tenth day, and about a 7 σ/ι τ on the twelfth day. ,] is (g/1 L·, brother 14 days is about 5·5 L, multi-biliary production can reach 6g/lL above 6g/lL in the sixth to twelfth days, then, first, fourteen Chinese herbal medicine (4) (Half-branched, Codonopsis, Wuzi, Yi, Xianyu, Huang #, dandelion, Hedyotis diffusa °, rose, Gynostemma, Rhodiola, Acanthopanax, ginseng) ^ water, The wave is dried at 60 ° C, and the Chinese herbal extract mulberry strain (2) is obtained under the optimal yield control conditions (3): 4) Uni- or compound co-fermentation experiment (please refer to the figure): (2) Adding a single Chinese herbal medicine (4) When dandelion is co-fermentation [= 4 and 5 of the bacteria] 'Controlling the optimal yield with the mulberry strain (2): Read 3) Separate fermentation as a pair, mulberry Yellow strain (2) ^ Shi pieces (herb (4) dandelion in the optimal yield control conditions (in the early middle ^ Bu total fermentation, 8 200825177 using three different dandelion concentrations as 〇. _, 0.05%, 0.1% as a real = Group 'observation soil , Eight, ten, twelve, fourteen days mycelium and production set to change polysaccharides, observation of changes in the yield of mycelium found that the concentration billion 〇1%, square 〇5%
第八天後皆可維持在丄〇 g/1 L以上〔請參閱第四圖 〕，其實驗組蒲公英濃度於請％、M5%比照對組佳， 而觀察多醋體產量之變化發現濃度〇 〇1%、〇 〇5% U 皆可維：在6 g/l L〔請參閱第五圖〕；將桑黃菌株（2° 鲁）“、、加單方中草樂（4 )枸把共發酵時〔請參閱第六、七 圖〕以桑θ S株（2)於最佳產量控制條件（3)單獨 發酵做爲照對組，將桑黃菌株（2)添加單方中草藥（4 )枸把於最佳產量控制條件（3 )下共發酵，使用三種不 翻句減度爲、㈣做爲實驗組，觀察第 + 十十一十四天囷絲及多酿體產量變化，觀察 菌、、糸產里之變化發現第十天後濃度0.01%、0 05%、〇 π 皆可維持在1〇g/1L以上〔請參閱第六圖〕·，其第十= 參十四天實驗組枸杞濃度於0.01%、0.05%、0.1%產量比照 對組佳，而觀察多醣體產量之變化發現〔請參閱第七圖〕 二其第六至十天實驗組枸杞濃度於0.05%、0.1%多醣體產 ，比照對組佳，且第六至十天組枸杞濃度於Q〇i%、〇.阳 %、0.1%多醣體產量在6 g/1 L以上；將桑黃菌株（2 ) t加複方中草藥（4)(玫瑰、黃耆、五味子、枸杞、黨 共發酵時〔請參閱第八、九圖〕，以桑黃菌株（2) =取佳產量控制條件（3 )單獨發酵做爲照對組，將桑黃 囷株（2)添加複方中草藥（4)(玫瑰、黃耆、五味子 9 200825177 、枸杞、黨參）於最佳產量控制條件（3)下共發酵，使 用三種不種複方中草藥（4)濃度爲0.01%、0.05%、 0· 1%做爲實驗組，觀察第六、八、十 '十二、十四天菌絲 及多醣體產量變化，觀察菌絲產量之變化發現第八天後濃 度0·01%、0·05%、〇·1%皆可維持在1 〇 g/1 L以上〔請 麥閱第八圖〕’其實驗組複方中草藥（4 )濃度於〇. 01% 、0.1%產量比照對組佳，而觀察多醣體產量之變化發現〔 鲁請參閱第九圖〕，其實驗組複方中草藥（4 )濃度於〇·〇1 %、0.05%、0.1%多醣體產量比照對組佳，且複方中草藥 (4 )濃度於0· 01%、〇· 〇5%、〇· 1%多醣體產量可以高達 在8 g/1 L以上；接著，做腫瘤細胞存活測定（mtt assay )〔請參閱第一圖〕，以比較桑黃菌株（2 )單獨 發酵及桑黃菌株（2)與中草藥（4)共發酵抗癌之效果 ，先將人類大腸癌細胞（SW4 8 0 )、人類皮膚癌細胞 (A43 1)、人類前列腺癌細胞（PC -3)、人類肝 馨癌細胞（HepaG2 )四種癌細胞分別接種於2 4孔培養盤内 乂’於3 7。C培養與5 % c〇2下2 4小時，將測定桑黃 菌株（2 )與中草藥（4)絞股籃共發酵液對抗人類大腸 癌細胞（SW4 8 0 )之效果〔請參閱第十圖〕，將單獨 桑黃發酵液四種不同劑量爲5 〇 # i、丄〇 〇 # i、2 〇 〇从1、4 Ο Ο // 1分別加入人類大腸癌細胞（$ w 4 8 〇 t内做爲照對組，將桑黃菌株（2 )與單方中草藥（4 )紅股監三種不同濃度〇 〇1%、〇 〇5%、〇1%共發酵，三 種不同濃度各有四種不同劑量爲5 〇 # ii 〇 〇 #丄、 200825177 2 0 0 // 1、4 G G // 1分別加人人類大腸癌細胞（s w 480)内做爲實驗組，於37。c培養與5 % c〇2下 培養7 2小枯後去除培養液以緩衝溶液p B s WASH，加 入乙醇UlediGl)使細胞蚊並以町伽viQlet染色，以 分光光度計分析儀在波長5 9 〇奈米（5 9 〇nm)測定癌 細胞的存活率，觀察桑黃絞股籃共發酵和桑黃單獨發酵對 抗人類大腸癌細胞（SW4 8 0 )之效果比較，桑黃絞股 φ 監共發酵的二個濃度、0.05%、〇·ι%對於抑制人類 大腸癌細胞（SW4 8 0 )皆優於桑黃單獨發酵，尤其是 剑里增加至1 〇 0 // 1以上時，可毒殺癌細胞使癌細胞存 活率於6 0 %以下，其抑制癌細胞效果明顯；將測定桑黃 菌株（2)與單方中草藥（4)黃耆共發酵液對抗人類皮 膚癌細胞（Α431)之效果〔請參閱第十一圖〕，將單 獨桑黃發酵液四種不同劑量爲5 0 // 1、1 〇 〇 μ 1、2 0 0 // 1、4 0 0 /ζ 1分別加入人類皮膚癌細胞（a 4 3 • 1 )内做爲照對組，將桑黃菌株（2 )與單方中草藥（4 )黃耆三種不同濃度0.01%、0.05%、0.1%共發酵，三種 不同濃度各有四種不同劑量爲5 〇 // 1、1 〇 〇 μ 1、2 〇〇// 1、400/Ζ 1分別加入人類皮膚癌細胞（Α43 1)内做爲實驗組，觀察桑黃黃耆共發酵和桑黃單獨發酵 對抗人類皮膚癌細胞（A 4 3 1 )之效果比較，桑黃黃耆 共發酵的三個濃度0.01%、0.05%、0.1%對於抑制人類皮 膚癌細胞（A 4 3 1 )皆優於桑黃單獨發酵，劑量在1 〇 〇 # 1時可使癌細胞存活率於6 0 %以下，且隨劑量增加 11 200825177 抑制癌細胞效果更佳，劑量於4 〇 〇 # 1可使癌細胞存活 率於20 %以下；將測定桑黃菌株（2)與單方中草藥（ 4 )仙渣共發酵液對抗人類前列腺癌細胞（p c _ 3)之 效果〔請參閱第十二圖〕，將單獨桑黃發酵液四種不同劑 *^50/^1 >l〇〇^i. 200^1. 400^1^ 別加入人類前列腺癌細胞（P C _ 3 )内做爲照對組，: 桑黃菌株（2)與單方中草藥（4)仙渣三種不同濃度 • 0.G1%、G.G5%、G.l%共發酵’三種不同濃度各有四種不 同劑量爲 50/zl、1〇〇“、2〇〇/zl、4〇〇“ 1分別加入人類前列腺癌細胞（P c_ 3 )内做 ，觀察桑黃仙渣共發酵和桑黃單獨發酵對抗人類前列腺癌 、、’田胞（P C 3 )之效果比杈，桑黃仙渣共發酵的三個濃 度0.01%、0.05%、0.1%對於抑制人類前列腺癌細胞（p C - 3 )皆優於桑黃單獨發酵，劑量在i 〇 〇 #丨時可使 癌細胞存活率於6 0 %以下，且隨劑量增加抑制癌細胞效 • 果更佳；將測定桑黃菌株（2)與單方中草藥（4)刺五 加共發酵液對抗人類肝癌細胞（HepaG2 )之效果〔請參閱 第十二圖〕，於第二天將單獨桑黃發酵液四種 5〇“、1〇 …、2〇0“、4〇〇 …別： 入人痛肝癌細胞（HepaG2 )内做爲照對組，蔣暴立键嫉Γ 2)與單方中草藥（4)刺五加三種不同濃度/〇〇1%、 0.05%、0.1%共發酵’三種不同共發酵濃度各有四種不同 劑量爲 50//1、l〇〇yl、2〇〇 从！ 、4〇〇//1 分別加入人類肝癌細胞（HepaG2 )内做爲實驗組，觀察桑 12 200825177 黃刺五加共發酵和桑黃單獨發酵對抗人類肝癌細胞（After the eighth day, it can be maintained at 丄〇g/1 L or above (please refer to the fourth figure). The concentration of dandelion in the experimental group is better than that of the group, and the concentration of dandelion is better than that of the group. 〇1%, 〇〇5% U can be dimensioned: at 6 g/l L [please refer to the fifth picture]; the mulberry strain (2° Lu) “, plus the unilateral Chinese grass music (4) At the time of fermentation (please refer to the sixth and seventh figures), the mulberry θ S strain (2) was used as the irradiation control group under the optimal yield control condition (3), and the mulberry yellow strain (2) was added to the single Chinese herbal medicine (4). Co-fermentation under the optimal yield control condition (3), using three kinds of non-collapse reduction, (4) as the experimental group, observing the change of the silk and multi-breast production at the +11-fourth day, observing the bacteria, After the tenth day, the concentration of 0.01%, 0 05%, and 〇π can be maintained above 1〇g/1L (please refer to the sixth figure), and the tenth = the fourteenth day experimental group. The concentration of strontium in the 0.01%, 0.05%, and 0.1% yields was better than that of the group, and the change in the yield of the polysaccharide was observed (see Figure 7). The sixth to ten days of the experimental group 枸杞 concentration 0.05%, 0.1% polysaccharide production, compared with the group, and the concentration of 枸杞 in the sixth to ten days group was Q〇i%, 〇.yang%, 0.1% polysaccharide yield was above 6 g/1 L; Strain (2) t plus compound Chinese herbal medicine (4) (rose, scutellaria, schisandra, wolfberry, party co-fermentation [please refer to the eighth and ninth maps], with mulberry strain (2) = good yield control conditions (3 Separate fermentation as a pair, and add the Chinese herbal medicine (4) (Rose, Astragalus, Schisandra 9 200825177, Radix, Codonopsis pilosula) to the best yield control conditions (3). Using three kinds of compound Chinese herbal medicine (4) concentration of 0.01%, 0.05%, 0.1% as the experimental group, observe the sixth, eighth, ten 'twelve, fourteen days of mycelial and polysaccharide yield changes, observe the bacteria The change of silk yield found that after the eighth day, the concentration of 0·01%, 0. 05%, and 〇·1% could be maintained above 1 〇g/1 L (please read the eighth picture] 'the experimental group compound Chinese herbal medicine ( 4) The concentration of 〇. 01%, 0.1% yield is better than that of the group, and the change of the polysaccharide yield is observed [L, please refer to the ninth figure], the experimental group compound Chinese herbal medicine 4) The concentration of 多糖·〇1%, 0.05%, 0.1% polysaccharides is better than that of the group, and the compound Chinese herbal medicine (4) concentration is 0.01%, 〇·〇5%, 〇·1% polysaccharide yield can be Up to 8 g / 1 L; then, do the tumor cell survival assay (mtt assay) (see the first image) to compare the fermentation of Phellinus igniarius (2) alone and the strain of mulberry (2) and Chinese herbal medicine (4) Co-fermentation anti-cancer effect, first human colon cancer cells (SW4 80), human skin cancer cells (A43 1), human prostate cancer cells (PC -3), human liver cancer cells (HepaG2) Inoculate in a 24-well culture dish, respectively, at 37. C culture and 5% c〇2 for 24 hours, will determine the effect of mulberry strain (2) and Chinese herbal medicine (4) stranded co-fermented liquid against human colon cancer cells (SW4 80) [see the tenth figure 〕, the four different doses of mulberry yellow fermentation broth are 5 〇 # i, 丄〇〇 # i, 2 〇〇 from 1, 4 Ο Ο / / 1 respectively added to human colon cancer cells ($ w 4 8 〇t) As a pairing group, the mulberry strain (2) and the unilateral Chinese herbal medicine (4) red stocks were co-fermented at three different concentrations of 〇〇1%, 〇〇5%, 〇1%, and four different doses for each of the three different concentrations. 5 〇# ii 〇〇#丄, 200825177 2 0 0 // 1, 4 GG // 1 Add human colon cancer cells (sw 480) as experimental group, culture at 37.c and 5% c〇2 After culturing 7 2 small dry, remove the culture solution to buffer solution p B s WASH, add ethanol UlediGl) to make the cell mosquito and stain with Machiya viQlet, and use spectrophotometer analyzer at wavelength 5 9 〇 nanometer (5 9 〇nm To determine the survival rate of cancer cells, observe the effect of co-fermentation of mulberry strands and mulberry yellow alone against human colorectal cancer cells (SW4 80), mulberry strands The two concentrations of φ super-fermentation, 0.05%, 〇·ι% are superior to mulberry yellow fermentation for inhibiting human colorectal cancer cells (SW4 80), especially when the sword increases to 1 〇 0 // 1 or more. It can poison cancer cells to make cancer cells survive below 60%, and its effect on inhibiting cancer cells is obvious; the geranium strain (2) and the single Chinese herbal medicine (4) scutellaria co-fermented liquid will be tested against human skin cancer cells (Α431). The effect [please refer to the eleventh figure], the four different doses of mulberry yellow fermentation broth are added to human skin, respectively, 50 0 / 1, 1 〇〇 μ 1, 2 0 0 / 1, 1 0 0 / ζ 1 The cancer cells (a 4 3 • 1 ) were used as a pairing group, and the mulberry strain (2) and the single Chinese herbal medicine (4) astragalus were co-fermented at different concentrations of 0.01%, 0.05%, and 0.1%. Four different doses were 5 〇// 1, 1 〇〇μ 1, 2 〇〇// 1, 400/Ζ 1 were added to human skin cancer cells (Α43 1) as experimental group, observe mulberry yellow jaundice Comparison of the effects of fermentation and mulberry yellow fermentation on human skin cancer cells (A 4 3 1 ), the three concentrations of mulberry yellow sassafras co-fermentation were 0.01%, 0.05%, 0.1% for the inhibition of human skin cancer cells (A 4 3 1 ) is superior to mulberry alone fermentation, the dose of 1 〇〇 # 1 can make the cancer cell survival rate below 60%, and increase with dose 11 200825177 inhibition of cancer The cell effect is better. The dose of 4 〇〇# 1 can make the cancer cell survival rate below 20%; the geranium strain (2) and the single Chinese herbal medicine (4) slag co-fermentation solution will be determined against human prostate cancer cells (pc _ 3) The effect (please refer to the twelfth figure), the four different agents of the mulberry yellow fermentation broth*^50/^1 >l〇〇^i. 200^1. 400^1^ are added to human prostate cancer The cells (PC _ 3 ) were used as the pairing group: three different concentrations of mulberry strain (2) and unilateral Chinese herbal medicine (4) slag • 0.G1%, G.G5%, Gl% co-fermentation 'three different concentrations Each of the four different doses was 50/zl, 1〇〇", 2〇〇/zl, 4〇〇" 1 was added to human prostate cancer cells (P c_ 3 ), and the mulberry slag co-fermentation and mulberry were observed. The effect of yellow fermentation alone against human prostate cancer, 'Tianji (PC 3 ) is better than 杈, and the three concentrations of mulberry slag co-fermentation are 0.01%, 0.05%, 0.1%. Human prostate cancer cells (p C - 3 ) are superior to mulberry alone in fermentation, and the dose of i 〇〇#丨 can make the cancer cell survival rate below 60%, and the effect of inhibiting cancer cells with increasing dose is better. The effect of the mulberry yellow strain (2) and the single Chinese herbal medicine (4) Acanthopanax senticosus fermentation solution against human hepatoma cells (HepaG2) will be determined (see Figure 12), and the next day, the mulberry yellow fermentation broth will be 5〇", 1〇..., 2〇0", 4〇〇... Don't: Enter the human liver cancer cell (HepaG2) as a photo group, Jiang violent key 嫉Γ 2) and unilateral Chinese herbal medicine (4) thorn Five plus three different concentrations / 〇〇 1%, 0.05%, 0.1% co-fermentation 'three different co-fermentation concentrations each have four different doses of 50 / / 1, l 〇〇 yl, 2 〇〇 from! 4〇〇//1 was added to human hepatocarcinoma cells (HepaG2) as experimental group, and observed mulberry 12 200825177 spurs five plus co-fermentation and mulberry yellow fermentation alone against human liver cancer cells (
HepaG2 )之效果比較，桑黃刺五加共發酵的三個濃度〇肩 %、0.G5%、0.1%對於抑制人類肝癌細胞（喊⑵）皆優 於桑黃單獨發酵，在較低劑量5 Q # i時癌細胞存活率二 6 0 /以下且卩过劑里增加抑制癌細胞效果更佳；將測定 桑黃菌株（2)與複方中草藥（玫瑰、黃耆、五味子^二 杞、蓋芩）共發酵對抗人類大腸癌細胞（s w 4 8 〇 )之 # =果〔請參閱第十四圖〕，將單獨桑黃發酵液四種不同劑 置爲 5CU1、i〇0/fl、2〇〇//1、4〇〇“p》 別加入人類大腸癌細胞（SW4 8 〇 )内做 ^ 桑黃菌株⑴與複方中草藥（玫瑰、黃考、五^子、: 把、黨參）三種不同濃度〇〇1%、〇〇5%、〇1%共發酵， 二種不同共發酵濃度各有四種不同劑量爲1、 20〇“i、4〇〇"i分別加入人類大腸癌細 胞（SW480)内做爲實驗組’觀察桑黃菌株（2)血 舉複方中草藥（4)(玫瑰、黃耆、五味子、拘杞、黨幻 共發酵和桑黃單獨發酵對抗人類大腸癌細胞（SW4 8 0 )之效果比較’桑黃與複方巾草藥（4 )共發酵Q.l%濃度 對於抑制人類大腸癌細胞（SW4 8 〇)優於桑黃單獨發 酵’且桑黃與複方中草藥（4 )共發酵0.1%濃度於較低劑 篁5 Ο β 1時癌細胞存活率於6 〇 %以下，且隨劑量增加 $制癌細胞效果更佳；將測定桑黃菌株（2)與複方中草 樂（4)(玫瑰、黃耆、五味子、枸杞、黨參）共發酵對 抗人類皮膚癌細胞（A4 3 1 )之效果〔請參閱第十五圖 13 200825177 〕，將單獨桑黃發酵液四種不同劑量爲、ι〇〇 …200…400“分別加入人類皮膚癌細胞 (A 4 3 1)内做爲照對組，將桑黃菌株（2)與複方中 草藥（4)(玫瑰、黃耆、五味子、枸杞、黨參）三種不 同濃度0·01%、0·05%、〇·1%共發酵，三種不同共；酵濃 度各有四種不同劑量爲5 〇从i、丄〇 〇 # 1、2 〇 〇/辰 1、4 0 0 // 1分別加入人類皮膚癌細胞（A 4 3 1 )内 做爲實驗組，觀察桑黃菌株（2 )與複方中草藥（4)( 玫瑰、黃耆、五味子、枸把、黨參）共發酵和桑黃單 酵對抗人類皮膚癌細胞（A4 3 1 )之效果比較，桑普二 複方中草藥（4 )共發酵〇. 1%、〇 〇5%濃度對於抑制 皮膚癌細胞（A4 3 1 )優於桑黃單獨發酵，且桑黃與= 方中草藥（4)共發酵濃度於較低劑量5 0/M時日士 活率於6 Q %以下’且隨劑量增加抑制癌細胞 土，將測定桑黃菌株（2)與複方中草藥（4 )(玫 :細=:味：、拘杞、黨參)共發酵對抗人類前列腺 :讀酵液四種不同劑量爲…丨'100…Π Q Λ U # 1勿別加入人類前列腺癌細胞（p C 一 0.01%、0 05% :。枸杞一黑茶）二種不同濃度 四種不同劑量^5 0酵，三種不同共發酵濃度各有 0 0二1 〇 0#1、2 0 0 …4 为別加入人類前列腺癌細胞（P C - 3 )内做爲 14 200825177 實驗組，觀察桑黃菌株（2)與複方中草藥（4)(玫瑰 只耆、五味子、枸杞、黨參）共發酵和桑黃單獨發酵對 抗人類前列腺癌細胞（PC_ 3)之效果比較，桑黃與複 方中草藥（4 )共發酵0.1%濃度對於人類前列腺癌細胞（ P c - 3 )抑制優於桑黃單獨發酵，且桑黃與複方中草藥 (4 )共發酵0.1%濃度於較低劑量5 〇 #丨時癌細胞存活 率於3 0 %以下，且劑量增加至丄〇 〇 # i以上時癌細胞 • 存活率於9 %以下，抑制癌細胞效果絶佳；將測定桑黃菌 株（2)與複方中草藥（4)(玫瑰、黃耆、五味子、枸 =[黨參）共發酵對抗人類肝癌細胞（HepaG2)之效果〔 °月參閱第十七圖〕，將單獨桑黃發酵液四種 “1、1〇〇 …2〇“1、4 0 0 …別力二 人類肝癌細胞（HepaG2 )内做爲照對組，將桑黃菌株（2 ^與複方中草藥（4 )(玫瑰、黃耆、五味子、枸杞、黨 :)—種不同濃度〇· 01%、〇· 05%、〇· 1%共發酵，三種不 同共發酵濃度各有四種不同劑量爲5 〇 # 1、1 〇 〇 #工 200//1、4〇〇#ι分別加入人類肝癌細胞（ HepaG2 )内做爲實驗組，觀察桑黃菌株（2 )與複方中草 藥if)(玫瑰、黃耆、五味子、枸杞、黨參）共發酵和 桑黃單獨發酵對抗人類肝癌細胞（HepaG2 )之效果比較， 桑黃與複方中草藥（4)共發酵〇.1%、〇屬濃度對於人 類肝癌細胞（HepaG2 )抑制優於桑黃單獨發酵，且桑黃與 複方中草藥（4 )共發酵〇· 1%濃度於較低劑量5 〇 # 1時 癌細胞存活率於4 〇 %以下，且隨劑量增加抑制癌細胞效 15 200825177 果更佳；以桑黃菌株（2)與複方中草藥（4)(玫瑰、 黃耆、五味子、枸杞、黨參）濃度0· 1%可使癌細胞存活率 於6 0 %以下，抑制癌細胞效果最佳，接著，做大型震叠 式液態培養發酵量產〔請參閱第十八圖〕，將桑黃菌株（ 2)與濃度0.1%複方中草藥（4)(玫瑰、黃耆、五味子 、枸杞、黨參）於葡萄醣（glucose)濃度4 %、玉米粉（ corn steep powder)濃度 1 %、KH2P04 濃度 0· 05%、環境酸Comparison of the effects of HepaG2), the three concentrations of scutellariae sinensis, three concentrations of 〇 shoulder, 0. G5%, 0.1% for the inhibition of human liver cancer cells (Cry (2)) are superior to mulberry alone fermentation, at lower doses 5 When Q # i, the survival rate of cancer cells is 260/s or less, and the effect of inhibiting cancer cells is better in the sputum-removing agent; the strain of mulberry yellow (2) and the compound Chinese herbal medicine (rose, scutellaria, schisandra, scorpion, scorpion, scorpion) Co-fermentation against human colorectal cancer cells (sw 4 8 〇)# = fruit [see Figure 14], the four different agents of mulberry yellow fermentation broth are set to 5CU1, i〇0/fl, 2〇〇 //1, 4〇〇 “p” Do not add human colorectal cancer cells (SW4 8 〇) to do three different concentrations of mulberry yellow strain (1) and compound Chinese herbal medicine (rose, yellow test, Wuzhizi,: put, Codonopsis) 〇1%, 〇〇5%, 〇1% co-fermentation, two different co-fermentation concentrations each have four different doses of 1, 20 〇 "i, 4 〇〇 " i respectively added to human colon cancer cells (SW480) As an experimental group, 'observation of mulberry strains (2) blood-supplement compound Chinese herbal medicine (4) (rose, scutellaria, schisandra, stagnation, party sci-fi fermentation and The effect of mulberry alone fermenting against human colorectal cancer cells (SW4 80) was compared with that of 'Sanghuang and compound towel herbs (4) co-fermentation Ql% concentration for inhibiting human colorectal cancer cells (SW4 8 〇) superior to mulberry yellow fermentation alone' And the mulberry yellow and the compound Chinese herbal medicine (4) co-fermented 0.1% concentration to the lower dose 篁5 Ο β 1 when the cancer cell survival rate is below 6 〇%, and the dose-increasing dose of cancer cells is better; the mulberry yellow will be determined The effect of co-fermentation of strain (2) with compound Zhongle (4) (rose, scutellaria, Schisandra, medlar, Codonopsis pilosula) against human skin cancer cells (A4 3 1 ) (please refer to Figure 15 Figure 13 200825177), The four different doses of mulberry yellow fermentation broth were ι〇〇...200...400" respectively added to human skin cancer cells (A 4 3 1) as a pairing group, and the mulberry yellow strain (2) and compound Chinese herbal medicine (4) (Rose, Astragalus, Schisandra, Poria, Codonopsis pilosula) three different concentrations of 0. 01%, 0. 05%, 〇 · 1% co-fermentation, three different total; yeast concentration each has four different doses of 5 〇 from i ,丄〇〇# 1,2 〇〇/辰 1, 4 0 0 // 1 respectively added to human skin cancer cells (A 4 3 1 ) As an experimental group, observe the mulberry strain (2) and the compound Chinese herbal medicine (4) (rose, scutellaria, schisandra, medlar, Codonopsis) co-fermentation and mulberry yellow single yeast against human skin cancer cells (A4 3 1) Comparison of the effects, Sangpu two compound Chinese herbal medicine (4) co-fermentation 1. 1%, 〇〇 5% concentration for inhibiting skin cancer cells (A4 3 1 ) is superior to mulberry yellow fermentation alone, and mulberry yellow and = Chinese herbal medicine (4) The co-fermentation concentration is lower than 6 Q% at the lower dose of 50/M, and the cancer cell soil is inhibited with increasing dose, and the mulberry strain (2) and the compound Chinese herbal medicine (4) will be determined. : Fine =: taste:, cautious, Codonopsis) co-fermentation against human prostate: four different doses of reading yeast solution...丨'100...Π Q Λ U # 1 Do not add human prostate cancer cells (p C -0.01%) , 0 05% :.枸杞一黑茶) Two different concentrations of four different doses of ^50 fermentation, three different co-fermentation concentrations of 0 0 2 1 〇 0 #1, 2 0 0 ... 4 for the addition of human prostate cancer cells (PC - 3 As an experimental group of 14 200825177, observe the germination of mulberry strain (2) with compound Chinese herbal medicine (4) (Rose scutellaria, Schisandra, Poria, Codonopsis pilosula) and mulberry alone to fight human prostate cancer cells (PC_ 3) Compared with the effect, the 0.1% concentration of mulberry yellow and compound Chinese herbal medicine (4) fermented for human prostate cancer cells (P c - 3 ) was better than that of mulberry yellow alone, and the mixture of mulberry and compound Chinese herbal medicine (4) was 0.1%. At lower doses of 5 〇#丨, the survival rate of cancer cells is below 30%, and when the dose is increased above 丄〇〇# i, the survival rate of cancer cells is below 9%, and the effect of inhibiting cancer cells is excellent; (2) The effect of co-fermentation of compound Chinese herbal medicine (4) (rose, scutellaria, schisandra, sorghum = [Dangshen) against human hepatocellular carcinoma cells (HepaG2) [°], see the seventeenth figure], separate mulberry yellow fermentation broth Kind of "1, 1〇〇...2〇" 1, 4 0 0 ... Human liver cancer cells (HepaG2) were used as a pairing group, and the strains of mulberry yellow (2 ^ and compound Chinese herbal medicine (4 ) (rose, scutellaria, schisandra, wolfberry, party:) were grown at different concentrations 〇·01%, 〇· 05%, 〇·1% co-fermentation, three different co-fermentation concentrations each have four different doses of 5 〇#1,1 〇〇#工200//1,4〇〇#ι respectively added to human hepatoma cells (HepaG2) As an experimental group, observe the effect of co-fermentation of mulberry strain (2) with compound Chinese herbal medicine if) (rose, radix, Schisandra, medlar, Codonopsis pilosula) and mulberry alone on human hepatoma cells (HepaG2), mulberry yellow Co-fermentation with compound Chinese herbal medicine (4) 1.1%, 〇 浓度 concentration for human hepatoma cells (HepaG2) inhibition is superior to mulberry yellow fermentation alone, and mulberry yellow and compound Chinese herbal medicine (4) co-fermentation 〇 · 1% concentration is lower When the dose is 5 〇# 1, the survival rate of cancer cells is below 4%, and the effect of cancer cells is inhibited with the increase of dose. 15 200825177 is better; the strain of mulberry (2) and the compound Chinese herbal medicine (4) (rose, scutellaria, schisandra , 枸杞, Codonopsis sinensis) concentration of 0·1% can make cancer cell survival rate of 60% Next, the best effect on inhibiting cancer cells, followed by large-scale earthquake-type liquid culture fermentation mass production [see Figure 18], the mulberry strain (2) and the concentration of 0.1% compound Chinese herbal medicine (4) (rose, yellow耆, schisandra, wolfberry, Codonopsis pilosula) in glucose concentration 4%, corn steep powder concentration 1%, KH2P04 concentration 0. 05%, environmental acid
驗值PH4、轉速每分鐘1 2 5轉（rpra)條件下與桑黃共 發酵六至十四天，觀察在第六、八、十、十二、十四天菌 矛八穴爲1 絲總重之變化，菌絲總重第六天爲7. 78g/1 L w 〇.20g/lL、第十天爲21.收/11^'第十二天19.89§/11_ 第十四天爲17.31g/l L，於發酵第八天時其菌絲總重可達 到1 0 g/1 L，且在其第八至十四天發酵期間菌絲總重皆 可維持在1 〇 g/1 L以上，其產量足夠生技量產者。 综上所述，當知本發明具有新穎性，且本發明未見之 於任何刊物，當符合專利法第21、22條之規定。 唯以上所述者，僅為本發明之一較佳實施例而已，當 不能以之限定本發明之_。即大凡依本發明申 =作之均㈣化與修飾，皆應仍屬本發明專利涵蓋之範 200825177 【圖式簡單說明】 ， 第一圖：係本發明之流程圖 第二圖：係本發明之桑黃於最佳條件下單獨發酵菌絲 總重曲線變化示意圖 第一圖·係本發明之桑黃於最佳條件下單獨發酵多醣 體產量曲線變化示意圖The test value was PH4, the rotation speed was 215 rpm (rpra), and the mulberry was fermented for six to fourteen days. The sixth, eighth, tenth, twelfth and fourteenth days were observed. The change in weight, the total weight of hyphae was 7.78g/1 L w 〇.20g/lL, the tenth day was 21. Received /11^' the twelfth day 19.89§/11_ The fourteenth day was 17.31 g/l L, the total weight of mycelium can reach 10 g/1 L on the eighth day of fermentation, and the total weight of hyphae can be maintained at 1 〇g/1 L during the eighth to fourteenth day of fermentation. Above, its output is sufficient for biotech producers. In summary, it is understood that the present invention is novel, and the present invention is not disclosed in any publication, and is in compliance with the provisions of Articles 21 and 22 of the Patent Law. The above description is only a preferred embodiment of the present invention, and the present invention cannot be limited thereto. That is to say, according to the invention, the average (four) and modification should still belong to the scope of the invention patent 200825177 [Simplified illustration], the first figure: the second diagram of the flow chart of the invention: the invention Schematic diagram of the change of the total weight curve of the fermented hyphae under the optimal conditions of the mulberry yellow. The first diagram is a schematic diagram showing the change of the yield curve of the single fermentation polysaccharide of the mulberry yellow of the present invention under the optimal conditions.
第四圖·係本發明之桑黃與蒲公英共發酵菌絲總重數 據及曲線變化示意圖 第圊係本發明之桑黃與蒲公英共發酵多醣體產量 數據及曲線變化示意圖 第六圖：係本發明之桑黃與枸杞共發酵菌絲總重數據 及曲線變化示意圖 第Θ係本發明之桑黃與枸杞共發酵多醣體產量數 據及曲線變化示意圖 第圖係、本發明之桑黃與複方中草藥共發酵菌絲總 重數據及曲線變化示意圖 第九圖係、本發明之桑黃與複方中草藥共發酵多釀體 產量數據及曲線變化示意圖 第十圖係、本發明之桑黃與絞股藍共發酵對人類大腸 癌細胞（SW4 8 0 )做癌細胞存活測定 ^ (MTT assay )數據及柱狀圖 第十圖·係本發明之桑黃與黃考共發酵對人類皮膚 癌、、、田胞（A 4 3 1 )做癌細胞存活測定（Μ τ T assay )數據及柱狀圖 17 200825177 笫十二圖：係本發明之桑黃與仙渣共發酵對人類前列 腺癌細胞（p C _ 3 )做癌細胞存活測定（M T T assay )數據及柱狀圖 第十二圖·係本發明之桑黃與刺加五共發酵對人類肝 癌細胞（HepaG2 )做癌細胞存活測定（M τ τ assay )數據及柱狀圖 第十四圖：係本發明之桑黃與複方中草藥共發酵對人 類大腸癌細胞（SW4 8 0 )做癌細胞存活測 定（Μ Τ T assay )數據及柱狀圖 第十五圖：係本發明之桑黃與複方中草藥共發酵對人 類皮膚癌細胞（A 4 3 1 )做癌細胞存活測定 (Μ Τ T assay )數據及柱狀圖 第十六圖：係本發明之桑黃與複方中草藥共發酵對人 類别列腺癌細胞（P C - 3 )做癌細胞存活測 定（Μ Τ T assay )數據及柱狀圖 第十七圖：係本發明之桑黃與複方中草藥共發酵對人 類肝癌細胞（HepaG2 )做癌細胞存活測定（M Τ T assay )數據及柱狀圖 第十八圖：係本發明之桑黃與複方中草藥共發酵以大型 震盪式液態培養其菌絲總重數據及曲線變化示 意圖 200825177 【主要元件符號說明】 1 液態培養基 3 最佳產量控制條件 桑黃菌株 中草藥The fourth figure is a schematic diagram of the total weight data and curve change of the co-fermented hyphae of the mulberry yellow and dandelion of the present invention. The third embodiment of the invention relates to the yield data and curve change of the polysaccharide of the mulberry yellow and dandelion of the present invention. Schematic diagram of the total weight data and curve change of the co-fermented hyphae of the mulberry yellow and medlar. The third embodiment of the mulberry yellow and sorghum co-fermented polysaccharide production data and curve change diagram of the present invention, the mulberry yellow and the compound Chinese herbal medicine co-fermentation of the present invention Schematic diagram of hyphae total weight data and curve change ninth figure, the mulberry yellow and compound Chinese herbal medicine co-fermentation multi-breast production data and curve change diagram of the present invention, the tenth figure, the invention of the mulberry yellow and Gynostemma co-fermentation on the human large intestine Cancer cells (SW4 8 0) for cancer cell survival assay ^ (MTT assay) data and histograms of the tenth figure · The mulberry yellow and yellow test co-fermentation of the present invention on human skin cancer, and field cells (A 4 3 1) Do cancer cell survival assay (Μτ T assay) data and histogram 17 200825177 笫12: The co-fermentation of mulberry yellow and slag of the present invention on human prostate cancer cells (p C _ 3 ) Cancer cell survival assay (MTT assay) data and histogram. Twelfth map. The cancer cell survival assay (M τ τ assay ) data of human hepatoma cells (HepaG2) and the sapphire and spur five co-fermentation of the present invention Histogram 14th view: The co-fermentation of the mulberry yellow and the compound Chinese herbal medicine of the present invention for the cancer cell survival assay (Μ Τ T assay ) of human colorectal cancer cells (SW4 8 0) and the histogram of the histogram: Co-fermentation of mulberry yellow and compound Chinese herbal medicine of the present invention for human skin cancer cells (A 4 3 1 ) for cancer cell survival assay (Μ Τ T assay ) data and histogram FIG. 16 : The mulberry yellow of the present invention Compound Chinese herbal medicine co-fermentation for human cancer cell lineage (PC - 3 ) for cancer cell survival assay (Μ Τ T assay ) data and histogram Figure 17: The invention of the mulberry yellow and compound Chinese herbal medicine co-fermentation to humans Hepatocarcinoma cells (HepaG2) for cancer cell survival assay (M Τ T assay ) data and histogram Figure 18: The total weight of hyphae in a large oscillating liquid culture by co-fermentation of the mulberry yellow and compound Chinese herbal medicine of the present invention Curve change diagram 20082517 7 [Key component symbol description] 1 Liquid medium 3 Optimal yield control conditions Mulberry strain Chinese herbal medicine
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