Submerged Fermentation and Biological Activity of Tremella fuciformis and Antrodia cinnamomea Co-culture in Different Su
何育澤、徐泰浩、林芳儀
E-mail: [email protected]
ABSTRACT
Tremella fuciformis Berk, more commonly known as white fungus contains poLysaccharides with a variety of biological activities such as: anti-allergy, anti-tumor, and skin whitening. Antrodia cinnamomea, which is more commonly known as niu-chang-chih or chang-chih, also possesses a variety of biological activities, such as anti-tumor and anti-oxidation properties. According to the Literature, however, there has not been a study with Antrodia cinnamomea and Tremella fuciformis co-cultured. In this study, different Beansand Grains were used to explore the effects of T. fuciformis and A. cinnamomea cultured on the bioactive component. In this study, the first shake flask test of red beans, green beans, oatmeal and job’s tears were employed as extract mediums both separately and co-cultured and biological activity was measured. Other variables that were explored were different temperatures, shock speed and feed rate difference. Finally, the optimal culture conditions were determined from the metabolites using an antioxidant test and from a HeLa anti-tumor test. The results show that to achieve higher biomass and intracellular polysaccharide production in the shake flask experiment the green bean extract provided the best yield among the single cultures (4.35mg/mL, 1.38mg/mL). The optimal temperatures and shock speed were 25℃ and 180rpm respectively; they were found to produce a high biomass and greater intracellular polysaccharide and crude triterpenoid production. In terms of feed rate difference a higher biomass and intracellular polysaccharide production was found at 2.0vvm (9.49mg/mL, 1.8mg/mL, respectively) but in crude triterpenoids production there was no significant difference. Anti-oxidation was measured by determining the ferrous ion cheLating ability, the DPPH scavenging capacity and the reducing capacity concentration increase. The ferrous ion chelating ability of the methanol extract of mycelium when 10mg/mL was 66%. The DPPH scavenging ability was 40% of the concentration when 10mg/mL, and the reducing ability was 1.51 of the concentration when 10mg/mL. An MTT assay of HeLa cervical cancer cell in a hot water extract of mycelium and extracellular polysaccharides in a 24 hour period found no significant effects on HeLa cells. The reLative survival rates of cells after 48 hours and 72 hours decreased gradually, however. The results showed that when the concentration of hot water in the extract of mycelium was increased from 100μg/mL to 500μg/mL, a downward trend in cells’
relative survival was found. Exopolysaccharides were also examined, and it was found when the concentration was 100μg/mL the cells’ relative survival rate would decline, but among concentrations of 300μg/mL to 700μg/mL the cells’ survival rate would increase. Concentrations of 900μg/mL to 1000μg/mL aLso displayed a decreased cellular relative survival rate. The results show that in the co-cuLtured Antrodia cinnamomea, polysaccharides and triterpenoids increase, an antioxidant capacity was found as well as the phenomenon of inhibition of HeLa cells. In the future, further research is warranted to study its specific immune activity in order to enhance its use in health food products.
Keywords : Tremella fuciformis、Antrodia cinnamomea、co-culture、biological activity Table of Contents
封面內頁 簽名頁 中文摘要 iii 英文摘要 v 誌謝 vii 目錄 viii 表目錄 xiv 1.前言 1 2.文獻回顧 2 2.1銀耳生物學特性 2 2.2銀耳生 物活性成份 2 2.2.1銀耳多醣 2 2.3樟芝簡介 3 2.3.1樟芝之特徵與形態 4 2.3.2樟芝活性成份 5 2.4豆類與穀類簡介 6 2.4.1綠豆 簡介 6 2.4.2紅豆之簡介 7 2.4.3麥片之簡介 7 2.4.4薏仁之簡介 8 2.5共培養 9 2.6影響真菌代謝之因素 10 2.6.1碳源 10 2.6.2氮 源 11 2.6.3培養溫度 11 2.6.4攪拌與通氣 12 2.6.5無機鹽類 13 2.6.6接種量 13 3.1實驗架構流程圖 14 3.2實驗菌株 15 3.2.1真菌 菌株 15 3.2.2細胞株 15 3.3實驗藥品 15 3.3.1農產品 16 3.4儀器設備 17 3.5實驗材料 18 3.5.1銀耳母株培養 18 3.5.2樟芝母株培 養 18 3.5.3菌種保存 19 3.5.4農產品培養基配製 19 3.6實驗方法 19 3.6.1液態共生培養接菌量試驗 19 3.6.2搖瓶試驗 20 3.6.3五 公升發酵槽 20 3.7分析方法 21 3.7.1菌絲體生物質量測定 21 3.7.2胞內多醣配製與測定 21 3.7.3胞外多醣配製與測定 22 3.7.4 總醣測定 22 3.7.5粗三?蓬?w 23 3.7.6抗氧化活性分析 24 3.7.7細胞實驗 26 4.1搖瓶培養試驗 29 4.1.1液態培養樟芝菌絲體之形 態變化 29 4.2液態培養銀耳菌絲體之形態變化 35 4.2.1以豆類與穀類萃取液培養銀耳探討生物活性成份之影響 38 4.3液態共 培養之菌絲體形態變化 41 4.4以最適豆類與穀類比較共培養與單一培養生物活性成份之響 44 4.5接菌量在共培養對生物質 量及胞內外多醣之影響 48 4.6震盪培養轉速對生物活性成份之影響 50 4.7共培養在不同溫度下對生物活性成份之影響 55 4.8 五公升發酵槽共培養試驗 60 4.8.1五公升發酵槽液態共培養 60 4.8.2以五公升發酵槽探討不同通氣量對生物性成份之影 響 62 4.9抗氧化試驗 64 4.9.1亞鐵螯合能力試驗 64 4.9.2清除DPPH能力試驗 67 4.9.3還原力試驗 70 4.10細胞毒性試驗 73 5.結論
80 參考文獻 82 圖4.1a-d複式顯微鏡觀察(1000X)Antrodia cinnmoamea菌絲生長之形態變化 30 圖4.2 a-d MIC-D觀
察(22X)Antrodia cinnamomea菌絲球生長之形態變化 31 圖4.3以豆類與穀類培養Antrodia cinnamomea對生產生質量及生物活 性成份之影響 34 圖4.4 a-d為複式顯微鏡觀察(1000X) Tremella fuciformis菌絲生長之形態變化 36 圖4.5 a-d以複式顯微 鏡(1000X)及MIC-D(22X)觀察Tremella fuciformis菌絲生長之形態變化 37 圖4.6以豆類與穀類培養Tremella fuciformis生產生質 量、生物活性成份之影響 40 圖4.7以複式顯微鏡(1000X)共培養第7天菌絲形態，白色鍵號為樟芝、黑色鍵號為銀耳 42 圖4.8 a-b為MIC-D(22X)觀察共培養菌絲球生長之形態變化 43 圖4.9以綠豆萃取液為培養基探討共培養與單一菌株培養對生物活 性成份之影響 46 圖4.10麥片萃取液為培養基探討共培養與單一菌株培養對生物活性成份之影響 47 圖4.11接菌量對共培養 後對生物活性成份之影響 49 圖4.12以麥片萃取液為培養基經共培養後比較不同溫度對生物活性成份之影響 57 圖4.13以綠 豆萃取液為培養基經共培養後比較不同溫度對生物活性成份之影響 58 圖4.14以麥片加綠豆萃取液為培養基經共培養後比較 不同溫度對生物活性成份之影響 59 圖4.15 a-d Tremella fuciformis、Antrodia cinnamomea共培養之五公升發酵槽生長形態變 化 61 圖4.16五公升攪拌式發酵槽經共培養後不同進氣量對生物活性成份之影響 63 圖4.17單一培養及共培養菌絲體甲醇萃 取物比較其亞鐵螯合力 66 圖4.18單一培養及共培養菌絲體甲醇萃取物比較其DPPH清除能力 69 圖4.19單一培養及共培養菌 絲體甲醇萃取物比較其還原能力 72 圖4.20比較單一與共培養菌絲體熱水萃取液於1mg/mL與HeLa細胞共培養24、48、72小 時對細胞存活率之影 響 76 圖4.21比較單一與共培養胞外多醣於1mg/mL與HeLa細胞共培養24、48、72小時對細胞存活率 之影響 77 圖4.22比較單一與共培菌絲體熱水萃取液不同濃度下與HeLa細胞共培養72小時對細胞存活率之影響 78 圖4.23比 較單一與共培胞外多醣不同濃度下與HeLa細胞共培養72小時對細胞存活率之影響 79 表4.1以綠豆萃取液為培養基經共培養 後比較不同震盪轉速對生物活性成份之影響 52 表4.2以麥片萃取液為培養基經共培養後比較不同震盪轉速對生物活性成份 之影響 53 表4.3以麥片加綠豆混合萃取液為培養基經共培養後比較不同震盪轉速生物活性成份之 54
REFERENCES
1.文博均。2004。探討深層發酵中環境因子對巴西洋菇生產多醣之影響。國立中央大學碩士論文。 2.王洪凱、林福呈、李德葆。2002。
真菌中溫度敏感型相關基因研究進展。微生物學報。42(5):634-638。 3.王慧杰、李自剛、辛婷、李欣。2011。多菌種混合發酵玉米秸稈 生產含?○J白飼料的研究。湖南農業科學。(7):125-128。 4.田雪梅、宋愛榮。2005。溫度對樟芝菌絲體生長的影響試驗。中國食用菌
。24(5):51-52。 5.任俊平。2001。巧種薏米深加工分路銷售效益高。中國種業。(1):38。 6.池建偉、張名位、徐志宏、魏振承、孫玲、王 志堅。2001。河東烏麥胚芽營養麥片的研制。華南師範大學學報。(4):63-67。 7.吳大康、張久春、楊曉麗。2002。銀耳深層發酵條件的 研究。食品科學。23(1):64-68。 8.吳玉鑾、羅海英、陳意光、冼燕萍、羅東輝。2011。?☆?高效液相色譜法檢測麥片和啤酒中β-葡聚糖 的含量。分析試驗室。30(1):66-69。 9.吳秋曄、梁志欽。2005。Antrodia camphorata洋薏仁固態醱酵萃取物之抑菌作用。台灣農業化學 與食品科學 43(4):295-303。 10.吳國榮、程光宇、陸長梅、周長芳、顧龔平。2000。豆類種子的超氧物歧化?′〝吽B同工?﹞弮鶱酋w性
。南京師大學報。23(4):88-91。 11.呂國英、潘慧娟、吳永志、范雷法。2009。蛹蟲草無性型菌絲體提取液體外抗氧化活性研究。菌物學 報。28(4):597-802。 12.李存芝、晏日安、歐仕益、黃才歡、劉柳、黃樺宏。2011。薏米及其蛋白?☆悛囿漣飫韙 坒遄C暨南大學學報
。32(1):53-56。 13.李敏。2001。綠豆化學成份及藥理作用的研究概況。上海中醫藥雜誌。35(5):47-79。 14.李會芬。2010。我國紅小豆的 利用及加工現狀。現代農村科技。(22):67-68。 15.李艷春、馬恩龍、王小龍、王敏偉。2008。銀耳孢糖合用氟尿嘧啶對肉瘤S180和肝 癌H22小鼠的抗腫瘤作用。中國醫院藥學雜誌。28(3):209-211。 16.汪少芸、葉秀云、饒平凡。2004。綠豆生物活性物質及功能的研究進 展。中國食品學報。4(1):98-102。 17.車長遠。2010。樟芝的人工培養及其友效成份提取的研究現狀。科技信息。35:50。 18.周素娟
。2010。對我國單一原料保健食品審批概況的調查及注冊管理的建議。中國食品衛生雜誌。22(2):143-147。 19.孟繁岳、車會蓮、杜杰、
李蓉。2005。樟芝抑瘤作用對荷瘤小鼠免疫功能的影響。中國公共衛生。21(10):1224。 20.林克融。2002。探討培養基之pH值與Xanthan gum的添加巴西蘑菇多醣體生成之影響。國立中央大學碩士論文。 21.林怡朱。2005。靈芝不同生長階段液態培養菌絲與子實體之水溶 性多醣特性。國立台灣大學碩士論文。 22.林芳珊。2007。保健佳品話銀耳。食品與健康。10:32-33 23.武守華。2009。四種子囊菌的抗 氧化活性及抗氧化成分研究。湖南師範大學碩士論文。 24.侯建明、藍進、高益槐。2008。銀耳多糖抗潰瘍作用實驗研究。中國療養醫 學。17(5):316。 25.姜瑞芝、陳懷永、陳英紅、高陽、高其品。2006。銀耳孢糖的化學結構初步研究及其免疫活性。中國天然藥物。4(1):
73-76。 26.胡勤星、席興文、魏永俠、劉長偉、李峰。2009。不同碳源條件下鏈格孢菌培養特征的差異。湖南農業科學。4:38-40。 27.
韋璐、許文濤、屈偉、梁志宏、王金愛、黃昆侖。2010。一種從薏仁種子中提取出來的抗真菌蛋白。農業生物技術學報。18(4):689-694
。 28.孫勇、蔣繼宏、陳玉芹、高甜慧、陳鳳美。2007。10種多孔類真菌體外抗腫瘤活性的篩選。微生物學通報。34 (1): 112-115。 29.徐 文清。2010。銀耳多糖抑制腫瘤作用及其機制的研究。第七屆全國生化與生物技術藥物學術年會 :76-80。10月1日。中國藥學會。綿陽
。 30.徐胤桓。2011。不同蛹蟲草菌株親源分析、生物活性成分含量及抗氧化特性探討。大葉大學研究所碩士論文。 31.徐華麗、于曉風
、曲紹春、張瑞、王迎慶、睢大員。2008。銀耳孢糖對小鼠體內移植性腫瘤及免疫功能的影響。中國現代應用藥學。25(2):93-95 32.崔節 泉。2012。有機氮源在青霉素發酵生產過程中的作用。科技創新與應用。1:41。 33.張宏海。2008。樟芝液體發酵及多糖提取工藝研究
。河南科技大學碩士論文。 34.張宏海。2008。樟芝深層液體發酵條件的試驗研究。食品與藥品。10(5):18-21。 35.張迅捷、陳紹軍、陳 冠敏。2011。樟芝多醣對小鼠免疫功能影響的研究。福建師範大學學報。27(5):79 36.張鐵英。2004。紅豆中多酚類物質的提取及其含量 測定的研究。中國食品添加劑。(5):99-100。 37.梁志欽。1991。松杉靈芝浸漬發酵生產的1,3-βglucanase 對菌體胞外多醣的影響。國立 台灣大學碩士論文。 38.郭美蘭、穆俊祥、高喜葉、韓海霞、周峰、姚嶺柏。2011。不同無機鹽離子對平菇菌絲體生長的影響。北方園 藝。4:192-193。 39.郭惠菁。2005。以固態醱酵製備蟲草米及其品質與抗氧化性質。國立中興大學碩士論文。 40.陳宏慧。2004，液態培
養環境對茯苓（WoLfiporia cocos）菌絲體生長及其多醣體成分之影響。東海大學碩士論文。 41.陳怡倩。2001。利用批示液態培養來探 討檸檬酸對裂摺菌生長及其多醣體生成影響之研究。中央大學碩士論文。 42.陳勁初、林文鑫、陳清農、許勝傑、黃仕政、陳炎鍊
。2001。台灣特有真菌—樟芝菌絲體之開發。中華真菌學會會刊。16(1,2):7-22 43.陳書豪。2006。探討樟芝的溫度變化對液態發酵與固 態發酵生產三?衛?P多醣體之影響。中央大學碩士論文。 44.陳崗。2011。銀耳多糖的功能特性及其應用。中國食品添加劑。4:144-148。
45.陳體強、方忠王。2003。珍稀藥用菌樟芝研究現狀與進展。食用菌學報。10(4):55-60。 46.陸震鳴。2006。樟芝菌粉抗氧化和預防大
鼠急性酒精性肝損傷的研究。江南大學碩士論文。 47.陸震鳴。2009。樟芝深層液態發酵及其三?衛? X物的研究。江南大學博士論文。
48.曾軍、石國榮。2008。天然產物抗氧化活性的測定方法和原理。安徽農學通報。14(22):35-36。 49.黃大斌、楊菁、黃進華、翁景華
。2001。樟芝生物學特性研究。食用菌學報。8(2):24-28 50.黃秀錦。2008。銀耳多醣的提取、分離、純化及其功能性質研究。食品科學
。29(1):134-136。 51.黃思齊。2011。發酵產程擴大化級不同培養基對雲芝胞外醣?忖 S性之影響。大葉大學碩士論文。 52.楊萍、張 震。2009。銀耳的功能性及發展前景。食品研究與開發。30(7):179-180。 53.葉昇達。2003。厭氧發酵產氫菌與光合產氫菌共培養之產氫 效率比較研究。台中師範學院碩士論文。 54.鄒水洋、肖凱軍、郭祀遠。2009。康寧木霉液態發酵高產纖維素?〝M木聚糖?﹛C華南理工 大學學報。37(6):69-73。 55.趙金賢。2010。結合纖維素分解菌與固定化產醇菌之共培養系統以提升乙醇產量之研究。大葉大學碩士論文
。 56.趙珮岑。2004。黃白木耳液體發酵的生理活性探討。南台科技大學碩士論文。 57.趙琳、黃元新、劉昌時。2000。營養麥片中磷脂
的測定。廣西預防醫學。6(5):301-302。 58.劉心義、徐二華、陳繼偉、劉元剛、汪少芸、葉秀云。2007。紅豆中幾丁質?〞滲瞻 P表征
。福州大學學報。35(4):625-630。 59.劉月好。2003。薏米的營養及其在食品中的開發應用。食品科技。(9):47-49。 60.劉佳茹。2005。油 脂與介面活性劑隊舞菇生長與多醣生產之影響。大葉大學碩士論文。 61.劉娟、馬愛民、盛桂華、陳立國、謝筆鈞。2007。銀耳二型態 細胞差異性的初步研究。為生物學通報。34(5):880-884。 62.劉華、賈薇、張勁松。2008。樟芝菌絲體的不同提取方法及其抗腫瘤活性的 研究。食品與發酵工業。34(12):171-174。 63.劉福楊。2010。銀耳與香灰菌互作關系的研究。福建農林大學碩士論文。 64.謝寶貴。1989
。銀耳及其伴生菌木質纖維素降解?〞漪膍s。華中農業大學碩士論文。 65.顏軍、徐光域、郭曉強、劉嵬、李曉光、苟小軍。2005。銀 耳粗多糖的純化及抗氧化活性研究。食品科學。26(9):169-172。 66.顏軍、郭曉強、鄔曉勇、徐光域、苟小軍。2006。銀耳多醣的提取及 其清除自由基作用。成都大學學報(自然科學板)。25(1):35-38。 67.魏宇昆。2010。內生真菌與禾本科植物的共生機制研究進展。中國農 學通報。26(12):295-298。 68.羅水忠、潘利華、羅建平。2008。綠豆β-葡萄糖??〞漱擢鰳瞻 P性質研究。糧油加工。4:92-94。 69.羅利 娟、習平根、姜子德、威佩坤。2004。純培養擬莖點霉屬真菌的產孢條件。菌物學報。23(2):219-225。 70.羅信昌。2008。中國銀耳研究 之歷史回顧。中國菌物學會第四屆會員代表大會暨全國第七屆菌物學學術討論會論文集。35(32):10202-10203。 71.Ao, Z. H., Xu, Z. M., Lu, Z. M., Xu, H. Y., Zhang, X. M. and Dou, W. F. 2009. Niuchangchih (Antrodia camphorata) and its potential in treating Liver diseases.
Journal of Ethnopharmacology. 121(2):194-212. 72.Bao, H. D., Ochiai, Y. and Ohshima, T. 2010. Antioxidative activities of hydrophilic extracts prepared from the fruiting body and spent cuLture medium of Flammulina velutipes. Bioresource Technology. 101(15):6248-6255. 73.Berovic, M., Habijanic, J., Zore, I., Wraber, B., Hodzar, D., Boh, B. and PohLeven, F. 2003. Submerged cultivation of Ganoderma lucidum biomass and immunostimulatory effects of fungal polysaccharides. Journal of BiotechnoLogy. 103(1): 77-86. 74.Brosnahan, J., Summers, A., Nguyen, V., ZuKley, L., PohL, R. and Rippe, J. M. 2007. Effects of Exercise with or without a Hypocaloric Diet Including Oatmeal on Cardiovascular Disease Risk Factors Following Structured Weight loss. Journal of the American Dietetic Association. 107(8):A39. 75.Chang, H. L., Chao, G. R., Chen, C.
C. and Mau, J. L. 2001. Non-volatile taste components of Agaricus blazei, Antrodia camphorata and Cordyceps militaris mycelia. Food Chemistry.
74(2):203-207. 76.Chang, T. T. and Chou, W. N. 1995. Antrodia cinnamomea sp. nov. on Cinnamomum kanehirai in Taiwan. Mycological Research. 99(6):756-758. 77.Chen, B. 2010. Optimization of extraction of Tremella fuciformis polysaccharides and its antioxidant and antitumour activities in vitro. Carbohydrate Polymers. 81(2):420-424. 78.Cheung, C. K. 1996. The hypochoLesteroLemic effect of two edible mushrooms:
AuricuLaria auricula (tree-ear) and Tremella fuciformis (white jelly-Leaf) in hypercholesterolemic rats1 . Nutrition Research. 16(10): 1721-1725.
79.Cho, E. J., Oh, J. Y., Chang, H. Y. and Yun, J. W. 2006. Production of exo-polysaccharides by submerged mycelia culture of a mushroom Tremella fuciformis. Journal Biotechnol. 127(1):129-140. 80.ConnoLLy, M. L., Lovegrove, J. A. and Tuohy, K. M. 2010. In vitro evaluation of the microbiota modulation abilities of different sized whole oat grain flakes. Anaerobe. 16(5):483-488. 81.Fang, Q. H. and Zhong, J. J. 2002.
Submerged fermentation of higher fungus Ganoderma lucidum for production of valuable bioactive metabolites-ganoderic acid and polysaccharide. Biochemical Engineering JournaL. 10(1):61-65. 82.Horn, L. V., Emidy, L. A., Liu, K., Liao, Y., BaLLew, C., King, J. and StamLer, J. 1988. Serum Lipid response to a fat-modified, oatmeal-enhanced diet. Preventive Medicine. 17(3):377-386. 83.Huang, B. M., Chiang, M. T., Yao, H. T. and Chiang, W. 2005. The effect of adLay oiL on plasma lipids, insulin and leptin in rat. Phytomedicine. 12(6-7):433-439.
84.Kim, S. W., Hwang, H. J., Xu, C. P., Sung, J. M., Choi, J. W. and Yun, J. W. 2003. Optimization of submerged culture process for the production of mycelial biomass and exo-polysaccharides by Cordyceps militaris C738. Journal of Applied Microbiology. 94:120-126. 85.Kumar, S., Verma, A.K., Misra, A., Tripathi, A., Chaudhari, B. P., Prasad, R., Jain, S. K., Das, M. and Dwivedi P. D. 2011. Allergenic responses of red kidney bean (Phaseolus vulgaris cv chitra) polypeptides in BALB/c mice recognized by bronchial asthma and allergic rhinitis patients. Food Research International. 44(9):2868-2879. 86.Lee, I. H., Hung, Y. H. and Chou, C. C. 2008. Solid-state fermentation with fungi to enhance the antioxidative activity, total phenolic and anthocyanin contents of black bean. International Journal of Food Microbiology. 121(2):150-156. 87.Li, P., Wang, H., Liu, G., Li, X. and Yao, J.2011. The effect of carbon source succession on laccase activity in the co-culture process of Ganoderma lucidum and a yeast. Enzyme and Microbial Technology. 48(1):1-6. 88.Liang, C. H., Syu, J. L. and Mau, J. L. 2009. Antioxidant properties of solid-state fermented adlay and rice by Phellinus linteus. Food Chemistry. 116(4):841-845. 89.Lin, E. S. and Sung, S. S. 2006. Cultivating
conditions influence exopolysaccharide production by the edibLe Basidiomycete Antrodia cinnamomea in submerged culture. International Journal
of Food Microbiology. 108(2):182-187. 90.Liu, C. J., Chiang, C. C. and Chiang, B. H. 2012. The elicited two-stage submerged cultivation of Antrodia cinnamomea for enhancing triterpenoids production and antitumor activity. Biochemical Engineering Journal. 64(15):48-54. 91.Lo, H.
C., Tsai, F. A., Wasser, S. P., Yang, J. G. and Huang, B. M. 2006. Effects of ingested fruiting bodies, submerged culture biomass, and acidic poLysaccharide glucuronoxylomannan of Tremella mesenterica Retz.:Fr. on glycemic responses in normal and diabetic rats . Life Sciences. 78(17):
1957-1966. 92.Mau, J. L., Chang, C. N., Haung, S. H. and Chen, C. C. 2004. Antioxidant properties of methanolic extracts from Grifola frondosa Morchella esculenta and Termitomyces albuminosus mycelia. Food Chemistry. 87(1):111-118. 93.Mau, J. L., Huang, P. N., Huang, S. J. and Chen, C. C. 2004. Antioxidant properties of methanolic extracts from two kinds of Antrodia camphorata mycelia. Food Chemistry. 86(1):25-31.
94.Mau, J. L., Huang, P. N., Huang, S. J. and Chen, C. C. 2004. Antioxidant properties of methanolic extracts from two kinds of Antrodia camphorata mycelia. Food Chemistry. 86(1):25-31. 95.Nebus, J. 2010. The rapid and lasting efficacy of an oatmeal lotion in improving the moisturization and skin barrier properties of extra dry, itchy skin. Journal of the American Academy of Dermatology. 62(3):1609. 96.Oh, J. Y., Cho, E. J., Nam, S. H., Choi, J. W. and Yun, J. W. 2007. Production of polysaccharide–peptide complexes by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala. Process Biochemistry. 42(3):352-362. 97.Oliveira, R. P. S., Torres, B. R., Perego, P., OLiveira, M. N. and Converti, A. 2012. Effect of inulin on growth and acidification performance of different probiotic bacteria in co-cultures and mixed culture with Streptococcus thermophilus. Biochemical Engineering Journal. 62(15):62-69. 98.Oyaizu, M. 1986. Antioxidative activity of browning products of glucosamine fractionated by organic solvent and thin-Layer chromatography. Journal of the Japanese Society for Food Science and Technology. 35:771-775. 99.Ragaee, S., AbdeL-AaL, E. S. and Noaman, M. 2006. Antioxidant activity and nutrient composition of selected cereals for food use. Food Chemistry. 98(1):32-38. 100.Ram?瀑ez, M., MoraLes, D., Mart?瀋ez-MoraLes, F., Okoh, A. I. and
Trejo-Hern?鴨dez , M. R. 2010. Benzo[a]pyrene removal by axenic- and co-cultures of some bacterial and fungal strains. International Biodeterioration & Biodegradation. 64(7): 538-544. 101.Randhir, R. and Shetty, K. 2007. Mung beans processed by solid-state bioconversion improves phenolic content and functionality relevant for diabetes and ulcer management. Nnovative Food Science & Emerging Technologies.
8(2):197-204. 102.Rau, U., Gura, E., Olszewski, E.and Wangner, F. 1992. Enhanced glucan formation of filamentous fungi by effective mixing.
Journal of Industrial Microbiology and Biotechnology. 9:12-26. 103.Shimada, K., Fujikakw, K., Yahara, K. and Nakamura, T. 1992.
Antioxidation of soybean oil in cyclodexztrin emulsion. Journal of Agricultural and Food Chemistry. 40:945-948. 104.Shu, C. H. and Lung, M. Y.
2008. Effect of culture pH on the antioxidant properties of Antrodia camphorata in submerged culture. Journal of the Chinese Institute of Chemical Engineers. 39(1):1-8. 105.Wagner, R., MitcheLL, D. A. and Sassaki, G. L. 2004. Links between morphoLogy and physioLogy of Ganoderma Lucidum in submerged cuLture for the production of exopoLysaccharide. JournaL of BiotechnoLogy. 114(1-2):153-164. 106.Wang, C.
Y., Wu, S. C., Ng, C. C. and Shyu, Y. T. 2010. Effect of Lactobacillus-fermented adlay-based milk on lipid metabolism of hamsters fed
cholesterol-enriched diet. Food Research International. 43(3):819-824. 107.Weintraub, N. S., Rhodes, K. D., Brook, R. D. and Rubenfire. 2007.
Outcomes Assessment for Patients in a Multi-Disciplinary lipid Management Program with an Initial Nutrition Run-in Period Shows Positive Results Journal of the American Dietetic Association. 107(8):A39. 108.Wen, C. L., Chang, C. C., Huang, S. S., Kuo, C. L., Hsu, S. L., Deng, G. J.
and Huang, G. J. 2011. Anti-in?ammatory effects of methanol extract of Antrodia cinnamomea mycelia both in vitro and in vivo. Journal of Ethnopharmacology. 137(1):575-584. 109.WiLLiams, W. B., CuveLier, M. E. and Berset, C. 1995. Use of a free radical method to evaluate antioxidant activity. LWT - Food Science and Technology. 28(1): 25-30. 110.Wu, J., Ding, Z. Y. and Zhang, K. C. 2006. Improvement of exopolysaccharide production by macro-fungus Auricularia auricula in submerged culture. Enzyme and Microbial Technology. 39(4):743-749.
111.Wu, S. H. and Chan, T. T. 1997. Antrodia camphorata(niu-chang-Chih) new combination of medicinal fungus in Taiwan. Bulletin of the Polish Academy of Sciences Chemistry. 38:273-27. 112.Yang, F. C. and Liau, C. B. 1998. The influence of environmental conditions on polysaccharide formation by Ganoderma lucidum in submerged culture. Process Biochem. 33(5): 547-553 113.Yang, F. C., Haung, H. C. and Yang, M. J. 2003. The influence of environmental conditions on the mycelial growth of Antrodia cinnamomea in submerged cuLtures. Enzyme and Microbial Technology. 33(4):395-402. 114.Yang, J. H., Lin, H. C. and Mau, J. L. 2002. Antioxidant properties of severaL commercial mushroom. Food Chemistry. 77(2):229-235. 115.Ye, X. Y. and Ng, T. B. 2001. Peptides from Pinto Bean and Red Bean with Sequence Homology to Cowpea 10-kDa Protein Precursor Exhibit Antifungal, Mitogenic, and HIV-1 Reverse Transcriptase-Inhibitory Activities. Biochemical and Biophysical Research Communications. 285(2):424-429. 116.Zhong, J. J., Fang, Q. H. and Tang, Y. J. 2002. Enhanced production of medicinal mushroom Ganoderma lucidum by manipulation of oxygen supply. Journal of Plant Biotechnology.4(3)109-115. 117.Zhu, H., Tian, B., Liu, W., Zhang, S., Cao, C., Zhang, Y. and Zou, W. 2012. A three-stage culture process for improved exopolysaccharide production by Tremella fuciformis. Bioresource Technology. 116:526-528.
