TWI689258B - 牛樟芝醱酵糙米，其製法及其組成物 - Google Patents
牛樟芝醱酵糙米，其製法及其組成物 Download PDFInfo
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- TWI689258B TWI689258B TW106146655A TW106146655A TWI689258B TW I689258 B TWI689258 B TW I689258B TW 106146655 A TW106146655 A TW 106146655A TW 106146655 A TW106146655 A TW 106146655A TW I689258 B TWI689258 B TW I689258B
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- Prior art keywords
- brown rice
- antrodia
- antrodia cinnamomea
- fermented
- cinnamomea
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Abstract
本發明係關於一種經牛樟芝醱酵之糙米，特徵在於將牛樟芝菌種接入由糙米作為培養基質進行醱酵而得。根據本發明製得之牛樟芝醱酵糙米除含有原糙米的特殊營養成分，亦包含牛樟芝的特有活性成分，可供直接做為食材及用於製備養生糙米產品，如糙米粉、糙米漿、玄米茶及作為其他食品之營養調配物等。
Description
本發明係關於一種經牛樟芝醱酵之糙米。更特別地，本發明係關於一種經由將牛樟芝菌種接入由糙米為培養基質進行醱酵而得的牛樟芝醱酵糙米。
依據歷史記載，1773年開蘭第一人吳沙從中國福建渡海來臺灣開墾宜蘭，與當地原住民進行貿易活動，交流頻繁，吳沙從原住民取得牛樟芝熬煮食用，直到1985年，牛樟芝始流傳到平地，發現其具有良好食用功效，於是各大學及政府研究機構均積極投入牛樟芝的基源、化學成分、藥理活性及人工培養等之研究，應用潛力深受學術界肯定。牛樟芝(Antrodia camphorata
、Antrodia cinnamomea
、Taiwanofungus camphoratus
) 屬於非褶菌目、多孔菌科、薄孔菌屬、樟芝種，乃多年生蕈菌類。牛樟芝，民間的稱呼尚有樟芝、樟菇、樟菰、樟內菇、牛樟菇、紅樟等等，是臺灣特有種的菇類，在天然狀況下只生長在臺灣特有的常綠闊葉大喬木牛樟樹(Cinnamomum kanehirai
)樹幹內中空的心材上，截至目前為止，世界上僅發現於臺灣，是故堪稱為世界的珍寶。
牛樟芝所含的化合物大致可歸類為多醣體(polysaccharides)、三萜類(triterpenoids)、雙萜類(diterpenes)、倍半萜烯類化合物(sesquiterpenes)、倍半萜內酯(sesquiterpene lactones)、苯類(benzenoids)、苯醌衍生物(benzoquinone derivatives)、超氧歧化酶(superoxide dismutase)、多炔類(polyacetylenes)、腺苷(adenosine)、馬來酸(maleic acid)和琥珀酸(succinic acid)的衍生物，蛋白質(含免疫蛋白)、酚類化合物(phenolic compounds)、固醇類(steroids)和揮發性物質(牛樟芝的香味)等。許多第一次被發現和命名的成分都以牛樟芝學名中之屬名開頭，例如antcin、antrocin、antrodin、antrocamphin及antroquinonol等。不論椴木栽培的牛樟芝子實體或固態和液態培養的牛樟芝菌絲體都曾被發現具有抗氧化、抗發炎、保肝、抗酒精性肝炎、抗過敏、降血脂、神經保護及抗癌等功效。
自1991年之後，牛樟芝的研究開始起步，2000年之後漸受重視，牛樟芝人工培植有牛樟椴木栽培法，其能獲得類似野生牛樟芝之成分。另外，亦有牛樟芝固態培養法及液體培養法，其中固態培養法又稱太空包栽培法，係將牛樟芝菌種以包含有纖維物、醣類、五穀雜糧類等之太空包進行菌絲體培養。利用固體培養基栽培牛樟芝，雖然能獲得與野生子實體相近的成分，但是培養時間需要約六個月，且培養成本稍高；而液體培養法係利用醱酵槽進行菌種液體醱酵以收取菌絲體，雖具有培養時間短(約14天)的優點，但缺點是無法取得野生牛樟芝特有的三萜類成分，且與野生牛樟芝子實體所共有的成分相去甚遠。目前市面上常見的牛樟芝產品，大多是利用液態及固態醱酵生產菌絲體或子實體，其目的在於不會造成砍樹的環保顧慮。
先前技術已有嚐試將牛樟芝菌種接種於果肉或瓜果食材中進行培養，例如臺灣專利I551683揭露一種，包含將牛樟芝接種於百香果果肉，以將該百香果果肉作為用於培養牛樟芝的一種基質，其宣稱可以提升牛樟芝的多醣體含量；以及臺灣專利I551686 揭露一種牛樟芝培養方法，包含將牛樟芝植入南瓜，於24~26℃之溫度下培養3~6週，使該牛樟芝利用該南瓜所含有之營養成分生長。然而，目前尚無文獻揭露僅利用單一種穀物，尤其是糙米，做為牛樟芝的培養基質。
稻米是世界過半數人口之主食，而超過90%的世界稻作面積是在米食國家「亞洲」，其在產米國家經濟方面扮演著相當重要之角色。糙米是稻米經過簡單加工脫殼後所產的一種米，其仍保留部分外層組織，如皮層、糊粉層和胚芽；稻米中60%-70%的維生素、礦物質和大量必需氨基酸都聚集在上述的外層組織內，且擁有比白米高出6倍的膳食纖維，可以預防、減緩便秘，所以糙米向來被視為是一種健康食材。糙米中含有獨特且豐富的生育醇(tocopherols, Toc)、生育三烯醇(tocotrienols, T3)及γ-谷維素(γ-oryzanol)、γ-氨基丁酸(γ-aminobutyric acid, GABA)、阿魏酸、多酚類化學物及膳食纖維等機能性成分，具有降血脂、降膽固醇、預防心血管疾病、抗氧化及抗癌等功效。流行病學或臨床研究上，均證明這些機能性成分具有不同程度的預防疾病作用，如改善體質、調節生理機能及增進健康。
臺灣專利I331506揭露一種牛樟芝人工栽培方法，將牛樟芝菌種接種於包含白米、小米、高梁或小麥的五穀雜糧、椰子水、根莖類作物及糖類的培養基中培養長出子實體，以取代牛樟樹作為培養牛樟芝的營養源。該專利之方法雖可藉由常見穀類資材作為牛樟芝的營養源來節省栽培時間與成本，但是其培養基質組成複雜，不能當做食材使用，且培養時間長(六個月以上)，子實體之生成不穩定。本發明係首先利用糙米做為牛樟芝培養的營養基質進行醱酵培養，於短時間內生產類似牛樟芝子實體，其含有多種牛樟芝的特有化學成分，以及糙米中的主要營養成分；此外，根據本發明方法製得的牛樟芝醱酵糙米，除了可用作為保健食品之素材外，也可當作直接烹煮的食材使用。
本發明基於以上之目的發現，將牛樟芝菌種接入僅由糙米作為營養基質之培養基進行醱酵，可生產牛樟芝類子實體，並進一步分析所得之牛樟芝醱酵糙米的組成分，證實其除保有原糙米的特殊營養成分如生育醇、生育三烯醇及γ-谷維素等之外，亦包含牛樟芝的特有活性成分，包括1,4-二甲氧基-2,3-亞甲氧基-5-甲基苯(1,4-dimethoxy-2, 3-methylenedioxy-5-methyl-benzene，非三萜類)、樟菇酸C (antcin C)、2,2’,5,5’-四甲氧基-3,4,3’,4’-雙亞甲二氧基-6,6’-二甲基雙酚(2,2’,5,5’-tetramethoxy-3,4,3’,4’-bimethylenedioxy-6,6’-dimethyl- biphenyl，酚類)、樟菇酸H (antcin H)、去氫硫色多孔菌酸 (dehydrosulphurenic acid)、安卓凱因C (antrocamphin C，非三萜多炔類) 、樟菇酸B (antcin B)、樟菇酸G (antcin G)、樟菇酸A (antcin A)、15-乙醯基去氫硫色多孔菌酸(15-acetyldehydrosulfurenic acid)及去氫齒孔菌酸(dehydroeburicoic acid)等。
於是，本發明之一方面係關於，一種牛樟芝醱酵糙米，其特徵在於係將牛樟芝菌種以糙米為營養基質進行醱酵培養而得，所述經牛樟芝醱酵之糙米包含糙米的特殊營養成分及牛樟芝的特有活性成分。
於本發明之一些具體實施態樣，所述之牛樟芝醱酵糙米包含生育醇、生育三烯醇及γ-谷維素等糙米的特殊營養成分。於本發明之另一項具體實施態樣，所述之牛樟芝醱酵糙米包含1,4-二甲氧基-2,3-亞甲氧基-5-甲基苯、樟菇酸C、2,2’,5,5’-四甲氧基-3,4,3’,4’-雙亞甲二氧基-6,6’-二甲基雙酚、樟菇酸H、去氫硫色多孔菌酸、安卓凱因C、樟菇酸B、樟菇酸G、樟菇酸A、15-乙醯基去氫硫色多孔菌酸及去氫齒孔菌酸等牛樟芝的特有活性成分。
本發明之另一方面，係關於一種製備牛樟芝醱酵糙米之方法，特徵在於包含將牛樟芝菌種接入僅由糙米作為培養基質，於溫度21~23℃、濕度65~70%下進行醱酵培養一至二個月，較佳為40天，至牛樟芝類子實體完全佈滿於該培養基；及收取該牛樟芝醱酵糙米。
於本發明之一些具體實施例，所述之方法進一步包含將所收取之牛樟芝醱酵糙米於70~80℃下進行乾燥20~30小時，較佳係於70~80℃下以熱風乾燥約24小時，或以冷凍乾燥約48小時。於本發明之其他具體實施例，所述之方法進一步包含將所述經過乾燥之牛樟芝醱酵糙米研磨成乾燥糙米粉末。於本發明之另一些具體實施例，所述之方法進一步包含將所述之牛樟芝醱酵糙米以水及酒精萃取及濃縮，而得一牛樟芝醱酵糙米萃取濃縮液。於本發明之其他具體實施例，所述之方法進一步包含將所述之牛樟芝醱酵糙米萃取濃縮液進行乾燥及研磨，得一牛樟芝醱酵糙米萃取物粉末。
本發明之又另一方面，係關於一種食品組成物，其特徵在於包含所述經過乾燥之牛樟芝醱酵糙米或糙米粉末或牛樟芝醱酵糙米萃取物。
本發明使用之牛樟芝菌株包括Antrodia cinnamomea BCRC 35396(ATCC 200183) ， Antrodia cinnamomea BCRC 36401(TARI SL-7) ， Antrodia cinnamomea KJ-HL 等
。
於本說明書中所稱的“糙米”(亦稱為玄米)，係指稻榖脫去外殼後，留下胚芽與米糠的部份。所稱“糙米的特殊營養成分”係指存在糙米中對身體有益的機能性成分，主要包含生育醇(tocopherols, Toc)、生育三烯醇(tocotrienols, T3)、γ-谷維素(γ-oryzanol)、γ-氨基丁酸 (γ-aminobutyric acid, GABA)、阿魏酸、多酚類化合物及膳食纖維等。
於本說明書中所稱的“牛樟芝特有活性成分”係指特有存在牛樟芝中具有藥用生理活性的成分，主要包含多醣體(polysaccharides)、三萜類(triterpenes)、雙萜類(diterpenes)、倍半萜烯類化合物(sesquiterpenes)、倍半萜內酯(sesquiterpene lactones)、苯類(benzenoids)、苯醌衍生物(benzoquinone derivatives)、超氧歧化酶(superoxide dismutase)、多炔類(polyacetylenes)、腺苷(adenosine)、馬來酸(maleic acid)和琥珀酸(succinic acid)的衍生物，蛋白質(含免疫蛋白)、酚類化合物(phenolic compounds)、固醇類(steroids)、維生素類(如維生素B、菸鹼酸等)、微量元素(如鈣、磷、鍺等)、核酸、胺基酸、木酚素、γ-胺基丁酸(GABA)和血壓穩定物質(如antrodia acid)等。
本發明之其他特色及優點將於下列實施範例中被進一步舉例與說明，而該實施範例僅作為輔助說明，並非用於限制本發明之範圍。
實施例一、牛樟芝醱酵糙米之製備
培養基之製備：將糙米與水以1：1.1比例混合，置入玻璃廣口瓶中，每瓶含培養基重約100~120克，於121℃下進行滅菌1小時。接菌：取約2 mm x 3 mm 方塊之牛樟芝菌種(Antrodia cinnamomea KJ-HL
)或使用液態及固態菌種，接種於前述之糙米培養基。培養條件：溫度：21~23℃，濕度：65~70%，換氣頻率：每隔12小時，換氣5分鐘，二氧化碳濃度：1000~2000 ppm，於無光照下培養1至2個月。
經40天醱酵培養後，於糙米培養基已佈滿呈現淡黃色的牛樟芝類子實體，收取牛樟芝醱酵糙米，以70~80℃之熱風乾燥約24小時，或以冷凍乾燥約48小時，得到經乾燥的牛樟芝醱酵糙米。所得之經乾燥牛樟芝醱酵糙米可直接做為食材，炊煮成糙米飯或壓製成穀片，或可用於製作玄米茶的材料；亦可進一步經研磨成牛樟芝醱酵糙米粉末，添加入其他食材或飲料中作為營養調和劑，製成養生食品組成物，如養生糙米漿、糙米牛奶、糙米餅等。
實施例二、牛樟芝醱酵糙米萃取濃縮液之製備
將如實施例一所製得之牛樟芝醱酵糙米，以水及酒精作為溶劑，於95℃下進行萃取5小時，得到牛樟芝醱酵糙米萃取液。再將牛樟芝醱酵糙米萃取液以不鏽鋼篩網(150 mesh)進行過濾。
接著將經過濾之萃取液移至真空濃縮機，於溫度52±2℃進行濃縮，待濾液濃縮至濃縮標準線後，取出濃縮液，置入流動層噴霧乾燥機內(溫度62±2℃、幫浦轉速35 rpm、風車轉速40 rpm)，進行噴霧乾燥。取出乾燥之萃取物，經磨粉(篩網孔徑90 mesh)及過篩(篩網孔徑60 mesh)後，得到牛樟芝醱酵糙米萃取物粉末。
實施例三、牛樟芝醱酵糙米之稻米特殊營養成分分析
為確定糙米經牛樟芝醱酵後是否仍保有其特殊的營養成分，遂針對醱酵糙米中的生育醇(tocopherols)、生育三烯醇(tocotrienols)及γ-谷維素(γ-oryzanol)等活性成分進行測試。
活性成分萃取
取0.5g牛樟芝醱酵糙米樣品於3 mL的正己烷，混和均勻，於60℃水浴振盪萃取20分鐘。然後以2000 rpm離心15分鐘後取上清液，以同樣方法抽取兩次。最後，合併三次萃取之上清液，以0.45 μM 濾膜過濾，取濾液，以備HPLC分析(參照，Huang & Ng,Journal of Chromatography A 1218
:4709-4713, 2011)。
活性成分分析
本實例所使用之高效液相層析儀(HPLC)為Hitachi系統裝置，包括：層析幫浦(Model L-2100/2130)、紫外-可見分光偵測器(Model U-1800)及營光偵測器(Model L-2485)。糙米活性成分的分析，係利用螢光偵測器於波長290 nm (激發波長)及330 nm (發射波長)進行。分析條件如下：移動相含正己烷、異丙醇、乙酸乙酯及醋酸；分離管柱為Inertsil SiL 100A，5 μm，4.6x250 mm；移動相流速為0.7 mL/min。
結果如圖1所示，本發明之牛樟芝醱酵糙米在醱酵後，經n-己烷萃取後，進行HPLC分析，仍保有糙米的特殊營養成分，包括生育醇(圖中α-Toc代表α-生育醇，β-Toc代表β-生育醇，γ-Toc代表γ-生育醇，δ-Toc代表δ-生育醇)、生育三烯醇(α-T3代表α-生育三烯醇，β-T3代表β-生育三烯醇，γ-T3代表γ-生育三烯醇，δ-T3代表δ-生育三烯醇)及γ-谷維素(γ-oryzanol代表γ-谷維素)等。
實施例四、牛樟芝醱酵糙米之牛樟芝活性成分分析
為確定本發明之牛樟芝醱酵糙米是否含有牛樟芝的特有活性成分，遂針對牛樟芝醱酵糙米中的三萜類、非三萜類安卓凱因C及酚類等活性成分進行測試。
萃取液製備步驟：取20 mg牛樟芝醱酵糙米樣品置於1.5 mL微量離心管，加入0.8 mL 95%酒精 (1:40, w/v)，混合均勻。於室溫下，將混合物於超音波水浴中震盪1小時，之後於6000 rpm下進行離心 5分鐘，取上清液以孔徑0.45 μm濾膜過濾至1.5 mL微量離心管中，以95%酒精潤洗並定量至1 mL，將萃取液貯存於-20°C以備進行HPLC分析(參照，Lin et al.,Journal of Agricultural and Food Chemistry 59
:7626-7635, 2011)。
HPLC分析條件：管柱為Luna C18(2) (5 μm, 4.6 x 250 mm, Phenomenex)；偵測波長為254 nm；移動相含A: 0.1%醋酸(H2
O含0.1% (v/v) acetic acid)、B：甲醇 (MeOH)、C：乙腈(acetonitrile, ACN)；移動相流速如下表。 
分析結果如圖2所示。由圖2A之HPLC圖譜顯示(圖2A中a代表1,4-二甲氧基-2,3-亞甲氧基-5-甲基苯(a: 1,4-Dimethoxy-2,3-methylenedioxy-5-methylbenzene)；b 代表樟菇酸C(b: Antcin C)；c 代表2,2’,5,5’-四甲氧基-3,4,3’,4’-雙亞甲二氧基-6,6’-二甲基雙酚(c: 2,2',5,5'-Tetramethoxy-3,4,3',4'- bimethylenedioxy-6,6'-dimethylbiphenyl)；d 代表樟菇酸H(d: Antcin H)；e 代表去氫硫色多孔菌酸(e: Dehydrosulphurenic acid)；f 代表安卓凱因C(f: Antrocamphin C)；g 代表樟菇酸B(g: Antcin B)；h 代表樟菇酸G(h: Antcin G)；I 代表樟菇酸A(i: Antcin A)；j 代表15-乙醯基去氫硫色多孔菌酸(j: 15-Acetyldehydrosulfurenic acid)及k 代表去氫齒孔菌酸(k: Dehydroeburicoic acid))，即本發明之牛樟芝醱酵糙米確實含有牛樟芝的特有活性指標成分，包括1,4-二甲氧基-2,3-亞甲氧基-5-甲基苯、樟菇酸C、2,2’,5,5’-四甲氧基-3,4,3’,4’-雙亞甲二氧基-6,6’-二甲基雙酚、樟菇酸H、去氫硫色多孔菌酸、安卓凱因C、樟菇酸B、樟菇酸G、樟菇酸A、15-乙醯基去氫硫色多孔菌酸及去氫齒孔菌酸。而沒有接種牛樟芝之醱酵糙米(對照組，圖2B)，及接種牛樟芝菌種而未經醱酵之糙米(未醱酵糙米，圖2C)，其HPLC分析圖譜與空白組(圖2D)類似，皆不含有任何牛樟芝特有的活性成分。顯示，本發明牛樟芝醱酵糙米含有的牛樟芝特有活性成分，是需經過糙米與牛樟芝菌種醱酵培養所產生。
本發明首創以富含營養價值的糙米做為培養牛樟芝的營養基質，經醱酵培養後獲得同時含有糙米的特殊營養成分及牛樟芝的特有活性成分之牛樟芝發酵糙米。由於本發明僅使用糙米作為牛樟芝之醱酵培養基，其成分單純，故所得的牛樟芝醱酵糙米可直接進行烹煮食用。且糙米本身即為已知具有高營養價值的食材，經與牛樟芝醱酵培養後，除保有原糙米特殊營養成分之外，更獲得牛樟芝的特有活性成分，可藉此進一步提升其利用於製備養生產品的產業價值及廣度。
無
圖1係顯示本發明實施例一所製得牛樟芝醱酵糙米的糙米特殊營養成分分析結果。
圖2A為本發明實施例一所製得牛樟芝醱酵糙米之牛樟芝活性成分分析結果。
圖2B為未接種牛樟芝之醱酵糙米樣本(作為對照組，其醱酵培養條件與圖2A之牛樟芝醱酵糙米相同)。
圖2C為接種牛樟芝菌種而未經醱酵之未醱酵糙米樣本(作為另一對照組，其牛樟芝菌接種條件與圖2A之牛樟芝醱酵糙米相同)
圖2D為空白組。
Claims (14)
- 一種牛樟芝醱酵糙米，其特徵在於係將牛樟芝菌種以由糙米與水以1：1.1比例混合並經滅菌而得之營養基質於溫度21~23℃、濕度65~70%，換氣頻率：每隔12小時，換氣5分鐘，二氧化碳濃度：1000~2000ppm及無光照下進行醱酵培養而得，其中該牛樟芝醱酵糙米包含糙米的特殊營養成分，包括生育醇、生育三烯醇及γ-谷維素；及牛樟芝的特有活性成分，包括多醣體、三萜類、雙萜類、倍半萜烯類、倍半萜內酯、苯類、苯醌衍生物、超氧歧化酶、多炔類及腺苷。
- 如請求第1項所述之牛樟芝醱酵糙米，其中該牛樟芝的特有活性成分包括1,4-二甲氧基-2,3-亞甲氧基-5-甲基苯、樟菇酸C、2,2’,5,5’-四甲氧基-3,4,3’,4’-雙亞甲二氧基-6,6’-二甲基雙酚、樟菇酸H、去氫硫色多孔菌酸、安卓凱因C、樟菇酸B、樟菇酸G、樟菇酸A、15-乙醯基去氫硫色多孔菌酸及去氫齒孔菌酸。
- 一種製備如請求第1項所述之牛樟芝醱酵糙米的方法，其特徵在於包含：將牛樟芝菌種接入由糙米與水以1：1.1比例混合並經滅菌而得之培養基質；於溫度21~23℃、濕度65~70%，換氣頻率：每隔12小時，換氣5分鐘，二氧化碳濃度：1000~2000 ppm及無光照下進行醱酵培養一至二個月，至牛樟芝類子實體完全佈滿於該糙米培養基；及收取該牛樟芝醱酵糙米。
- 如請求第3項所述之方法，其中該牛樟芝菌種係以2mm x 3mm方塊接入該培養基，或以液態及固態菌種接入培養基。
- 如請求第3項所述之方法，其係於醱酵培養40天收取該牛樟芝醱酵糙米。
- 如請求第3項所述之方法，其進一步包含將該收取之牛樟芝醱酵糙米於70~80℃下進行乾燥20~30小時或經冷凍乾燥約48小時，而得一乾燥之牛樟芝醱酵糙米。
- 如請求第6項所述之方法，其係將該收取之牛樟芝醱酵糙米於70~80℃下以熱風乾燥約24小時。
- 如請求第6項所述之方法，其進一步包含將該乾燥之牛樟芝醱酵糙米進行研磨成一牛樟芝醱酵糙米粉末。
- 如請求第3項所述之方法，其進一步包含將該收取之牛樟芝醱酵糙米以水及酒精萃取並過濾，而得一牛樟芝醱酵糙米萃取液。
- 如請求第9項所述之方法，其進一步包含將該牛樟芝醱酵糙米萃取液進行濃縮，而得一牛樟芝醱酵糙米萃取濃縮液。
- 如請求第10項所述之方法，其進一步包含將所述之牛樟芝醱酵糙米萃取濃縮液經噴霧乾燥及研磨，得一牛樟芝醱酵糙米萃取物粉末。
- 一種食品組成物，其特徵在於包含如請求第1項所述之牛樟芝醱酵糙米。
- 一種食品組成物，其特徵在於包含如請求第8項所述之方法製得的牛樟芝醱酵糙米粉末。
- 一種食品組成物，其特徵在於包含如請求第11項所述之方法製得的牛樟芝醱酵糙米萃取物粉末。
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| TW106146655A TWI689258B (zh) | 2017-12-29 | 2017-12-29 | 牛樟芝醱酵糙米，其製法及其組成物 |
| JP2018075501A JP6546678B1 (ja) | 2017-12-29 | 2018-04-10 | ベニクスノキタケ発酵玄米の製法及びその組成物 |
| US16/039,487 US20190200653A1 (en) | 2017-12-29 | 2018-07-19 | Antrodia cinnamomea fermented brown rice, preparing method and composition thereof |
| US17/136,540 US20210112832A1 (en) | 2017-12-29 | 2020-12-29 | Antrodia cinnamomea fermented brown rice, preparing method and composition thereof |
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