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1 發酵時間對猴頭菌固態發酵燕麥之活性成分影響陳淑德教授國立宜蘭大學食品科學系兩岸穀物與雜糧健產業研討會 1

2 Health foods oat ( 台灣的燕麥健康食品有十四種 ) 調節血脂功能產品 : (1) 桂格原片原味大燕麥片 (2) 桂格三寶燕麥 (3) 桂格黃金麩片燕麥片 (4) 機能燕麥麵 (5) 桂格養生燕麥麵 (6) 桂格美味大燕麥片 - 楓糖口味 (7) 統一陽光高纖燕麥穀奶 (8) 愛之味純濃燕麥 ( 天然原味 ) (9) 高野家燕麥片調節血脂功能, 不易形成體脂肪功能 : (1) 桂格即沖即食大燕麥片 (2) 桂格 100 % 喝的燕麥調節血糖功能 : (1) 桂格機能燕麥片調節血脂功能, 免疫調節功能 : (1) 桂格美味大燕麥片 ( 水果優格口味 ) 2

愛之味純濃燕麥 ( 天然原味 ) (9) 高野家燕麥片調節血脂功能, 不易形成體脂肪功能 : (1) 桂格即沖即食大燕麥片 (2) 桂格

3 高齡化社會 65 歲的老年人口只要超過全國總人口的 7% 即為高齡化社會台灣自 1993 年起邁入高齡化社會以來,65 歲以上老人所占比率持續攀升,2012 年底已達 11.2% 2012 年神經退化性疾病 : 阿滋海默症 ( 失智症 ) 和巴金森氏症分別佔台灣 65 歲以上老年人口的 0.6% 和 1-2 % 健康食品延緩衰老評估功能 (2013 年 11 月新修正草案 ) 已增加神經退化性疾病的評估方法, 如 : 誘導腦部組織退化的阿滋海默症 ( 失智症 ) 和巴金森氏症的動物模式 3

4 Neurodegenerative disease Endoplasmic reticulum stress (ER stress) Parkinson s disease Alzheimer s disease (Alzheimer's Association 2008) 4

5 Hericium erinaceum ( 猴頭菇 ) classification Fungi ( 真菌界 ) Eumycota ( 真菌門 ) Basidiomycotina ( 擔子菌亞門 ) Hymenomycetes ( 層菌綱 ) Aphyllophorales ( 非褶菌目 ) Hydnaceae ( 齒菌科 ) Hericioideae ( 猴頭菌亞科 ) Hericieae ( 猴頭菌 ) Hericium ( 猴頭菌屬 ) 5 Fresh fruiting body Dried fruiting body

Aphyllophorales ( 非褶菌目 ) Hydnaceae ( 齒菌科 ) Hericioideae ( 猴頭菌亞科

6 Hericium sp. alternate name China name: 刺蝟菇白髮菇花頭菌花菜菌 Japan name: 獅子茸獅子菇 yamabushitake( 山伏茸 ) English name:tree hedgehog, lion s mane (Pegler, 2003) 中國四大名菜 : 海蔘猴頭菇燕窩熊掌 6

7 Bioactive components of H. erinaceum Polysaccharides 多醣 Proteins 蛋白質 Terpenoids 三萜 Hericenone 猴頭酮 Erinacine 猴頭素 (Herre et al., 2004; Hazekawa et al., 2010) 7

8 Immunological stimulation of H. erinaceus polysaccharide Mushroom s polysaccharide could increase IL-1 and IL-2 secretion, increase T-cells content. (Wang et al., 2001; 吳等,2005) H. erinaceus and H. laciniatum polysaccharide could increase total T-cells CD4 + cells CD8 + cells and macrophages content. (Wang et al., 2001) H. erinaceus hot water extracts induced the IFN-g and IL-12, NO expression, by the activation of NK cell. (Yim et al., 2007) H. erinaceus hot water extracts induced the NO production and IL-1-b expression. (Son et al., 2006) 8

9 Bio-functions of hericenone and erinacines Promoting nerve growth factor (NGF) synthesis. Treatment and prevention of dementia. (Mori et al., 2008) Neuroprotective effects. (Hazekawa et al., 2010) Erinacine can stimulate mouse astrogliai cells to secrete NGF more than hericenone. (Kawagishia et al., 1991, 1994)

, 2008) Neuroprotective effects. (Hazekawa et al.

10 NGF ( 神經生長因子 ) synthesis by erinacine Concentration NGF (pg/ml) Erinacine A 1.0 mm ± 36.2 Erinacine B 1.0 mm ± 6.5 Erinacine C 1.0 mm ± 59.6 (Kawagishi et al., 1994) Epinephrine (Control) 1.0 mm 69.2 ± 17.2 Erinacine E 5.0 mm 105 ± 5.2 (Kawagishi et al.,1996) Erinacine F 5.0 mm 175 ± 5.2 Epinephrine (Control) 5.0 mm 70.2 ± 5.4 Erinacine H 33.3 mg/ml 31.5 ± 1.7 (Lee et al., 2000) 10

, 1994) Epinephrine (Control) 1.0 mm 69.2 ± 17.2 Erinacine E 5.0 mm 105 ± 5.2 (Kawagishi et al.

11 Effects of mushroom extracts on the secretion of NGF from 1321N1 cells. H. erinaceus (H) ( 猴頭菇 ), P. eryngii (P)( 杏鮑菇 ), G. frondosa (G) ( 舞茸 ), A. blazei (A)( 巴西蘑菇 ) (Mori et al., 2008) 11

12 0 hpf 24 hpf 48 hpf HEE treatment 1% EtOH staining a a b b cd c cd d HEE (ppm) control EtOH (%) 1% 1% 1% 1% 1% 1% 1% Effect of pretreatment with different concentrations of ethanol extracts from H. erinaceum solid-state fermented product (HEE) on the number of died brain neuron cells in zebrafish embryos. (mean ± S. D., n = 6) 12

different concentrations of ethanol extracts from H.

13 0 hpf 24 hpf 48 hpf water 1% EtOH HEE staining Effect of post-treatment with 200 ppm H. erinaceum fermented product ethanol extracts (HEE) on the number of died brain neuron cells in zebrafish embryos. (mean ± S. D., n = 6) 13

14 Cultivation of H. erinaceum Solid-state cultivation Submerged fermentation Fruiting body Mycelium Mycelium 14

15 Preparation of the Hericium erinaceus liquid and solid-state fermented product Hericium erinaceus BCRC Liquid PDA dish, cultivation at 25 o C 5% glucose, 0.1%Yeast extracts, 0.1% K 2 HPO 4, 0.05% MgSO 4 Pre-activation (ph 5-6, 150rpm, at 25 o C ) Oat solid substrate, cultivation at 25 o C 121 o C, 20 min sterilization 50 o C hot air drying and milling H erinaceus solid-state oat powder product 15

05% MgSO 4 Pre-activation (ph 5-6, 150rpm, at 25 o C ) Oat solid substrate, cultivation at 25 o C 121

16 Change of mycelia dry weight during 14 days H. erinaceus submerged fermentation (pre-activation) mg/ml Biomass (mg/ml) Time (day) 16

17 Change of polysaccharide content during 14-days H. erinaceus submerged fermentation (pre-activation) mg/ml Polysaccharide (mg/ml) Time (days) 17

18 Crude polysaccharide (%) Effects of cultivation time on crude polysaccharide of H. erinaceus solid-state fermented oat product % ab a b b Days b b 18

19 Crude triterpenoids (%) Effects of cultivation time on crude triterpenoids of H. erinaceus solid-state fermented oat product. 2.14% a 1.5 b c e d d Days 19

20 Erinacine A (%) Effect of cultivation time on erinacine A of H. erinaceus solid-state fermented oat product b 0.11% ab a a c d Days 20

21 NGF (pg) Effect of adding different concentration H. erinaceum fermentation product ethanol extracts on PC12 cells producing NGF after 72 h cultivation. Control a a a b H. erinaceum 10 ppm Blank Concentration (ppm) 21

22 Conclusions Pre-activated H. erinaceus conditions: 5% glucose medium, ph 6.0 and 5-days fermentation The optimal crude polysaccharides, triterpenoids, and erinacine A contents were in Hericium erinaceus 35-days solid-state fermented oat product. The addition ethanol extracts from Hericium erinaceus fermented products significantly improved secretion of NGF and neurite outgrowth of PC12 cells. 22

23 誌謝農糧署的 2008~2010 的猴頭菌子實體和發酵產物萃取液化學組成和生理功能之探討科技計畫經費支助參與此計畫的宜蘭大學食品科學系研究生 : 林玉婷呂宛儒何純誼和顏威揚 23

24 Thanks! 歡迎聆聽謝謝大家! 24

壹、摘要

壹、摘要中華民國第四十五屆中小學科學展覽會作品說明書高職組農業及生物科技科最佳團隊合作獎 091404 當菜遇到牛奶國立苗栗高級農工職業學校作者姓名 : 職二吳韋璇職二李婌玉職二陳雨暄職二陳思涵指導老師 : 許智凱 1 ..P.2..P.2..P.3..P.3..P.3..P.12..P.21..P.22..P.22