建立安全有效的神麴發酵製程最佳化的研究

鄧正賢
亞州大學 保健營養生技學系
本計畫之目的在於完整建立中藥材神麴規格化發酵製程，並與古典籍收載傳統發酵條件比較，以液相層析法比較指標成份苦杏仁苷 (Amygdalin)、蒼耳總酚酸 (Phenolic acids)、青蒿素(artemisinin)差異，並以蛋白酶 (Protease)、澱粉酶 (Amylase) 與脂解酵素 (lipase) 為活性指標，以黃麴毒素 (AflatoxinB1 B2 G1 G2) 污染為負指標，篩選優良發酵菌組，建立安全有效的神麴發酵製程最佳化研究。本計畫為期一年，以不同領域研究團隊的方式，分三大部分進行進行。(一) 子計畫ㄧ：神麴發酵菌種分離及純化：著重於建立神麴菌種分離及純化，並進行菌數檢測，包括生菌數、酵母菌菌數、黴菌菌數等。(二) 子計畫二：神麴傳統發酵條件與單一菌種發酵的技術評估：以蛋白質酶 (Protease)、澱粉酶 (Amylase)、脂解酵素 (lipase) 活性測定為指標，並以黃麴毒素污染為負指標，篩選獲得優良發酵菌組，包括接種量、水含量、發酵溫度與發酵時間，並比較指標成份苦杏仁苷 (Amygdalin)、蒼耳總酚酸 (Phenolic acids)、青蒿素(artemisinin) 差異。(三) 子計畫三：神麴優良菌株基因序列分析：進行核糖體基因 ITS 序列分析及粒線體基因限制片段多型性分析，並視情況進行微衛星體序列大小(SSR)、擴大複製片段長度多型性 (AFLP)、逢機複製片段長度多型性 ( RAPD) 等，以確定優良菌種的基因序列，確定種原並保存技術。
關鍵字：神麴、發酵、基因序列

The optimal fermentation process for developing Massa Medicata Fermentata

Jeng-Shyan Deng Ph.D
Department of Health and Nutrition Biotechnology, Asia University
Evidence has suggested that mycotoxins contaminated in Shenqu produced in traditional processing, related to exposure fungal in fermented preparation. Shenqu was naturally contaminated by fungi that may become toxic for the human organism if the total amount ingested through consumption exceeds a certain tolerable dose. The quality management of Shenqu materials is very important for protectting consumer health. Therefore, a new process for developing Massa Medicata Fermentata was recommended in order to assess the optimal conditions for microbial fermentation of Shenqu.1.The sub-subject I: To culture and determine the kinds of beneficial microorganism in Shenqu fermentation process. The first part comprises the basic research dealing with identifing and classifing the microorganisms in Shenqu fermentation process. Beside, total plate count test were performed and yeast and mold testing represents the total fungi in a sample. 2.The sub-subject II: To evaluate the effect of isolated unidentified Lactobacillus strains and pure inoculation fermentation. The objective of sub-subject II was to evaluate the effect of isolated unidentified Lactobacillus strains on promoting digestion. The active ingredients evaluated were lipase, protease and amylase, and screen out the microorganisms for the presence of aflatoxins. Improving the traditional fermentation process was followed at various conditions including the inoculum amount, fermentation time, temperature and water content. 3.The sub-subject III: DNA sequencing analysis for the species identification of isolated yeast. Identification of Shenqu important yeasts by ITS sequencing, especially using the ITS region, is reliable and can be used as an accurate alternative to conventional identification methods. Microsatellite PCR, AFLP and RAPD techniques were further utilized for yeast isolates subtyping.
關鍵字：Massa Medicata Fermentata, fermentation, DNA sequence