生產治黴色基素之方法

本發明係一種生產治黴色基素之方法，其係提供一培養基，包括約0.01至2.0%脫脂黃豆粉、約0.01至2.0%葡萄糖及約0.01至1.0%碳酸鈣，將鏈黴菌(Streptomyces padanus PMS－702)培養於該培養基質中，設定培養條件為轉速約100至400rpm、通氣量約0.5至1.5vvm及培養基質呈弱酸性，並於培養過程中添加油脂或脂肪酸或其混合物於培養基質中，進行大量液態培養，以獲取治黴色基素

應用液態培養StreptomycespadanusPMS-702生產fungichromin─醱酵槽培養條件探討

In this study, a novel strain named Streptomyces padanus PMS-702, harbouring antifungal activity, was used to produce polyene macrolide antibiotic, fungichromin in various cultivation conditions. The antibiotic fungichromin was found to be an active ingredient for the control of Rhizoctonia damping-off of cabbage and tomato late blight. It was also found useful in the clinical cancer treatment. The main purpose of this research was to study the effect of different cultivation conditions on the production of fungichromin in submerged culture using 5-L fermentor. Under the cultivation conditions of agiation rate 300rpm、aeration 1 vvm、and without pH control,the highest fungichromin production of 231.27 mg/L was obtained after 4.5 day’s cultivation.To conduct the nutrition addition study in flask , it was found the supplement with oil could enhance the production of fungichromin enormously.本研究中，使用具有抗真菌活性的鏈黴菌菌株Streptomyces padanus PMS-702，進行多烯大環內酯類抗生素fungichromin 之醱酵生產研究。此抗生素對於立枯絲核菌、蕃茄晚疫病菌有良好的抑制效果，也可應用於如腫瘤等之臨床醫療上。 本研究主要探討於醱酵槽中液態培養中，不同培養條件對Streptomyces padanus PMS-702生產fungichromin之影響。由醱酵槽試驗結果發現，以轉速300rpm、通氣量1vvm、不控制pH值的條件下，經培養4.5天後，fungichromin產量可達231.27mg/L。而以饋料方式饋入碳源，則無明顯提升fungichromin產量結果出現。此外在搖瓶添加物試驗結果發現，添加油脂可大為提升fungichromin產量。目錄 中文摘要 I 英文摘要 II 目錄 III 表目錄 VII 圖目錄 VIII 第一章、緒論 1 1-1、前言 1 第二章 文獻回顧 4 2-1、抗生素 4 2-1-1、抗生素分類與作用機制 4 2-1-2、抗生素之應用 7 2-1-3、抗生素產生方式 7 2-1-4、抗生素製程與一般醱酵製程之比較 10 2-1-5、提高抗生素產量的方法 11 2-2、鏈黴菌 13 2-2-1、鏈黴菌屬在生物防治上之應用 15 2-2-2、鏈黴菌屬產物之特性 16 2-3、Fungichromin之介紹 17 2-3-1、Fungichromin之結構及特性 17 2-3-2、Fungichromin的相關研究 19 2-4、饋料批次式醱酵之程序 21 第三章 材料與方法 32 3-1-1、菌種 32 3-2、菌種保存 32 3-3、基礎培養基搖瓶實驗 32 3-4、醱酵槽批次式液態醱酵 33 3-4-1、不同轉速對S. padanus PMS-702於醱酵槽中批次醱酵之影響 33 3-4-2、不同通氣量對S. padanus PMS-702於醱酵槽中批次醱酵之影響 33 3-4-3、調控不同pH值對S. padanus PMS-702於醱酵槽中批次醱酵之影響 33 3-5、醱酵槽饋料批次式醱酵 34 3-5-1、探討葡萄糖一次饋料添加對S. padanus PMS-702於醱酵槽中批次醱酵之影響 34 3-5-2、探討葡萄糖持續饋料添加對S. padanus PMS-702於醱酵槽中批次醱酵之影響 34 3-6、搖瓶添加物實驗 35 3-6-1、添加不同離子 35 3-6-2、添加不同脂肪酸 36 3-6-3、添加不同油脂 36 3-6-4、饋料方式添加油脂 37 3-7、最佳醱酵條件於醱酵槽中應用探討 38 3-8、分析方法 38 3-8-1、溶氧電極（DO sensor）之校正 38 3-8-2、酸鹼電極（pH sensor）之校正 39 3-8-4、基質殘糖（glucose）量分析 40 3-8-5、Fungichromin定量分析 41 第四章 結果與討論 46 4-1、醱酵槽批次式液態醱酵 46 4-1-1、不同轉速對S. padanus PMS-702於醱酵槽中批次醱酵之影響 46 4-1-2、不同通氣量對S. padanus PMS-702之影響 52 4-1-3、控制不同pH值對S. padanus PMS-702批次醱酵之影響 56 4-2、醱酵槽饋料批次式醱酵 61 4-2-1、探討葡萄糖一次饋料添加之影響 61 4-2-2、探討葡萄糖連續饋料添加對S. padanus PMS-702於醱酵槽中批次醱酵之影響 65 4-3、搖瓶添加物探討 69 4-3-1、探討離子添加對Streptomyces padanus PMS-702於搖瓶醱酵之影響 69 4-3-2、探討油脂添加對Streptomyces padanus PMS-702於搖瓶醱酵之影響 75 第五章 結論與展望 89 5-1、結論 89 5-2、未來展望 91 參考文獻 92 附錄 96 表目錄 表2-1、培養基中解除碳源、氮源回饋抑制作用之方法 22 表2-2、不同抗生素之純化方法 23 表2-3、去除副產物之方法 24 表2-4、大環內酯類化合物之共軛雙鍵數目及最大紫外光吸收光譜值 25 表2-5、fungichromin之理化性質 26 表2-6、fungichromin與lagosin及cogomycin的分子式及物理性質 27 表2-7、各種不同饋料策略整理表格 29 表2-8、Streptomyces padanus PMS-702菌株之抑菌族譜 30 表2-9、1990年後應用放線菌防治植物病害之案例 31 表3-1、International Streptomyces Project Medium (ISP4) 42 表3-2、活化培養基組成 42 表3-3、醱酵槽培養基組成 42 圖目錄 圖2-1、Fungichromin之結構式 28 圖3-1、葡萄糖濃度分析流程 43 圖3-2、Fungichromin定量分析流程 44 圖3-3、本實驗所使用之醱酵槽系統 45 圖4-1-1-1、醱酵槽批次培養S. padanus PMS-702不同轉速對溶氧量之影響 49 圖4-1-1-2、醱酵槽批次培養S. padanus PMS-702不同轉速對pH值之影響 49 圖4-1-1-3、不同轉速對醱酵槽批次培養S. padanus PMS-702葡萄糖利用之影響 50 圖4-1-1-4、不同轉速對醱酵槽批次培養S. padanus PMS-702 fungichromin產量之影響 50 圖4-1-1-5、不同培養時間下，S.padanus PMS-702之菌體型態 51 圖4-1-2-1、不同通氣量對醱酵槽批次培養S. padanus PMS-702 DO之影響 54 圖4-1-2-2、不同通氣量對醱酵槽批次培養S. padanus PMS-702 pH值之影響 54 圖4-1-2-3、不同通氣量對醱酵槽批次培養S. padanus PMS-702葡萄糖利用之影響 55 圖4-1-2-4、不同通氣量對醱酵槽批次培養S. padanus PMS-702 fungichromin 產量之影響 55 圖4-1-3-1、醱酵槽批次培養S. padanus PMS-702不同pH值對DO之影響 59 圖4-1-3-2、醱酵槽批次培養S. padanus PMS-702不同pH值對葡萄糖利用之影響 59 圖4-1-3-3、醱酵槽批次培養S. padanus PMS-702不同pH值對fungichromin 產量之影響 60 圖4-2-1-1、葡萄糖一次饋料添加對S. padanus PMS-702之溶氧值與pH值之影響 63 圖4-2-1-2、葡萄糖一次饋料添加對S. padanus PMS-702之葡萄糖利用與fungichromin產量之影響 64 圖4-2-2-1、葡萄糖連續饋料添加對S. padanus PMS-702之溶氧值與pH值之影響 67 圖4-2-2-2、葡萄糖連續饋料添加對S. padanus PMS-702之葡萄糖利用與fungichromin產量之影響 68 圖4-3-1-1、基礎培養基中添加Fe2+對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 72 圖4-3-1-2、基礎培養基中添加Mg2+對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 72 圖4-3-1-3、基礎培養基中添加Zn2+對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 73 圖4-3-1-4、基礎培養基中添加Cu2+對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 73 圖4-3-1-5、基礎培養基中添加PO42-對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 74 圖4-3-1-6、基礎培養基中添加SO42-對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 74 圖4-3-2-1、基礎培養基中添加oleic acid對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 80 圖4-3-2-2、基礎培養基中添加stearic acid對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 80 圖4-3-2-3、基礎培養基中添加linoleic oil對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 81 圖4-3-2-4、基礎培養基中添加corn oil對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 81 圖4-3-2-5、基礎培養基中添加cottonseed oil對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 82 圖4-3-2-6、基礎培養基中添加soybean oil對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 82 圖4-3-2-7、饋料添加oleic acid對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 83 圖4-3-2-8、饋料添加soybean oil對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 83 圖4-3-2-9、饋料添加cottonseed oil對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 84 圖4-3-2-10、饋料添加corn oil對S. padanus PMS-702生產fungichromin及菌絲乾重之影響 8

高敏感特質中高齡學習者面對數位學習壓力及疫情焦慮之學習滿意度與後疫情調適：復原力之中介效果 Learning Satisfaction and Post-Pandemic Adaptation in Highly Sensitive Middle-aged and Elderly Learners in the Face of Digital Learning Stress and Pandemic Anxiety

人口老化趨勢及近年疫情大流行加速了中高齡數位學習之推動與普及，在此環境改變下，對高敏感特質的長輩是一大考驗。本研究旨在探討高敏感特質之中高齡者於疫情變化使數位學習環境更為普及之狀況下，所產生的數位學習壓力、心理憂鬱及學習滿意度的關聯，並以復原力作為中介變項，最後亦探討疫情趨緩後中高齡者在後疫情之感受。本研究採用問卷調查法進行資料蒐集，研究工具為高敏感特質、疫情焦慮、數位學習壓力、憂鬱情緒量表、學習滿意度、復原力及後疫情正向感受等7份量表；以45歲以上有數位學習經驗的中高齡者為研究對象，共回收208份有效問卷（男72位，女136位，平均年齡57.93，標準差6.79），調查所得資料採用結構方程模型（Structural Equation Modeling, SEM）中Bootstrap統計方法進行中介模式分析。研究結果顯示：一、中高齡者高敏感特質程度愈高，疫情焦慮愈高、數位學習壓力愈大；二、復原力在疫情焦慮與心理憂鬱間具完全中介效果，疫情焦慮愈高，使復原力愈差，造成心理憂鬱愈高；復原力在數位學習壓力與心理憂鬱、學習滿意度間具部分中介效果，數位學習壓力愈大，使復原力愈差，造成心理憂鬱愈高、學習滿意度愈不佳；三、中高齡者心理憂鬱愈高、學習滿意度愈不佳，後疫情正向感受愈低。根據結果，本研究對未來的研究者提出相關建議，作為日後研究之參考。 Introduction Population aging trends and the accelerated promotion and popularization of digital learning in recent years, exacerbated by the global pandemic, have posed significant challenges and impacts to middle-aged and elderly learners on mental health, learning satisfaction, and psychological adoption, especially for middle-aged and elderly individuals with highly sensitive traits. This study aims to investigate the relationship between digital learning stress, pandemic anxiety, depression, learning satisfaction and post-pandemic experiences in highly sensitive middle-aged and elderly learners in the context of the widespread adoption of digital learning due to the pandemic, and explore the role of resilience as a mediating variable. Literature Review Highly Sensitive People (HSP) was first defined by Aron & Aron (1997) as “the in-depth processing (including unconsciousness) of internal and external stimuli under various circumstances, resulting in higher physiological and emotional responses.” The scientific term is “Sensory Processing Sensitivity (SPS)” People with high sensitivity are more susceptible to stimuli and are less likely to have adverse effects under stress than in the past. However, most studies on highly sensitive trait populations focus on children (Pluess & Boniwell, 2015) and adolescents (Iimura, 2022) as research subjects, there is a lack of research focusing on the elderly as the main research subjects. Organizations and nations worldwide recognize the critical importance of addressing population aging and actively respond to this challenge. Taiwan officially entered an ‘aging society’ in 1993, as the proportion of elderly population exceeded 7%. Sadan et al. (2021) observed that older adults exhibit a weaker adaption capacity to environmental changes. During environmental change, older adults suffered higher pressure. Highly sensitive trait populations are more likely to have negative reactions when stimulated by negative stressful environments (Aron & Aron, 1997). The trait is positively correlated with stress (Andresen et al., 2018; Gearhart & Bodie, 2012; Meyerson et ai., 2020). Systematic review studies indicate that SPS increases the risk of stress-related problems when responding to negative environments, but can also gain greater benefits from positive and supportive experiences (Greven, 2019). Therefore, this study aims to investigate middle-aged and elderly with highly sensitive traits, to explore if this population manifests similarly. In 2020, due to the COVID-19 pandemic, the number of people suffering from depression increased by 25.6% globally (Santomauro et al., 2021). Many research have highlighted the negative impact of the epidemic on mental health. With psychological depression interventions, the life satisfaction and health status of middle-aged and elderly people can be improved, and loneliness can be alleviated (Jo & Kim, 2008). When life satisfaction is significantly improved, depression will decrease (Lee, 2020; MAltun & Yazici, 2015). Reviewing the meta-analysis and literature review related to depression and anxiety, studies have pointed out that brief passive psychoeducation intervention for depression and psychological distress can reduce symptoms (Donker, 2009), and physical activity can reduce depression and anxiety in non-clinical populations (Duncan, 2015). Furthermore, relevant research has pointed out that resilience can serve as a mediating factor that affects the relationship between environmental stress and mental health. (Chang, 2012; Egeland et al., 1993; Faircloth, 2017; Kim et al., 2018; Liu et al., 2017). Resilience also mediates the relationship between Covid-19 stress and mental health (including depression and anxiety). The higher the stress related to the epidemic, the lower the resilience, and the higher the depression and anxiety (Ho et al., 2022; Kumar et al., 2022). As discussed above, resilience is one of the key factors for individuals facing negative environmental stress (including pandemic anxiety, and digital learning stress), which in turn affects their psychological adaptation. Methodology Data collection for this study was conducted through a questionnaire survey targeting individuals aged 45 and above who have experience with digital learning. The questionnaire included 7 scales: highly sensitive people traits, pandemic anxiety, digital learning pressure, depression, learning satisfaction, resilience, and post-epidemic positive moods. The pre-test was conducted in Dec. 2022 by digital questionnaires. A total of 106 valid responses were collected (41 male, 65 female, mean age = 60.00, SD = 1.41). All scales were subjected to exploratory factor analysis after the pre-test. Internal consistency was confirmed and the questions in these constructs were stable and consistent. A total of 208 valid responses were collected (72 male, 136 female, mean age = 57.93, SD = 6.79). The collected data were analyzed using structural equation modeling (SEM) with the Bootstrap statistical method for mediation analysis. Finding The results of the study revealed the following key findings: 1. Middle-aged and elderly individuals with higher levels of highly sensitive traits experienced higher levels of pandemic anxiety and digital learning stress. 2. Resilience played a significant mediating role in the relationship between pandemic anxiety and depression. Higher pandemic anxiety was associated with lower resilience, leading to increased depression. Additionally, resilience partially mediated the relationship between digital learning stress and depression as well as learning satisfaction. Greater digital learning stress was linked to reduced resilience, resulting in higher depression and lower learning satisfaction. 3. Elevated levels of depression and lower learning satisfaction among elderly individuals were associated with reduced post-pandemic positive experiences. Discussion and Suggestion Finally, several suggestions were proposed for future studies. The current study shows that the role of resilience plays in mediating pandemic anxiety and psychological depression. As well as the relationship between digital learning stress, psychological depression, and learning satisfaction. This suggests that for highly sensitive middle-aged and elderly learners, the higher the pandemic anxiety, the worse resilience, and the psychological depression are. The higher the digital learning stress, the worse resilience, and the psychological depression is, the worse learning satisfaction. In addition, after the peak of the pandemic, the post-pandemic positive experiences are still poor. The study’s findings provide the psychological state insights of middle-aged and elderly learners with highly sensitive traits during the pandemic and e-learning environment. Future studies may conduct further research to develop more effective support strategies to enhance the resilience of the middle-aged and elderly. Which may help people face the challenges, reduce psychological depression, and increase their learning satisfaction. Also, improve the post-pandemic positive experiences, and mental health