類別:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/88
2024-03-29T07:02:36Z龍鬚菜之酵素降解
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/33203
標題: 龍鬚菜之酵素降解; Enzymatic Degradation of Gracilaria
作者: Yung-Te Hou; 侯詠德
摘要: 本研究探討以纖維素水解酶(Cellulase)水解醣類生質(蔗渣、龍鬚菜)的最佳條件(溫度50℃,pH 4.6)。並且比較醣類生質未經前處理及經過酸處理(基質加入0.2 M H2SO4,以殺菌釜加溫至121℃處理半小時)、冷凍前處理(基質加入去離子水浸泡,並於-10℃下靜置20分鐘)後的酵素水解速率差異。醣類生質經酵素水解成單醣、雙醣及寡醣後,再利用DNS (Dinitrosalicylic acid)定量還原端產生的量,以推估酵素水解效率。另外,利用酚-硫酸法以及葡萄糖感測器來定量酵素水解後產生的總醣濃度及葡萄糖濃度。本研究所開發的海藻生質,在經過冷凍前處理後確實提高了酵素水解速率以及醣類增加量,可望有效應用於生質能源領域。; Lignocellulosic biomass has been utilized to produce ethanol in the last two decades. The main process involved in this conversion is hydrolysis of cellulose in the lignocellulosic materials to produce reducing sugars. Due to the recalcitrant nature, the biomass can not be easily converted to monomeric sugars. Therefore, the low yield and high cost of the hydrolysis process are the major challenges in biomass energy development. In our research, marine seaweeds Gracilaria is used as the biomass. Owing to their hydration ability, an “Ice-crystal pretreatment” is developed to enhance the hydrolysis of cellulosic biomass.
The enzymatic hydrolysate of biomass was measured by three methods. Reducing sugars is determined by the dinitrosalicylic method (DNS). Total soluble sugars is determined by the phenol-sulphuric acid method. Glucose is determined by the glucose biosensor. In our result, the validity of “Ice-crystal pretreatment” were confirmed. The effect of “Ice-crystal pretreatment” was also compared with acid pretreatment. In conclusion, marine seaweeds Gracilaria with “Ice-crystal pretreatment” may be a new approach to biomass energy development in the future.2006-01-01T00:00:00Z鼓勵銀髮族做肢體運動與認知能力訓練之娛樂系統
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77853
標題: 鼓勵銀髮族做肢體運動與認知能力訓練之娛樂系統; An Entertainment System to Motivate the Elderly to Do Physical and Cognitive Hybrid Activities
作者: Yu-Hsiang Lin; 林毓祥
摘要: 在地老化的長照理念在全世界已經越來越盛行,為了讓長者可以在自己的居住地安全地、舒適地且獨立地生活,本研究希望能減緩長者在運動與認知能力上的退化。為了達到這個目的,本研究利用感測器、微控制器以及遊戲開發的技術開發了一個適合長者的娛樂系統。這個系統可以輕易的讓長者採用,並且可以取得長者在認知能力上的重要資訊,這些資訊可以做為未來相關專家分析之用。這個娛樂系統包含一個特殊的體感控制器、一個節奏遊戲以及遊戲資料分析的方法。首先,長者玩家要利用拍打功的動作例如搖擺雙手與經穴拍打來激發這個體感遊戲控制器。統計研究結果顯示這樣的動作對於玩過我們娛樂系統的長者來說具有吸引力。另外,長者還必須要克服節奏遊戲中的各項挑戰。每項挑戰都是由三種遊戲情境所構成,每個遊戲情境都可以特別針對長者的反應時間、短期記憶以及工作記憶等認知能力加以刺激。在遊戲互動的過程中,本娛樂系統會將長者的遊戲行為資料記錄下來。最後,本研究提出一套資料分析的方法來評鑑長者在遊戲中的反應時間、短期記憶與工作記憶這三種認知能力,進行量化分析。此外,本系統除了成本低廉、操作親和外,模組化的結構容許在軟硬體上做快速的擴充,易於大量的推廣。; In this research, we attempt to assist the elderly to live in their personal house safely, comfortably and independently by retaining their cognitive and physical abilities. We utilize sensors, a microcontroller, gaming technologies to create a cost effective entertainment system. The system can be easily adopted by the elderly and can acquire useful information of their cognitive skills for further analysis by the specialists or diagnosis systems. The system consists of a special motion-sensing controller, a rhythm game and a gameplay data analysis approach. Firstly, to interact with our system, the elderly players are asked to perform physical actions like hands flapping and acupressure points’ slapping to activate the motion sensing controller. Statistical results show this kind of physical motion is attractive to the elderly having played our system. Secondary, the elderly players need to overcome challenges from the rhythm game. The challenges are composed of three game scenarios. Each game scenario can activate a specific cognitive skills like response time, short-term memory or working memory. During the game’s playing, the system will record the elderly’s gameplay data. With the gameplay data, we propose a data analysis approach to evaluate the elderly players’ performances in response time, short-term memory and working memory. The approach can quantify each cognitive skill mentioned above. Finally, the system is available and friendly for the elderly to use. Besides, the modular design allows the system to be extended or modified easily and rapidly. Therefore, it can be widely promoted to the public in future.2017-01-01T00:00:00Z黏蛋白-1適體篩選與分析
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/54128
標題: 黏蛋白-1適體篩選與分析; Mucin-1 Aptamer Selection and Analysis
作者: Wei-Chen Huang; 黃惟甄
摘要: 黏蛋白-1 (mucin-1,簡稱MUC1)是一種在多數腺癌細胞表面上過表現的穿膜醣蛋白,可作為一癌症指標,不論在診斷或治療上皆具有重要意義。由於目前仍未有以MUC1部分開放閱讀框 (ORF)之序列 (315 a.a. - 420 a.a.) 為目標的適體。根據其未來之應用性,本研究欲篩選一可適用於生物感測的適體,在修飾之下仍可與MUC1結合。因此,本研究將以此序列片段作為目標,進行MUC1之適體篩選。篩選利用的方法為系統性配位子指數增益演繹技術,以單顆玻璃珠作為反應基材,人類血清白蛋白作為反向篩選的目標,藉由多回合的擇選與擴增,得到與MUC1具親和性與選擇性的特定單股 DNA群;並以MEME分析軟體將其定序結果進行分析與挑選,最後利用表面電漿共振 (surface plasmon resonance,簡稱SPR) 之親和力測試,找出與MUC1親和力最高的序列,也就是DNA適體。篩選以演化樹來表示其序列演化的過程,其中包含K1、K2、K3三個分支。篩選過程中發現非專一性序列於序列群中的數量,可以作為演化過程中優勢序列的預測指標。根據SPR中每一個序列與MUC1反應的訊號及其組成,發現GC/AT比與序列之親和性具有中度相關 (r = 0.499)。本研究成功篩選出一個MUC1適體,研究中將其命名為K1R4.2,此適體不會因為加上修飾物而改變其結合能力,於含有Fe(CN)63-/4之條件下仍可與MUC1結合。此外,經由SPR動態分析得到適體K1R4.2與MUC1結合之kon¬ (1.33×10-4¬¬ sec-1nM-1)、koff¬ (7.05×10-3¬ sec-1)¬,並計算求得其解離常數 (KD¬) 為53.0 nM;且於穩態時根據不同濃度的適體K1R4.2結合於MUC1上的量,可計算得其KD值為55.1 nM。此適體與MUC1具有高親和力,且序列中GC含量高 (65.6%),表示其DNA密度及對熱與鹼的穩定性較高,根據這些特性優勢,未來可將其應用於適體感測器的開發,甚至是醫學上生物性治療的發展。; Mucin-1 (MUC1) is a transmembrane glycoprotein that is overexpressed on the surface of most adenocarcinoma cells, and it is served as a tumor marker. It is of great significance in both diagnosis and treatment. There is currently no aptamer selected targeting the MUC1 partial ORF (315 a.a. - 420 a.a.). It has MUC1 binding functionality after modification. According to its future application, this study intends to select a suitable aptamer for electrochemical sensing. Therefore, this sequence fragment is used as the target for MUC1 aptamer selection in this study. The selection method is called the systematic evolution of ligands by exponential enrichment (SELEX), with the use of single glass bead as the reaction substrate and human serum albumin as the target of reverse selection. Through multiple rounds of selection and amplification, a specific single-strand DNA pool with selectivity and affinity to MUC1 is obtained. For sequence analysis and selection, an analysis software MEME Suite is used. Finally, to find a sequence with the highest affinity to MUC1, which is the DNA aptamer, the affinity test was conducted by surface plasmon resonance (SPR). The evolution process of the sequences during the selection is shown by an evolution tree, which includes three branches, K1, K2, and K3. The number of non-specific sequences found in the pool during the selection can be used as a predictor of the dominant sequences during the evolution process. According to the SPR binding signal of the sequences to MUC1, and the composition of each sequence, it was found that the GC/AT ratio is moderately related to the affinity of the sequence (r = 0.499). In this study, a MUC1 aptamer was successfully found, which is named K1R4.2. This aptamer will not change its binding ability after modification, and it can also bind to MUC1 in the presence of Fe(CN)63-/4. Besides, the kon¬ (1.33×10-4¬¬ sec-1nM-1) and the koff¬ (7.05×10-3¬ sec-1) of aptamer K1R4.2 binding to MUC1 were obtained through SPR dynamic analysis, and the calculated KD is 53.0 nM. According to the amount of aptamer K1R4.2 bound to MUC1 at steady state, the obtained KD is 55.1 nM. This aptamer has a strong affinity to MUC1, and the GC content in the sequence is high (65.6%). This indicates that its DNA density and stability to heat and alkali are both high. Based on these characteristics and advantages, it can be applied to the development of aptasensors in the future, and even the progress of medical biological treatment.2020-01-01T00:00:00Z鹼前處理向日葵葉柄產製生質乙醇之研究
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40785
標題: 鹼前處理向日葵葉柄產製生質乙醇之研究; Alkaline Pretreatment of Sunflower Stalks on the Bioethanol Production
作者: Che-Wei Yang; 楊哲維
摘要: 本研究以向日葵葉柄作為原料本研究以台灣台南田間產的農業廢棄物之向日葵葉柄作為原料,其原料之組成分析為32.09% 纖維素、15.13% 半纖維素、18.83%木質素、21.98% 其他成分、7.29% 灰分、3.02% 醋酸與1.66%之水分,在前處理階段先以0.5%與4% ( w/v )之氫氧化鈉(NaOH)溶液,於1大氣壓下、溫度95℃等條件下,對向日葵葉柄進行前處理90分鐘,判斷其較佳之鹼處理濃度之後,以4%氫氧化鈉(NaOH)溶液為鹼處理濃度,提升其處理溫度為103℃與延長浸泡時間為3小時,此製程本文中以4.0% NaOHIII表示,接著將 4.0% NaOHIII之前處理物進行二階段過氧化氫處理,並分為有無添加氫氧化鈉之方式,使其pH值分別為7.5與11,再利用NREL Methods進行此五種前處理製程之前處理物進行組成分析,除此之外再利用單一酵素之纖維水解酶Cellulclast® 1.5L劑量為21 FPU/g,對此五種前處理物進行酵素水解72小時,並由其組成分析以及水解效果來判斷最佳之鹼前處理製程。在酵素水解階段使用最佳前處理製程4.0% NaOHIII+H2O2V之前處理物於後續酵素水解進行最佳水解製程之探討,並使用混合酵素纖維水解酶Cellulclast® 1.5L搭配葡萄糖苷酶Novozyme® 188,於溫度50℃、pH值4.5與90 rpm等條件下,對酵素活性比例、酵素劑量、基質濃度、批次式與饋料式水解等方式進行酵素水解72小時,由水解液中纖維雙醣、葡萄糖、木糖、還原醣濃度、水解產率與葡萄糖產率等數據來判斷最佳酵素水解製程。醱酵階段中,使用市售酵母發粉,以1 g酵母發粉配置事先滅菌處理之營養液 ( broth ),以1 g︰100 mL比例完成酵母接種物,於恆溫相中控制溫度37℃與120 rpm等條件下活化至少6小時,接著取90mL酵素水解製程之水解液,選擇葡萄糖濃度較高之水解液,也就是批次式基質15%與20%之水解液,以及兩種饋料式基質15%之水解液,搭配10mL之已活化之酵母接種物於溫度37℃與120 rpm等條件下,進行醱酵24小時,探討計算其葡萄糖、木糖、醋酸、乙醇濃度以及其理論值,最後獲致下列結論:
研究結果顯示:最佳前處理製程為以4.0% NaOHIII加上二階段過氧化氫以及添加氫氧化鈉之處理,本文中4.0% NaOHIII+H2O2V表示,其組成分析成分為50.74%纖維素、8.87%半纖維素、19.90%木質素、16.33%其他成分、1.66%水分與2.50%之灰分,前處理後利用濾袋固液分離後之前處理液則利用高效能液相層析儀檢測含有葡萄糖濃度0.43 mg/mL、木糖濃度 0.30 mg/mL與醋酸2.72 mg/mL之濃度,若將前處理液作醱酵處理,由於水解液中可醱酵醣濃度過低,加上有醋酸等會抑制醱酵之因素,顯示鹼前處理液並不適用做醱酵處理。在水解酵素階段,最佳酵素活性比例為FPU/g:CBU/g=10:2,而在此最佳酵素活性比例下之最佳劑量為21 FPU/g 、4.2 CBU/g之混合酵素,可於基質濃度5%、溫度50℃、90 rpm酵素水解72小時下,獲得13.03 mg/mL之葡萄糖濃度、60.96%之水解產率。除此之外若提高水解基質濃度,其葡萄糖產率以及水解產率反而呈現下降狀態,因此在高基質濃度下水解需採用饋料式之酵素水解方式,以同樣為基質濃度15%為例,以饋料式之酵素水解其最終葡萄糖濃度與水解產率分別為31.86 mg/mL與45.66%,而相對的批次式酵素水解基質濃度15%者之葡萄糖濃度與水解產率分別為28.15 mg/mL與42.25%,顯現在高基質濃度下,饋料式酵素水解為較理想之製程。在醱酵階段,將四種高葡萄糖濃度之水解液,以未添加葡萄糖、排毒、滅菌等方式進行醱酵24小時,則可在24小時內消耗大部分葡萄糖,而最終乙醇濃度為13.14~15.40 mg/mL,其理論值約為89.82~94.79%。; In this study, sunflower stalks which contained 32.09% cellulose, 15.13% semicellulose, 18.83% lignin, 21.98% others, 7.29% ash , 3.02% acetic acid and 1.66% moisture, was first pretreated by 0.5% and 4.0% NaOH solution at 95℃for 90 mins under atmosphere pressure. In the next step, we change the pretreatment parameters such as temperature, immerse time and second hydrogen peroxide pretreatment. In order to find the best pretreatment process, dried solid fraction was hydrolyzed by enzymes of cellulase from Trichoderma reesei C2730 ( Celluclast 1.5L ) under conditions of pH 4.5 and 50℃ for 72 hours with shaking water bath. In hydrolysis step, different mixing enzyme proportion, loadings and substrate ratios were studied to find out the optimum hydrolysis parameters. And then the hydrolysate was fermented with Saccharomyces cerevisiae under conditions at 37℃ and 120 rpm for 24 hours. The glucose, xylose, acetic acid and ethanol concentration were investigated in fermentation step.
As result, 0.12~0.43 mg/mL of glucose and 0.16~0.30 mg/mL of xylose were observed in liquid fraction. In hydrolysis step, the enzyme loading of 21 FPU/g Celluclast 1.5L plus 4.2 CBU/g Novozyme 188 represented the best balance between economy and efficiency. 260.60 mg/mL of glucose yield and 60.96% of conversion ratio were obtained under this enzyme loading with 5% substrate ratio and rising the substrate ratio did help improving the concentration of glucose and reducing sugar in the hydrolysate. In fermentation step, the four kind of hydrolysates without sterilization and detoxification were fermented for 24 hours and comsuming the most part of glucose. The ethanol concentration was measured around 13.14~15.40 mg/mL, corresponding to 89.82~94.79% of theoretical ethanol yield.2008-01-01T00:00:00Z