生物化學

We invented the Midori which means green Bioreactor (MBR), beads of euglena and other microalgae fixed in calcium alginate that absorbs carbon dioxide (CO2). We examined the effect of 19 different solutions and two different organisms on MBR cultivation. Surprisingly, when the MBR was supplied with carbon dioxide or cultured with yeast, they became drastically darker green. Chromatography revealed this green color to be that of microalgae such as green algae or Euglena because chlorophyll a and chlorophyll b were detected. Under sunlight, MBR absorbed CO2 and the absorption rate was 1.5 L CO2/day/1L of MBR. Furthermore, when we put MBR in the water tank, they increased the amount of dissolved oxygen without polluting the environment. These results indicate that MBR can absorb CO2 by photosynthesizing without leaking out the inside microalgae.

為研究花青素在不同因素影響下的抗 UV 能力，選用矮仙丹作為研究對象。以矮仙丹花瓣的花青素，研究不同濃度及 pH 值的環境下，花青素的抗 UV 能力，以及在照射 UV 後， 矮仙丹 SOD、POD 之濃度變化。由實驗得知，紅花的花青素含量大於粉花。對比了紅花及粉花的 UV-Vis 光譜圖後發現：在相同濃度下，紅花的吸光度大於粉花且兩者的最大吸收高峰皆位於約 510nm~520nm 處，由此推知矮仙丹的花瓣含有大量的花青素。而花青素在不同pH 值環境下，會因為結構改變而呈現不同的顏色。我們測試在不同 pH 值緩衝液的花青素是否具有不同的抗 UV 能力，發現鹼性高於酸性。植物在代謝過程中容易產生具有毒害作用的活性氧，由於活性氧會對細胞造成傷害，於是演化出抗氧化酶，如 SOD、POD 等，用以清除活性氧，降低對植物的傷害。

In the first part of this project, distilled water and 95% ethanol were used to extract ultra-violet(UV)-absorbing and anti-oxidizing compounds from different types of algae including kelp, wakame, sea grape and nori. Activated charcoal was used in attempt to purify the extracts and removed excessive pigments. It was found that the charcoal was more effective in adsorbing pigments from ethanol extract in which up to 80 to 100% pigments could be removed. The UV-absorbing and anti-oxidizing properties of the algae extracts were also studied. All algae extracts showed significant UV-absorbing and anti-oxidizing properties. In particular, extract formed by using 3 g kelp powder in distilled water could significantly reduce 50% UV intensity and react 96.5% DPPH solution which acts as a source of free radicals. In the second part of the project, three applications of algae were explored in details. Firstly, it was found that kelp, wakame and nori extracts by using over 2 g of algae in 30 mL olive oil could absorb UVA and UVB by over 90%, which is comparable to the performance of zinc oxide, a common ingredient in commercial sunscreen products. Costs of preparing the sunscreens were also compared. Except for wakame extract, all other extracts were cheaper than using zinc oxide. Moreover, the kelp extract was found to maintain its UV- absorbing and anti-oxidizing abilities after at least 30 days of storage under room conditions. Lastly, sodium alginate was successfully extracted from kelp with a product yield up to 30%. The alginate solution was then used to form a calcium-alginate protective coating on plastic slides to reduce UV intensity by up to 50%. This aims to apply on nails or fingers during UV nail gel polish to protect against UVR.

This study primarily investigates the components of food residues in the stomach of flying squirrels and the metabolism of the intestinal bacteria Floricoccus tropicus. Using nuclear magnetic resonance (NMR) hydrogen spectrum analysis, the signals of long-chain fatty acids were detected in both n-hexane and 75% ethanol extracts of flying squirrel gastric residues and acorns, suggesting that acorns may be a primary food source for the flying squirrels. The study also identified Floricoccus tropicus, one of the lactic acid bacteria, from the intestines of flying squirrels and discovered its role in the metabolism of fatty acids in acorns. Results showed that polyunsaturated fatty acids significantly decreased during fermentation, indicating that they were converted into short-chain fatty acids with anti-inflammatory properties. In the antibacterial activity experiments, the acorn grease showed no inhibitory effects before fermentation, but after fermentation, the acorn grease exhibited inhibitory effects against E. coli. Furthermore, in anti-inflammatory tests, fermented acorn grease samples significantly suppressed the production of NO and TNF-α in LPSinduced RAW 264.7 cells, with greater inhibition at higher concentrations. In conclusion, the lactic acid bacteria Floricoccus tropicus was found to metabolize fatty acids of acorns into compounds with antibacterial and anti-inflammatory effects.

Durians’ seeds have potential as a food source due to their content and nutrients. Durians’ seeds contain fiber, minerals, vitamins A, B1, B2, C, carbohydrates, folate, potassium and copper. Nutrients are needed for the body's health and growth and development process. Durians’ seeds have the potential as a nutrient-rich food alternative. Researchers made an innovation in the form of cereal, Healthy Cereal Organic (HCO). Analysis of durians’ seed content through two stages. First, wet lab examination and second, in silico method. The wet lab examination shows the results that durians’ seeds contain 10.17 Kcal of fat energy, 4.09% ash content, 11.25% water content, 72.79% carbohydrates, 1.13% total fat and 10.74% protein and the in silico method shows the content of carotenoid compounds (vitamin A, quercetin, beta-carotene, zeaxanthin) as a drug delivery system which means that this compound is able to be absorbed by the body with the help of albumin as a carrier that maintains stability and increases its activity. Feasibility analysis based on toxicity tests, Durians’ seed compounds show inactive (non-toxic) results. Allergenicity test showed non-allergen durians’ seed content. Hedonic test was conducted on 20 panelists dominant to the HCO1 sample for aroma by 60%, texture 90%, taste 40%. It can be concluded that durians’seeds can be used as a basic ingredient for making nutrient-rich Healthy Cereal Organic (HCO).

硫磺菇（Laetiporus sulphureus）和桑黃菇（Sanghuangporus sanghuang）是東亞，特別是台灣森林中的兩種真菌。這些真菌的次級代謝物，特別是多醣，具有抗炎和抗癌的生物效應；其地面子實體長期被當地人作為傳統藥物使用。然而，這些藥用特性及其機制尚未充分研究。本研究旨在分析和量化這些真菌多醣的抗炎效果。從硫磺菇中提取硫酸化多醣，從桑黃菇中提取非硫酸化多醣，並使用水和乙醇進行多步純化。隨後，將純化後的產品餵給巨噬細胞進行體外測試以檢查其抗炎性。硫酸化多醣的最佳濃度為150 ppm，能夠最大程度地降低自由基濃度21.6%，且不影響細胞活力。相比之下，桑黃菇的所有多醣濃度均顯示出增強的細胞炎症，顯示其作為藥物無效，因為沒有去除真菌毒素。相比之下，硫磺菇的硫酸化多醣顯示出其藥用潛力，對生物醫學和生物探索領域具有新啟示。

Previous research observed increased TNTs formation between hypoxic and normoxic neuroblastoma cells, aiding hypoxic cell survival. CHGA was identified as a potential factor in this process. This study compared CHGA expression and whether CHGA exists in TNTs in five cell lines, with SH-SY5Y showing the highest levels, followed by SK-N-BE(2)C, while the other three showed lower expression. Future studies will focus on the impact of CHGA on cell survival and its mechanisms.

This study primarily investigates the components of food residues in the stomach of flying squirrels and the metabolism of the intestinal bacteria Floricoccus tropicus. Using nuclear magnetic resonance (NMR) hydrogen spectrum analysis, the signals of long-chain fatty acids were detected in both n-hexane and 75% ethanol extracts of flying squirrel gastric residues and acorns, suggesting that acorns may be a primary food source for the flying squirrels. The study also identified Floricoccus tropicus, one of the lactic acid bacteria, from the intestines of flying squirrels and discovered its role in the metabolism of fatty acids in acorns. Results showed that polyunsaturated fatty acids significantly decreased during fermentation, indicating that they were converted into short-chain fatty acids with anti-inflammatory properties. In the antibacterial activity experiments, the acorn grease showed no inhibitory effects before fermentation, but after fermentation, the acorn grease exhibited inhibitory effects against E. coli. Furthermore, in anti-inflammatory tests, fermented acorn grease samples significantly suppressed the production of NO and TNF-α in LPSinduced RAW 264.7 cells, with greater inhibition at higher concentrations. In conclusion, the lactic acid bacteria Floricoccus tropicus was found to metabolize fatty acids of acorns into compounds with antibacterial and anti-inflammatory effects.

據 WHO 於 2023 的統計，全球約有四千萬人罹患阿茲海默症，且近年來患病率逐年上升，已成為當今無法避免的疾病。造成阿茲海默症的因子眾多，目前著重研究 Tau 蛋白毒性與其的關係。前人研究發現真核生物缺乏嘌呤時會形成嘌呤體，而嘌呤體始於 PAICS 泛素化，Cullin 在其中擔任重要角色，然而現今仍未得知哪些 Cullin 與 PAICS 泛素化較有關聯，故本研究以黑腹果蠅為實驗模型，用 T- test 統計數據並探討不同 Cullin 在阿茲海默症毒性下對於 PAICS 泛素化形成嘌呤體的影響。根據實驗結果，本研究發現 Cullin3 的複眼面積顯著變小，推測其最可能和 PAICS 泛素化有關聯；而與預期相反的是 Cullin5、6，結果顯示其果蠅複眼面積皆顯著大於對照組。本研究結果可提供 Cullin3 對阿茲海默症研究扮演重要角色。

An exploration of Pachyrhizus erosus as an option for plant-based milk in the 印尼n market for people with lactose intolerance. With its prevalence in tropical climates, Pachyrhizus erosus is an affordable crop in 印尼. Pachyrhizus erosus is a root vegetable containing a calcium content of 15.6 mg per cup (130g) in its unprocessed form, concentrated in its white flesh. Based on its affordability and calcium content, Pachyrhizus erosus can be transformed into a beverage product with nutritional qualities on par with existing plant-based milk, therefore being a solution for calcium sustenance that is more accessible due to its high capability to be locally grown in 印尼. However, this may be a partial case as Pachyrhizus erosus only thrives in regions with long warm seasons. In this research, three trials of Pachyrhizus erosus-based milk recipe have been conducted Trial 1 consists of a 1:1 ratio of Pachyrhizus erosus to water, Trial 2 consists of a 2:1 ratio of Pachyrhizus erosus to water, and Trial 3 consisting of a 10:10:1 ratio of Pachyrhizus erosus to water, and to a small amount of soybean. Based on the results of 14 organoleptic test respondents, it is concluded that the best ratio of ingredients is 10:10:1 (water: Pachyrhizus erosus: soybean) due to an overall preference of the third trial with this ratio, in terms of taste, aroma, color, and consistency. Pachyrhizus erosus is the dominant ingredient in developing alternate plant-based milk. However, findings from the most preferred trial in the organoleptic test suggest that implementing a minor amount of soybean would stabilize the milk-like consistency and flavor. Pachyrhizus erosus’ ability to retain calcium in its water content has been additionally proven in a calcium test using a reagent solution of ammonium oxalate, as even with the trials’ step of straining the liquid content of Pachyrhizus erosus that had been blended with added water, all three trials tested positive based on the high level of the precipitate. Other tests that tackle the quality of each trial include In Silico Testing, biuret protein test, alcohol test, COB test, and pH level testing.