葉怡均Yeh, Yi-Chun石家宜Shih, Chia-I2020-10-192025-07-162020-10-192020http://etds.lib.ntnu.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=id=%22G060742012S%22.&%22.id.&http://rportal.lib.ntnu.edu.tw:80/handle/20.500.12235/111128左旋多巴經由全細胞生成之4,5-多巴雙加氧酶能將其催化成甜菜醛胺酸，透過加入2-胺基對苯二甲酸 (2-AIPA) ，可進一步生成具有紅螢光的甜菜色素 (2-AIPA-BX)。本研究開發了一種簡單且經濟的方法，可大量合成甜菜色素用於二價銅離子檢測，在銅離子濃度為25~500 μM的範圍內，有隨著濃度提升而紅螢光下降的趨勢，呈現出良好的線性 (R = 0.99) 且最低偵測極限為21.92 μM，也成功將2-AIPA-BX應用於人工尿液樣品及紙基材上的檢測，以簡單、低樣品耗量、低成本且便攜式的設置進行二價銅離子的檢測。此外，本研究也經由4,5-多巴雙加氧酶上特定的胺基酸位置突變以改變其反應的選擇性，成功使其反應選擇性從左旋多巴變成多巴胺，證實此方法的可行性及未來發展性。In the human body, copper ions participate in the catalysis of many enzymes. In this study, we developed a simple and economical method for the green synthesis of Cu2+ sensors based on 2-aminoterephthalic acid–betaxanthin (2-AIPA–BX). 2-AIPA–BX was synthesized by coupling 2-aminoterephthalic acid and betalamic acid produced from DOPA-extradiol-4,5-dioxygenase in situ. It has a high degree of selectivity for Cu2+ over interfering metal ions. In the range of Cu2+ concentration of 25~500 μM, 2-AIPA-BX had a tendency to lessen the fluorescence as the concentration increases, the result showed a good linearity and the limit of detection was 21.92 μM. Additionally, we successfully incorporated 2-AIPA–BX into paper-based analytical devices to facilitate simple, inexpensive, and portable setup with lower sample consumption for onsite monitoring of environmental and biological samples. Dopamine is a neurotransmitter that plays an important role in the function of the central nervous system and cardiovascular system. In this study, the specific amino acid mutation on DOPA 4,5-dioxygenase was used to change the selectivity of its reaction, and its reaction activity was successfully changed from L-DOPA to dopamine, confirming the feasibility and future development of this method.二價銅離子多巴胺螢光感測器4,5-多巴雙加氧酶綠色合成甜菜色素全細胞生物感測器Cu2+dopaminefluorogenic sensorsDOPA 4,5-dioxygenasegreen synthesisbetaxanthinwhole-cell biosensors