Development of highly sensitive multi-biotip by using thermostable enzymes of thermophilic microorganism.

利用嗜热微生物的耐热酶开发高灵敏度的多生物探针。

• 批准号: 17590038
• 负责人: HASEBE Yasushi
• 金额: $ 2.46万
• 依托单位: Saitama Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17590038/
• 关键词: Thermostable enzyme; Biotip; Multi-sensing; highly sensitive sensing; flow-detector; biosensor; uric acid; glutamic acid; 高感度化; バイオセンサ; グルコース; 乳酸; NADH

Primary aim of this study is a development of flow-type highly sensitive multi-biotip using several thermo-stable enzymes to detect simultaneously several kinds of analytes.1) Uric acid sensor using thermo-stable uricase and peroxidase-hybrid carbon felt-based flow-through bioelecterocatalytic detectorThis sensor system allowed continues detection of uric acid over the concentration range from 0.1 to 100□M. Moreover this uric acid sensor allowed highly sensitive and accurate analysis of highly diluted human urine sample without any interference, and analytical results is well consistent with that of spectrophotometry. The sensor is much stable and kept its original activity during more than 3 month storage.2) Glutamate biosensor using thermo-stable glutamate oxidase and DNA-Cu(II) complex.Novel type glutamate biosensor was developed, in which H_2O_2 liberated during the glutamate oxidase was cathodically detected by the novel electrocatalytic activity of DNA-Cu(II) complex. This sensor allowed amperometric detection of glutamate over the concentration range from 0.1 to 50□M. Based on the permselectivity of DNA-film, interference-free detection was possible even in the case of body fluid such as brain liquids.On future work, if other thermostable oxidases are adopted in place of uricase and glutamate oxidase, the establishment of multi-biotip would be possible.

Primary aim of this study is a development of flow-type highly sensitive multi-biotip using several thermo-stable enzymes to detect simply several kinds of analytes.1) Uric acid sensor using thermo-stable uricase and peroxidase-hybrid carbon felt-based flow-through bioelecterocatalytic detectorThis sensor system allowed continues detection of uric acid over the concentration range from 0.1 to 100□M.此外，该尿酸传感器允许对高度稀释的人类尿液样品进行高度敏感和准确的分析，而不会进行任何干扰，并且分析结果与分光光度计的结果非常一致。该传感器非常稳定，并在3个月的存储期间保持了其原始活性。2）使用热稳定的氧化物和DNA-CU（II）复合物的谷氨酸生物传感器。开发了谷氨酸型谷氨酸生物传感器，其中H_2O_2在氧化物中释放了H_2O_2在氧化物中释放了谷氨酸氧化物在氧化含量的过程中被天然化的电动性电动性活性（II）II（II II）。该传感器允许在浓度上对谷氨酸的安培检测范围为0.1至50□m。基于DNA-FILM的允许选择性，即使在体液（例如脑液体）的情况下，也可能进行无干扰检测。未来的工作，如果采用其他热稳定氧化物代替尿液酶和氧化谷氨酸氧化物，则可以建立多生物蛋白。

项目成果

「研究成果報告書概要(和文)」より

摘自《研究结果报告摘要（日文）》

• 发表时间: 2005
• 作者: Kawauchi;et. al.;Nishimura et al.;Dezawa et al.;Yoshizawa et al.;星野 幹雄;星野 幹雄
• 通讯作者: 星野 幹雄