Development of surface-engineered yeast cells displaying phytochelatin synthase and their application to cadmium biosensors by the combined use of pyrene-excimer fluorescence

Hideyuki Matsuura, Yosuke Yamamoto, Misa Muraoka, Kenji Akaishi, Yasuhisa Hori, Kanoko Uemura, Naoki Tsuji, Kazuo Harada, Kazumasa Hirata, Takeshi Bamba, Hitoshi Miyasaka, Kouichi Kuroda, Mitsuyoshi Ueda

研究成果: Contribution to journalArticle査読

12 被引用数 (Scopus)

抄録

The development of simple, portable, inexpensive, and rapid analytical methods for detecting and monitoring toxic heavy metals are important for the safety and security of humans and their environment. Herein, we describe the application of phytochelatin (PC) synthase, which plays a critical role in heavy metal responses in higher plants and green algae, in a novel fluorescent sensing platform for cadmium (Cd). We first created surface-engineered yeast cells on which the PC synthase from Arabidopsis (AtPCS1) was displayed with retention of enzymatic activity. The general concept for the sensor is based on the Cd level-dependent synthesis of PC2 from glutathiones by AtPCS1-displaying yeast cells, followed by simple discriminative detection of PC2 via sensing of excimer fluorescence of thiol-labeling pyrene probes. The intensity of excimer fluorescence increased in the presence of Cd up to 1.0 μM in an approximately dose-dependent manner. This novel biosensor achieved a detection limit of as low as 0.2 μM (22.5 μg/L) for Cd. Although its use may be limited by the fact that Cu and Pb can induce cross-reaction, the proposed simple biosensor holds promise as a method useful for cost-effective screening of Cd contamination in environmental and food samples. The AtPCS1-displaying yeast cells also might be attractive tools for dissection of the catalytic mechanisms of PCS.

本文言語英語
ページ(範囲)1197-1202
ページ数6
ジャーナルBiotechnology Progress
29
5
DOI
出版ステータス出版済み - 9 2013
外部発表はい

All Science Journal Classification (ASJC) codes

  • バイオテクノロジー

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