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Development of surface‐engineered yeast cells displaying phytochelatin synthase and their application to cadmium biosensors by the combined use of pyrene‐excimer fluorescence
Author(s) -
Matsuura Hideyuki,
Yamamoto Yosuke,
Muraoka Misa,
Akaishi Kenji,
Hori Yasuhisa,
Uemura Kanoko,
Tsuji Naoki,
Harada Kazuo,
Hirata Kazumasa,
Bamba Takeshi,
Miyasaka Hitoshi,
Kuroda Kouichi,
Ueda Mitsuyoshi
Publication year - 2013
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.1789
Subject(s) - phytochelatin , biosensor , pyrene , yeast , chemistry , fluorescence , excimer , biochemistry , nanotechnology , enzyme , glutathione , materials science , organic chemistry , physics , quantum mechanics
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 PC 2 from glutathiones by AtPCS1‐displaying yeast cells, followed by simple discriminative detection of PC 2 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. © 2013 American Institute of Chemical Engineers Biotechnol. Prog ., 29:1197–1202, 2013