Biosensor for acrylamide based on an ion-selective electrode using whole cells of Pseudomonas aeruginosa containing amidase activity

被引:25
作者
Silva, Nelson [1 ]
Gil, Dulce [1 ]
Karmali, Amin [1 ]
Matos, Manuel [1 ]
机构
[1] Inst Super Engn Lisboa, Chem Engn & Biotechnol Res Ctr, P-1950062 Lisbon, Portugal
关键词
ion selective electrode; wild-type amidase; whole cells of Pseudomonas aeruginosa; biosensor; acrylamide; polyethersulfone membrane; TANDEM MASS-SPECTROMETRY; KINETIC-PROPERTIES; FOODS; ENZYME; GEL;
D O I
10.1080/10242420802604964
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The aim of this work was to develop a biosensor for toxic amides using whole cells of Pseudomonas. aeruginosa containing amidase activity, which catalyses the hydrolysis of amides such as acrylamide producing ammonia and the corresponding organic acid. Whole cells immobilized in several types of membrane in the presence of glutaraldehyde and an ammonium ion-selective electrode, were used for biosensor development. This biosensor exhibited a linear response in the range of 0.1-4.010-3 M of acrylamide, a detection limit of 4.4810-5 M acrylamide, a response time of 55 s, a sensitivity of 58.99 mV mM-1 of acrylamide and a maximum t1/2 of 54 days. The selectivity of this biosensor towards other amides was investigated, which revealed that it cross-reacted with acetamide and formamide, but no activity was detected with phenylacetamide, p-nitrophenylacetamide and acetanilide. It was successfully used for quantification of acrylamide in real industrial effluents and recovery experiments were carried out which revealed an average substrate recovery of 93.3%. The biosensor is cheap since whole cells of P. aeruginosa can be used as source of amidase activity.
引用
收藏
页码:143 / 151
页数:9
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