標題: Enhancing ATP-based bacteria and biofilm detection by enzymatic pyrophosphate regeneration
作者: Lee, Hui-Ju
Ho, Ming-Rong
Bhuwan, Manish
Hsu, Ching-Yi
Huang, Meng-Shun
Peng, Hwei-Ling
Chang, Hwan-You
生物科技學系
Department of Biological Science and Technology
關鍵字: ATP regeneration system;ADP-glucose pyrophosphorylase;Luminescence
公開日期: 15-四月-2010
摘要: The manufacturing processes of many electronic and medical products demand the use of high-quality water. Hence the water supply systems for these processes are required to be examined regularly for the presence of microorganisms and microbial biofilms. Among commonly used bacteria detection approaches, the ATP luminescence assay is a rapid, sensitive, and easy to perform method. The aim of this study is to investigate whether ATP regeneration from inorganic pyrophosphate, a product of the ATP luminescence assay, can stabilize the bioluminescence signals in ATP detection. ADPglc pyrophosphorylase (AGPPase), which catalyzes the synthesis of ATP from PP(i) in the presence of ADPglc, was selected because the system yields much lower luminescence background than the commercially available ATP sulfurylase/adenosine 5'-phosphosulfate (APS) system which was broadly used in pyrosequencing technology. The AGPPase-based assay could be used to measure both PPi and ATP quantitatively and shows 1.5- to 4.0-fold slight increases in a 10-min assay. The method could also be used to stabilize the luminescence signals in detection of Escherichia coli, Pseudo monas aeruginosa, and Bacillus cereus in either broth or biofilm. These findings suggest that the AGPPase-based ATP regeneration system will find many practical applications such as detection of bacterial biofilm in water pipelines. (C) 2009 Elsevier Inc. All rights reserved.
URI: http://dx.doi.org/10.1016/j.ab.2009.12.032
http://hdl.handle.net/11536/5509
ISSN: 0003-2697
DOI: 10.1016/j.ab.2009.12.032
期刊: ANALYTICAL BIOCHEMISTRY
Volume: 399
Issue: 2
起始頁: 168
結束頁: 173
顯示於類別:期刊論文


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