Title: Enhancing ATP-based bacteria and biofilm detection by enzymatic pyrophosphate regeneration
Authors: Lee, Hui-Ju
Ho, Ming-Rong
Bhuwan, Manish
Hsu, Ching-Yi
Huang, Meng-Shun
Peng, Hwei-Ling
Chang, Hwan-You
Department of Biological Science and Technology
Keywords: ATP regeneration system;ADP-glucose pyrophosphorylase;Luminescence
Issue Date: 15-Apr-2010
Abstract: 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
ISSN: 0003-2697
DOI: 10.1016/j.ab.2009.12.032
Volume: 399
Issue: 2
Begin Page: 168
End Page: 173
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