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Related Concept Videos

Hydrolysis of ATP01:08

Hydrolysis of ATP

The bonds of adenosine triphosphate (ATP) can be broken through the addition of water, releasing one or two phosphate groups in an exergonic process called hydrolysis. This reaction liberates the energy in the bonds for use in the cell—for instance, to synthesize proteins from amino acids.
If one phosphate group is removed, a molecule of ADP—adenosine diphosphate—remains, along with inorganic phosphate. ADP can be further hydrolyzed to AMP—adenosine monophosphate—by the removal of a second...

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Related Experiment Video

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Use of Stopped-Flow Fluorescence and Labeled Nucleotides to Analyze the ATP Turnover Cycle of Kinesins
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A microfluidic in situ analyzer for ATP quantification in ocean environments.

Tatsuhiro Fukuba1, Yusuke Aoki, Noriyuki Fukuzawa

  • 1Institute of Industrial Science, University of Tokyo, Tokyo, Japan. bafuk@iis.u-tokyo.ac.jp

Lab on a Chip
|September 1, 2011
PubMed
Summary

We developed a new in situ analyzer for measuring microbial activity via total adenosine triphosphate (ATP) in ocean waters. This system accurately quantifies ATP in diverse marine environments, from coastal bays to deep-sea hot springs.

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Area of Science:

  • Environmental Science
  • Biotechnology
  • Analytical Chemistry

Background:

  • Microbial activity is crucial for ocean ecosystem health.
  • Accurate quantification of adenosine triphosphate (ATP) is essential for assessing microbial biomass and activity.
  • Existing methods for ATP measurement in marine environments often lack in situ capabilities or require complex sample processing.

Purpose of the Study:

  • To develop and test a functionally integrated in situ analyzer (IISA-ATP) for microbial activity assays.
  • To enable quantitative determination of total (particulate and dissolved) ATP in ocean environments.
  • To validate the system's performance using standard solutions and environmental samples.

Main Methods:

  • Utilized a PDMS-glass hybrid microfluidic device for cell lysis and ATP quantification.
  • Employed a luciferin-luciferase bioluminescence assay for sensitive ATP detection.
  • Integrated transparent heaters and a temperature sensor for precise temperature control.
  • Incorporated a miniature pumping module and control module for automated operation.

Main Results:

  • Demonstrated a linear correlation between bioluminescence intensity and ATP concentrations from 2 × 10⁻¹² to 2 × 10⁻⁸ M.
  • Achieved a detection limit of 1.1 × 10⁻¹¹ M with the completed IISA-ATP system.
  • Successfully quantified total ATP in laboratory analysis of Tokyo Bay seawater (2.7 × 10⁻¹⁰ M).
  • Successfully operated the system in situ at a submarine hot spring in Okinawa, determining total ATP to be 3.4 × 10⁻¹⁰ M.

Conclusions:

  • The IISA-ATP system provides a reliable and sensitive method for in situ measurement of total ATP in marine environments.
  • The developed microfluidic device and integrated system enable accurate microbial activity assessment in diverse oceanic settings.
  • The successful field deployment demonstrates the system's potential for real-time monitoring of marine microbial ecosystems.