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Related Experiment Videos

Liquid nitrogen cryo-impacting: a new concept for cell disruption

R A Smucker, R M Pfister

    Applied Microbiology
    |September 1, 1975
    PubMed
    Summary

    Liquid nitrogen cryo-impacting efficiently disrupts bacterial cells, including Bacillus megaterium endospores and Escherichia coli. This novel method releases dipicolinic acid-Ca2+ and shows superior lactate dehydrogenase activity compared to sonic treatment.

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    Area of Science:

    • Microbiology
    • Biotechnology
    • Biophysics

    Background:

    • Bacterial cell disruption is crucial for accessing intracellular components.
    • Existing methods like sonication can be energy-intensive and less efficient for certain structures like endospores.
    • There is a need for novel, high-efficiency bacterial lysis techniques.

    Purpose of the Study:

    • To introduce and evaluate a new method for bacterial cell disruption: liquid nitrogen cryo-impacting.
    • To assess the efficiency of cryo-impacting in breaking bacterial endospores and vegetative cells.
    • To compare the efficacy of cryo-impacting with conventional sonication.

    Main Methods:

    • Bacterial cells, including Bacillus megaterium endospores and Escherichia coli, were subjected to liquid nitrogen cryo-impacting.
    • Cell breakage was assessed by measuring the release of dipicolinic acid-Ca2+ chelate from endospores.
    • Lactate dehydrogenase activity in the supernatant was quantified to evaluate cell lysis efficiency for E. coli.

    Main Results:

    • Liquid nitrogen cryo-impacting achieved high-efficiency bacterial cell disruption in two minutes or more.
    • The release of dipicolinic acid-Ca2+ chelate from Bacillus megaterium endospores correlated with cell breakage.
    • Cryo-impacting yielded significantly higher lactate dehydrogenase activity from Escherichia coli cells compared to sonication.

    Conclusions:

    • Liquid nitrogen cryo-impacting is a highly efficient method for disrupting bacterial cells and endospores.
    • This technique offers a promising alternative to traditional cell lysis methods.
    • Cryo-impacting demonstrates superior efficiency in releasing intracellular enzymes compared to sonication.

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