Protease

Protease is a general term for enzymes that catalyze the hydrolysis of peptide bonds (-CO-NH-), and is also called peptidase or proteinase. Proteases are used in biological experiments for a variety of purposes, including cell wall lysis, cell detachment, and protein fragmentation.

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Discovery and utilization of Achromopeptidase®

When extracting plasmid DNA from bacteria with rigid cell walls, such as Gram-positive bacteria, it is necessary to break down the bacterial cell wall using bacteriolytic enzymes.

The most commonly used bacteriolytic enzyme is lysozyme, which hydrolyzes peptidoglycan, a component of cell walls.1) However, some bacteria, such as Staphylococcus aureus, are resistant to the lytic action of lysozyme, making plasmid extraction difficult.2)

In 1977, plasmid DNA was successfully extracted from Staphylococcus aureus using achromopeptidase, a bacteriolytic enzyme from Achromobacter lyticus.2) Achromopeptidase has a broad bacteriolytic spectrum and stronger bacteriolytic activity than egg white lysozyme. It is now used for bacteriolysis of various bacteria, including Gram-positive bacteria, and for the preparation of protoplasts of actinomycetes.

References

  1. Principles and protocols for nucleic acid experiments by purpose” ed. by Hirao, I., Kurumizaka, H., Yodosha, Japan, (2011). (Japanese)
  2. Horinouchi, S., et al.: Agricultural and Biological Chemistry, 41(12), 2487(1977).
    A new isolation method of plasmid deoxyribonucleic acid from Staphylococcus aureus using a lytic enzyme of Achromobacter lyticus.

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