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Enzymes02:34

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Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
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Updated: Feb 3, 2026

Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds
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Staphylococcus aureus Cas9 is a multiple-turnover enzyme.

Paul Yourik1, Ryan T Fuchs1, Megumu Mabuchi1

  • 1RNA and Genome Editing, New England Biolabs Inc., Ipswich, Massachusetts 01938, USA.

RNA (New York, N.Y.)
|October 24, 2018
PubMed
Summary

Staphylococcus aureus Cas9 (SauCas9) is a multiple-turnover enzyme, unlike SpyCas9. This finding offers new insights for CRISPR/Cas9 genome editing applications.

Keywords:
CRISPRCas9S. aureusS. pyogenesgene editingsgRNA

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Cas9 nuclease is central to CRISPR adaptive immunity in bacteria.
  • Cas9, guided by sgRNA, enables sequence-specific DNA cleavage, making it a vital genome editing tool.
  • While Cas9 homologs share catalytic features, sequence diversity may cause mechanistic variations.

Purpose of the Study:

  • To directly compare the enzymatic activities of Staphylococcus aureus Cas9 (SauCas9) and Staphylococcus pyogenes Cas9 (SpyCas9).
  • To investigate potential mechanistic differences, particularly regarding reaction turnover and DNA cleavage products.

Main Methods:

  • In vitro biochemistry assays.
  • Enzyme kinetics studies comparing SauCas9 and SpyCas9.

Main Results:

  • SauCas9 functions as a multiple-turnover enzyme, a characteristic not previously reported for Cas9 homologs.
  • Unlike SpyCas9, SauCas9-mediated DNA cleavage does not result in detectable single-stranded DNA degradation post-double-strand break.

Conclusions:

  • SauCas9 exhibits distinct enzymatic properties compared to SpyCas9, notably its multiple-turnover capability.
  • These findings provide crucial insights for optimizing and designing future CRISPR/Cas9-based genome editing technologies.