Purification and preliminary crystallization of alanine racemase from Streptococcus pneumoniae
2007

Purification and Crystallization of Alanine Racemase from Streptococcus pneumoniae

publication Evidence: moderate

Author Information

Author(s): Strych Ulrich, Davlieva Milya, Longtin Joseph P, Murphy Eileen L, Im Hookang, Benedik Michael J, Krause Kurt L

Primary Institution: University of Houston

Hypothesis

The study investigates the alanine racemase enzyme from Streptococcus pneumoniae as a potential target for new antibiotic therapies.

Conclusion

The alanine racemase gene from S. pneumoniae was successfully isolated and characterized, showing potential for drug design.

Supporting Evidence

  • The enzyme was purified to electrophoretic homogeneity and preliminary crystals were obtained.
  • Specific activities of 87.0 and 84.8 U mg-1 were determined for the conversion of D- to L-alanine and L- to D-alanine.
  • The cloned S. pneumoniae gene fully restored the wild-type phenotype in an E. coli D-alanine auxotroph.
  • Sequence analysis showed high similarity with other alanine racemases.

Takeaway

Scientists found a way to isolate and study an important enzyme from a bacteria that can cause pneumonia, which could help in creating new medicines.

Methodology

The alr gene was amplified, cloned, expressed in E. coli, and the enzyme was purified and crystallized.

Digital Object Identifier (DOI)

10.1186/1471-2180-7-40

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