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Related Experiment Videos

Gene inactivation in Lactococcus lactis: histidine biosynthesis

C Delorme1, J J Godon, S D Ehrlich

  • 1Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique, Jouy en Josas, France.

Journal of Bacteriology
|July 1, 1993
PubMed
Summary

Most dairy Lactococcus lactis strains need histidine for growth, unlike nondairy strains. Genetic mutations in the histidine operon explain this requirement in dairy bacteria, impacting histidine biosynthesis.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacterial Genetics

Background:

  • Lactococcus lactis is crucial in dairy fermentation.
  • Bacterial auxotrophy, the inability to synthesize essential nutrients, can arise from genetic alterations.
  • Histidine biosynthesis is a well-studied metabolic pathway in bacteria.

Purpose of the Study:

  • To investigate the prevalence of histidine auxotrophy in Lactococcus lactis.
  • To identify the genetic basis of histidine auxotrophy in dairy strains.
  • To understand the impact of adaptation to milk on bacterial metabolism.

Main Methods:

  • Screening of dairy and nondairy Lactococcus lactis strains for histidine auxotrophy.
  • Histidinol growth tests to assess functional histidine precursor utilization.

Related Experiment Videos

  • Southern and Northern hybridization to detect and analyze the histidine operon.
  • Gene cloning, sequencing, and complementation analysis in Escherichia coli.

    • Main Results:

    • 56 out of 60 dairy strains required histidine, while only 1 out of 11 nondairy strains did.
    • Mutations, including frameshifts in histidine biosynthesis genes (hisC, hisG, hisH) and regulatory regions, were identified in an auxotrophic strain.
    • Reduced histidine operon transcript levels were observed in auxotrophic strains compared to prototrophic ones.
    • Dairy strains showed varying degrees of auxotrophy, with some capable of reversion.

    Conclusions:

    • Adaptation to dairy environments frequently leads to histidine auxotrophy in Lactococcus lactis.
    • Specific mutations within the histidine operon are responsible for histidine auxotrophy in these strains.
    • The genetic basis for auxotrophy can vary, influencing reversibility and phenotype.