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Bioengineered nisin derivatives with enhanced activity in complex matrices.

Susan Rouse1, Des Field, Karen M Daly

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Bioengineered nisin A variants show improved antimicrobial properties by better diffusion through complex food matrices, effectively controlling Listeria monocytogenes in chocolate milk.

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Area of Science:

  • Microbiology
  • Biotechnology
  • Food Science

Background:

  • Nisin A is a well-characterized lantibiotic with antimicrobial properties.
  • Bioengineering ribosomally synthesized peptides like nisin A allows for variant generation.
  • Previous work showed hinge region engineering enhances antimicrobial activity against Gram-positive pathogens.

Purpose of the Study:

  • To create and screen a larger bank of nisin A hinge variants for enhanced bioactivity.
  • To investigate the mechanisms behind enhanced bioactivity in nisin A variants.
  • To evaluate the efficacy of promising nisin A variants in a food matrix.

Main Methods:

  • Generation of a library of nisin A hinge variants.
  • Screening of variants using agar-based diffusion assays against target pathogens.
  • Characterization of lead variants focusing on specific activity and diffusion properties.
  • Inhibition assays using nisin A variants against Listeria monocytogenes in carrageenan-stabilized chocolate milk.

Main Results:

  • Screening identified multiple nisin A hinge variants with enhanced bioactivity.
  • Enhanced bioactivity was often linked to improved diffusion through complex polymers, not increased specific activity.
  • Two variants, SVA and NAK, demonstrated superior control of Listeria monocytogenes in chocolate milk.
  • These variants outperformed native nisin A in the complex food system.

Conclusions:

  • Bioengineering the hinge region of nisin A can yield variants with improved diffusion characteristics.
  • Enhanced diffusion is a key factor for improved antimicrobial efficacy in complex food environments.
  • Nisin A variants with enhanced diffusion show significant potential for controlling Listeria monocytogenes in food products like chocolate milk.