Applied Microbiology and Biotechnology

, Volume 93, Issue 2, pp 671–678 | Cite as

Enhanced production of nukacin D13E in Lactococcus lactis NZ9000 by the additional expression of immunity genes

  • Tijo Varghese Puramattathu
  • Mohammad R. Islam
  • Mami Nishie
  • Sae Yanagihara
  • Jun-ichi Nagao
  • Ken-ichi Okuda
  • Takeshi Zendo
  • Jiro Nakayama
  • Kenji SonomotoEmail author
Applied genetics and molecular biotechnology


Nukacin D13E (D13E) is a variant of type-A(II) lantibiotic nukacin ISK-1 produced by Staphylococcus warneri ISK-1. D13E exhibited a twofold higher specific antimicrobial activity than nukacin ISK-1 against a number of Gram-positive bacteria. We previously reported the heterologous production of D13E in Lactococcus lactis NZ9000 under the control of nisin-controlled gene expression system. In this study, we demonstrated enhanced production of D13E by the additional expression of immunity genes, nukFEG. The nukacin ISK-1 immunity, conferred by the ABC transporter complex, NukFEG, and the lantibiotic-binding protein, NukH, was not overwhelmed by D13E. The additional NukFEG resulted in a fourfold increase in the immunity level of the strain and a 5.2-fold increase in D13E production. The additional NukFEGH-expressing strain with the highest D13E immunity showed reduced level of production. Further improvement in D13E production was achieved by using pH-controlled batch fermentation.


Lantibiotic Nukacin ISK-1 Nisin-controlled gene expression Immunity genes Fermentation 



The authors would like to thank Dr. Shun Iwatani for his helpful suggestions. This work was partially supported by grants from the following organizations: the Japan Society for the Promotion of Science (JSPS), the Japan Science Society, the Novartis Foundation (Japan) for the Promotion of Science, the Novozymes Japan Research Fund, and the Nagase Science and Technology Foundation.

Supplementary material

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Tijo Varghese Puramattathu
    • 1
  • Mohammad R. Islam
    • 1
  • Mami Nishie
    • 1
  • Sae Yanagihara
    • 1
  • Jun-ichi Nagao
    • 2
  • Ken-ichi Okuda
    • 1
  • Takeshi Zendo
    • 1
  • Jiro Nakayama
    • 1
  • Kenji Sonomoto
    • 1
    • 3
    Email author
  1. 1.Laboratory of Microbial Technology, Division of Applied Molecular Microbiology and Biomass Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate SchoolKyushu UniversityFukuokaJapan
  2. 2.Section of Infection Biology, Department of Functional BioscienceFukuoka Dental CollegeFukuokaJapan
  3. 3.Laboratory of Functional Food Design, Department of Functional Metabolic Design, Bio-Architecture CenterKyushu UniversityFukuokaJapan

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