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Archives of Microbiology

, Volume 181, Issue 4, pp 314–323 | Cite as

Bacillus amyloliquefaciens strains isolated from moisture-damaged buildings produced surfactin and a substance toxic to mammalian cells

  • Raimo MikkolaEmail author
  • Maria A. Andersson
  • Pavel Grigoriev
  • Vera V. Teplova
  • Nils-Erik L. Saris
  • Frederick A. Rainey
  • Mirja S. Salkinoja-Salonen
Original Paper

Abstract

Fungicidic Bacillus amyloliquefaciens strains isolated from the indoor environment of moisture-damaged buildings contained heat-stable, methanol-soluble substances that inhibited motility of boar spermatozoa within 15 min of exposure and killed feline lung cells in high dilution in 1 day. Boar sperm cells lost motility, cellular ATP, and NADH upon contact to the bacterial extract (0.2 μg dry wt/ml). Two bioactive substances were purified from biomass of the fungicidal isolates. One partially characterized substance, 1,197 Da, was moderately hydrophobic and contained leucine, proline, serine, aspartic acid, glutamic acid and tyrosine, in addition to chromophore(s) absorbing at 365 nm. In boar sperm and human neural cells (Paju), the compound depolarized the transmembrane potentials of mitochondria (ΔΨ m) and the plasma membrane (ΔΨ p) after a 20-min exposure and formed cation-selective channels in lipid membranes, with a selectivity K+:Na+:Ca2+ of 26:15:3.5. The other substance was identified as a plasma-membrane-damaging lipopeptide surfactin. Plate-grown biomass of indoor Bacillus amyloliquefaciens contained ca. 7% of dry weight of the two substances, 1,197 Da and surfactin, in a ratio of 1:6 (w:w). The in vitro observed simultaneous collapse of both cytosolic and mitochondrial ATP in the affected mammalian cell, induced by the 1,197-Da cation channel, suggests potential health risks for occupants of buildings contaminated with such toxins.

Keywords

Bacillus amyloliquefaciens Black lipid membrane Cation channel Ionophore Membrane-damage Mitochondria Moisture-damaged building Plasma membrane Surfactin Toxin 

Abbreviations

RP-HPLC

Reversed-phase high-performance liquid chromatography

BLM

Black lipid membrane

DAD

Diode-array detector

ΔΨm

Mitochondrial membrane potential

ΔΨp

Plasma membrane potential

JC-1

5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethylbenz-imidazolo carbocyanine iodide

AM

Calcein acetoxymethyl ester

PI

Propidium iodide

MALDI-TOF-MS

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry

ESI-IT-MS

Electrospray ionization ion trap mass spectrometry

EC50

Endpoint concentration which caused ≥50% change in the viability parameters

FCCP

Carbonyl cyanide 4-trifluoromethoxyphenylhydrazone

Notes

Acknowledgments

The study was supported by grants from Academy of Finland grant 50733, the grant for Center of Excellence “Microbial Resources,” Magnus Ehrnrooth Foundation and the Finnish Medical Society. We would like to thank Leif C. Andersson for collaboration, Anu Harju for growing Paju cells, the Viikki Science Library for excellent information services, Hannele Tukiainen, Leena Steininger and Tuula Suortti for many kinds of help.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Raimo Mikkola
    • 1
    Email author
  • Maria A. Andersson
    • 1
  • Pavel Grigoriev
    • 1
    • 2
  • Vera V. Teplova
    • 1
    • 3
  • Nils-Erik L. Saris
    • 1
  • Frederick A. Rainey
    • 4
  • Mirja S. Salkinoja-Salonen
    • 1
  1. 1.Department of Applied Chemistry and MicrobiologyUniversity of HelsinkiHelsinkiFinland
  2. 2.Institute of Cell BiophysicsRussian Academy of SciencesMoscow RegionRussia
  3. 3.Institute of Theoretical and Experimental BiophysicsRussian Academy of SciencesMoscow RegionRussia
  4. 4.Department of Biological SciencesLouisiana State UniversityBaton RougeUSA

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