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Biofilm formation by a Bacillus subtilis strain that produces -polyglutamate

Masaaki Morikawa^1,2,

Mitsuru Haruki^1,

¹ Department of Material and Life Science, Graduate School of Engineering, Osaka University, Osaka 565-0871, Japan
² Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA

Correspondence
Masaaki Morikawa
morikawa{at}ees.hokudai.ac.jp

The extracellular matrix produced by Bacillus subtilis B-1, an environmental strain that forms robust floating biofilms, was purified, and determined to be composed predominantly of -polyglutamate (-PGA), with a molecular mass of over 1000 kDa. Both biofilm formation and -PGA production by B. subtilis B-1 increased with increasing Mn²⁺ or glycerol concentration. -PGA was produced in a growth-associated manner in standing culture, and floating biofilms were formed. However, -PGA was produced in a non-growth-associated manner in shaking culture conditions. When B. subtilis B-1 was grown in a microaerated culture system, floating biofilm formation and -PGA production were significantly retarded, suggesting that oxygen depletion is involved in the initial steps of floating biofilm formation in standing culture. Proteomic analysis of membrane proteins demonstrated that flagellin, oligopeptide permease and Vpr protease precursor were the major proteins produced by cells in a floating biofilm and a colony.

Present address: Division of Biosphere Science, Graduate School of Environmental Science, Hokkaido University, Sapporo 060-0810, Japan.

Present address: Department of Materials Chemistry and Engineering, College of Engineering, Nihon University, Koriyama, Fukushima 963-8642, Japan.

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