Insights into Clostridium phytofermentans biofilm formation

Aggregation, microcolony development and the role of extracellular DNA

Trevor R. Zuroff, Weimin Gu, Rachel L. Fore, Susan B. Leschine, Wayne R. Curtis

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Biofilm formation is a critical component to the lifestyle of many naturally occurring cellulose-degrading microbes. In this work, cellular aggregation and biofilm formation of Clostridium phytofermentans, a cellulolytic anaerobic bacterium, was investigated using a combination of microscopy and analytical techniques. Aggregates included thread-like linkages and a DNA/protein-rich extracellular matrix when grown on soluble cellobiose. Similar dense biofilms formed on the surface of the model cellulosic substrate Whatman no. 1 filter paper. Following initially dispersed attachment, microcolonies of ~500 μm diameter formed on the filter paper after 6 days. Enzymic treatment of both the biofilm and cellular aggregates with DNase and proteinase resulted in significant loss of rigidity, pointing to the key role of extracellular DNA and proteins in the biofilm structure. A high-throughput biofilm assay was adapted for studying potential regulators of biofilm formation. Various media manipulations were shown to greatly impact biofilm formation, including repression in the presence of glucose but not the β(1→4)-linked disaccharide cellobiose, implicating a balance of hydrolytic activity and assimilation to maintain biofilm integrity. Using the microtitre plate biofilm assay, DNase and proteinase dispersed ~60 and 30 % of mature biofilms, respectively, whilst RNase had no impact. This work suggests that Clostridium phytofermentans has evolved a DNA/protein-rich biofilm matrix complementing its cellulolytic nature. These insights add to our current understanding of natural ecosystems as well as strategies for efficient bioprocess design.

Original languageEnglish (US)
Pages (from-to)1134-1143
Number of pages10
JournalMicrobiology (United Kingdom)
Volume160
Issue numberPART 6
DOIs
StatePublished - Jan 1 2014

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Clostridium
Biofilms
DNA
Cellobiose
Deoxyribonucleases
Peptide Hydrolases
Anaerobic Bacteria
Disaccharides
Extracellular Matrix Proteins
Ribonucleases
Cellulose
Ecosystem
Life Style
Microscopy
Proteins

All Science Journal Classification (ASJC) codes

  • Microbiology

Cite this

Zuroff, Trevor R. ; Gu, Weimin ; Fore, Rachel L. ; Leschine, Susan B. ; Curtis, Wayne R. / Insights into Clostridium phytofermentans biofilm formation : Aggregation, microcolony development and the role of extracellular DNA. In: Microbiology (United Kingdom). 2014 ; Vol. 160, No. PART 6. pp. 1134-1143.
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Insights into Clostridium phytofermentans biofilm formation : Aggregation, microcolony development and the role of extracellular DNA. / Zuroff, Trevor R.; Gu, Weimin; Fore, Rachel L.; Leschine, Susan B.; Curtis, Wayne R.

In: Microbiology (United Kingdom), Vol. 160, No. PART 6, 01.01.2014, p. 1134-1143.

Research output: Contribution to journalArticle

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