Bacterial Cellulose as a Substrate for Microbial Cell Culture

ABSTRACT Bacterial cellulose (BC) has a range of structural and physicochemical properties that make it a particularly useful material for the culture of bacteria. We studied the growth of 14 genera of bacteria on BC substrates produced by Acetobacter xylinum and compared the results to growth on the commercially available biopolymers agar, gellan, and xanthan. We demonstrate that BC produces rates of bacterial cell growth that typically exceed those on the commercial biopolymers and yields cultures with higher titers of cells at stationary phase. The morphology of the cells did not change during growth on BC. The rates of nutrient diffusion in BC being higher than those in other biopolymers is likely a primary factor that leads to higher growth rates. Collectively, our results suggest that the use of BC may open new avenues in microbiology by facilitating bacterial cell culture and isolation.

[1]  Y. Chang,et al.  Agar degradation by microorganisms and agar-degrading enzymes , 2012, Applied Microbiology and Biotechnology.

[2]  Dieter Klemm,et al.  Bacterial synthesized cellulose — artificial blood vessels for microsurgery , 2001 .

[3]  Paul Gatenholm,et al.  Engineering microporosity in bacterial cellulose scaffolds , 2008, Journal of tissue engineering and regenerative medicine.

[4]  K. Zengler,et al.  Cultivating the uncultured , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[5]  B. Weissberger,et al.  GELRITE as an Agar Substitute in Bacteriological Media , 1983, Applied and environmental microbiology.

[6]  Katrin Frankenfeld,et al.  Loading of Bacterial Cellulose Aerogels with Bioactive Compounds by Antisolvent Precipitation with Supercritical Carbon Dioxide , 2010 .

[7]  D. Weibel,et al.  Polyacrylamide hydrogels as substrates for studying bacteria. , 2012, Chemical communications.

[8]  S. Babbar,et al.  Xanthan Gum: An Economical Partial Substitute for Agar in Microbial Culture Media , 2006, Current Microbiology.

[9]  M. Sadowsky,et al.  Inheritance of host-controlled restriction of nodulation by Bradyrhizobium japonicum strain USDA 110 , 1996 .

[10]  A. Belarbi,et al.  Use of a new gelling agent (Eladium©) as an alternative to agar-agar and its adaptation to screen biofilm-forming yeasts , 2010, Applied Microbiology and Biotechnology.

[11]  P. Janmey,et al.  Tissue Cells Feel and Respond to the Stiffness of Their Substrate , 2005, Science.

[12]  Julian H. George,et al.  Exploring and Engineering the Cell Surface Interface , 2005, Science.

[13]  Richard C Boucher,et al.  Effects of reduced mucus oxygen concentration in airway Pseudomonas infections of cystic fibrosis patients. , 2002, The Journal of clinical investigation.

[14]  S. Babbar,et al.  Guar gum: a cheap substitute for agar in microbial culture media , 2005, Letters in applied microbiology.

[15]  Albert J. Keung,et al.  Substrate modulus directs neural stem cell behavior. , 2008, Biophysical journal.

[16]  A. Hitchens,et al.  The Introduction of Agar-agar into Bacteriology , 1939, Journal of bacteriology.

[17]  A. Oliver,et al.  High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. , 2000, Science.

[18]  Paul Gatenholm,et al.  Bacterial cellulose as a potential meniscus implant , 2007, Journal of tissue engineering and regenerative medicine.

[19]  Molly M Stevens,et al.  Exploring and engineering the cell surface interface. , 2011, Science.

[20]  S. Ribeiro,et al.  Thermal characterization of bacterial cellulose–phosphate composite membranes , 2007 .

[21]  Masatoshi Iguchi,et al.  Bacterial cellulose—a masterpiece of nature's arts , 2000 .

[22]  S. Anderson,et al.  Biogenesis of bacterial cellulose. , 1991, Critical reviews in microbiology.

[23]  Paul Gatenholm,et al.  Bacterial cellulose-based materials and medical devices: current state and perspectives , 2011, Applied Microbiology and Biotechnology.

[24]  K. V. Van Vliet,et al.  Substrata mechanical stiffness can regulate adhesion of viable bacteria. , 2008, Biomacromolecules.

[25]  L. Poulsen,et al.  Substratum-induced morphological changes in a marine bacterium and their relevance to biofilm structure , 1994, Journal of bacteriology.

[26]  Yoshihito Osada,et al.  High Mechanical Strength Double‐Network Hydrogel with Bacterial Cellulose , 2004 .

[27]  Glyn O. Phillips,et al.  Handbook of hydrocolloids. , 2009 .

[28]  L. Hamoen,et al.  Membrane potential is important for bacterial cell division , 2010, Proceedings of the National Academy of Sciences.

[29]  D. Weibel,et al.  Bacterial Swarming: A Model System for Studying Dynamic Self-assembly. , 2009, Soft matter.

[30]  Karen A Robinson,et al.  Cystic fibrosis pulmonary guidelines: chronic medications for maintenance of lung health. , 2007, American journal of respiratory and critical care medicine.

[31]  D. Weibel,et al.  Bacteria-surface interactions. , 2013, Soft matter.

[32]  T. Matsuyama,et al.  Dimorphic transition in Escherichia coli and Salmonella typhimurium: surface-induced differentiation into hyperflagellate swarmer cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.