Suppression of toxin production in Clostridium difficile VPI 10463 by amino acids.
The impact of various growth conditions on the expression of toxins and other proteins by Clostridium difficile VPI 10463 was studied. During non-starved conditions, the rate of toxin synthesis paralleled that of total protein during both exponential growth and stationary phase, and in both defined and complex media. Biotin limitation reduced growth rate and bulk protein synthesis, whereas toxin expression continued, leading to a 50- to 200-fold increase in intracellular toxin levels. Concomitantly, several 22 kDa proteins were up-regulated as revealed by two-dimensional PAGE analysis. The toxin yield was 30-fold higher in peptone yeast extract (PY) than in PY containing glucose (PYG). By contrast, glucose limitation reduced toxin yields by 20- to 100-fold in defined media. By elevating the buffering capacity and bicarbonate concentration, toxin yields were increased by 10-fold in PY and PYG. The high toxin production by C. difficile during growth in PY was lowered 100-fold by adding a blend of nine amino acids and several 60-100 kDa proteins were concomitantly down-regulated. It was concluded that toxin expression in C. difficile VPI 10463 was not affected by growth rate, growth phase, catabolite repression or the stringent response. Instead the co-expression of toxins and a few specific additional proteins appeared to be influenced by metabolic pathways involving CO2 assimilation, carboxylation reactions and metabolism of certain amino acids.
Karlsson
Sture
S
Microbiology and Tumourbiology Center, Karolinska Institute, S-17177 Stockholm, Sweden.|Department of Bacteriology, Swedish Institute for Infectious Disease Control, S-17182 Solna, Sweden.
Burman
Lars G
LG
Department of Bacteriology, Swedish Institute for Infectious Disease Control, S-17182 Solna, Sweden.
Åkerlund
Thomas
T
Department of Bacteriology, Swedish Institute for Infectious Disease Control, S-17182 Solna, Sweden.
D000596:Amino Acids / Q000378:metabolism* / Q000494:pharmacology; D001427:Bacterial Toxins; D001639:Bicarbonates / Q000378:metabolism / Q000494:pharmacology; D001710:Biotin / Q000378:metabolism / Q000494:pharmacology; D016360:Clostridioides difficile / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D003470:Culture Media; D004768:Enterotoxins / Q000096:biosynthesis*; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics
D000596:Amino Acids; D001427:Bacterial Toxins; D001639:Bicarbonates; D003470:Culture Media; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; D001710:Biotin; D005947:Glucose