Cell wall composition and surface properties in Bacillus subtilis: anomalous effect of incubation temperature on the phage-binding properties of bacteria containing varied amounts of teichoic acid.

Adsorption of bacteriophage SP50 to walls and heat-killed cells of Bacillus subtilis 168 appeared to be irreversible at both 37 and 0 degree C. Few, if any, active phage were desorbed when phage-wall complexes, formed at either temperature, were suspended in fresh medium. Bacteria rich in wall teichoic acid (TA) bound phage rapidly at both 0 and 37 degrees C, binding at the higher temperature being approximately twice as fast. Bacteria containing diminished proportions of TA showed less rapid phage adsorption but the reduction in rate was greater at 37 than at 0 degree C and bacteria containing only small proportions of TA bound phage more rapidly at 0 degree C than they did at 37 degrees C. These findings show that at low phage receptor density the temperature affects some component(s) involved in the phage-bacterium interaction such that the collision efficiency is increased at the lower temperature. The possible effect of temperature on the organization of bacterial surface components is discussed.

Relation between wall teichoic acid content of Bacillus subtilis and efficiency of adsorption of bacteriophages SP 50 and phi 25.

Efficient adsorption of bacteriophages SP 50 and phi 25 occurred only to bacilli that contained wall teichoic acid and neither phage bound to phosphate limited bacilli that contained teichuronic acid instead of teichoic acid. Though both phages require the presence of teichoic acid, their receptors are not identical. Efficient binding of phage phi 25 required the presence of greater proportions of teichoic acid in the wall and the receptor for this phage was destroyed when bacteria or isolated walls were heated at pH 4 whereas the ability of these samples to bind phage SP 50 was unaffected by such treatment. Efficient binding of phage SP 50 was not highly dependent on the presence of glucosyl substituents on the teichoic acid. Such substituents were required for phage phi 25 binding though their anomeric configuration appeared to be unimportant since the phages bound well to both strains W23 and 168, the wall teichoic acids of which carry glucosyl substituents of opposite anomeric configuration. The differences in the nature of the receptors may be of value in the use of the phages as probes for the location and distribution of teichoic acid in the wall.

Influence of phosphate supply on teichoic acid and teichuronic acid content of Bacillus subtilis cell walls.

Bacillus subtilis 168 was grown in chemostat culture in fully defined media containing a constant concentration of magnesium and concentrations of phosphate that varied from those giving phosphate-limited growth to those in which phosphate was present in excess and magnesium was limiting. Phosphate-limited bacteria were deficient in wall teichoic acid and contained less than half as much cellular phosphate as did bacteria grown in excess of phosphate. Approximately 70% of the additional phosphate in the latter bacteria was present as wall teichoic acid, indicating that the ability of the bacteria to discontinue teichoic acid synthesis when grown under phosphate limitation permits a substantial increase in their growth yield. Since not all of the additional phosphate is present as wall teichoic acid other cellular phosphates may also be present in reduced amounts in the phosphate-limited bacteria. The content of phosphate groups in walls of magnesium-limited bacteria was similar to the content of uronic acid groups in walls of phosphate-limited bacteria, and walls of bacteria grown in media of intermediate composition contained intermediate proportions of the two anionic polymers. Phage SP50, used as a marker for the presence of teichoic acid, bound densely to nearly all of the bacteria in samples containing down to 22% of the maximum content of teichoic acid. Apparently, therefore, nearly all of these bacteria contain teichoic acid, and the population does not consist of a mixture of individuals having exclusively one kind of anionic polymer. Bacteria containing less than 22% of the maximum content of teichoic bound in a nonuniform manner, and possible explanations for this are discussed.