A thermophilic bacillus (minimal growth temperature 41 C, optimal 55 to 58 C, and maximal 65 C) was isolated from a manure pile. It is very similar to Bacillus stearothermophilus, but it differs in its inability to hydrolyze starch. The thermophilic isolate is a prototroph which grows in a minimal medium consisting of glucose, ammonium salt, phosphate buffer, and inorganic salts. At all temperatures studied (low to high), the same minimal nutritional requirements prevailed. The Arrhenius constant for growth was found to be 15,000 and 13,500 cal/mole in the minimal and rich media, respectively.

A new procedure for production and purification of the thermophilic bacteriophage TP-84 in high yields is described. Cultures of Bacillus stearothermophilus strain 10, enriched with nutrients to obtain heavy growth and to prevent sporulation and maintained at a pH of 6.5, were infected with the phage in a 100-liter fermentor. Addition of magnesium chloride (0.01 M) and a temperature of 58 C were essential for maximal phage production. Phage (5 × 10 11 infective particles/ml) was precipitated with polyethylene glycol (molecular weight 6,000) in the presence of sodium chloride and was further purified by cesium chloride density centrifugation.

A 250- to 300-fold purification of a nicotinamide adenine denucleotide phosphate (NADP)-dependent glutamate dehydrogenase (GDH, E.C. 1.4.1.4) with a yield of 60% from a thermophilic bacillus is described. More than one NADP-specific GDH was detected by polyacrylamide gel electrophoresis. The enzyme is of high molecular weight (approximately 2 X 10-6), similar to that of the beef and frog liver GDH. The pI of the thermophilic GDH is at pH 5.24. The enzyme is highly thermostable at the pH range of 5.8 to 9.0. The purified GDH, unlike the crude enzyme, was very labile at subzero temperatures. An unidentified factor(s) from the crude cell-free extract prevented the inactivation of the purified GDH at -70 C. Various reactants of the GDH system and D-glutamate also protected, to some extent, the enzyme from inactivation at -70 C. From the Michaelis constants for glutamate (1.1 X 10-2M), NADP (3 X 10-4M), ammonia (2.1 X 10-2M), alpha-ketoglutarate (1.3 X 10-3M), and reduced NADP (5.3 X 10-5M), it is suggested that the enzyme catalyzes in vivo the formation of glutamate from ammonia and alpha-ketoglutarate. The amination of alpha-ketoglutarate and deamination of glutamate by the thermophilic GDH are optimal at the pH values of 7.2 and 8.4, respectively.

Protein breakdown of 5 to 7% per hr was found in nitrogen-starved cells of an unclassified prototrophic thermophilic bacillus; a similar protein-breakdown rate (6.5% per hr) was found in resting cells of Escherichia coli. In the thermophile, the rate of protein breakdown was markedly influenced by the temperature; it was maximal between 45 and 55 C, and it decreased considerably at 35 and 75 C, temperatures which are only slightly below or above the minimal and maximal growth temperatures. Growing cultures of the thermophile showed little, if any, protein breakdown, a finding similar to that of others with E. coli.