Although germinated conidia ofNeurospora crassa transport adenine through two different systems, only one of these, namely, the general purine transport system, which transports adenine, hypoxanthine, guanine, and 6-methylpurine, is present in freshly harvested conidia of the wild type. The second system develops during germination. The latter system can transport adenine and 6methylpurine. Time course and kinetic studies of adenine transport in freshly harvested conidia of an ad-8 mutant indicated that, in contrast to the wild type, the general purine transport activity is very low in this strain and that the second adenine transport system is possibly present in the ungerminated conidia. A study of adenine and hypoxanthine uptake in ad-8 and ad-4 mutants, both of which cannot utilize hypoxanthine for growth, indicated that the two transport systems may be under different metabolic controls.

Twelve cultures of Hyphomycetes capable of growth upon the ethane component of natural gas were isolated from raw sewage by a continuous enrichment technique. In addition, all isolates also grew on propane and n-butane but not methane. These cultures are described and their taxonomic position discussed. Four were tentatively placed in Graphium; three were identified as Phialophora jeanselmei (Langeron) Emmons; the remaining five are similar to Acremonium sensu Gams.

A fungus which grows well on a mineral salts solution with natural gas as the carbon source is described and provisionally identified as a Graphium species. Its taxonomic relation to several genera is presented. This organism was isolated from sewage after selection by enrichment techniques and continuous culture. The fermentor was operated at ambient temperatures, 28 °C ± 2, at a volume of 10 liters with a dilution rate of 10 liters/4 days to 10 liters/1.7 days. Coty's mineral salts medium gave the highest tissue yield. When the pH of the incoming mineral salts medium was decreased stepwise from 7.0 to 5.0 the pH of the reactor became self-adjusting, varying from around 2.7 to 3.5, and the dry weight of microbial tissue obtained varied from 65 to 275 mg/h. Also present in the continuous culture was an acid tolerant bacterium, which, when isolated, grew well on natural gas, methanol, and ethanol, and a strain of Trichoderma, which, when isolated, did not use natural gas as a carbon source. In mixed culture the Trichoderma is thought to grow on metabolites produced by either or both the Graphium and the acid-tolerant bacterium during oxidation of natural gas. The nature of the relationship is being investigated. The mixed culture has been under continuous cultivation for 18 months.

6-Methylpurine, an analog of adenine, inhibits the growth of Neurospora cra88a. From Idnetic studies it was found that 6-methylpurine is converted to its nucleotide form by adenine phosphorbosyltransferase (EC 2.4.2.7), and inhibits the de novo purine biosynthesis. Adenine relieves the growth inhibition caused by 6-methylpurine, whereas hypoxanthine is not very effective. Studies dealing with hypoxanthine utilization in the presence of 6-methylpurine indicated a severely reduced uptake of hypoxanthine and a general slowdown in its further metabolism. Two mutants (Mepr-3 and MepT-10) which are resistant to 6-methylpurine were characterized. Studies of purine base uptake and the in vivo and in vitro conversion to nucleotides indicated that Mepr-10 may be an adenine phosphoribosyltranserase-defective mutant, whereas MepT-3 may be a mutant with altered feedback response to 6-methylpurine. Both mutants showed a severely lowered hypoxanthine phosphoribosyltransferase activity, but because 6-methylpurine did not have any effect on the conversion of hypoxanthine to IMP in the wild type, it was concluded that 6-methylpurine resatance in these mutants cannot be due to lowered hypoxanthine phosphoribosyltransferase activity, but rather that the lowering of enzyme activity may be a secondary effect.

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