Kinetics of Utilization of Organic Compounds in the Growth of Mycobacterium tuberculosis.

Bowles, Jean A. (University of Colorado School of Medicine, Denver), and William Segal. Kinetics of utilization of organic compounds in the growth of Mycobacterium tuberculosis. J. Bacteriol. 90:157-163. 1965.-To obtain a workable system for a study of the kinetics of nutrient utilization (based on specific quantitative assay) by Mycobacterium tuberculosis, several cultural refinements were introduced: the use of shake culture, a 40-fold increase in the size of inoculum, substitution of glutamate for asparagine as nitrogen source, and elimination of glucose from the medium with glycerol remaining as carbon source. These modifications resulted in reduction to a tenth of the lag phase of glycerol utilization (from 40 to 4 days), and in a greatly increased rate of growth. Both coordinate and sequential patterns of nutrient utilization were in evidence, except in the case of citrate, which was never utilized under a variety of conditions of culture. The coordinate pattern of glucose-glutamate and glucose-glycerol utilization would appear to rule out catabolite repression by glucose. However, elimination of glucose from the medium resulted in elimination of the 4-day lag period before glutamate utilization was initiated, leaving open to question the role of glucose in this system. Evidence is presented for the hypothesis that the sequential pattern of glutamate-glycerol utilization is a function of glutamate repression of glycerol oxidation in the growth of M. tuberculosis, although no diauxie effect is apparent. In a determination of which nutrient-utilization systems were regulated by induction, only in the case of glycerol was evidence obtained for an inducible system. The enzymatic mechanisms underlying these patterns of nutrient utilization are presently being investigated.     

Nitrogenous Compounds and Nitrogen Metabolism in the Liliaceae: II. The Nitrogenous Compounds of Leaves of the Genus Tulipa: Environmental Effects on the Composition of Tulipa gesneriana

Because of the predominance of -methyleneglutamine in Tulipagesneriana a quantitative, chromatographic investigation hasbeen made to show the factors which determine the compositionof the alcohol-soluble nitrogen fraction in Tulipa A comparative study of the amino-acids that occur free in theleaves of fourteen different species of Tulipa has been made,-methyleneglutamine varied from as much as 60–70 per cent,of the amino-acid nitrogen down to levels below those whichpermit its chromatographic detection. The effects of temperature during growth and of diurnal variationon the nitrogenous compounds of the leaves of T. getnerianaare described. Excised shoots of T. gesneriana were exposed to light and darknessto affect the balance of protein synthesis and breakdown sothat the concomitant effects on the -methyleneglutamine acidcompounds and other main components of the soluble nitrogenfraction could be determined. Lastly, the relative ease of entry of C¹⁴ from C¹⁴O₂ into thesugars and certain other compounds of T. gesneriana is contrastedwith the relative difficulty with which it enters the -methyleneglutamineof the mature leaves on excised shoots. The occurrence and metabolism of -methyleneglutamine in T. gesnerianaleaves may, therefore, be regarded as additional to and superimposedupon an otherwise typical pattern of nitrogen metabolism: inthese leaves -methylene-glutamine does not assume the role usuallydischarged by glutamine.     

Studies in the Genus Elaphoglossum Schott: II. The Root and Bud Traces

Further investigations upon some Jamaican species of Elaphoglossumhave shown that, parallel to a series of stelar forms demonstratedearlier, there is another series showing a progressive reductionof the lateral bud trace. The anatomical differences betweenthe root and bud traces are indicated and the development ofthe stele in a young outgrown bud is described.     

Continuous Culture of Ruminal Microorganisms in Chemically Defined Medium: II. Culture Medium Studies

Ruminal ciliates have been grown in continuous culture in chemically defined media and in the absence of viable bacteria. Oligotrichic ruminal ciliates seem to require insoluble carbohydrates for growth; the holotrichic ciliates require soluble carbohydrates, but at low concentrations. Both groups of ciliates utilize amino acids as their principal nitrogen source when these are supplied in micromolar concentrations; at millimolar concentrations, amino acids are toxic, possibly from excessive ammonia formation arising from ciliate deaminase activity. Holotrichic ruminal ciliates are destroyed by overdeposition of amylopectin when glucose is present above 0.1% concentration in the medium. Ecological requirements of ruminal ciliates are also described.