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Abstract: An aldolase was partially purified from fermenter grownMycobacterium tuberculosis H37Rv cells. The aldolase has a molecular weight of 150,000, possesses a tetrameric structure and cleaves both fructose diphosphate and fructose-1-phosphate, the former being cleaved 17 times faster. The enzyme was inactivated by treatment with NaBH4 in the presence of fructose diphosphate or dihydroxyacetone, phosphate suggesting Schiff base formation during its catalytic function. Thiol reagents, EDTA and metal ions had no apparent effect on the aldolase activity. These results show that aldolase is of Class I type. However, this enzyme, unlike the mammalian Class I aldolase, was unaffected by carboxypeptidase A. N-ethylmaleiniide and dithionitrobenzoic acid.

Uptake of radioactive tryptophan by Mycobacterium tuberculosisH37Rv grown in vitro and in vivo was investigated. Kmvalues indicated low affinity, and sodium azide inhibited uptake. Rifampin at the minimal inhibitory concentration had no effect, whereas ethambutol inhibited uptake only in the bacilli grown under in vitro conditions. The significance of these results is discussed.

1. Activation of glucose 6-phosphate is one of the unique properties of pyruvate kinase from Mycobacterium smegmatis. 2. Pyruvate kinase, partially purified from ultrasonic extracts of the mycobacteria by (NH4)2SO4 fractionation, exhibited sigmoidal kinetics at various concentrations of phosphoenolpyruvate, with a high degree of co-operativity (Hill coefficient, h = 3.7) and S0.5 value of 1.0 mM. 3. In the presence of glucose 6-phosphate, the degree of co-operativity shown by the phosphoenolpyruvate saturation curve was decreased to h = 2.33 and the S0.5 value was lowered to 0.47 mM. 4. The enzyme was activated by AMP and ribose 5-phosphate also, but the activation constant was lowest with glucose 6-phosphate (0.24 mM). 5. The enzyme was strongly inhibited by ATP at all phosphoenolpyruvate concentrations. The concentrations of ATP required to produce half-maximal inhibition of enzyme activity at non-saturating (0.2 mM) and saturating (2 mM) phosphoenolpyruvate concentrations were 1.1 mM and 3 mM respectively. 6. The inhibition of ATP was partially relieved by glucose 6-phosphate. 7. The enzyme exhibited Michaelis-Menten kinetics with ADP as the variable substrate, with an apparent Km of 0.66 mM. 8. The enzyme required Mg2+ or Mn2+ ions for activity. It was not activated by univalent cations. 9. The kinetic data indicate that under physiological conditions glucose 6-phosphate probably plays a significant role in the regulation of pyruvate kinase activity.

Some biochemical properties ofTrigonella foenum-groecuminfected withPeronospora trifoliorumhave been investigated. Qualitative changes were observed in lipids, coumarins, amino acids and indole compounds only. A quantitative study of the phosphate content was also carried out. Neutral lipids did not show any significant changes. In addition to the fourteen phospholipid bands, the infected leaves contained a new band corresponding in Rfto phosphatidic acid or polyglycerophosphate. Seven coumarins were present in both the extracts. In addition 3-hydroxy coumarin, 5-hydroxy-4-methylcoumarin and 7-hydroxy-4-methyl-coumarin were detected in infected leaves. Higher levels of indole acetic acid and three of its derivatives were observed due to infection. Infection altered the free amino acid content. Tryptophan appeared and the total phosphate content increased. Increase in phenols and indoles suggest a higher respiratory rate whereas a higher content of inorganic phosphate suggests uncoupling of phosphorylation from respiration.

Aspergillus parasiticusATCC 15517 produced 28 to 30 mg of aflatoxin per 100 ml of a medium containing sucrose, asparagine, and salts in stationary and shaken cultures. In the absence of asparagine in the medium, the toxin yields fell drastically, and the thin-layer chromatograms of the chloroform extracts of the cultures indicated the total absence of aflatoxin G1and the presence of new intense blue and green fluorescent bands having RFvalues lower than aflatoxins. Initial pH was critical and had to be around 4.5 for good growth and high toxin production on this medium. Optimum concentrations of KH2PO4and MgSO4·7H2O in the medium were much lower than those normally used in fungal growth media.