Biochemical and physiological characterization of the efrotomycin fermentation

Summary An efrotomycin fermentation was characterized through physical, chemical and biochemical studies. Growth of the actinomycete,Nocardia lactamdurans occurred during the first 50 h of the fermentation cycle at the expense of glucose, protein, and triglycerides. The initiation of efrotomycin biosynthesis was observed when glucose dropped to a low concentration. Upon glucose depletion, cell growth ceased and a switch in the respiratory quotient occurred. Efrotomycin biosynthesis was supported by the utilization of soybean oil and starch. Analysis of triglyceride metabolism showed that no diglycerides or monoglycerides accumulated during the fermentation. The activity of extracellular enzymes (lipase, protease, and amylase) increase during the cell growth phase and decreased significantly after 150 h. The concentrations of DNA, tetrahydro-vitamin K₂ (a membrane component), and free amino acids in the supernatant increased dramatically late in the fermentation cycle (225 h), indicating massive cell lysis. During this same time period, a reduction in cellular respiratory activity and efrotomycin biosynthesis were observed.     

Chemical composition of gas vesicles isolated from Anabaena flos-aquae

Summary Chemical analysis of purified preparations of gas vesicles isolated from the filamentous blue-green alga Anabaena flos-aquae has shown that they are similar in composition to those isolated from the unicellular alga Microcystis aeruginosa by Jones and Jost (1970), being proteinaceous structures, free of lipid and carbohydrate. The gas vesicle protein from Anabaena contains the same 14 amino acids, in broadly the same proportions; in addition there is a small proportion of proline. No sulphur-containing amino acids are present. The empirical formula, suggested by the amino acid ratios, indicates a molecule of 15000 MW.     

Synthesis of Two Natural Salmon Calcitonins

The structure of calcitonin isolated from salmon¹ (SCT 1) has been recently established² and confirmed by synthesis⁸. It possesses an exceptionally high level of activity, exhibiting about 20–50 times the hypocalcaemic potency of any other calcitonin isolated from mammalian species.     

The role of phospholipids in the modulation of enzyme activities in the chromaffin granule membrane

(1) 93% of protein of chromaffin granule membranes can be solubilized by 1.3% (w/v) sodium cholate. The solubilized material can be substantially delipidated by ammonium sulphate precipitation. After three such cycles less than 2% of the endogenous phospholipids remain. (2) The chromaffin granule membrane Mg2+-ATPase depends on the presence of phospholipids for retention of its full activity. Soybean and extracted chromaffin granule phospholipids fully reactivate the delipidated enzyme provided only one delipidation step is used. (3) Successive ammonium sulphate precipitation steps result in a delipidated, and deactivated ATPase preparation which can be only partially reactivated on re-addition of phospholipids. (4) The phospholipid specificity for reactivation of the Mg2+-ATPase is broad. Although acidic phospholipids allow higher activities than neutral phospholipids, the main requirement appears to be the hydrophobic environment provided by the phospholipid hydrocarbon chains. (5) Correlations between changes in slope in the Arrhenius plot of the Mg2+-ATPase, and phase transitions in the phospholipid used for reactivation suggest that the 'fluidity' of the hydrocarbon chains modulates the activity of the enzyme.