DE NOVO SYNTHESIS OF ALPHA-AMYLASE BY BACILLUS STEAROTHERMOPHILUS.
Welker, N. E. (Western Reserve University, Cleveland, Ohio and University of Illinois, Urbana), and L. Leon Campbell. De novo synthesis of alpha-amylase by Bacillus stearothermophilus. J. Bacteriol. 86:1202-1210. 1963.-The pH optimum for the synthesis of alpha-amylase by washed-cell suspensions was 6.7. alpha-Amylase synthesis began soon after the addition of the inducer (maltose, methyl-beta-d-maltoside, or phenyl-alpha-d-glucoside, at 10(-3)m), proceeded at a linear rate for 60 min, and then leveled off. Cell suspensions without inducer produced small amounts of alpha-amylase. The addition of glucose (2 x 10(-3)m), sucrose (10(-3)m), or glycerol (4 x 10(-3)m) to washed-cell suspensions failed to stimulate the production of alpha-amylase. Nitrogen starvation of washed cells for 60 min with fructose as a carbon source or by induction with pure maltose showed that the ability to produce alpha-amylase was lost. Examination of the amino acid pool at this time showed a general depletion of amino acids and the complete disappearance of tyrosine, phenyl-alanine, proline, and valine. Replenishment of the amino acid pool with casein hydrolysate (0.5%) restored the ability of the cells to produce alpha-amylase. Chloramphenicol and 8-azaguanine were shown to inhibit alpha-amylase synthesis. Inhibition was observed immediately upon the addition of chloramphenicol to cell suspensions preinduced for varying periods of time. Actinomycin D and mitomycin C also inhibited alpha-amylase synthesis when added to induced washed-cell suspensions. The amino acid analogues, norvaline, norleucine, and ethionine, inhibited alpha-amylase formation by 72, 53, and 38%, respectively. p-Fluorophenylalanine inhibited the synthesis of active alpha-amylase by 92% and the incorporation of proline-C(14) into alpha-amylase and cellular proteins by 95 and 74%, respectively.
D000596:Amino Acids; D000681:Amylases; D001407:Bacillus; D050260:Carbohydrate Metabolism; D002244:Carbon; D002701:Chloramphenicol; D005632:Fructose; D001411:Geobacillus stearothermophilus; D005947:Glucose; D006863:Hydrogen-Ion Concentration; D008320:Maltose; D008660:Metabolism; D009584:Nitrogen; D011392:Proline; D011506:Proteins; D012106:Research; D013395:Sucrose; D014633:Valine / Q000031:analogs & derivatives; D000516:alpha-Amylases
D000596:Amino Acids; D011506:Proteins; D005632:Fructose; D013395:Sucrose; D002701:Chloramphenicol; D008320:Maltose; D002244:Carbon; D011392:Proline; C005313:norvaline; D000681:Amylases; D000516:alpha-Amylases; D014633:Valine; D005947:Glucose; D009584:Nitrogen
AMINO ACIDS; AMYLASE; BACILLUS; CARBOHYDRATE METABOLISM; CHLORAMPHENICOL; EXPERIMENTAL LAB STUDY; GLUCOSE; HYDROGEN-ION CONCENTRATION; MALTOSE; METABOLISM; NITROGEN; SUCROSE