Comparison of acetohydroxy-acid synthetases from two extreme thermophilic bacteria.
The acetohydroxy-acid synthetases from two extreme bacterial thermophiles, Thermus aquaticus and Bacillus sp., have been studied. The two enzymes have different pH optima, 8 and 6, respectively, and both are feedback inhibited by valine. The inhibition is of interest because it is not expressed below 60 C, but only at higher temperatures which are optimal for catalytic activity. Valine inhibition in T. aquaticus was noncompetitive, whereas in Bacillus sp., it was competitive. Isoleucine (10(-3) M) also inhibited the two enzymes, whereas leucine (10(-3) M) did not. There was no concerted feedback when the amino acids were added in together. The sensitivity of the enzymes to valine could not be removed by HgCl(2). Both enzymes required Mg(2+) and thiamine pyrophosphate for optimal activity, whereas only the enzyme from T. aquaticus required flavine adenine dinucleotide in addition. None of these cofactors was essential for the feedback inhibition caused by valine. The enzymes from both bacteria could be repressed, but only in the presence of all three branched-chain amino acids indicating that, as in Escherichia coli and Salmonella typhimurium, the repression system is multivalent.
D001407:Bacillus / Q000201:enzymology* / Q000254:growth & development; D001419:Bacteria / Q000201:enzymology* / Q000254:growth & development; D002474:Cell-Free System; D004794:Enzyme Repression; D005246:Feedback; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007532:Isoleucine / Q000494:pharmacology; D007773:Lactates / Q000096:biosynthesis; D007930:Leucine / Q000494:pharmacology; D008190:Lyases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D008628:Mercury / Q000494:pharmacology; D011773:Pyruvates; D013045:Species Specificity; D013835:Thiamine Pyrophosphate / Q000378:metabolism; D014633:Valine / Q000494:pharmacology
D007773:Lactates; D011773:Pyruvates; D007532:Isoleucine; D005182:Flavin-Adenine Dinucleotide; D008190:Lyases; D008628:Mercury; D007930:Leucine; D014633:Valine; D008274:Magnesium; D013835:Thiamine Pyrophosphate