An active center tryptophan residue in liquefying alpha-amylase from Bacillus amyloliquefaciens.
Liquefying alpha-amylase from Bacillus amyloliquefaciens was inactivated on treatment with N-bromosuccinamide. Preincubation of the enzyme with either of the substrate, or competitive inhibitor provided significant protection against inactivation. The relationship between activity loss and the number of tryptophan residues modified, as well as presence of substrate/inhibitor in the reaction mixture, demonstrated that only one of three modifiable tryptophan residues is at or near the active center. The apparent Km of the modified enzyme for soluble starch increased manifold, thus implicating the sensitive tryptophan residue in the substrate binding region of the enzyme.
D001407:Bacillus / Q000201:enzymology*; D001665:Binding Sites; D001974:Bromosuccinimide / Q000494:pharmacology; D055598:Chemical Phenomena; D002621:Chemistry; D006868:Hydrolysis; D013053:Spectrophotometry; D013213:Starch / Q000378:metabolism; D014364:Tryptophan / Q000032:analysis*; D014443:Tyrosine / Q000032:analysis; D000516:alpha-Amylases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*
D014443:Tyrosine; D014364:Tryptophan; D013213:Starch; D000516:alpha-Amylases; D001974:Bromosuccinimide