Further evidence for a partially folded intermediate in penicillinase secretion by Bacillus licheniformis.
Protoplasis of Bacillus licheniformis 749/C (a mutant constitutive for penicillinase production) continued to synthesize and release penicillinase in hypertonic growth medium in the presence of trypsin and chymotrypsin at 25 mug each per ml. When the protoplasts were stripped of about half of their membrane-bound penicillinase by pretreatment at pH 9.5 or with a higher level of trypsin, penicillinase activity no longer increased in the presence of the proteases. This effect was immediately eliminated after addition of soybean trypsin inhibitor. These proteases do not significantly inhibit general protein synthesis. Stripped protoplasts of strain 749/C and of uninduced strain 749 (unable to synthesize penicillinase) were incubated with 50 mug of chymotrypsin per ml, and the supernatent fluids were examined immunochemically for peptides derived from the penicillinase chain. Such fargments were found only with the protoplasts capable of synthesizing penicillinase (strain 749/C). The direct detection of the products of protease degradation of a susceptible form of penicillinase provides strong evidence that, in stripped protoplasts of B. licheniformis 749/C, penicillinase synthesis continues in the presence of trypsin or chymotrypsin and that, in these modified membranes, the protease is able to act on an early form of the enzyme that has not yet attained the protease-resistant conformation characteristic of the membrane-bound and exopenicillinases. This finding is discussed in terms of the current models of penicillinase secretion.
D000818:Animals; D001407:Bacillus / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D002918:Chymotrypsin / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D010405:Penicillinase / Q000096:biosynthesis / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D011487:Protein Conformation; D011523:Protoplasts / Q000201:enzymology*; D011817:Rabbits / Q000276:immunology; D011863:Radioimmunoassay; D014357:Trypsin / Q000378:metabolism
D010446:Peptide Fragments; D002918:Chymotrypsin; D014357:Trypsin; D010405:Penicillinase