Trypsin-like protease of Streptomyces exfoliatus SMF13, a potential agent in mycelial differentiation.
Streptomyces exfoliatus SMF13 sequentially produced leupeptin, leupeptin-inactivating enzyme (LIE) and trypsin-like protease (TLP). TLP was produced upon exhaustion of glucose. Autolysis of mycelium was accompanied by an increase in TLP activity. However, in three bld mutants isolated from S. exfoliatus SMF13 after UV-mutagenesis, mycelium autolysis did not occur, and neither LIE nor TLP was produced, although leupeptin was produced. Production of both LIE and TLP was restored in a spontaneous Spo+ revertant of a bld mutant. In contrast, two whi mutants sequentially produced leupeptin, LIE and TLP. The molecular mass of TLP produced during morphological differentiation was estimated to be 31.8 kDa by SDS-PAGE. The N-terminal amino acid sequence was RVGGTxAAQGNFPFQQxLSM. TLP was competitively inhibited by leupeptin; the inhibition constant was 0.015 microM. TLP effectively hydrolysed the mycelial protein extract of S. exfoliatus SMF13, but the hydrolytic activity was inhibited by leupeptin. It was concluded that morphological differentiation and production of TLP are coordinately regulated, that TLP may function as an enzyme in the metabolism of mycelial proteins, and that the hydrolytic activity of TLP is regulated by autogenous leupeptin in S. exfoliatus SMF13.
Kim
I S
IS
Department of Microbiology, College of Natural Sciences, Seoul National University, Korea.
Lee
K J
KJ
D000595:Amino Acid Sequence; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology; D007700:Kinetics; D007976:Leupeptins / Q000096:biosynthesis / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D009154:Mutation; D009842:Oligopeptides / Q000737:chemistry; D011480:Protease Inhibitors / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D013379:Substrate Specificity; D014357:Trypsin / Q000235:genetics / Q000378:metabolism*
D007976:Leupeptins; D009842:Oligopeptides; D011480:Protease Inhibitors; D010450:Endopeptidases; D014357:Trypsin; D005947:Glucose; C032854:leupeptin