Characterization of the Mutant of Streptomyces sp. SL-387(KCTC 0102BP) Producing Aminopeptidase M Inhibitors

Aminopeptidase M 저해제를 생산하는 Streptomyces sp. SL-387 (KCTC 0102BP) 변이주의 특성

  • Chung, Myung-Chul ;
  • Chun, Hyo-Kon ;
  • Lee, Ho-Jae ;
  • Lee, Choong-Hwan ;
  • Kho, Yung-Hee
  • 정명철 (한국과학기술연구원 유전공학연구소 미생물화학연구그룹) ;
  • 전효곤 (한국과학기술연구원 유전공학연구소 미생물화학연구그룹) ;
  • 이호재 (한국과학기술연구원 유전공학연구소 미생물화학연구그룹) ;
  • 이충환 (한국과학기술연구원 유전공학연구소 미생물화학연구그룹) ;
  • 고영희 (한국과학기술연구원 유전공학연구소 미생물화학연구그룹)
  • Published : 1995.02.01


Since the original productivity of new aminopeptidase M inhibitors MR-387A and B by Streptomyces sp. SL-387 (KCTC 0102BP) was not enough for further chemical and biological evaluation, mutation of parent strain by the treatment of N-methyl-N'-nitro-N-nitrosoguanidine was performed in order to obtain a clone with greater inhibitory activity. Mutant N-3 was selected due to a 6-fold greater productivity (40 $\mu$g/ml) than that of the wild type(6.7 $\mu$g/ml). This mutant was resistant to 3,4-dehydro-DL-proline, an antimetabolite of proline, with 25 $\mu$g/ml of minimum inhibitory concentration. Furthermore, the characteristic morphological change from spiral spore chain in wild type to straight in mutant was observed. An aminopeptidase M nhibitor different from MR-387A and B was isolated from the culture broth of the mutant. This inhibitor was composed of 2 proline, 1 valine, and an unknown amino acid which is presumably 3-amino-4-phenylbutanoic acid. IC$_{50}$ value (89.1 $\MU$g/ml) of the purified inhibitor was lower than that of other inhibitors, which may be due to the absence of 2(S)-hydroxyl group within the structure of 3-amino-4-phenyl- butanoic acid.


  1. J. Antibiotics. v.38 Production of actinonin, an inhibitor of aminopeptidase M, by actinomycetes Umezawa,H.;T.Aoyagi;T.Tanaka;H.Suda;A.Okuyama;H.Naganawa;M.Hamada;T.Takeuchi
  2. Kor. J. Appl. Microbiol. Biotechnol. v.22 Taxonomic characteristics of strain producing MR-387A and B, new inhibitors of aminopeptidase M, and their production Chung,M.C.;H.K.Chun;H.J.Lee;Y.H.Kho
  3. J. Antibiotics. v.43 Probestin, a new inhibitor of aminopeptidase M, produced by Streptomyces azureus MH663-2F6 Aoyagi,T.;S.Yoshida;Y.Nakamura;Y.Shgihara;M.Hamada;T.Takeuchi
  4. J. Med. Chem. v.20 Synthesis and structure-activity relationships of bestatin analogues, inhibitors of aminopeptidase B Nishzawa,R.;T.Saino
  5. J. Biochem. Mol. Biol. v.28 The novel synthetic substance MR-387 C(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl-L-valyl-L-prolyl-L-leucine as an aminopeptidase M inhibitor Cheng,M.C.;H.K.Chun;H.J.Lee;Y.H.Kho
  6. Biochim. Biophys. Acta. v.40 N-terminal degradation of low molecular weight opioid peptide in human cerebrospinal fluid Benterm,I.F.;E.M.Hirsh;A.J.Tuchman;P.E.Ward
  7. Biochim. Biophys. Acta. v.452 Aminopeptidase activities on the surface of mammalian cells Aoyagi,T.;H.Suda;M.Nagai;K.Ogawa;J.Suzuki;T.Takeuchi;H.Umezawa
  8. J. Pharmacol. Exp. Ther. v.261 Inhibition of the enkephalin-metabolizing enzymes by the first systemically active mixed inhibitor prodrug RB101 induces potent analgesic responces in mice and rats Noble,F.;J.M.Soleilhac;E.Soroca-Lucas;S.Turcaud;M.C.Fournie-Zaluski;B.P.Roques
  9. J. Antibiotics. v.29 Bestatin, an inhibitor of aminopeptidase B, produced by actinomycetes Umezawa,H.;T.Aoyagi;H.Suda;M.Hamada;T.Takeuchi
  10. Mol. Pharmacol. v.29 Characterization of aminopeptidases responsible for inactivating endogenous (Met) enkephalin in brain slices using peptidase inhibitors and aminopeptidase M antibodies Giros,B.;C.Gros;B.Solhonne;J.C.Schwartz
  11. J. Antibiotics. v.31 Amastatin, an inhibitor of aminopeptidase A, produced by actinomycetes Aoyagi,T.;H.Tobe;F.Kojima;M.Hamada;T.Takeuchi;H.Umezawa
  12. Mammalian Ectoenzymes. Kenny,A.J.;S.L.Stephenson;A.J.Turner
  13. Degradation of bioactive substances;Physiology and pathology Peptidases involved in the metabolism of bioactive peptides Kenny,A.J.;N.M.Hooper;J.H.Henrickson(ed.)
  14. J. Antibiotics. v.44 Leuhistin, a new inhibitor of aminopeptidase M, produced by Bacillus laterosporus BMII56-14F1 Aoyagi,T.;S.Yoshida;N.Matsuda;T.Ikeda;M.Hamada;T.Takeuchi
  15. Agric. Biol. Chem. v.46 Synthesis and structure-activity relationships of amastatin analogues, inhibitors of aminopeptidase A Tobe,H.;H.Morishima;T.Aoyagi;H.Umezawa;K.Ishiki;K.Nakamura;T.Yoshioka;Y.Shimauchi;T.Inui
  16. Biochemistry. v.24 Identification of aminopeptidase M as an enkephaline-inactivating enzyme in rat cerebral membranes Gros,C.;B.Giros;J.C.Schwartz