1887

Abstract

was investigated for extracellular proteinases. The major extracellular proteinase activity detected under all conditions tested was -leucine aminopeptidase activity. Slight extracellular -proline aminopeptidase activity was also detected. No clear evidence for the presence of serine proteinases in culture broths was found using several different methods. The major extracellular proteinase of , i.e. the -leucine aminopeptidase, was purified 33-fold to homogeneity. The purified enzyme was found by SDS-PAGE to have an of 34000. The purified enzyme had a final specific activity of 3·6 units mg, a for -leucine--nitroanilide of 300 μ and a of 4·2 μmol min mg. The pure enzyme did not exhibit proteolytic activity on azocasein or -proline--nitroanilide, substrates for other proteinase activities observed in crude extracts of .

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1993-03-01
2024-12-06
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References

  1. Aretz W., Koller K. P., Riess G. 1989; Proteolytic enzymes from recombinant Streptomyces lividans TK24. FEMS Microbiology Letters 65:31–36
    [Google Scholar]
  2. Bascaran V., Hardisson C., Braña A. 1990; Regulation of extracellular protease production in Streptomyces clavuligerus. Applied Microbiology and Biotechnology 34:208–213
    [Google Scholar]
  3. Bender E., Vogel R., Koller K. -P., Engles J. 1990; Synthesis and secretion of hirudin by Streptomyces lividans. Journal of Bacteriology 34:203–207
    [Google Scholar]
  4. Bradford M. M. 1976; A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  5. Butler M. J., Bergeron A., Krygsman P., Soostmeyer G., Zimny T., Malek L. T. 1991; Cloning and characterization of an aminopeptidase P gene from S. lividans 66. Abstracts, 1991 International Symposium on Biology of Actinomycetes P 1–123
    [Google Scholar]
  6. Butler M. J., Davey C. C., Krygsman P., Walczyk E., Malek L. T. 1992; Cloning of genetic loci involved in endoprotease activity in Streptomyces lividans 66: a novel neutral protease gene with an adjacent divergent putative regulatory gene. Canadian Journal of Microbiology 38:912–920
    [Google Scholar]
  7. Chandrasekaran S., Dhar S. C. 1987; Multiple proteases from Streptomyces moderatus. I. Isolation and purification of five extracellular proteases. Archives of Biochemistry and Biophysics 257:395–401
    [Google Scholar]
  8. Colletta P. L., Miller P. G. G. 1986; The extracellular proteases of Myxococcus xanthus. FEMS Microbiology Letters 37:203–207
    [Google Scholar]
  9. Delmar E. G., Largman C., Brodrick J. W., Geokas M. C. 1979; A sensitive new substrate for chymotrypsin. Analytical Biochemistry 99:316–320
    [Google Scholar]
  10. Dreyfuss O., Adam S. A., Choi Y. D. 1984; Physical change in cytoplasmic messenger ribonucleoproteins in cells treated with inhibitors of mRNA transcription. Molecular and Cell Biology 4:415–423
    [Google Scholar]
  11. Erlanger B. F., Kokowski N., Cohen W. 1961; The preparation and properties of two new chromogenic substrates of trypsin. Archives of Biochemistry and Biophysics 95:271–278
    [Google Scholar]
  12. Feder J. 1968; A spectrophotometric assay for neutral protease. Biochemical and Biophysical Research Communications 32:326–332
    [Google Scholar]
  13. Gibb G. D., Ordaz D. E., Strohl W. R. 1989; Overproduction of extracellular proteinase activity by Streptomyces C5-A13 in fed-batch fermentations. Applied Microbiology and Biotechnology 31:119–124
    [Google Scholar]
  14. Krest’yanova I. N., Vasil’eva L. I., Bartoshevich I. E., Akparov Vkh., Nakhapetian L. A. 1983; Isoelectric focusing of a preparation of proteolytic enzymes from Streptomyces 771. Prikladnaya Biokhimyia i Mikrobiologiya 19:217–225 (English translation: Prikladnaya Biokhimiya i Mikrobiologiya 19 175–183).
    [Google Scholar]
  15. Lampel J. S., Aphale J. S., Lampel K. A., Strohl W. R. 1992; Cloning and sequencing of a gene encoding a novel extracellular neutral proteinase from Streptomyces sp. strain C5 and expression of the gene in Streptomyces lividans 1326. Journal of Bacteriology 174:2797–2808
    [Google Scholar]
  16. Lichenstein H., Brawner M. E., Miles L. M., Myers C. A., Young P. R., Simon P. L., Eckardt T. 1988; Secretion of interleukin-1β and Escherichia coli galactokinase by Streptomyces lividans. Journal of Bacteriology 170:3924–3929
    [Google Scholar]
  17. Lichenstein H. S., Busse L. A., Smith G. A., Narhi L. O., McGinley M. O., Rohde M. F., Katzowitz J. L., Zukowski M. M. 1992; Cloning and characterization of a gene encoding extracellular metalloprotease from Streptomyces lividans. Gene 111:125–130
    [Google Scholar]
  18. Malek L. T., Soostmeyer G., Davey C., Krygsman P., Compton J., Gray J., Zimny T., Stewart D. 1990; Secretion of granulocyte macrophage-colony stimulating factor (GM-CSF) in Streptomyces lividans. Journal of Cell Biochemistry Supplement 14A:127
    [Google Scholar]
  19. Morihara K., Oka T., Tsuzuki H. 1967; Multiple proteolytic enzymes of Streptomyces fradiae. Biochimica et Biophyisca Acta 139:382–387
    [Google Scholar]
  20. Nakamura T., Kiuchi N., Fujiyama S. 1984; Purification and partial characterization of leucine aminopeptidase from Actinomyces viscosus. Matsumoto Shigaku 10:130–135
    [Google Scholar]
  21. Narahashi Y. 1970; Pronase. Methods in Enzymology 19:651–664
    [Google Scholar]
  22. Pokorny M., Vitale Lj., Turk V., Renko M., Zuvanic J. 1979; Streptomyces rimosus extracellular proteinases. 1. Characterization and evaluation of various crude preparations. European Journal of Applied Microbiology and Biotechnology 8:81–90
    [Google Scholar]
  23. Roberts D. V., Elmore D. T. 1974; Kinetics and mechanism of catalysis by proteolytic enzymes. Biochemical Journal 141:545–554
    [Google Scholar]
  24. Spungin A., Blumberg S. 1989; Streptomyces griseus aminopeptidase is a calcium-activated zinc metalloprotein. Purification and properties of the enzyme. European Journal of Biochemistry 183:471–477
    [Google Scholar]
  25. Taguchi S., Kumagai I., Nakayama J., Suzuki A., Miura K. 1989; Efficient extracellular expression of a foreign protein in Streptomyces using secretory proteinase inhibitor (SSI) gene fusions. Bio/Technology 7:1063–1066
    [Google Scholar]
  26. Uwajima T., Yoshikawa N., Terada O. 1973; A crystalline aminopeptidase from Streptomyces peptidofaciens: physio-chemical properties and characteristics as a Ca-metalloprotease. Agricultural and Biological Chemistry 37:2727–2733
    [Google Scholar]
  27. Vinci V. A. 1988 Biochemical and genetic analysis of serine proteases of Streptomyces spp PhD Dissertation The Ohio State University, Columbus, USA:
    [Google Scholar]
  28. Vitale Lj., Renko M., Lenarcic B., Turk V., Pokorny M. 1986; Streptomcyes rimosus extracellular proteinases. 3. Isolation and characterization of leucine aminopeptidase. Applied Microbiology and Biotechnology 23:449–455
    [Google Scholar]
  29. Vosbeck K. D., Chow K., Awad W. M. 1973; The proteolytic enzymes of the K-l strain of Streptomyces griseus obtained from a commercial preparation (pronase). Journal of Biological Chemistry 248:6029–6034
    [Google Scholar]
  30. Wagner F. W., Ray L. E., Ajabnoor M. A., Ziemba P. L., Hall R. L. 1979; Bacillus subtilis aminopeptidase: purification, characterization and some enzymatic properties. Archives of Biochemistry and Biophysics 197:63–72
    [Google Scholar]
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