1887

Abstract

A high -asparaginase (-asparagine amidohydrolase; EC 3.5.1.1) activity was found under conditions of lysine overproduction in cultures of . -Asparaginase was purified 98-fold by protamine sulphate precipitation, DEAE-Sephacel anion exchange, ammonium sulphate precipitation and Sephacryl S-200 gel filtration. The asparaginase protein was subjected to PAGE under non-denaturing conditions, identified by an reaction and eluted from the gel in an active form. The estimated from gel filtration and SDS-PAGE was 80000. The -asparaginase activity was inhibited by the -asparagine analogue 5-diazo-4-oxo--norvaline. Neither -asparagine nor -glutamine was a substrate for the enzyme. -Asparaginase was produced constitutively; its role may be that of an overflow enzyme, converting excess asparagine into aspartic acid, the direct precursor of lysine and threonine.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-3-515
1990-03-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/3/mic-136-3-515.html?itemId=/content/journal/micro/10.1099/00221287-136-3-515&mimeType=html&fmt=ahah

References

  1. Abe S., Takayama K. I., Kinoshita K. S. 1967; Taxonomical studies on glutamic acid-producing bacteria. Journal of General and Applied Microbiology 13:279–301
    [Google Scholar]
  2. Bachmann B. J. 1983; Linkage map of Escherichia coli K-12.Edition 7. Microbiological Reviews 47:180–230
    [Google Scholar]
  3. Del Casale T., Sollitti P., Chesney R. H. 1983; Cytoplasmic l-asparaginase: isolation of a defective strain and mapping of asnA. Journal of Bacteriology 154:513–515
    [Google Scholar]
  4. Handschumacher R. E., Bates C. J., Chang P. K., Andrews A. T., Fischer G. A. 1968; 5-Diazo-4-oxo-l-norvaline: reactive asparagine analog with biological specificity. Science 161:62–63
    [Google Scholar]
  5. Hjerten S. 1962; Molecular sieve chromatography on polyacrylamide gels, prepared according to a simplified method. Archives of Biochemistry and Biophysics suppl. 1:147–151
    [Google Scholar]
  6. Ishino S., Yamaguchi K., Shirahata K., Araki K. 1984; Involvement of meso-α, ɛ-diaminopimelate d-dehydrogenase in lysine biosynthesis in Corynebacterium glutamicum. Agricultural and Biological Chemistry 48:2557–2560
    [Google Scholar]
  7. Jones B. N., Paabo S., Stein S. 1981; Amino acid analysis and enzymatic sequence determination of peptides by an improved o-phthaldialdehyde precolumn labeling procedure. Journal of Liquid Chromatography 4:565–586
    [Google Scholar]
  8. Kafkewitz D., Goodman D. 1974; l-Asparaginase production by the rumen anaerobe Vibrio succinogenes. Applied Microbiology 21:206–209
    [Google Scholar]
  9. Kase H., Nakayama K. 1974; Mechanism of l-threonine and l-lysine production by analog-resistant mutants of Corynebacterium glutamicum. Agricultural and Biological Chemistry 38:993–1000
    [Google Scholar]
  10. Kinoshita S., Nakayama K., Akita S. 1958; Production of l-glutamic acid. Bulletin of the Agricultural Chemical Society of Japan 22:176–185
    [Google Scholar]
  11. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  12. Martín J. F. 1989; Molecular genetics of amino acid-producing corynebacteria. Symposia of the Society for General Microbiology 44:25–59
    [Google Scholar]
  13. Martín J. F., Santamaria R., Sandoval M., Mateos L. M., Gil J. A., Aguilar A. 1987; Cloning systems in amino acid-producing corynebacteria. Bio/ Technology 5:137–146
    [Google Scholar]
  14. Roberts J., Holcenberg J. S., Dolowy W. C. 1972; Isolation, crystallization, and properties of Achromobacteriaceae glutaminase-asparaginase with antitumour activity. Journal of Biological Chemistry 241:84–90
    [Google Scholar]
  15. Vuillet S., Spinnler H. E., Blachere H. 1986; Analysis of amino acid requirements of Clostridium thermocellum. Applied Microbiology and Biotechnology 23:496–498
    [Google Scholar]
  16. Willis R. C., Woolfolk R. 1974; Asparagine utilization in Escherichia coli. Journal of Bacteriology 118:231–241
    [Google Scholar]
  17. Wriston J. C. 1970; Asparaginase. Methods in Enzymology 17A:732–742
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-3-515
Loading
/content/journal/micro/10.1099/00221287-136-3-515
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error