Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1988 Apr;54(4):984–989. doi: 10.1128/aem.54.4.984-989.1988

Production and Purification of d-Aminoacylase from Alcaligenes denitrificans and Taxonomic Study of the Strain

Ying-Chieh Tsai 1,*, Ching-Ping Tseng 1, Kuang-Ming Hsiao 1, Ling-Yun Chen 1
PMCID: PMC202584  PMID: 16347618

Abstract

A d-aminoacylase-producing microorganism, strain DA181, isolated from soil was identified as Alcaligenes denitrificans subsp. denitrificans. This strain produced about 29,300 units (micromoles of product formed per hour) of d-aminoacylase and 2,300 units of l-aminoacylase per gram of cells (wet weight) when cultivated in a medium containing 1% N-acetyl-dl-leucine as the carbon source. The d-aminoacylase was purified 345-fold. The specific activity of the purified enzyme was 108,600 units per mg of protein when N-acetyl-d-methionine was used as a substrate. The apparent molecular weight was 58,000, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. N-Acetyl-d-methionine was the favored substrate, followed by N-acetyl-d-phenylalanine. This enzyme had a high stereospecificity, and its hydrolysis of N-acetyl-l-amino acids was almost negligible.

Full text

PDF
984

Images in this article

986Image
on p.986
988Image
on p.988

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bodanszky M., Perlman D. Peptide antibiotics. Science. 1969 Jan 24;163(3865):352–358. doi: 10.1126/science.163.3865.352. [DOI] [PubMed] [Google Scholar]
  2. Chibata I., Tosa T., Sato T., Mori T. Production of L-amino acids by aminoacylase adsorbed on DEAE-Sephadex. Methods Enzymol. 1976;44:746–759. doi: 10.1016/s0076-6879(76)44053-3. [DOI] [PubMed] [Google Scholar]
  3. Fukagawa Y., Kubo K., Ishikura T., Kouno K. Deacetylation of PS-5, a new beta-lactam compound, I. Microbial deacetylation of PS-5. J Antibiot (Tokyo) 1980 Jun;33(6):543–549. doi: 10.7164/antibiotics.33.543. [DOI] [PubMed] [Google Scholar]
  4. KAMEDA Y., TOYOURA E., KIMURA Y. Occurrence of D-acylase in soil bacteria. Nature. 1958 Apr 26;181(4617):1225–1225. doi: 10.1038/1811225a0. [DOI] [PubMed] [Google Scholar]
  5. KAMEDA Y., TOYOURA E., YAMAZOE H., KIMURA Y., YASUDA Y. Hydrolysis and metabolism by soil bacteria of benzoyl derivatives of D- and L-forms of some amino-acids. Nature. 1952 Nov 22;170(4334):888–889. doi: 10.1038/170888a0. [DOI] [PubMed] [Google Scholar]
  6. Kubo K., Ishikura T., Fukagawa Y. Deacetylation of PS-5, a new beta-lactam compound. II. Separation and purification of L- and D-amino acid acylases from Pseudomonas sp. 1158. J Antibiot (Tokyo) 1980 Jun;33(6):550–555. doi: 10.7164/antibiotics.33.550. [DOI] [PubMed] [Google Scholar]
  7. Kubo K., Ishikura T., Fukagawa Y. Deacetylation of PS-5, a new beta-lactam compound. III. Enzymological characterization of L-amino acid acylase and D-amino acid acylase from Pseudomonas sp. 1158. J Antibiot (Tokyo) 1980 Jun;33(6):556–565. doi: 10.7164/antibiotics.33.556. [DOI] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. SARGES R., WITKOP B. GRAMICIDIN A. VI. THE SYNTHESIS OF VALINE- AND ISOLEUCINE-GRAMICIDIN A. J Am Chem Soc. 1965 May 5;87:2020–2027. doi: 10.1021/ja01087a028. [DOI] [PubMed] [Google Scholar]
  11. SARGES R., WITKOP B. GRAMICIDIN. VII. THE STRUCTURE OF VALINE- AND ISOLEUCINE-GRAMICIDIN B. J Am Chem Soc. 1965 May 5;87:2027–2030. doi: 10.1021/ja01087a029. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES