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. 1973 Nov;26(5):709–718. doi: 10.1128/am.26.5.709-718.1973

Purification and Properties of an Aryl Acylamidase of Bacillus sphaericus, Catalyzing the Hydrolysis of Various Phenylamide Herbicides and Fungicides

G Engelhardt 1,2, P R Wallnöfer 1,2, R Plapp 1,2
PMCID: PMC379889  PMID: 4762392

Abstract

From Bacillus sphaericus ATCC 12123 an aryl acylamidase (EC 3.5.1.13) was purified to homogeneity by ion exchange chromatography, gel filtration, and polyacrylamide gel electrophoresis. The enzyme is inducible by various phenylamides of the acylanilide, phenylcarbamate, and methoxysubstituted phenylurea type. It has a molecular weight of 75,000. Enzyme activity was inhibited by sulfhydryl reagents, several metal ions, and 3,4-dichloroaniline (a product of linuron degradation). A requirement for divalent metal ions in enzyme activity could not be demonstrated. In the presence of 6 M urea an irreversible inactivation of the enzyme occurred. The hydrolysis of L-alanine-4-nitroanilide was competitively inhibited by puromycin.

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Selected References

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  1. ALLEN M. B. The thermophilic aerobic sporeforming bacteria. Bacteriol Rev. 1953 Jun;17(2):125–173. doi: 10.1128/br.17.2.125-173.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aubert J. P., Millet J. Etude d'une L-leucyl-beta-naphtylamide hydrolase en relation avec la sporulation chez Bacillus megaterium. C R Acad Sci Hebd Seances Acad Sci D. 1965 Nov 15;261(20):4274–4277. [PubMed] [Google Scholar]
  4. Behal F. J., Asserson B., Dawson F., Hardman J. A study of human tissue aminopeptidase components. Arch Biochem Biophys. 1965 Aug;111(2):335–344. doi: 10.1016/0003-9861(65)90194-3. [DOI] [PubMed] [Google Scholar]
  5. Behal F. J., Carter R. T. Naphthylamidases of Sarcina lutea. Can J Microbiol. 1971 Jan;17(1):39–45. doi: 10.1139/m71-007. [DOI] [PubMed] [Google Scholar]
  6. Behal F. J., Cox S. T. Arylamidase of Neisseria catarrhalis. J Bacteriol. 1968 Oct;96(4):1240–1248. doi: 10.1128/jb.96.4.1240-1248.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Behal F. J., Folds J. D. A comparative study of bacterial alanine aminohydrolases. Biochem Biophys Res Commun. 1967 May 5;27(3):344–349. doi: 10.1016/s0006-291x(67)80104-9. [DOI] [PubMed] [Google Scholar]
  8. Behal F. J., Folds J. D. Arylamidase of Neisseria catarrhalis. Arch Biochem Biophys. 1967 Aug;121(2):364–371. doi: 10.1016/0003-9861(67)90089-6. [DOI] [PubMed] [Google Scholar]
  9. Burton G., Blenden D. C., Goldberg H. S. Naphthylamidase activity of Leptospira. Appl Microbiol. 1970 Apr;19(4):586–588. doi: 10.1128/am.19.4.586-588.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dennen D. W., Allen C. C., Carver D. D. Arylamidase of Cephalosporium acremonium and its specificity for cephalosporin C. Appl Microbiol. 1971 May;21(5):907–915. doi: 10.1128/am.21.5.907-915.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ellis S., Nuenke J. M. Dipeptidyl arylamidase III of the pituitary. Purification and characterization. J Biol Chem. 1967 Oct 25;242(20):4623–4629. [PubMed] [Google Scholar]
  12. Engelhardt G., Wallnöfer P. R., Plapp R. Degradation of linuron and some other herbicides and fungicides by a linuron-inducible enzyme obtained from Bacillus sphaericus. Appl Microbiol. 1971 Sep;22(3):284–288. doi: 10.1128/am.22.3.284-288.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Engelhardt G., Wallnöfer P. R., Plapp R. Identification of N, O-dimethylhydroxylamine as a microbial degradation product of the herbicide, linuron. Appl Microbiol. 1972 Mar;23(3):664–666. doi: 10.1128/am.23.3.664-666.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Lanzilotta R. P., Pramer D. Herbicide transformation. II. Studies with an acylamidase of Fusarium solani. Appl Microbiol. 1970 Feb;19(2):307–313. doi: 10.1128/am.19.2.307-313.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mahadevan S., Tappel A. L. Arylamidases of rat liver and kidney. J Biol Chem. 1967 May 25;242(10):2369–2374. [PubMed] [Google Scholar]
  17. NIMMO-SMITH R. H. Aromatic N-deacylation by chick-kidney mitochondria. Biochem J. 1960 May;75:284–293. doi: 10.1042/bj0750284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. ORNSTEIN L. DISC ELECTROPHORESIS. I. BACKGROUND AND THEORY. Ann N Y Acad Sci. 1964 Dec 28;121:321–349. doi: 10.1111/j.1749-6632.1964.tb14207.x. [DOI] [PubMed] [Google Scholar]
  19. Riley P. S., Behal F. J. Amino acid- -naphthylamide hydrolysis by Pseudomonas aeruginosa arylamidase. J Bacteriol. 1971 Nov;108(2):809–816. doi: 10.1128/jb.108.2.809-816.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sharabi N. E., Bordeleau L. M. Biochemical decomposition of the herbicide N-(3,4-dichlorophenyl)-2-methylpentanamide and related compounds. Appl Microbiol. 1969 Sep;18(3):369–375. doi: 10.1128/am.18.3.369-375.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tjeder A. The occurrence of amino acid naphthylamidase in baker's yeast. Acta Chem Scand. 1966;20(5):1442–1444. doi: 10.3891/acta.chem.scand.20-1442. [DOI] [PubMed] [Google Scholar]
  22. Tweedy B. G., Loeppky C., Ross J. A. Metabolism of 3-(p-bromophenyl)-1-methoxy-1-methylurea (metobromuron) by selected soil microorganisms. J Agric Food Chem. 1970 Sep-Oct;18(5):851–853. doi: 10.1021/jf60171a031. [DOI] [PubMed] [Google Scholar]
  23. WOLFE R. G., NEILANDS J. B. Some molecular and kinetic properties of heart malic dehydrogenase. J Biol Chem. 1956 Jul;221(1):61–69. [PubMed] [Google Scholar]
  24. Wallnöfer P. R., Bader J. Degradation of urea herbicides by cell-free extracts of Bacillus sphaericus. Appl Microbiol. 1970 May;19(5):714–717. doi: 10.1128/am.19.5.714-717.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wallnöfer P., Engelhardt G. [The degradation of phenylamides by Bacillus sphaericus]. Arch Mikrobiol. 1971;80(4):315–323. [PubMed] [Google Scholar]

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