Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 Mar;113(3):1170–1176. doi: 10.1128/jb.113.3.1170-1176.1973

Characterization of Esterases Produced by a Ruminal Bacterium Identified as Butyrivibrio fibrisolvens1

Wayne W Lanz a,2, Phletus P Williams a
PMCID: PMC251678  PMID: 4734862

Abstract

An obligately anaerobic ruminal bacterial isolate was selected from 18 tributyrin-degrading isolates and identified as Butyrivibrio fibrisolvens strain 53. The culture in late exponential phase contained enzymes which could be released by sonic disruption. These enzymes degraded substrates at a rate in the order 1-naphthyl acetate (NA) > 1-naphthyl butyrate > 1-naphthyl propionate but did not degrade 1-naphthyl palmitate or 1-naphthyl phosphate. The enzymes on NA were neither stimulated nor inhibited by CoCl2, MgCl2, and MnCl (each varied from 10−6 to 10−4 M). CaCl at 10−3 M stimulated esterase activity by 16%. Aliphatic substrates were hydrolyzed at a rate in the order triacetin > tributyrin > tripropionin, and ethyl acetate > ethyl formate. Similarly, aromatic fluorescein diesters were degraded at a rate in the order acetyl > propionyl > caproyl > butyryl > capryl > lauryl. Polyacrylamide gel electrophoretic zymograms indicated that the enzyme composite contained cathodally migrating bands. By column chromatography, these enzymes were separated into six NA-degrading fractions. Fraction V contained an esterase which had an optimal temperature of 39 C, a Km of 7.6 × 10−4 on NA, and a molecular weight of about 66,000. This enzyme was inhibited by paraoxon (41%, 10−4 M), eserine (17%, 10−2 M), NaF (17%, 10−2 M), and diisopropyl fluorophosphate (62%, 10−4 M) but not by 1-naphthyl N-methyl carbamate at 8.4 × 10−4 M.

Full text

PDF
1170

Selected References

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

  1. ALDRIDGE W. N. Serum esterases. I. Two types of esterase (A and B) hydrolysing p-nitrophenyl acetate, propionate and butyrate, and a method for their determination. Biochem J. 1953 Jan;53(1):110–117. doi: 10.1042/bj0530110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ahearn D. G., Meyers S. P., Nichols R. A. Extracellular proteinases of yeasts and yeastlike fungi. Appl Microbiol. 1968 Sep;16(9):1370–1374. doi: 10.1128/am.16.9.1370-1374.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BERGMANN F., SEGAL R., RIMON S. A new type of esterase in hog-kidney extract. Biochem J. 1957 Nov;67(3):481–486. doi: 10.1042/bj0670481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. BRYANT M. P., SMALL N. The anaerobic monotrichous butyric acid-producing curved rod-shaped bacteria of the rumen. J Bacteriol. 1956 Jul;72(1):16–21. doi: 10.1128/jb.72.1.16-21.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baillie A., Thomson R. O., Batty I., Walker P. D. Some preliminary observations on the location of esterases in Bacillus cereus. J Appl Bacteriol. 1967 Aug;30(2):312–316. doi: 10.1111/j.1365-2672.1967.tb00302.x. [DOI] [PubMed] [Google Scholar]
  6. Blackburn T. H. The protease liberated from Bacteroides amylophilus strain H18 by mechanical disintegration. J Gen Microbiol. 1968 Aug;53(1):37–51. doi: 10.1099/00221287-53-1-37. [DOI] [PubMed] [Google Scholar]
  7. Brown J. P. Cytochemical electron microscopic localization o esterase activity in Lactobacillus casei. Appl Microbiol. 1970 Jun;19(6):1001–1004. doi: 10.1128/am.19.6.1001-1004.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clarke D. G., Hawke J. C. Studies on rumen metabolism. VI. In vitro hydrolysis of triglyceride and isolation of a lipolytic fraction. J Sci Food Agric. 1970 Sep;21(9):446–452. doi: 10.1002/jsfa.2740210902. [DOI] [PubMed] [Google Scholar]
  9. DESNUELLE P., SAVARY P. SPECIFICITIES OF LIPASES. J Lipid Res. 1963 Oct;4:369–384. [PubMed] [Google Scholar]
  10. Fulghum R. S., Baldwin B. B., Williams P. P. Antibiotic susceptibility of anaerobic ruminal bacteria. Appl Microbiol. 1968 Feb;16(2):301–307. doi: 10.1128/am.16.2.301-307.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GARTON G. A., HOBSON P. N., LOUGH A. K. Lipolysis in the rumen. Nature. 1958 Nov 29;182(4648):1511–1512. doi: 10.1038/1821511a0. [DOI] [PubMed] [Google Scholar]
  12. GARTON G. A., LOUGH A. K., VIOQUE E. Glyceride hydrolysis and glycerol fermentation by sheep rumen contents. J Gen Microbiol. 1961 Jun;25:215–225. doi: 10.1099/00221287-25-2-215. [DOI] [PubMed] [Google Scholar]
  13. GOMORI G. Human esterases. J Lab Clin Med. 1953 Sep;42(3):445–453. [PubMed] [Google Scholar]
  14. HOBSON P. N., MANN S. O. The isolation of glycerol-fermenting and lipolytic bacteria from the rumen of the sheep. J Gen Microbiol. 1961 Jun;25:227–240. doi: 10.1099/00221287-25-2-227. [DOI] [PubMed] [Google Scholar]
  15. Henderson C. A study of the lipase produced by Anaerovibrio lipolytica, a rumen bacterium. J Gen Microbiol. 1971 Jan;65(1):81–89. doi: 10.1099/00221287-65-1-81. [DOI] [PubMed] [Google Scholar]
  16. Holmes R. S., Masters C. J. The developmental multiplicity and isoenzyme status of cavian esterases. Biochim Biophys Acta. 1967 Mar 15;132(2):379–399. doi: 10.1016/0005-2744(67)90157-x. [DOI] [PubMed] [Google Scholar]
  17. KIMMEL J. R., SMITH E. L. Crystalline papain. I. Preparation, specificity, and activation. J Biol Chem. 1954 Apr;207(2):515–531. [PubMed] [Google Scholar]
  18. Kepler C. R., Tove S. B. Biohydrogenation of unsaturated fatty acids. 3. Purification and properties of a linoleate delta-12-cis, delta-11-trans-isomerase from Butyrivibrio fibrisolvens. J Biol Chem. 1967 Dec 25;242(24):5686–5692. [PubMed] [Google Scholar]
  19. 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]
  20. Langenberg K. F., Bryant M. P., Wolfe R. S. Hydrogen-oxidizing methane bacteria. II. Electron microscopy. J Bacteriol. 1968 Mar;95(3):1124–1129. doi: 10.1128/jb.95.3.1124-1129.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. MARKERT C. L., HUNTER R. L. The distribution of esterases in mouse tissues. J Histochem Cytochem. 1959 Jan;7(1):42–49. doi: 10.1177/7.1.42. [DOI] [PubMed] [Google Scholar]
  22. Murphy P. M., Masterson C. L. Determination of multiple forms of esterases in rhizobium by paper electrophoresis. J Gen Microbiol. 1970 Apr;61(1):121–129. doi: 10.1099/00221287-61-1-121. [DOI] [PubMed] [Google Scholar]
  23. Oberhofer T. R., Maddox L. Combined medium to determine deoxyribonuclease activity and phenylalanine deamination by Enterobacteriaceae. Appl Microbiol. 1970 Feb;19(2):385–386. doi: 10.1128/am.19.2.385-386.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Okuda H., Fujii S. Lipase and esterase in adipose tissue and liver. J Biochem. 1967 Mar;61(3):313–319. doi: 10.1093/oxfordjournals.jbchem.a128550. [DOI] [PubMed] [Google Scholar]
  25. Smith P. B., Hancock G. A., Rhoden D. L. Improved medium for detecting deoxyribonuclease-producing bacteria. Appl Microbiol. 1969 Dec;18(6):991–993. doi: 10.1128/am.18.6.991-993.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Smith R. F., Willett N. P. Rapid plate method for screening hyaluronidase and chondroitin sulfatase-producing microorganisms. Appl Microbiol. 1968 Sep;16(9):1434–1436. doi: 10.1128/am.16.9.1434-1436.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Williams P. P., Dinusson W. E. Composition of the ruminal flora and establishment of ruminal ciliated protozoal species in isolated calves. J Anim Sci. 1972 Mar;34(3):469–474. doi: 10.2527/jas1972.343469x. [DOI] [PubMed] [Google Scholar]
  28. Williams P. P., Feil V. J. Identification of trifluralin metabolites from rumen microbial cultures. Effect of trifluralin on bacteria and protozoa. J Agric Food Chem. 1971 Nov-Dec;19(6):1198–1204. doi: 10.1021/jf60178a033. [DOI] [PubMed] [Google Scholar]
  29. Williams P. P., Stolzenberg R. L. Ruminal bacterial degradation of benzo(b)-thien-4-yl methylcarbamate (Mobam) and effect of Mobam on ruminal bacteria. Appl Microbiol. 1972 Apr;23(4):745–749. doi: 10.1128/am.23.4.745-749.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES