Skip to main content
SpringerLink
Log in

Human β-glucuronidase

Measurement of its activity in gallbladder bile devoid of intrinsic interference

  • Original Articles
  • Published:
Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

Our purpose is to develop a standard method for preparing the bile for β -glucuronidase determination by removal of bile acids and conjugated bilirubin which interfere with its activity. The bile acids and conjugated bilirubin in their purified solutions and in the diluted gallbladder biles could be extracted completely with cholestyramine in powder form or tetrahexylammonium chloride (THAC) in chloroform or ethyl acetate. The enzyme was, however, partially precipitated with cholestyramine and denatured by chloroform but not by ethyl acetate. A standard procedure, therefore, includes extraction of the diluted gallbladder bile with THAC in ethyl acetate, followed by determination of the maximal velocity (V max ) of the enzyme by a kinetic method employing phenolphthalein glucuronide as the substrate. The average V max of β-glucuronidase in the 20 normal gallbladder biles was 165±86 nmol/min/ml (mean±SD), a 23.5- fold increase over the activity before extraction. The measured activity represented the true activity of the enzyme in the bile for recovery of activity of the enzyme added to the bile was practically complete.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Ho KJ, Ho LHC, Kruger OR: Characterization and determination of the activity of biliary β-glucuronidase in rats. J Lab Clin Med 93:916–925, 1979

    PubMed  Google Scholar 

  2. Ho YC, Ho LHC, Ho KJ: Human hepatic β-glucuronidase: An enzyme kinetic study. Enzyme 33:9–17, 1985

    PubMed  Google Scholar 

  3. Ho KJ, Hsu SC, Chen JS, Ho LHC: Human biliary β-glucuronidase: Correlation of its activity with deconjugation of bilirubin in the bile. Eur J Clin Invest 16:361–367, 1986

    PubMed  Google Scholar 

  4. Ho KJ, Ho LHC: Inhibitory effect of bile acids on the activity of human β-glucuronidase at its optimal pH. Proc Soc Exp Biol Med 167:304–309, 1981

    PubMed  Google Scholar 

  5. Ho KJ: Human β-glucuronidase: Studies on the effect of pH and bile acids in regard to its role in the pathogenesis of cholelithiasis. Biochim Biophys Acta 827:197–206, 1985

    PubMed  Google Scholar 

  6. Sakamoto W, Nishikaze O, Sakakibara E: Isolation of an inhibitor of β-glucuronidase from porcine sublingual gland. Biochim Biophys Acta 329:72–80, 1973

    PubMed  Google Scholar 

  7. Matsushiro T, Suzuki N, Sato T, Maki T: Effects of diet on glucaric acid concentration in bile and the formation of calcium bilirubinate gallstones. Gastroenterology 72:630–633, 1977

    PubMed  Google Scholar 

  8. Simmons CJ, Davis M, Dordoni B, Williams R: Urinaryd-glucaric acid assay by an improved enzymatic procedure. Clin Chim Acta 51:47–51, 1974

    PubMed  Google Scholar 

  9. Hofmann AF: Efficient extraction of bile acid conjugates with tetraheptylammonium chloride, a liquid ion exchanger. J Lipid Res 8:55–57, 1967

    PubMed  Google Scholar 

  10. Tennent DM, Siegel H, Zanetti ME, Kuron GW, Ott WH, Wolf FJ: Plasma cholesterol lowering action of bile acid binding polymers in experimental animals. J Lipid Res 1:469–473, 1960

    PubMed  Google Scholar 

  11. Hashim SA, Van Itallie TB: Use of bile acid sequestrant in treatment of pruritus associated with biliary cirrhosis. J Invest Dermatol 35:253–254, 1960

    PubMed  Google Scholar 

  12. Gordon ER, Chan TH, Samodai K, Goresky CA: The isolation and further characterization of the bilirubin tetrapyrroles in bile-containing human duodenal juice and dog gall-bladder bile. Biochem J 167:1–8, 1977

    PubMed  Google Scholar 

  13. Kurtin WE, Schwesinger WH: Assay of β-glucuronidase in bile following ion-pair extraction of pigments and bile acids. Anal Biochem 147:511–516, 1985

    PubMed  Google Scholar 

  14. Jendrassik L, Grof P: Vereinfachte photometrische Methoden zur Bestimmung des Blutbilirubins. Biochem Z 297:81–86, 1938

    Google Scholar 

  15. Sigma Technical Bulletin No. 605: The quantitative colorimetric determination of bilirubin. Sigma Chemical Co., St. Louis, Missouri, 1978

  16. Talalay P: Enzymatic analysis of steroid hormones. Methods Biochem Anal 8:119–143, 1960

    PubMed  Google Scholar 

  17. Ho YC, Ho KJ: Differential quantitation of urinary β-glucuronidase of human and bacterial origins. J Urol 134: 1227–1230, 1985

    PubMed  Google Scholar 

  18. Oviedo MA, Ho KJ, Biss K, Liu LB, Taylor CB: Gallbladder bile composition in different ethnic groups. Arch Pathol Lab Med 101:208–212, 1977

    PubMed  Google Scholar 

  19. Marsh CA: Metabolism ofd-glucuronolactone in mammalian systems: Identification ofd-glucaric acid as a normal constituent of urine. Biochem J 86:77–86, 1963

    Google Scholar 

  20. Lineweaver H, Burk D: The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666, 1934

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, University of Alabama at Birmingham Medical Center and Veterans Administration Hospital, 35233, Birmingham, Alabama

    Yen-Ching Ho & Kang-Jey Ho

Authors
  1. Yen-Ching Ho
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kang-Jey Ho
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ho, YC., Ho, KJ. Human β-glucuronidase. Digest Dis Sci 33, 435–442 (1988). https://doi.org/10.1007/BF01536028

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01536028

Key words

Search

Navigation