Enzyme Characteristics of β-d-Galactosidase- and β-d-Glucuronidase-Positive Bacteria and Their Interference in Rapid Methods for Detection of Waterborne Coliforms andEscherichia coli

ABSTRACT Bacteria which were β-d-galactosidase and β-d-glucuronidase positive or expressed only one of these enzymes were isolated from environmental water samples. The enzymatic activity of these bacteria was measured in 25-min assays by using the fluorogenic substrates 4-methylumbelliferyl-β-d-galactoside and 4-methylumbelliferyl-β-d-glucuronide. The enzyme activity, enzyme induction, and enzyme temperature characteristics of target and nontarget bacteria in assays aimed at detecting coliform bacteria and Escherichia coli were investigated. The potential interference of false-positive bacteria was evaluated. Several of the β-d-galactosidase-positive nontarget bacteria but none of the β-d-glucuronidase-positive nontarget bacteria contained unstable enzyme at 44.5°C. The activity of target bacteria was highly inducible. Nontarget bacteria were induced much less or were not induced by the inducers used. The results revealed large variations in the enzyme levels of different β-d-galactosidase- and β-d-glucuronidase-positive bacteria. The induced and noninduced β-d-glucuronidase activities ofBacillus spp. and Aerococcus viridans were approximately the same as the activities of induced E. coli. Except for some isolates identified asAeromonas spp., all of the induced and noninduced β-d-galactosidase-positive, noncoliform isolates exhibited at least 2 log units less mean β-d-galactosidase activity than induced E. coli. The noncoliform bacteria must be present in correspondingly higher concentrations than those of target bacteria to interfere in the rapid assay for detection of coliform bacteria.

[1]  K. Wallenfels,et al.  20 β-Galactosidase , 1972 .

[2]  R. Colwell,et al.  Effect of seawater on Escherichia coli beta-galactosidase activity. , 1996, The Journal of applied bacteriology.

[3]  J. Hobbie,et al.  Use of nuclepore filters for counting bacteria by fluorescence microscopy , 1977, Applied and environmental microbiology.

[4]  J. D. Berg,et al.  Rapid detection of total and fecal coliforms in water by enzymatic hydrolysis of 4-methylumbelliferone-beta-D-galactoside , 1988, Applied and environmental microbiology.

[5]  N. Hasan,et al.  Induction of β-Galactosidase in Lactobacillus plantarum , 1974 .

[6]  S. Apte,et al.  Possible interference of lactose-fermenting marine vibrios in coliform beta-D-galactosidase assays. , 1995, The Journal of applied bacteriology.

[7]  Cheryl M. Davies,et al.  Rapid detection of faecal coliforms in sewage using a colorimetric assay of β-d-galactosidase , 1995 .

[8]  W. Sandine,et al.  β-Galactosidase of Streptococcus lactis , 1965 .

[9]  E. W. Frampton,et al.  Methods for Escherichia coli identification in food, water and clinical samples based on beta-glucuronidase detection. , 1993, The Journal of applied bacteriology.

[10]  R. Colwell,et al.  Effect of seawater on Escherichia coli β-galactosidase activity , 1996 .

[11]  윤현수,et al.  생쥐자궁의 β-galactosidase의 발현 , 1998 .

[12]  A. E. Greenberg,et al.  Standard methods for the examination of water and wastewater : supplement to the sixteenth edition , 1988 .

[13]  H. Nelis,et al.  Rapid detection of coliform bacteria and influence of non-target bacteria , 1997 .

[14]  S. M. Peterson,et al.  β‐D‐galactosidase activity of viable, non‐culturable coliform bacteria in marine waters , 1995, Letters in applied microbiology.

[15]  H. Nelis,et al.  Development of a sensitive chemiluminometric assay for the detection of beta-galactosidase in permeabilized coliform bacteria and comparison with fluorometry and colorimetry , 1995, Applied and environmental microbiology.

[16]  B. Drasar,et al.  Intestinal bacteria and the hydrolysis of glycosidic bonds. , 1971, Journal of medical microbiology.

[17]  S. Kjelleberg.  Starvation in Bacteria , 1993, Springer US.

[18]  R. Jacox,et al.  STREPTOCOCCAL β-GLUCURONIDASE , 1953 .

[19]  W. Kneifel,et al.  Fluorogenic and chromogenic substrates used in bacterial diagnostics , 1991 .

[20]  S. Apte,et al.  Plant andAlgal Interference inBacterial 3-D-Galactosidase and,3-D-Glucuronidase Assays , 1994 .

[21]  W. Sandine,et al.  BETA-GALACTOSIDASE OF STREPTOCOCCUS LACTIS. , 1965, Journal of bacteriology.

[22]  Awwa,et al.  Standard Methods for the examination of water and wastewater , 1999 .

[23]  E. Rosenblum,et al.  INDUCTION OF LACTOSE UTILIZATION IN STAPHYLOCOCCUS AUREUS , 1963, Journal of bacteriology.

[24]  S. Apte,et al.  Rapid detection of sewage contamination in marine waters using a fluorimetric assay of β-d-galactosidase activity , 1994 .

[25]  H. Nelis,et al.  Limitations of highly sensitive enzymatic presence-absence tests for detection of waterborne coliforms and Escherichia coli , 1997, Applied and environmental microbiology.

[26]  M. Pommepuy,et al.  Monitoring of fecal pollution in coastal waters by use of rapid enzymatic techniques , 1994, Applied and environmental microbiology.

[27]  R. E. Buchanan,et al.  Bergey's Manual of Determinative Bacteriology. , 1975 .

[28]  G. Dahlén,et al.  Screening Plate Method for Detection of Bacterial β-Glucuronidase , 1973 .

[29]  F. Stoeber,et al.  Inducibility of β-Glucuronidase in Wild-Type and Hexuronate-Negative Mutants of Escherichia coli K-12 , 1974 .

[30]  S. T. Cowan.  Bergey's Manual of Determinative Bacteriology , 1948, Nature.

[31]  C. Harwood,et al.  Detection of induced β‐galactosidase activity in individual non‐culturable cells of pathogenic bacteria by quantitative cytological assay , 1995, Molecular microbiology.

[32]  J. Monod,et al.  Genetic regulatory mechanisms in the synthesis of proteins. , 1961, Journal of molecular biology.

[33]  R. Y. Morita.  Bioavailability of Energy and the Starvation State , 1993 .

[34]  S. Apte,et al.  Plant and algal interference in bacterial beta-D-galactosidase and beta-D-glucuronidase assays , 1994, Applied and environmental microbiology.

[35]  R. Goldschmidt,et al.  Induction and general properties of beta-galactosidase and beta-galactoside permease in Pseudomonas BAL-31 , 1977, Journal of bacteriology.