Characterization of Enterobacteria by Esterase Specific-Activity Profiles

Goullet, P; Picard, Bertrand

doi:10.1099/00221287-136-3-431

Cited by 30 publications

(30 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…1 from the mother strain, with only one band and a migration front (M r) value of 55. Since the latter enzymes have been described as among the most discriminating enzymes in various taxonomic studies [11,25], this analysis suggests that there were little or no differences between the 22 single-spore strains and the mother strain ORS 140102 which can be therefore considered as a pure strain if the pigmentation and colony morphology variations can be ascribed to physiological variations (such as inoculum effects, even though the inoculation techniques were as standardized as possible) but not genetic factors. 1) Consequently, the 23 strains tested had the same zymotype for malate dehydrogenase and esterases (Fig.…”

Section: Enzyme Electrophoretic Comparison Of the Single-spore Culturesmentioning

confidence: 89%

“…Electrophoretic profiles (after horizontal acrylarnideagarose gel electrophoresis) of fl-naphthyl propionate esterase from the mother strain (M) and three of the single-spore strains [11][12][13], after cultivation on BAP medium. Electrophoretic profiles (after horizontal acrylarnideagarose gel electrophoresis) of fl-naphthyl propionate esterase from the mother strain (M) and three of the single-spore strains [11][12][13], after cultivation on BAP medium.…”

Section: Enzyme Electrophoretic Comparison Of the Single-spore Culturesmentioning

confidence: 99%

“…Since analysis of enzyme profiles is considered to be one of the most discriminating techniques in bacterial taxonomy [10,11], this approach was used to answer the two following questions: (i) were all these cultures identical? Since analysis of enzyme profiles is considered to be one of the most discriminating techniques in bacterial taxonomy [10,11], this approach was used to answer the two following questions: (i) were all these cultures identical?…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Electrophoretic comparison of enzymes from 22 single-spore cultures obtained from Frankia strain ORS 140102

Prin¹

1991

FEMS Microbiology Letters

View full text Add to dashboard Cite

Summary Individual spores of Frankia strain ORS 140102 were allowed to germinate on Qmod medium. Germination percentages were approximately 30%. Colonies from germinated spores were individually picked off and subcultured on two media giving 22 single‐spore cultures. Although colony morphology and pigmentation of the single‐spore cultures differed, the study of the electrophoretic patterns of enzymes showed that malate dehydrogenase and esterase profiles of the 22 strains and the mother‐strain were very similar.

show abstract

Section: Enzyme Electrophoretic Comparison Of the Single-spore Culturesmentioning

confidence: 89%

Section: Enzyme Electrophoretic Comparison Of the Single-spore Culturesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Electrophoretic comparison of enzymes from 22 single-spore cultures obtained from Frankia strain ORS 140102

Prin¹

1991

FEMS Microbiology Letters

View full text Add to dashboard Cite

show abstract

“…Electrophoretic characterization of enzyme allelic polymorphism is generally performed using starch gel electrophoresis [1], or polyacrylamide gel electrophoresis [5]. However, the electrophoretic resolution in starch gels is not always optimal, and the analysis of sizeable numbers of enzymes in large numbers of bacterial strains, as required in population genetics, may be rather time‐consuming and tedious with such methods.…”

Section: Introductionmentioning

confidence: 99%

Multilocus enzyme analysis in aerobic and anaerobic bacteria using gel electrophoresisânitrocellulose blotting

Combe¹,

Lemeland²,

Pestel-Caron³

et al. 2000

FEMS Microbiology Letters

View full text Add to dashboard Cite

An optimized multilocus enzyme electrophoresis method, which involves polyacrylamide-agarose gel electrophoresis followed by electrophoretic transfers on nitrocellulose sheets, was developed for the analysis of enzyme polymorphism in several aerobic and anaerobic bacterial species including Staphylococcus aureus, Streptococcus pneumoniae, S. agalactiae, Klebsiella pneumoniae and K. oxytoca, Clostridium bifermentans and C. sordellii, and Prevotella bivia. Serial electrophoretic transfers (during 5-15 min each) from a single polyacrylamide gel could be achieved for most enzymes studied, and allowed an increased definition of enzyme bands on nitrocellulose as compared to migration gels. Four enzymes, which could not be blotted in such conditions, could still be stained in gels after blotting. Thus, the method allowed the combined analysis of several enzymes after a single gel electrophoresis separation. The analysis of enzyme polymorphism in the various species studied raised the interest of polymorphic loci such as esterase or glutamic-oxaloacetic transaminase for epidemiologic studies. The method characterized a genetic diversity of enzyme loci of S. pneumoniae higher than previously reported, and is thus convenient for the analysis of genetic relationships between related isolates. Since the present method reduces the tediousness of multilocus enzyme electrophoresis and requires experimental conditions that are not specific for the bacterial population studied, it may be proposed for rapid population genetics analysis of a wide variety of bacteria.

show abstract

“…2). Cocaine esterase was induced by cocaine, since specific activities of the enzyme in crude extracts from cells of P. maltophilia [14]). Sections of an unstained gel run under identical conditions were assayed for cocaine esterase activity by incubation with cocaine (2 mM in 1 ml of 50 mM MOPS buffer, pH 7.0, 30°C), and the reaction was monitored by HPLC.…”

mentioning

confidence: 99%

Identification of a cocaine esterase in a strain of Pseudomonas maltophilia

1992

View full text Add to dashboard Cite

A strain of Pseudomonas maltophilia (termed MB11L) which was capable of using cocaine as its sole carbon and energy source was isolated by selective enrichment. An inducible esterase catalyzing the hydrolysis of cocaine to ecgonine methyl ester and benzoic acid was identified and purified 22-fold. In the presence of the solubilizing agent cholate, cocaine esterase had a native Mr of 110,000 and was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be a monomer. In the absence of cholate, cocaine esterase had a native Mr of 410,000 and probably existed as a tetramer. The pH optimum of the enzyme was 8.0, and the Km values for cocaine, ethyl benzoate, and ethyl 2-hydroxybenzoate were 0.36, 1.89, and 1.75 mM, respectively.Inhibition studies indicated that the enzyme was a serine esterase, possibly possessing a cation-binding site similar to those of mammalian acetylcholinesterase and the atropine esterase of Pseudomonas putida PMBL-1.The cocaine esterase ofP. maltophilia MBl1L showed no activity with atropine, despite the structural similarity of cocaine and atropine.Microbial enzyme activities against alkaloids have been investigated as models of mammalian metabolism and as potential sources of new therapeutic compounds (16,17,38). Of the tropane alkaloids, the microbial metabolism of atropine has received the most significant attention. Corynebacterium belladonae catabolizes atropine via esterolytic hydrolysis of the tropic acid moiety followed by dehydrogenation, ring opening, and deamination of the tropane ring (27,28). A wide range of pseudomonads were also found to utilize atropine as their sole source of carbon and nitrogen (34). Further studies with nine of these strains identified two distinct classes of atropine esterase which possessed different physical and chemical properties (31). The atropine esterase from Pseudomonas putida PMBL-1 has been purified and extensively characterized (15,40,(43)(44)(45). Probing of the active site of the enzyme indicated an active serine residue and a classical charge relay system (43,44), although sequence analysis (15) and structure prediction (42) implied little homology to the serine protease families. Atropine esterase showed stereospecificity toward the (-)-isomer of atropine, (-)-hyoscyamine, and appeared to favor esters of tropic acid over those of acetic acid (34). Interestingly, the enzyme displayed no activity with the structurally related tropane alkaloid cocaine (34). In this paper, we describe the isolation of a strain of Pseudomonas maltophilia capable of using cocaine as its sole carbon and energy source and the partial purification and characterization of a cocaine esterase from this organism. Solutions and buffers were prepared by using water purified by a Milli-RO-60 system (Millipore Waters UK Ltd., Watford, United Kingdom). Ecgonine hydrochloride was prepared from cocaine by using the method of Findlay (11). The purity of the product was established by thin-layer chromatography (TLC), gas chromatography (GC), and HPLC analyses. Ecg...

show abstract

Characterization of Enterobacteria by Esterase Specific-Activity Profiles

Cited by 30 publications

References 45 publications

Electrophoretic comparison of enzymes from 22 single-spore cultures obtained from Frankia strain ORS 140102

Electrophoretic comparison of enzymes from 22 single-spore cultures obtained from Frankia strain ORS 140102

Multilocus enzyme analysis in aerobic and anaerobic bacteria using gel electrophoresisânitrocellulose blotting

Identification of a cocaine esterase in a strain of Pseudomonas maltophilia

Contact Info

Product

Resources

About

Characterization of Enterobacteria by Esterase Specific-Activity Profiles

Cited by 30 publications

References 45 publications

Electrophoretic comparison of enzymes from 22 single-spore cultures obtained from Frankia strain ORS 140102

Electrophoretic comparison of enzymes from 22 single-spore cultures obtained from Frankia strain ORS 140102

Multilocus enzyme analysis in aerobic and anaerobic bacteria using gel electrophoresisânitrocellulose blotting

Identification of a cocaine esterase in a strain of Pseudomonas maltophilia

Contact Info

Product

Resources

About

Multilocus enzyme analysis in aerobic and anaerobic bacteria using gel electrophoresisânitrocellulose blotting