ENZYME SUBSTRATES IN MICROBIOLOGY


COMMON CHROMOGENIC AND FLUOROGENIC METHODS - TARGET MICROORGANISMS AND ENZYMES

MethodEnzyme Dye/substrateMicroorganismSample*
Enumeration and isolation plate medium Glycosidase α-Galactosidase Indoxyl Salmonella C-F
      Indoxyl E. coli O157:H7 C-F
    β-Galactosidase Indoxyl Escherichia coli C
      Indoxyl E. coli O157:H7 C-F
      Indoxyl Coliforms F-W
      Indoxyl Staphylococcus saprophyticus C
      Alizarin Vancomycin-resistant enterococci C
    α-Glucosidase Indoxyl Cronobacter sakazakii F
      Indoxyl Staphylococcus aureus C-F
      Indoxyl Methicillin-resistant Staphylococcus aureus C
      Indoxyl Vancomycin-resistant enterococci C
    β-Glucosidase Indoxyl Klebsiella, Enterobacter, Serratia, Citrobacter C
      Indoxyl Extended spectrum β-lactamase producing enterobacteria C
      Indoxyl Vibrio C-F
      Indoxyl Enterococci C-W
      Indoxyl Vancomycin-resistant enterococci C
      Indoxyl Listeria spp F
      Indoxyl Candida kefyr, Candida lusitaniae, Candida tropicalis C
    Cellobiosidase Indoxyl Cronobacter sakazakii F
    β-Glucuronidase 4-MU, Indoxyl E. coli C-F-W
      Indoxyl Extended spectrum β-lactamase producing E. coli C
      Indoxyl Streptococcus agalactiae C
    β-Ribofuranosidase Indoxyl, 3',4'-Dihydroxyflavone Yersinia enterocolitica C
    Hexosaminidase Indoxyl, VLPA Candida albicans, Candida dubliniensis C
  Esterase Lipase Indoxyl Salmonella C-F
    PI-PLC Indoxyl Listeria monocytogenes F
    PC-PLC Indoxyl Bacillus F
    Phosphatase Indoxyl S. aureus C-F
      Indoxyl S. agalactiae C
      4-MU, Indoxyl Clostridium perfingens F
      Indoxyl Candida spp. C
  Aramidase/peptidase β-Alanyl arylamidase 7-Amido-1-pentyl-phenoxazin-3-one Pseudomonas aeruginosa C
  Miscellaneous Deoxyribonuclease Indoxyl S. aureus F
Spot test Esterase Lipase 4-Mu Salmonella C-F
  Arylamidase/peptidase L-Alanyl arylamidase AMC, p-Nitroanilide Gram+/Gram- F
  Miscellaneous β-Lactamase Nitrocefin, PADAC, CENTA, Cefesone β-Lactamase producing bacteria C
Most probable number Glycosidase β-Glucuronidase 4-MU E. coli F
    β-Galactosidase Nitrophenol, 4-MU Coliforms F
Cytometry Glycosidase β-Glucuronidase Fluorescein E. coli W
    β-Galactosidase Fluorescein Coliforms W

* C, clinical; F, food; W, water

Reference: Orenga, S.; James, A. L.; Manafi, M.; Perry, J. D.; Pincus, D. H., J. Microbiol. Methods 2009, 79 (2), 139-155.





BIOSYNTH ENZYME SUBSTRATES FOR MICROBIAL DETECTION - BY TARGET MICROORGANISMS




E. coli
by β-Glucuronidase

Cat. No. SubstrateShort name / abbreviationColorFluorescence ExcitationFluorescence Emission
B-7300 5-Bromo-4-chloro-3-indoxyl-beta-D-glucuronic acid, cyclohexylammonium salt monohydrate X-beta-D-GlcA blue-green    
B-7398 5-Bromo-4-chloro-3-indoxyl-beta-D-glucuronic acid, sodium salt anhydrous X-beta-D-GlcA blue-green    
B-7400 5-Bromo-4-chloro-3-indoxyl-beta-D-glucuronic acid, sodium salt trihydrate X-beta-D-GlcA blue-green    
B-7402 5-Bromo-4-chloro-3-indoxyl-beta-D-glucuronic acid, sodium salt trihydrate X-beta-D-GlcA blue-green    
B-8902 5-Bromo-3-indoxyl-beta-D-glucuronic acid, cyclohexylammonium salt Lapis-beta-D-GlcA deep blue    
I-6100 3-Indoxyl-beta-D-glucuronic acid, cyclohexylammonium salt Y-beta-D-GlcA dark blue    
I-6140 3-Indoxyl-beta-D-glucuronic acid, sodium salt Y-beta-D-GlcA dark blue    
B-7350 5-Bromo-6-chloro-3-indoxyl-beta-D-glucuronic acid, cyclohexylammonium salt Magenta-beta-D-GlcA magenta to lilac    
C-5050 6-Chloro-3-indoxyl-beta-D-glucuronic acid, cyclohexylammonium salt Salmon-beta-D-GlcA salmon orange    
H-7650 8-Hydroxyquinoline-beta-D-glucuronic acid   brown    
R-2150 Resorufin-beta-D-glucuronic acid, sodium salt   bright pink Ex. max. 572 nm Em. max. 585 nm
N-4450 4-Nitrophenyl-beta-D-glucuronic acid PNP-beta-D-GlcA yellow    
M-5700 4-Methylumbelliferyl-beta-D-glucuronic acid dihydrate MUG, 4-MU-beta-D-GlcA none Ex. max. 364 nm Em. max. 448 nm
R-2155_P00 RUG®
Resorufin-beta-D-glucuronic acid methyl ester, Biosynth Patent applied: EP11169147
  bright pink Ex. max. 572 nm Em. max. 585 nm
A-4735_P00 Aldol® 518 beta-D-glucuronic acid, Biosynth Patent applied: PCT/EP2010/056212   dark red Ex. max. 500 nm Em. max. 620 nm


E. coli / coliforms
by β-Galactosidase

Cat. No. SubstrateShort name / abbreviationColorFluorescence ExcitationFluorescence Emission
B-7150 5-Bromo-4-chloro-3-indoxyl-beta-D-galactopyranoside X-beta-D-Gal blue-green    
B-7200 5-Bromo-6-chloro-3-indoxyl-beta-D-galactopyranoside Magenta-beta-D-Gal magenta to lilac    
C-4990 4-Chloro-3-indoxyl-beta-D-galactopyranoside        
C-5000 6-Chloro-3-indoxyl-beta-D-galactopyranoside Salmon-beta-D-Gal salmon orange    
F-5500 6-Fluoro-3-indoxyl-beta-D-galactopyranoside F-beta-D-Gal red    
I-6000 3-Indoxyl-beta-D-galactopyranoside Y-beta-D-Gal dark blue    
I-7600 5-Iodo-3-indoxyl-beta-D-galactopyranoside Iodo-beta-D-Gal purple    
M-4100 N-Methylindoxyl-beta-D-galactopyranoside monohydrate Green-beta-D-Gal green    
B-8900 5-Bromo-3-indoxyl-beta-D-galactopyranoside Lapis-beta-D-Gal deep blue    
C-5248 Chlorophenol red beta-D-galactopyranoside sodium salt   dark red    
H-7500 8-Hydroxyquinoline-beta-D-galactopyranoside   brown    
M-5550 4-Methylumbelliferyl-beta-D-galactopyranoside 4-MU-beta-D-Gal none Ex. max. 364 nm Em. max. 448 nm
N-4300 4-Nitrophenyl-beta-D-galactopyranoside PNP-beta-D-GlcA yellow    
N-4190 2-Nitrophenyl-beta-D-galactopyranoside ONP-beta-D-Gal, ONPG yellow    
R-2110 Resorufin-beta-D-galactopyranoside   bright pink Ex. max. 572 nm Em. max. 585 nm
W-201644 3,4-Cyclohexenoesculetin beta-D-galactopyranoside   black    
A-4675_P00 Aldol® 518 beta-D-galactopyranoside, Biosynth Patent applied: PCT/EP2010/056212    dark red Ex. max. 500 nm Em. max. 620 nm
A-4676_P00 Aldol® 467 beta-D-galactopyranoside, Biosynth Patent applied: PCT/EP2010/056212    yellow-orange Ex. max. 370 + 460 nm Em. max. 530 nm
A-4679_P00 Aldol® 470 beta-D-galactopyranoside, Biosynth Patent applied: PCT/EP2010/056212    yellow-orange Ex. max. 370 + 460 nm Em. max. 530 nm
A-4691_P00 Aldol® 458 beta-D-galactopyranoside, Biosynth Patent applied: PCT/EP2010/056212    yellow Ex. max. 380 + 480 nm Em. max. 540 nm
A-4697_P00 Aldol® 484 beta-D-galactopyranoside, Biosynth Patent applied: PCT/EP2010/056212    red-orange Ex. max. 490 nm Em. max. 540 nm
A-4701_P00 Aldol® 495 beta-D-galactopyranoside, Biosynth Patent applied: PCT/EP2010/056212   orange Ex. max. 490 nm Em. max. 540 nm

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Listeria
by PI-PLC

Cat. No. SubstrateShort name / abbreviationColorFluorescence ExcitationFluorescence Emission
A-4702_P00 Aldol® 495 inositol-phosphate, ammonium salt, Biosynth Patent applied: PCT/EP2010/056212   orange Ex. max. 490 nm Em. max. 540 nm
B-7404_P00 5-Bromo-4-chloro-3-indoxyl myo-inositol-1-phosphate, ammonium salt, Biosynth patent protection by WO 98 / 38332 X-phos-inositol, X-IP blue-green    
A-4749_P00 Aldol® 514 inositol-phosphate, ammonium salt, Biosynth Patent applied: PCT/EP2010/056212   dark red Ex. max. 500 nm Em. max. 610 nm

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Salmonella
by Lipase

Cat. No. SubstrateShort name / abbreviationColorFluorescence ExcitationFluorescence Emission
B-7410  5-Bromo-4-chloro-3-indoxyl nonanoate X-nonanoate blue-green    
B-7411  5-Bromo-6-chloro-3-indoxyl nonanoate Magenta-nonanoate magenta to lilac    
B-8910  5-Bromo-3-indoxyl nonanoate Lapis-nonanoate deep blue    
C-5075  6-Chloro-3-indoxyl nonanoate Salmon-nonanoate salmon orange    
I-6150  3-Indoxyl nonanoate Y-nonanoate dark blue    
M-5744  4-Methylumbelliferyl nonanoate 4-MU-nonanoate none Ex. max. 364 nm Em. max. 448 nm
B-7102  5-Bromo-6-chloro-3-indoxyl caprylate Magenta-caprylate magenta to lilac    
C-4820  6-Chloro-3-indoxyl caprylate Salmon-caprylate salmon orange    
I-5690  3-Indoxyl caprylate Y-caprylate dark blue    
M-5526  4-Methylumbelliferyl caprylate 4-MU-caprylate none Ex. max. 364 nm Em. max. 448 nm
W-200568 4-Nitrophenyl caprylate PNP-caprylate yellow    
W-200727 2-Nitrophenyl caprylate ONP-caprylate yellow    
B-7100 5-Bromo-4-chloro-3-indoxyl caprylate X-carprylate blue-green    
B-8850 5-Bromo-3-indoxyl caprylate Lapis-caprylate deep blue    
A-4683_P00 Aldol® 470 nonanoate, Biosynth Patent applied: PCT/EP2010/056212    yellow-orange Ex. max. 370 + 460 nm Em. max. 530 nm
A-4714_P00 Aldol® 458 nonanoate solution, 0.75 M in DMSO, Biosynth Patent applied: PCT/EP2010/056212    yellow Ex. max. 380 + 480 nm Em. max. 540 nm
A-4715_P00 Aldol® 495 nonanoate solution, 0.75 M in DMSO, Biosynth Patent applied: PCT/EP2010/056212    orange Ex. max. 490 nm Em. max. 540 nm

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Staphylococcus aureus
by Phosphatase

Cat. No. SubstrateShort name / abbreviationColorFluorescence ExcitationFluorescence Emission
B-7420  5-Bromo-4-chloro-3-indoxyl phosphate, bis(2-amino-2-methyl-1,3-propanediol) salt X-phos, BCIP blue-green    
B-7430  5-Bromo-4-chloro-3-indoxyl phosphate, dilithium salt hydrate X-phos, BCIP blue-green    
B-7440  5-Bromo-4-chloro-3-indoxyl phosphate, dipotassium salt X-phos, BCIP blue-green    
B-7450  5-Bromo-4-chloro-3-indoxyl phosphate, disodium salt sesquihydrate X-phos, BCIP blue-green    
B-7452  5-Bromo-6-chloro-3-indoxyl phosphate, disodium salt monohydrate Magenta-phos magenta to lilac    
B-7455  5-Bromo-6-chloro-3-indoxyl phosphate, disodium salt, micronized Magenta-Phos magenta to lilac    
B-7458  5-Bromo-6-chloro-3-indoxyl phosphate, disodium salt Magenta-Phos magenta to lilac    
B-7470  5-Bromo-4-chloro-3-indoxyl phosphate, potassium salt X-phos, BCIP blue-green    
B-7500  5-Bromo-4-chloro-3-indoxyl phosphate, p-toluidine salt, high grade X-phos, BCIP blue-green    
B-7505  5-Bromo-4-chloro-3-indoxyl phosphate, p-toluidine salt, micronized X-phos, BCIP blue-green    
B-7508  5-Bromo-4-chloro-3-indoxyl phosphate, p-toluidine salt X-phos, BCIP blue-green    
B-7550  5-Bromo-6-chloro-3-indoxyl phosphate, p-toluidine salt Magenta-phos magenta to lilac    
B-8912  5-Bromo-3-indoxyl phosphate, disodium salt Lapis-phos deep blue    
B-8913  5-Bromo-3-indoxyl phosphate, p-toluidine salt Lapis-Phos deep blue    
C-5100  6-Chloro-3-indoxyl phosphate, p-toluidine salt      Salmon-phos salmon orange    
I-6200  3-Indoxyl phosphate, bis(2-amino-2-methyl-1,3-propanediol) salt Y-phos dark blue    
I-6300  3-Indoxyl phosphate, p-toluidine salt Y-phos dark blue    
M-5800 4-Methylumbelliferyl phosphate, free acid 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
M-5850 4-Methylumbelliferyl phosphate, bis(2-amino-2-methyl-1,3-propanediol) salt 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
M-5900 4-Methylumbelliferyl phosphate, bis(cyclohexylammonium) salt, trihydrate 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
M-5920 4-Methylumbelliferyl phosphate, dilithium salt 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
M-5950 4-Methylumbelliferyl phosphate, disodium salt trihydrate 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
N-4700 4-Nitrophenyl phosphate, bis(cyclohexylammonium) salt PNP-phos yellow    
N-4800 4-Nitrophenyl phosphate mono(2-amino-2-ethyl-1,3-propanediol) salt monohydrate PNP-phos yellow    
N-4990 4-Nitrophenyl phosphate, disodium salt PNP-phos yellow    
N-5001 4-Nitrophenyl phosphate, disodium salt hexahydrate, inclusive stability test PNP-phos yellow    
N-5015 4-Nitrophenyl phosphate, magnesium salt PNP-phos yellow    
N-5020 4-Nitrophenyl phosphate, potassium salt PNP-phos yellow    
B-7230 5-Bromo-4-chloro-3-indoxyl-alpha-D-glucopyranoside X-alpha-D-Glc blue-green    
C-5015 6-Chloro-3-indoxyl-alpha-D-glucopyranoside Salmon-alpha-D-Glc salmon orange    
N-4350 4-Nitrophenyl-alpha-D-glucopyranoside PNP-alpha-D-Glc yellow    
M-5600 4-Methylumbelliferyl-alpha-D-glucopyranoside 4-MU-alpha-D-Glc none Ex. max. 364 nm Em. max. 448 nm
A-4678_P00 Aldol® 470 phosphate, disodium salt, Biosynth Patent applied: PCT/EP2010/056212   yellow-orange Ex. max. 370 + 460 nm Em. max. 530 nm
A-4704_P00 Aldol® 495 phosphate, disodium salt, Biosynth Patent applied: PCT/EP2010/056212   red-orange Ex. max. 490 nm Em. max. 540 nm
A-4721_P00 Aldol® 515 phosphate, disodium salt, Biosynth Patent applied: PCT/EP2010/056212   dark red Ex. max. 500 nm Em. max. 610 nm
A-4688_P00 Aldol® 455 alpha-D-glucopyranoside, Biosynth Patent applied: PCT/EP2010/056212    yellow Ex. max. 380 + 480 nm Em. max. 540 nm
A-4696_P00 Aldol® 484 alpha-D-glucopyranoside, Biosynth Patent applied: PCT/EP2010/056212    yellow-orange Ex. max. 490 nm Em. max. 540 nm
A-4713_P00 Aldol® 518 alpha-D-glucopyranoside, Biosynth Patent applied: PCT/EP2010/056212    dark red Ex. max. 500 nm Em. max. 620 nm

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Bacillus cereus
by Phosphatase / PC-PLC

Cat. No. SubstrateShort name / abbreviationColorFluorescence ExcitationFluorescence Emission
B-7404_P00 5-Bromo-4-chloro-3-indoxyl myo-inositol-1-phosphate, ammonium salt, Biosynth patent protection by WO 98 / 38332 X-phos-inositol, X-IP blue-green    
A-4749_P00 Aldol® 514 inositol-phosphate, ammonium salt, Biosynth Patent applied: PCT/EP2010/056212   dark red Ex. max. 500 nm Em. max. 610 nm
A-4702_P00 Aldol® 495 inositol-phosphate, ammonium salt, Biosynth Patent applied: PCT/EP2010/056212   red-orange Ex. max. 490 nm Em. max. 540 nm
B-7116_P00 5-Bromo-4-chloro-3-indoxyl choline phosphate, Biosynth patent protection by WO 02 / 051853 X-phosphocholine blue-green    
B-7118_P00 5-Bromo-6-chloro-3-indoxyl choline phosphate, Biosynth patent protection by WO 02 / 051853 Magenta-phosphocholine magenta to lilac    
B-8855_P00 5-Bromo-3-indoxyl choline phosphate, Biosynth patent protection by WO 02 / 051853 Lapis-phosphocholine deep blue    
A-4672_P00 Aldol® 470 choline phosphate, Biosynth Patent applied: PCT/EP2010/056212    yellow-orange Ex. max. 370 + 460 nm Em. max. 530 nm
M-5528 4-Methylumbelliferyl cholin phosphate 4-MU-phosphocholine none Ex. max. 364 nm Em. max. 448 nm

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Pseudomonas aeruginosa
by PC-PLC

Cat. No. SubstrateShort name / abbreviationColorFluorescence ExcitationFluorescence Emission
B-7116_P00 5-Bromo-4-chloro-3-indoxyl choline phosphate, Biosynth patent protection by WO 02 / 051853 X-phosphocholine blue-green    
B-7118_P00 5-Bromo-6-chloro-3-indoxyl choline phosphate, Biosynth patent protection by WO 02 / 051853 Magenta-phosphocholine magenta to lilac    
B-8855_P00 5-Bromo-3-indoxyl choline phosphate, Biosynth patent protection by WO 02 / 051853 Lapis-phosphocholine deep blue    
A-4672_P00 Aldol® 470 choline phosphate, Biosynth Patent applied: PCT/EP2010/056212    yellow-orange Ex. max. 370 + 460 nm Em. max. 530 nm
M-5528 4-Methylumbelliferyl cholin phosphate 4-MU-phosphocholine none Ex. 366 nm  

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Clostridium perfringens
by Phosphatase

Cat. No. SubstrateShort name / abbreviationColorFluorescence ExcitationFluorescence Emission
A-4678_P00 Aldol® 470 phosphate, disodium salt, Biosynth Patent applied: PCT/EP2010/056212   yellow-orange Ex. max. 370 + 460 nm Em. max. 530 nm
A-4704_P00 Aldol® 495 phosphate, disodium salt, Biosynth Patent applied: PCT/EP2010/056212   red-orange Ex. max. 490 nm Em. max. 540 nm
A-4721_P00 Aldol® 515 phosphate, disodium salt, Biosynth Patent applied: PCT/EP2010/056212   dark red Ex. max. 500 nm Em. max. 610 nm
M-5800 4-Methylumbelliferyl phosphate, free acid 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
M-5850 4-Methylumbelliferyl phosphate, bis(2-amino-2-methyl-1,3-propanediol) salt 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
M-5900 4-Methylumbelliferyl phosphate, bis(cyclohexylammonium) salt, trihydrate 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
M-5920 4-Methylumbelliferyl phosphate, dilithium salt 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
M-5950 4-Methylumbelliferyl phosphate, disodium salt trihydrate 4-MU-phos, MUP none Ex. max. 364 nm Em. max. 448 nm
N-4700 4-Nitrophenyl phosphate, bis(cyclohexylammonium) salt PNP-phos yellow    
N-4800 4-Nitrophenyl phosphate mono(2-amino-2-ethyl-1,3-propanediol) salt monohydrate PNP-phos yellow    
N-4990 4-Nitrophenyl phosphate, disodium salt PNP-phos yellow    
N-5001 4-Nitrophenyl phosphate, disodium salt hexahydrate, inclusive stability test PNP-phos yellow    
N-5015 4-Nitrophenyl phosphate, magnesium salt PNP-phos yellow    
N-5020 4-Nitrophenyl phosphate, potassium salt PNP-phos yellow    


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