Innovation in Microbiology

While we keep our proprietary processes as trade secrets, we file patents for our unique application developments. Examples are:

A. Aldol® Novel Indicator Platform: A novel, versatile and customizable indicator platform

The Aldol® indicator platform is designed to visualize chemical, biochemical or even physical changes in the environment. As such it represents a modern version of traditional indicator systems. However, the Aldol® indicator platform is modular and can be tailored to many applications. For example, it can stain bacteria or change the color of a solution. It can produce fluorescence and a wide range of colors both in solution or as percipitate. Some types of Aldol® dyes readily dissolve in polyethylene or paraffin oil thereby creating bright and permanent fluorescence. Yet another important feature of the Aldol® indicator platform is that the signal (color or fluorescence) is generated without help of oxygen or any other auxiliary agent. It simply responds to a stimulus such as the activation through an enzyme by an intramolecular rearrangement which results in the structure of the dye.

Modular built Wide range of applications e.g. chromogenic & fluorogenic indicators
No O2 needed for staining Staining under anaerobic conditions
No auxilliary reagents needed No interference with cell growth
Wide color range complement indigo stains (yellow, organge, red)
Stable against unspecific hydrolysis Aldol ® acetates and other esters can be used in hydrolytic matrices such as ready to use plating media
Stain dissolves in hydrophobic phase Signal can be seperated from sample matrix for instance by staining pipette tip used in assay


Application Example:

Blood agar plate containing an Aldol® indicator to enhance specificity and sensitivity and allow for fluorescence read-out.



The Aldol® Patent

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Product information

Watch the Aldol® teaser video on YouTube


B. Economic, ultra-sensitive detection of beta-D-glucuronidase and E. coli

Resorufin is one of the most intensely fluorescent materials known until today. Hence Resorufin-beta-D-glucuronide represents the perfect agent to detect E. coli bacteria at ultra-low concentrations. The reason why the reagent is not widely used is its high manufacturing cost. However, we have discovered that the methyl ester analog, Resorufin-beta-D-glucuronide methyl ester, performs exactly the same as the free acid reagent Resorufin-beta-D-glucuronide. All the bacteria tested rapidly remove the methyl group and thus generate the desired free acid reagent Resorufin-beta-D-glucuronide in vivo. Since the manufacturing of the methyl ester analog Resorufin-beta-D-glucuronide methyl ester is economically feasible it is the reagent of choice for microbial water testing.

Comparison of Resorufin and the common MUG substrate.
From left to right: Resorufin-beta-D-glucuronide, control, Resorufin-beta-D-glucuronide methyl ester, and MUG.



Related Products:

Resorufin-beta-D-glucuronic acid, sodium salt


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Resorufin-beta-D-glucuronic acid methyl ester

Biosynth Patent applied:

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Download the Beta-D-Glucuronidase Patent here.


C. Detection of Phospholipase C

Many kinds of bacteria produce Phospholipase C (PLC) enzymes. Pathogenic Listeria species for instance secrete inositol specific PLC into the environment. We have developed a whole range of enzyme indicators derived from inositol phosphate including traditional H-MeU, Indolyl as well as advanced Aldol® types.

Choline specific PLC on the other hand is indicative of many Bacillus species. We have developed and patented a number of choline specific PLC substrates using traditional chromophores and fluorogens as well as modern Aldol® versions. These substrates have been used to detect and differentiate Bacillus species as well as to stain B. antracis. Lately, new interest in cholin specific indicators has arisen in the context of Pseudomonas quantification.

Application Example:

Staining of L. monocytogenes (turquois color colony) with X-IP on agar plates.
(Image courtesy of R&F labs)



Related Products:

Aldol® 495 inositol-phosphate, ammonium salt, Biosynth Patent applied: PCT/EP2010/056212


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Aldol® 470 choline phosphate, Biosynth Patent applied: PCT/EP2010/056212


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D. Beta-Lactamase Substrates

Beta-Lactamase (BLA) activity is a crucial indicator of bacterial resistance against cephalosporins which are still one of the most important classes of antibiotics. We have developed a simple indication system which is based on the colometric detection of metabolites resulting from the action of beta-Lactamase on the ENTIRE range of cephalosporins.



E. Indoxyl Derived Indicators

Indicators based on the release of indoxyl as a response to an enzymatic stimulus are in wide use. However, current use is limited to a few types of chloro- and bromoindoxyls. Did you know that the fluorinated or iodinated versions of indoxyl indicators offer alternative color schemes (purple) and that you can mix different types of indolyl substrates to produce hetero-indigo dyes yet further extending the color options and intensity?



Related Products:



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1-Pyrenyl b-D-glucuronide


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F. Tri-Aza-Cyclophane Substrates

Tri-Aza-Cyclophane (TAC) substrates are based on a phenolic azo dye that is freely soluble in buffers and produces intense color over a wide pH range. TAC based enzyme indicators are robust and available at low cost and represent a superior alternative to common substrates such as Phenolphthalein derivatives.


Related Product:

[2-(2-Thiazolylazo)-4-cresyl]-beta-D-glucuronic acid methyl ester, Biosynth
Patent applied: EP11169147


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G. Indoloindole Dyes: Novel Generic Indication Systems for Assays

This novel type of dye is produced in an aldol reaction, specifically by the response of an Aldol® indicator to an external stimulus such as enzymatic activity. indoloindole dyes cover the range from yellow to dark red. Some exhibit fluorescence in aqueous solution, some in hydrophobic environments and others even in solid state. The spectrum of fluorescence is often matrix dependent and spans from yellow to red and green. The dyes can be applied as soluble precursors and generated in situ or on target (staining). Some dyes display a strong affinity to polymeric materials such as polypropylene, diffuse deep into the material and create permanent and strong fluorescence. This effect can be made useful to separate signals from a sample matrix. If the dye for instance is produced in an assay it is capable of dyeing well plates, pipette tips or beads to make permanent fluorescence. The read out of such an assay can therefore even take place after discharging the sample.

Application Example:

The presence of bacteria in a sample broth is indicated by turning regular PE beads, added to the sample broth, permanently fluorescent.