Ultrapure (Type 1) Water And Characterizing Bacterial Signaling Molecules

Bacterial signaling is also called ‘Quorum sensing’. This whole process is about bacterial communication and coordination among them. In this, the bacterial species coordinate certain behaviors like virulence, biofilm formation, antibiotic resistance, etc.  They coordinate by this quorum sensing or bacterial signaling. To perform the coordination among themselves, they recruit a certain type of molecule known as signaling molecules.  These molecules are helpful in the identification and characterization of the signals. The phenomenon is essential for bacterial cells to assess their population density. All secreted molecules collectively regulate the gene expression in bacteria. This eventually helps in the whole process of bacterial sensing or quorum sensing.

The quorum sensing can be achieved artificially by the process of Ultra-high performance chromatography(UHPLC),  with high-resolution tandem mass spectrometry(HRMS/MS). This application is used to monitor the functions of bacteria regarding the signaling process. The researchers and scientists, in a lab, perform this application to obtain good results. And the results are obtained through the use of pure water. The above application needs pure water in continuous supply from the water purification system for the laboratory.  Normal tap water cannot be used because of the high level of impurities present in it.  It is mandatory to use only pure water from a lab water system to perform the bacterial signaling process.

Let’s see now how Ultrapure (Type 1) water from a lab water purification system is used in the bacterial signaling process :

Process Of Bacterial Signalling 

There are several signaling molecules present in bacteria for quorum sensing. The most common one is the N-acyl homoserine lactones(AHLs), which are used by many bacteria for the communication process among related species. This molecule is not alone. It is accompanied by certain messenger compounds called autoinducers to exert their effect. These autoinducers are also called pheromones.

In quorum sensing, the AHLs play a vital role. These are called the prime members of the signaling molecules family which helps in setting up communication among closely related species of bacteria family. The bacteria that show quorum sensing, secrete the autoinducers into their surroundings. In the nearby surroundings, these molecules subsequently bind to their target receptors within other bacterial species. After binding of autoinducer and target receptor, the autoinducers activate the transcription factor of certain genes. These genes can be of the autoinducer also.In the case of a small number of bacteria, the autoinducer concentration is approx. zero due to the diffusion process. But in a larger amount of bacteria, the autoinducer concentration passes to a threshold level that causes more secretion of transcription factors. And eventually, more synthesis of inducers in the surroundings. This process leads to a positive feedback loop.  With this process,  more transcription factors get activated, which causes the other bacterial cells to begin their transcription procedures. The purpose of bacterial sensing or quorum sensing is to maximize cell efficiency.  As the bacterial cells are of very small dimensions, identification of cells becomes tedious here. To get a better vision of bacterial cells and to get the results of their transcription rates, quorum sensing is used.

To perform the quorum sensing perfectly, the two main sub-processes should be done. These processes are quantitation and characterization of autoinducer molecules. This is done to understand quorum sensing. The only limitation to this process is the concentration of AHLs which is secreted in small amounts. But when the bacterial population is large, these inducers can be of high concentrations. In the marine environment, direct analysis can be difficult.  The solution to this problem can be over-pass by isolation and identification of AHL-producing strains and later on the subsequent characterization of compounds.

The analysis is done by high-performance liquid chromatography(HPLC) with some appropriate standards.  Pure water is used in this process which is obtained from the laboratory water purifier.  This technique is used with the combination of thin-layer chromatography to obtain good results. Although, other techniques can be also used for the identification of AHLs, like gas chromatography, mass spectrometry, nuclear magnetic resonance, typical methods of chemical identification, etc.

The techniques are used for the study of AHLs in the quorum sensing process.  In the whole method,  pure water is used in ample amounts. In other words, we can say that pure water is playing an important role in establishing the results of the above techniques.  All investigation processes are successful with the help of pure water only. This pure water can be obtained from the water purification system for laboratory use in less time.  As per the need for pure water in the process,  the lab water system is installed. Before installing, you can also check a water purification system for laboratory price online as well as offline mediums.

Pure Water In Chromatographic conditions In The Process

In the liquid-liquid extraction process of chromatographic conditions, water is needed to set the standard.  The organic phase is evaporated to reach the dry state and the extract is suspended into the HPLC system.  The extracts in this process are prepared through pure water obtained from a lab water purification system.  Further, the mobile phase in chromatographic conditions is prepared through the usage of pure water or lab water.

Pure Water In HPLC And Mass Spectrometry Process

These techniques fall into the category of analytical methods in the laboratory. So, they are more sensitive to other external parameters. In this case,  the techniques require a proficient water supply in the lab. This is achieved by setting a water purification system for laboratory use.  Type I water (Ultrapure Water) is required. The lab water system should be of great efficiency that can purify the water in minute ways also. The removal of impurities and other nucleases should also be done in a lab water purifier. 

Type I water is necessary for the purpose to achieve clean removal of other impurities and to obtain good spectrometric data.  The pure water further protects the instrumentation damage and thus, makes the experiment more reliable.  To execute the spectrometric analysis, pure water is used in abundance. If poor water or normal tap water is used in the process, then it can impair the results of UHPLC and mass spectrometry analysis. It can also damage the instruments and can also hamper the resources used.  The organic contaminants can also decrease the efficiency of the UHPLC chromatogram. They can stick to the surface of the chromatogram and further complicate the results.  Metal ion impurities can form adducts in the analytes that would decrease the result’s reliability.

Feedwater Recirculation In Technique

To achieve good results,  a suitable grade of feedwater is also used. This feedwater is circulated in the laboratory water purifier for the cleaning and pretreatment process.  The build-up of the recirculation reservoir helps in preventing bacterial build-up in the system. This would further help in achieving a good result.

Conclusion

Bacterial sensing is the collective effort of various genes. The autoinducers and other messengers help in the process.  To achieve the quorum sensing artificially in the laboratory, the use of UHPLC is done. And to get more good and reliable results, we use pure water from the water purification system for the laboratory. The pure water is free of any kind of impurities which makes it ideal for use in the above technique of identification of bacteria in the quorum sensing process.