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Contenuto archiviato il 2022-12-23

Novel biosensors and analysis kits based on genetically engineered biomolecules for formaldehyde assay

Obiettivo

Summary Control of environmental pollutants and toxic compounds in environment and food is of great importance for European countries. Formaldehyde is one of them. Formaldehyde is broadly used in industry, pharmacology as well as it is a natural metabolite. The existing formaldehyde assays are laborious and thus the development of the new selective and sensitive methods of its analysis is of a great importance. Development of biosensors is a modern trend of analytical technology. The essential advantages of biosensors are excellent selectivity and high sensitivity, short analysis time, and compatibility with computers. The main objective of this project is thus to develop enzyme- and cell-based biosensors and enzymatic analysis kits for assay of formaldehyde. Several biosensor and assay designs will be compared to optimise formaldehyde analysis in different real samples. First, cell based biosensors will be developed. Recently discovered formaldehyde-induced acidification of a medium around yeast cells due to efflux of formic acid will be exploited as a specific cell response to formaldehyde. For this genetically engineered yeast cells overproducing formaldehyde dehydrogenase will be prepared and attached to potentiometric or amperometric electrodes. Secondly, several enzymatic assay formats will be developed including enzyme-modified electrodes and homogeneous assay kits. Formaldehyde dehydrogenase and alcohol oxidase enzymes will be isolated from yeasts overproducers. Formaldehyde dismutase (FdDM) will be isolated from Pseudomonas sp. The performance of the biosensors and homogeneous assay kits will be evaluated using real chemical wastes, food and pharmaceutical samples. Research activities will be focused on the following six main research tasks: screening, selection and genetic engineering of microbial overproducers of formaldehyde-metabolising enzymes, isolation, purification, and characterisation of formaldehyde-selective enzymes, synthesis of novel redox polymeric mediators for development of reagentless biosensors, development of enzyme-based kits and biosensors for formaldehyde monitoring, development of an automated analysis system integrating the new enzyme kits and biosensors for formaldehyde, validation of the formaldehyde assays in industrial wastes, food and pharmaceuticals. The outcome of the project will be (i) novel enzymes, (ii) selection of yeasts overproducers of formaldehyde metabolising enzymes, (iii) automated analysis system for rapid analysis of formaldehyde. The expected results (a) will enable decreased costs of formaldehyde analysis, and (b) will offer simple and rapid assay procedures. Collected selection of microorganisms will be useful in remediation of chemical wastes. This issue will be addressed through other projects where rapid and cost effective analysis of formaldehyde is essential.

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Coordinatore

Research, management, and consultancy SciTech Link HB
Contributo UE
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Indirizzo
Borgäslingan 31
S-224 72 Lund
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Partecipanti (6)

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