Sensors for Detection and Quantification of Microbiological Protein Secretion

Simple SummaryContent extracted from patent full text and abstract with AI.

This invention describes a genetically engineered cell equipped with a gene for a fluorescent protein whose expression is directly linked to the secretion of proteins across the cell membrane. As the amount of secreted protein increases, the cell produces more fluorescent protein, resulting in increased fluorescence. This system enables rapid and easy identification or selection of cells or culture conditions that enhance protein secretion, simplifying screening processes in biotechnology and antibiotic research.

Use CasesContent extracted from patent full text and abstract with AI.

• High-throughput screening to identify bacterial strains with improved secretory capacities for industrial protein production (e.g., enzymes, antibodies).
• Optimization of fermentation media and culture conditions for maximizing protein yields in recombinant production systems.
• Detection and selection of beneficial genetic modifications (mutations or metabolic engineering) that enhance protein secretion.
• Screening for antibiotic compounds that disrupt bacterial membranes by monitoring changes in fluorescence as an indicator of membrane damage.
• Quality control and troubleshooting in bioprocesses by identifying cells with secretion bottlenecks or defects.
• Accelerated development of strains for production of pharmaceuticals, food enzymes, and industrial proteins.

BenefitsContent extracted from patent full text and abstract with AI.

• Enables fast, efficient, and non-destructive identification of high-secreting cells using simple fluorescence measurements.
• Allows generic screening methods independent of the type of protein being expressed, making the process broadly applicable.
• Reduces labor, time, and costs compared to traditional enzyme activity-based or product quantification assays.
• Facilitates high-throughput and single-cell resolution analyses, including automated cell sorting by flow cytometry.
• Improves optimization of both genetic constructs and environmental/culture parameters to maximize protein output.
• Enables efficient identification of novel antibiotics targeting bacterial membranes and assessment of their potency.
• Provides a versatile tool usable across a wide range of cell types (e.g., bacteria such as Escherichia, Bacillus, Corynebacterium) and production contexts.

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Patent Abstract

The present invention relates to a cell which is genetically modified with respect to its wild type and which comprises a gene sequence coding for a fluorescent protein, wherein the expression of the fluorescent protein depends on the amount of protein that is secreted across the cytoplasmic membrane into the extracytosolic space. The present invention also relates to a method for the identification of a cell having an increased secretion of protein across the cytoplasmic membrane into the extracytosolic space, a method for the identification of a culture medium composition that is optimized for the recombinant production of protein, a method for the identification of culture conditions that are optimized for the recombinant production of protein, a method for the identification of a compound that is characterized by an antibiotic activity due to its property to damage the membrane of a bacterial cell or to analyse the effect of such a compound on a population of genetically different bacterial cells or genetically identical cells in different physiological states or different growths phases, a method for the production of a cell which is genetically modified with respect to its wild type with optimized secretion of protein across the cytoplasmic membrane into the extracytosolic space, a cell obtained by this method, a method for the production of proteins and a method for the preparation of a mixture.