Green Fluorescent Protein in the Design of a Living Biosensing System for l-Arabinose

Abstract

Analysis of monosaccharides is typically performed using analytical systems that involve a separation step followed by a detection step. The separation step is usually necessary because of the high degree of structural similarity between different monosaccharides. A novel sensing system for monosaccharides is described here in which living bacteria were designed to detect a model monosaccharide, L-arabinose, without the need for a separation step. In such sensing systems, analytes are detected by employing the selective recognition properties found in certain bacterial proteins. These systems are designed so that a reporter protein is expressed by the bacteria in response to the analyte. The concentration of the analyte can be related to the signal generated by the reporter protein. In the sensing system described here, the green fluorescent protein (GFP) was used as the reporter protein. L-Arabinose concentrations can be determined by monitoring the fluorescence emitted by the bacteria at 509 nm after excitation of GFP at 395 nm. The system can detect L-arabinose at concentrations as low as 5 x 10(-7) M and is selective over D-arabinose, the stereoisomer of the analyte, as well as over a variety of pentose and hexose sugars.

Predictive and Interpretive Simulation of Green Fluorescent Protein Expression in Reporter Bacteria

“…In this capacity, it is generally being used in one of two ways: either to establish the conditional expression of a gene in response to a specific substance, growth condition, or habitat (5,6,15,21,23,40,45,48) or to analyze a sample or habitat for a substance or condition to which a particular gene is known to be responsive (3,8,10,16,17,22,26,35). In both cases, the assessment of whether or not a particular gene or its promoter is responsive is based on the comparison of GFP fluorescence in cells from two populations: for example, one grown in the presence of a substance and the other grown in its absence or one grown in vivo compared to the other in vitro.…”

Reporter Genes lucFF, luxCDABE, gfp, and dsred Have Different Characteristics in Whole-Cell Bacterial Sensors

“…The first specific sensor bacteria using bioluminescence reporter technology were used for the measurement of naphthalene (9). After that, many specific sensor bacteria have been developed for inorganic ions (10,11) and organic compounds (12,13). For use in sensor bacteria, reporter response should be fast, easy to measure, and sensitive.…”

Construction of green fluorescent protein based bacterial biosensor for heavy metal remediation

“…GFP has become a popular reporter for gene activity in bacteria. Therefore, it is generally being used either to establish the conditional expression of a gene in response to a specific substance, growth condition, or habitat (Barker et al, 1998;Shetty et al, 1999). The heavy metals such as Cd, Pb, nickel (Ni), zinc (Zn) and chromium (Cr) represent a serious problem to the environment and public health when they contaminate superficial and ground water (Barros et al, 2008).…”