Facile monitoring and evaluation of bacteria in a fermentation process using perfusion chromatography and polarimetry detection

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Abstract
A 670 nm diode laser polarimeter was used as a detector for living cells in perfusion chromatography. It was found that all microorganisms examined, including vegetal organisms such as algae and fungi as well as several strains of different bacteria, gave a levorotatory signal, producing negative peaks in the chromatograms. Most naturally occurring chiral molecules, such as sugars and amino acids are dextrorotatory and give positive polarimetric peaks. A hydrophilic OH‐functionalized polystyrene divinylbenzene spherical porous particle packing was used as a sieving and size exclusion material and packed in 27×0.95 cm columns. The theory of the close‐packing of spheres is summarized to show that common bacteria should be able to perfuse in the interparticle voids of the 50 μm packing. It is shown that bacteria are eluted with the exclusion volume; a selective permeation of polymer with molecular weight ranging from 103 to 2×106 is obtained, and total permeation occurs for small molecules. The restricted access columns are used to follow the changes in cell concentration in bacteria cultures. The growth of three different bacteria was studied using calibrated polarimetric signals.