caa a22 4500 605503923 CHVBK 20210128100613.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s00253-015-6517-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6517-1 A novel approach to monitor stress-induced physiological responses in immobilized microorganisms [Elektronische Daten] [Saúl Alonso, Manuel Rendueles, Mario Díaz] Microbial cell immobilization has long been considered as a potential bioprocessing strategy to increase both microorganisms' tolerance and fitness in fermentation systems. To date, little emphasis has been put on how the entrapped cells respond to the bioprocessing stresses encountered during the cultivation. The present work presents for the first time a methodology to decipher the real health status of the entrapped microorganisms by combining multiparameter flow cytometry with confocal fluorescence microscopy as monitoring tools. Comparison between resting free and immobilized cell-based systems enabled to characterize the spatial-temporal physiological response of entrapped Pseudomonas taetrolens cells during lactobionic acid production in submerged cultivation. Whereas cellular leakage from beads led to planktonic cells that faced a progressive loss of membrane integrity, immobilized cells underwent a prompt stress-induced physiological response featured by the predominance of cellular damaging. Moreover, visualization without matrix de-entrapment through confocal fluorescence microscopy revealed the overtime formation of cellular micro-colonies inside the beads. These micro-colonies comprised a shell made of dead cells, whereas the inward cells remained metabolically active. The proposed approach herein raises the possibility of using flow cytometry and confocal fluorescence microscopy as indicators of microbial cell immobilization, providing further key information on the health status and robustness of entrapped microorganisms. Springer-Verlag Berlin Heidelberg, 2015 Microbial immobilization nationallicence Stress-induced physiological responses nationallicence Flow cytometry nationallicence Confocal microscopy nationallicence Physiological status nationallicence Pseudomonas taetrolens nationallicence Alonso Saúl Department of Chemical and Environmental Engineering, Faculty of Chemistry, University of Oviedo, C/Julián Clavería s/n, 33071, Oviedo, Spain aut Rendueles Manuel Department of Chemical and Environmental Engineering, Faculty of Chemistry, University of Oviedo, C/Julián Clavería s/n, 33071, Oviedo, Spain aut Díaz Mario Department of Chemical and Environmental Engineering, Faculty of Chemistry, University of Oviedo, C/Julián Clavería s/n, 33071, Oviedo, Spain aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/8(2015-04-01), 3573-3583 0175-7598 99:8<3573 2015 99 253 https://doi.org/10.1007/s00253-015-6517-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6517-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Alonso Saúl Department of Chemical and Environmental Engineering, Faculty of Chemistry, University of Oviedo, C/Julián Clavería s/n, 33071, Oviedo, Spain aut NATIONALLICENCE 700 1- Rendueles Manuel Department of Chemical and Environmental Engineering, Faculty of Chemistry, University of Oviedo, C/Julián Clavería s/n, 33071, Oviedo, Spain aut NATIONALLICENCE 700 1- Díaz Mario Department of Chemical and Environmental Engineering, Faculty of Chemistry, University of Oviedo, C/Julián Clavería s/n, 33071, Oviedo, Spain aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/8(2015-04-01), 3573-3583 0175-7598 99:8<3573 2015 99 253