A novel microbial biosensor based on cells of Gluconobacter oxydans for the selective determination of 1,3-propanediol in the presence of glycerol and its application to bioprocess monitoring.
Novel and selective microbial amperometric biosensors that use Gluconobacter oxydans cells to monitor the bacterial bioconversion of glycerol (Gly) to 1,3-propanediol (1,3-PD) are described. Two different mediators, ferricyanide and flexible polyvinylimidazole osmium functionalized polymer (Os-polymer), were employed to prepare two different microbial biosensors, both of which gave high detection performance. The good operational stabilities of both types of biosensor were underlined by the ability to detect 1,3-PD throughout 140 h of continuous operation. Both microbial biosensor systems showed excellent selectivity for 1,3-PD in the presence of a high excess of glycerol [selectivity ratios (1,3-PD/Gly) of 118 or 245 for the ferricyanide and Os-polymer systems, respectively]. Further, the robustness of each microbial biosensor was highlighted by the high reliability of 1,3-PD detection achieved (average RSD of standards<2%, and well below 4% for samples). The biosensor implementing the Os-polymer mediator exhibited high selectivity towards 1,3-PD detection and allowed moderate sample throughput (up to 12 h-1) when integrated into a flow system. This system was used to monitor the concentration of 1,3-PD during a real bioprocess. Results from biosensor assays of 1,3-PD in bioprocess samples taken throughout the fermentation were in a very good agreement with results obtained from reference HPLC assays (R2=0.999).
Katrlík
Jaroslav
J
Department of Pharmaceutical Analysis and Nuclear Pharmacy, Faculty of Pharmacy, Comenius University, Odbojárov 10, 832 32, Bratislava, Slovakia. katrlik@fpharm.uniba.sk
Vostiar
Igor
I
Sefcovicová
Jana
J
Tkác
Ján
J
Mastihuba
Vladimír
V
Valach
Milan
M
Stefuca
Vladimír
V
Gemeiner
Peter
P
D015374:Biosensing Techniques / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004563:Electrochemistry; D005285:Fermentation; D005292:Ferricyanides / Q000737:chemistry; D020600:Gluconobacter oxydans / Q000201:enzymology / Q000254:growth & development*; D005990:Glycerol / Q000737:chemistry*; D007093:Imidazoles / Q000737:chemistry; D007700:Kinetics; D009992:Osmium / Q000737:chemistry; D011145:Polyvinyls / Q000737:chemistry; D011409:Propylene Glycols / Q000032:analysis*; D012680:Sensitivity and Specificity
D005292:Ferricyanides; D007093:Imidazoles; D011145:Polyvinyls; D011409:Propylene Glycols; C007931:hexacyanoferrate III; C072974:polyvinylimidazole; D009992:Osmium; C041787:1,3-propanediol; D005990:Glycerol