ABSTRACT
Enzymatic electrodes were fabricated by using three different immobilizations of glucose oxidase (GOx): covalent enzyme attachment (CA), enzyme coating (EC), and enzyme precipitate coating (EPC), here referred to as CA-E, EC-E, and EPC-E, respectively. When additional carbon nanotubes (CNTs) were introduced from 0 to 75wt% for the EPC-E design, its initial biosensor sensitivity was improved from 2.40×10(-3) to 16.26×10(-3) AâM(-1)âcm(-2), while its electron charge transfer rate constant was increased from 0.33 to 1.47s(-1). When a fixed ratio of CNTs was added for three different electrode systems, EPC-E showed the best glucose sensitivity and long-term thermal stability. For example, when 75wt% of additional CNTs was added, the initial sensitivity of EPC-E was 16.26×10(-3) AâM(-1)âcm(-2), while those of EC-E and CA-E were only 6.42×10(-3) and 1.18×10(-3) AâM(-1)âcm(-2), respectively. Furthermore, EPC-E retained 63% of its initial sensitivity after thermal treatment at 40°C over 41days, while EC-E and CA-E showed only 12% and 1% of initial sensitivities, respectively. Consequently, the EPC approach with additional CNTs achieved both high sensitivity and long-term stability, which are required for continuous and accurate glucose monitoring.
Subject(s)
Biosensing Techniques/methods , Electrochemical Techniques/instrumentation , Glucose Oxidase/chemistry , Glucose/analysis , Nanotubes, Carbon/chemistry , Bioelectric Energy Sources , Biosensing Techniques/instrumentation , Electrochemical Techniques/methods , Electrodes , Enzyme Stability , Enzymes, Immobilized/chemistry , Equipment Design , Glucose Oxidase/metabolism , Oxidation-Reduction , Sensitivity and SpecificityABSTRACT
The production of 1,3-propanediol (1,3-PD) was investigated with Klebsiella pneumoniae DSM 4799 using raw glycerol without purification obtained from a biodiesel production process. Fed-batch cultures with suspended cells revealed that 1,3-PD production was more effective when utilizing raw glycerol than pure glycerol (productivity after 47 h of fermentation, 0.84 g L(-1) 1 h(-1) versus 1.51 g L(-1) h(-1) with pure and raw glycerol,respectively). In addition, more than 80 g/L of 1,3-PD was produced using raw glycerol;this is the highest 1,3-PD concentration reported thus far for K. pneumoniae using raw glycerol. Repeated fed-batch fermentation with cell immobilization in a fixed-bed reactor was performed to enhance 1,3-PD production. Production of 1,3-PD increased with the cycle number (1.06 g L(-1) h(-1) versus 1.61 g L(-1) h(-1) at the first and fourth cycle, respectively)due to successful cell immobilization. During 46 cycles of fed-batch fermentation taking place over 1,460 h, a stable and reproducible 1,3-PD production performance was observed with both pure and raw glycerol. Based on our results, repeated fed batch with immobilized cells is an efficient fermentor configuration, and raw glycerol can be utilized to produce 1,3-PD without inhibitory effects caused by accumulated impurities.
