Development of a gas diffusion FIA system for on-line monitoring of ethanol.

A flow injection analysis (FIA) system for on-line monitoring of ethanol in cultivation media was developed, which combines the selectivity of a gas diffusion membrane with the substrate specificity of immobilized alcohol oxidase (AOD). The optimization of membrane material and immobilized enzyme was performed using different FIA modes such as dual detection and dual injection. A simple modification of a polypropylene membrane with silicone enabled a very flexible adjustment of the linear range for alcohol detection between 0.0006 and 60% (v v-1). The ethanol content of cultivation media could be determined continuously with a frequency of 120-180 samples per hour with an excellent correlation to gas chromatographic analysis (r = 0.9996). The relative standard deviation for 10 successive injections was lower than 0.5%.     

Utilization of a palladium-metal oxide semiconductor (Pd-MOS) sensor for on-line monitoring of dissolved hydrogen in anaerobic digestion

The use of a hydrogen-sensitive palladium-metal oxide semiconductor (Pd-MOS) sensor in combination with a membrane for liquid-to-gas transfer for the detection of dissolved hydrogen was investigated. The system was evaluated with known concentrations of dissolved hydrogen in water. The lowest concentration detected with this set-up was 160 nM. The method was applied to monitoring of a laboratory-scale anaerobic digestion process employing mixed sludge containing mainly food/industrial waste. Pulse loads of glucose were added to the system at different levels of microbial activity, and the microbial status of the culture was reflected in the dissolved hydrogen response. Simultaneous headspace hydrogen measurements were performed, and at the lower levels of dissolved hydrogen no corresponding headspace hydrogen could be detected. When glucose was added to a resting culture the dissolved hydrogen response was rapid and the first response could be detected 9 min after addition of glucose, whereas headspace hydrogen concentrations increased only after 80 to 110 min. This indicates limitations in the liquid-to-gas hydrogen transfer and illustrates the importance of hydrogen monitoring in the liquid. The sensor system developed is flexible, the membrane is easily replaceable, and the probe for liquid-to-gas hydrogen transfer can be adjusted easily to large-scale applications.     

An algorithmic approach to constructing the on-line estimation system for the specific growth rate

The objective of this article is to propose an algorithm for the on-line estimation of the specific growth rate in a batch or a fed-batch fermentation process. The algorithm shows the practical procedure for the estimation method utilizing the macroscopic balance and the extended Kalman filter. A number of studies of the on line estimation have been presented. However, there are few studies discussing about the selection of the observed variables and for the tuning of some parameters of the extended Kalman filter, such as covariance matrix and initial values of the state.The beginning of this article is devoted to explain the selection of the observed variable. This information is very important in terms of the practical know-how for using technique. It is discovered that the condition number is a practically useful and valid criterion for number is a practically useful and valid criterion for choosing the variable to be observed.Next, when the extended Kalman filter in applied to the online estimation of the specific growth rate, which is directly unmeasurable, criteria for judging the validity of the estimated value from the observed data are proposed. Based on the proposed criterial, the system equation of the specific growth rate is selected and initial value of the state variable and covariance matrix of the system noises are adjusted. From many experiments, it is certified that the specific growth rate in the batch or fed -batch fermentation can be estimated accurately by means of the algorithm proposed here. In these experiments, that is, when the cell concentration is measured directly, the extended Kalman filter using the convariance matrix with a constant element can estimate more accurately values of the specific growth rate than the adaptive extended Kalman filter does.     

An enzyme electrode for on-line determination of ethanol and methanol

Since a stable alcohol oxidase with a high specific activity is not commercially available, we propose to produce and purify this enzyme from a strain of the yeast Hansenula polymorpha. This alcohol oxidase was immobilized into a gelatin matrix and its activity was estimated by a pO(2) sensor. The enzyme electrode obtained was then used in a continuous flow system to measure methanol or ethanol concentrations. The sample oxygen content dependence of the signal was minimized by the support properties. Measuring time for each sample were less than two minutes including response data treatment and rinsing step. The enzyme electrode response was set for ethanol from 0.5mM to 15mM and for methanol from 10mM to 300mM. On repeated use, the electrode signal for 10mM of ethanol was stable for at least 500 assays. Analysis have been performed in different beverages such as wine and beer, and the results compared to those obtained with classical methods of analysis.     

Automatic on-line growth estimation method for outdoor algal biomass production

An on-line measuring procedure for estimating productivity in outdoor algal cultures was developed and tested experimentally. The procedure is based on a previously described method for on-line measuring net O(2) production rate (OPR). The data obtained by this method was found to correlate well with the conventional procedures for estimation productivity by measuring the changes in biomass concentration in the culture. The new procedure seems to be superior to the latter since it can be carried out in an almost continuous way and can give immediate indication on the productivity. OPR could be used to monitor on-line the photosynthetic and/or respiration activity in small research fermentors or in large-scale open systems outdoors.     

Fed-batch culture for L-lysine production via on-line state estimation and control

Computer application for fed-batch culture of Brevibacterium flavum for L-lysine production has been developed. The organisms are auxotrophic mutants for L-homoserine and are resistant to S-(2-aminoethyl)-L-cysteine. Adaptive control is applied for substrate addition. The sugar concentration is estimated using online respiratory measurement. During the period of fed-batch culture, the total sugar concentration is maintained at a given value. The cultivation strategy results in high productivity and high conversion yield.     

A nonlinear approach for the on-line estimation of the kinetic rates in bioreactors

The objective of this paper is to propose simple nonlinear observer-based estimators which allow the on-line estimation of kinetic rates inside a bioreactor from the measurements of components' concentrations. The design of these estimators does not require any model for the kinetic rates and the corresponding gains do not necessitate the resolution of any dynamical system. One of the main characteristics of these estimators lies in the easiness of their implementation and in particular their calibration. Their performances are firstly illustrated in simulation through a typical example. Then, estimation results corresponding to a real-life experiment are given in order to confirm the theoretical results and to demonstrate the usefulness of the proposed estimators in practical situations.     

A system for the on-line determination of glucose concentration

An off-line glucose analyzer, Yellow Springs Instrument (YSI) model 27 was modified and coupled to various peripheral components to produce a fast, fully automated system for the online determination of glucose concentration. The amount of time required to accomplish each measurement was in the order of two minutes. To demonstrate the utility of this system, various tests were performed. First, a stream containing known amounts of glucose was monitored on-line and the system was calibrated. The calibration curve was shown to be described by Michaelis-Menten kinetics. Once the system was properly calibrated, it was used to monitor the glucose concentration in the effluent stream of two different enzyme reactor systems. The glucose concentrations were within experimental error of those obtained via standard off-line techniques.     

Fermentation of biomass-generated producer gas to ethanol

The development of low-cost, sustainable, and renewable energy sources has been a major focus since the 1970s. Fuel-grade ethanol is one energy source that has great potential for being generated from biomass. The demonstration of the fermentation of biomass-generated producer gas to ethanol is the major focus of this article in addition to assessing the effects of producer gas on the fermentation process. In this work, producer gas (primarily CO, CO(2), CH(4), H(2), and N(2)) was generated from switchgrass via gasification. The fluidized-bed gasifier generated gas with a composition of 56.8% N(2), 14.7% CO, 16.5% CO(2), 4.4% H(2), and 4.2% CH(4). The producer gas was utilized in a 4-L bioreactor to generate ethanol and other products via fermentation using a novel clostridial bacterium. The effects of biomass-generated producer gas on cell concentration, hydrogen uptake, and acid/alcohol production are shown in comparison with "clean" bottled gases of similar compositions for CO, CO(2), and H(2). The successful implementation of generating producer gas from biomass and then fermenting the producer gas to ethanol was demonstrated. Several key findings following the introduction of producer gas included: (1) the cells stopped growing but were still viable, (2) ethanol was primarily produced once the cells stopped growing (ethanol is nongrowth associated), (3) H(2) utilization stopped, and (4) cells began growing again if "clean" bottled gases were introduced following exposure to the producer gas.     

An automatic method for on-line estimation of the photosynthetic rate in open algal ponds

An analytical model for dissolved oxygen concentration in an algal minipond was used to develop a new method for estimating, on-line, the net O(2) production rate (OPR) of the biological process. The method was tested experimentally and was found to provide crucial information on the vitality of the biological process and to provide an early warning of a possible forthcoming collapse of the ecosystern. It is suggested that the newly developed model and measurement method could provide investigators with useful tools for optimization of algal cultivation in the laboratory and plant.     

Discrete-time nonlinear observer-based estimators for the on-line estimation of the kinetic rates in bioreactors

Simple discrete-time estimators which allow the on-line estimation of the kinetic rates from the measurements of components' concentrations inside a bioreactor are proposed. In fact, the proposed estimators are obtained by a direct forward Euler discretization of continuous-time estimators. The design of the estimators in the continuous as well as in the discrete-time does not require or assume any model for the kinetic rates. One of the main characteristics of these estimators lies in the easiness of their calibration. We here emphasize on the performances of the discrete version of these estimators, whose stability and convergence are proved under the same conditions as in the continuous case with an additional mild assumption on the sampling time. Simulation and real-life experiments results corresponding to the discrete estimation are given. The accuracy of the obtained estimates as well as the easiness of the estimators' implementation do constitute reliable and powerful arguments for their use, in particular in adaptive control schemes.     

ATR-FTIR sensor development for continuous on-line monitoring of chlorinated aliphatic hydrocarbons in a fixed-bed bioreactor

This article describes the continuous on-line monitoring of a dechlorination process by a novel attenuated total reflection-Fourier transform infrared (ATR-FTIR) sensor. This optical sensor was developed to measure noninvasively part-per-million (ppm) concentrations of trichloroethylene (TCE), tetrachloroethylene (PCE), and carbon tetrachloride (CT) in the aqueous effluent of a fixed-bed dechlorinating bioreactor, without any prior sample preparation. The sensor was based on an ATR internal reflection element (IRE) coated with an extracting hydrophobic polymer, which prevented water molecules from interacting with the infrared (IR) radiation. The selective diffusion of chlorinated compound molecules from aqueous solution into the polymer made possible their detection by the IR beam. With the exclusion of water the detection limits were lowered, and measurements in the low ppm level became possible. The best extracting polymer was polyisobutylene (PIB) in the form of a 5.8-microm thick film, which afforded a detection limit of 2, 3, and 2. 5 mg/L (ppm) for TCE, PCE, and CT, respectively. Values of the enrichment factors between the polymer coating and the water matrix of these chloro-organics were determined experimentally and were compared individually with predictions obtained from the slopes of absorbance/concentration curves for the three analytes. Before coupling the ATR-FTIR sensor to the dechlorinating bioreactor, preliminary spectra of the chlorinated compounds were acquired on a laboratory scale configuration in stop-flow and flow-through closed-loop modes. In this way, it was possible to study the behavior and direct response of the optical sensor to any arbitrary concentration change of the analytes. Subsequently, the bioreactor was monitored with the infrared sensor coupled permanently to it. The sensor tracked the progression of the analytes' spectra over time without perturbing the dechlorinating process. To calibrate the ATR-FTIR sensor, a total of 13 standard mixtures of TCE, PCE and CT at concentrations ranging from 0 to 60 ppm were selected according to a closed symmetrical experimental design derived from a 3(2) full-factorial design. The above range of concentrations chosen for calibration reflected typical values during normal bioreactor operation. Several partial least squares (PLS) calibration models were generated to resolve overlapping absorption bands. The standard error of prediction (SEP) ranged between 0.6 and 1 ppm, with a relative standard error of prediction (RSEP) between 3 and 6% for the three analytes. The accuracy of this ATR-FTIR sensor was checked against gas chromatography (GC) measurements of the chlorocompounds in the bioreactor effluents. The results demonstrate the efficiency of this new sensor for routine continuous on-line monitoring of the dechlorinating bioreactor. This strategy is promising for bioprocess control and optimization.     

Microprocessor-based system for on-line analysis of respiratory responses to exercise

A microprocessor-based system was developed for the measurement and on-line calculation of values and derivatives of expiratory variables and their response to exercise. The system accepts analog signals from gas analysers, ECG electrodes and flow transducer, digitizes these signals, calculates values of required parameters and presents results on a video display. The system offers the user various options of logging and data processing. Plots of the values of calculated parameters versus exercise time and correlation between variables can be easily produced by the system. A mass storage unit was available with the system to enable the storage of calculated variables for off-line analysis. Hardcopy of displayed results and plots can be provided by the system.     

Evaluation of cell recycling in continuous fermentation of enzymatic hydrolysates of spruce with Saccharomyces cerevisiae and on-line monitoring of glucose and ethanol

The maximum growth rate of Saccharomyces cerevisiae ATCC 96581, adapted to fermentation of spent sulphite liquor (SSL), was 7 times higher in SSL of hardwood than the maximum growth rate of bakers' yeast. ATCC 96581 was studied in the continuous fermentation of spruce hydrolysate without and with cell recycling. Ethanol productivity by ATCC 96581 in continuous fermentation of an enzymatic hydrolysate of spruce was increased 4.6 times by employing cell recycling. On-line analysis of CO₂, glucose and ethanol (using a microdialysis probe) was used to investigate the effect of fermentation pH on cell growth and ethanol production, and to set the dilution rate. Cell growth in the spruce hydrolysates was strongly influenced by fermentation pH. The fermentation was operated in continuous mode for 210 h and a theoretical ethanol yield on fermentable sugars was obtained. Received: 25 May 1998 / Received revision: 11 August 1998 / Accepted: 12 August 1998