Estimation of fermentation biomass concentration by measuring oxygen uptake off-line with an oxygen electrode

Summary An indirect method for biomass determination was developed for aerobic cultivation in media containing solid substrates. This method involved off-line estimation of oxygen uptake with an oxygen electrode using a calibration factor and was applied to penicillin fermentation with Penicillium chrysogenum. The volumetric oxygen uptake rate was independent of the amount of sample sealed in the measuring cell. Storage of the sample for a period up to 2.5 h also had no influence on the rate of O₂ uptake. The specific O₂ uptake rate at 70 to 75 h fermentation time was used as the calibration factor to calculate the respiration-active part of the biomass. Specific penicillin V formation rates related to net dry weight, respiration-active biomass (X _A )and RNA content were compared. Only the penicillin V production rate related to X _A showed a similar behaviour as the penicillin V production rate related to RNA content. For this reason, it is assumed that X _A ,like RNA content, is more closely related to the metabolic state of the cells than the net dry weight. This method, therefore, provides a more suitable reference parameter for fermentation control than the generally used net dry weight.Offprint requests to: D. Siegmund     

Über die chemische Natur der Reserve-Polysaccharide in Acetabularia-Chloroplasten

Summary The reserve polysaccharides in chloroplasts of several species of Acetabularia have been identified as starch.The starch granules in situ as well as the isolated and purified particles stain with Lugol-reagent. They show the characteristic birefringence.Acid hydrolysis and degradation by -amylase followed by acid hydrolysis showed that the starch is composed only of glucose units.The statements of Vanden Driessche and Bonotto (1967) concerning the inulin character of the reserve polysaccharides in Acetabularia chloroplasts need correction.     

Protoplasmic streaming inAcetabularia

Summary A compilation of characteristics of the two different systems of intracellular transport inAcetabularia (Koop andKiermayer 1980 a and b) is given.The presence of microfilaments-presumably F-actin-in the cytoplasm ofAcetabularia is demonstrated by electron microscopy.The evidence for an involvement of microtubules in streaming is strengthened by the induction of birefringent vinblastine crystals in the stalk of vegetative cells.Isolated portions of cytoplasm formin vitro more than 100 m long filopodium-like processes, which are highly birefringent. The processes show intensive immunofluorescent staining with both, anti-actin and anti-tubulin as a primary antibody.A perfusion buffer is presented, which after replacing the vacuolar sap does not lead to a change in cytoplasmic morphology or streaming pattern and velocities.     

Determination of the apparent K mfor oxygen of Beneckea natriegens using the respirograph technique

The affinity for oxygen of the marine bacterium Beneckea natriegens was mesured using the Respirograph technique of Degn and Wohlrab. Values between 0.15 and 0.25 M oxygen were obtained for the apparent K _mfor oxygen regardless of the nature of the respiratory substrate. These values are an order of magnitude lower than those previously reported for B. natriegens using the conventional closed oxygen electrode system.     

Determination of the overall volumetric mass transfer coefficient kLa,by a temperature dependent change of gas solubility

Summary The solubility of oxygen in the liquid phase of a bioreactor was changed by a ramp change of temperature, and k_La was determined from the resulting return to equilibrium of dissolved oxygen activity. The maximum k_La that can be measured by this method in a standard laboratory scale bioreactor is 145 h^–1 corresponding to a temperature change rate of 320°C h^–1.Nomenclature p Difference between p_G and p_L (% saturation) - T Ramp change of temperature (°C) - E Temperature-compensated output from the oxygen electrode (A) - E_u Uncompensated output from the oxygen electrode (A) - k_La Overall volumetric mass transfer coefficient (h^–1) - k_La_Tm Overall volumetric mass transfer coefficient at temperature T_m (h^–1) - P_G Dissolved oxygen activity in equilibrium with the gas phase (% saturation) - p_L Dissolved oxygen activity (% saturation) - p_Lm Dissolved oxygen activity at time t_m (% saturation) - t Time (h) - t_m Time of maximum p (h) - T Temperature (°C) - T_cal Calibration temperature of the oxygen electrode (°C) - T_m Final temperature after a temperature shift (°C) - T_n Temperature at time t_n     

Oxygen mass transfer in a bubble column bioreactor containing lysed yeast suspensions

Summary Liquid-phase volumetric oxygen transfer coefficients were evaluated in a bubble column containing yeast suspensions, using the instationary oxygen absorption method and a polarographic oxygen electrode. The electrode time lag was found to be independent of both the system studied and the operating conditions. The volumetric oxygen mass transfer coefficients k _L a could be reasonably predicted by calculating k _L from the equation derived by Bhavaraju et al. or the empirical equation of Calderbank and Moo-Young and a from the experimental gas hold-up values.Nomenclature a Exponent in Eq.6 or specific gas-liquid interfacial area based on reactor volume m - b Exponent in Eq. 6 - C Constant in Eq 6 or oxygen concentration in the liquid phase g/ml - C ^* Equilibrium oxygen concentration g/ml - C ₀ Oxygen concentration in the liquid phase at t=0 g/ml - C _E Oxygen concentration as determined by the polarographic electrode g/ml - D _B Bubble equivalent diameter mm - D _l Oxygen diffusivity in the liquid phase m²/s - g Acceleration of gravity m/s² - K Consistency index Pasⁿ - K _L Liquid-phase mass transfer coefficient m/s - n Power law exponent - Pe _sw Peclet number based on bubble swarm velocity - S _C Schmidt number - Sh Sherwood number - i Time s - U _B Bubble rise velocity in infinite medium m/s - U _g Superficial air velocity based on column cross-sectional area m/s - U _sw Bubble swarm velocity defined by Eq.15 m/s - Y _MSW Mass transfer coeficient correction factor for mobile interfaces in pseudo-plastic fluids Eq. 7 - Y _MSW Mass transfer coefficient correction factor for immobile interface in pseudo-plastic fluids Eq. 8 Greek letters _l Density of liquid g/ml - _sus Density of unaerated suspension g/ml - _{wet cell} Density of yeast wet cells g/ml - _l Viscosity of the liquid Pas - _app Apparent viscosity of power law fluid Pas - _E Electrode time lag s - _l Time lag due to resistance of the gas-liquid interface s - _g Gas hold-up, volume fraction occupied by the gas phase - _l Liquid hold-up - _c Wet cell volume fraction     

Auslösung der circadianen Photosynthese-Rhythmik beiAcetabularia durch Blaulicht

Summary Additional irradiation with blue light of low intensity ofAcetabularia mediterranea cells, pretreated by prolonged irradiation with red, induces an increase of photosynthetic activity. This induction is accompanied by the appearance of rhythmic changes of the O₂ production. Maxima are found about 6, 30, 54, 78, ... hours after the onset of blue light irradiation. Thus blue light not only induces an increase of the rate of photosynthesis but also acts as a Zeitgeber for the circadian rhythm of photosynthesis inAcetabularia.

     

Endoplasmic reticulum,calciosomes and their possible roles in signal transduction

Summary Recent fluorescence, AVEC-DIC, and confocal laser scanning microscopic studies have revealed the dynamic nature and structural extent of a calcium-sequestering endoplasmic reticulum (ER) in plant cells. Various investigators have proposed different roles for the ER in cell motility. One, the ER plays a direct role in the generation of intracellular particle motions or two, the ER regulates particle motions indirectly. We show that the ER can be extruded fromAcetabularia cells, stains brightly with the fluorescent dye DiOC₆(3), and small (ca. 100 nm diameter) fluorescent vesicles are observed to move in or along the ER tubules. Intracellular particle movements in the giant algal cellAcetabularia can be transiently inhibited by IP₄, IP₃, and IP₂, compounds which in animal cells are known to cause the release of free calcium ions. A model is proposed which clarifies the possible relationships between the ER, calciosomes, calcium ions, and the microfilament-generated intracellular particle movements observed in plant cells.Abbreviations AVEC-DIC video microscopy in differential interference contrast - CFLSM confocal laser scanning microscope - DiOC₆(3) 3,3-dihexyloxacarbocyanine iodide - ER endoplasmic reticulum - IP₃ inositol triphosphate - N.A. numerical aperture - SIT silicon intensified target video camera - SR sarcoplasmic reticulum     

Bound indoles inAcetabularia

Zusammenfassung A. Chromatographische Analysen des Alkali-Hydrolysates von äthanolextrahiertenAcetabularia-Zellen ergaben, daß große Mengen noch nicht näher identifizierter, Salkowski-positiver Indole (rötlich) freigesetzt werden. Ihre Menge entspricht ungefähr der des freigesetzten Tryptophans. Die Indolylderivate sind bei der alkalischen Hydrolyse jedoch nicht aus Tryptophan entstanden.B. DieAcetabularia-Kristalle wurden als Indolylderivate identifiziert. Ultraviolettmikrospektrophotometrische Untersuchungen und eine Reihe von Färbetesten weisen auf ein 3-Indolylderivat hin, das keine Doppelbindungen in der Seitenkette und ein nichtsubtituiertes C in 2-Positon des Indolringes besitzt.Die intensive dauerhafte Rotfärbung durch das Salkowski-Reagens—modifiziert für cytologische Anwendungen—legt die Annahme nahe, daß Tryptophan in den Kristallen nicht die Hauptkomponente ist.Die Unlöslichkeit in sauren Medien, konzentrierten Ammoniumhydroxydlösugnen, organischen Lösungsmitteln und—nach Formolbehandlung—in starken Alkalilaugen läßt auf eine relativ hochmolekulare Substanz schließen.Das kristalline Indolylderivat wurde auch in anderen Arten der Dasycladaceen gefunden. Es ist hauptsächlich in der zentralen Vacuole dieser Zellen lokalisiert.Die Möglichkeit, daß die Salkowski-positiven Indole, die beiAcetabularia-Hydrolysaten gefunden wurden, Komponenten der Indolyl-Kristalle sind, muß durch weitere Untersuchungen an isolierten Kristallen geprüft werden.

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Summary A. Chromatographic analysis of alkaline hydrolyzates of ethanol-extractedAcetabularia whole cells shows appreciable amounts of unindentified Salkowski reactive indoles (red-pink) in quantities roughly comparable to tryptophane. These indolyl derivatives are not artefacts derived from tryptophane.B. TheAcetabularia crystals have been identified as an indolyl derivative. Analysis by ultraviolet microspectrophotometry and a number of colour tests indicate a 3-indolyl derivative without a double bound in the side chain and an unsubstituted C in the 2-position of the indole ring.The intense persistent pink colour given by Salkowski's reagent—modified for application at a cytological scale—strongly suggests that the bulk of the indoles in the crystals is not tryptophane.Insolubility in acid media, concentrated ammonia, organic solvents and—after formaldehyde treatment—in strong alkali hydroxides probably indicate a relatively high molecular weight compound.The crystalline indolyl derivative has been also found in several other genera of theDasycladaceae. Its localization resides mainly in the central vacuole.The tentative identification of the Salkowski-reactive indoles detected inAcetabularia hydrolyzates as components of the crystals must await confirmation by isolation and analysis of the crystals themselves.

     

Enhancement of oxygen mass transfer in stirred bioreactors using oxygen-vectors. 1. Simulated fermentation broths

Oxygen mass transfer represents the most important parameter involved in the design and operation of mixing-sparging equipment for bioreactors. It can be described and analyzed by means of the mass transfer coefficient, k_La. The k_La values are affected by many factors such as geometrical and operational characteristics of the vessels, media composition, type, concentration and microorganism morphology, and biocatalysts properties. The efficiency of oxygen transfer could be enhanced by adding oxygen-vectors in broths, such as hydrocarbons or fluorocarbons, without increasing the energy consumption for mixing or aeration. The experimental results obtained for simulated broths indicated a considerable increase of k_La in the presence of n-dodecane, and the existence of a certain value of n-dodecane concentration that corresponds to a maximum mass transfer rate of oxygen. The magnitude of the positive effect of n-dodecane depends both on the broths characteristics and operational conditions of the bioreactor.Notation d stirrer diameter, mm - d oxygen electrode diameter, mm - D bioreactor diameter, mm - h distance from the inferior stirrer to the bioreactor bottom, mm - H bioreactor height, mm - k_La oxygen mass transfer coefficient, s^-1 - l impeller blade length, mm - I oxygen electrode immersed length, mm - P power consumption for mixing of non-aerated broths, W - P_a power consumption for mixing of aerated broths, W - (P_a/V) specific power input, W/m³ - s baffle width, mm - v_S superficial air velocity, m/s - V volume of medium, m³ - w impeller blade height, mm - volumetric fraction of oxygen-vector - _a apparent viscosity, Pa*s - density, kg/m³     

Laccase electrode for the continuous-flow determination of phenolic compounds

Summary Laccase (p-diphenol, O₂ oxido-reductase, E.C. 1.10.3.2) from Botrytis cinerea was immobilized in a gelatin support on an O₂ sensing electrode. The enzyme was copolymerized with the inert protein using glutaraldehyde (1.25 % w/w) on the hydrophobic selective gas membrane of a pO₂ sensor and this was covered with a Nuclepore polycarbonate microporous film (0.03 m). The enzyme electrode was used in a continuous-flow system to measure the concentration of a wide range of phenolic substrates. The measuring time of each sample was about 1.5 min including response and rinsing times. The electrode response was set for hydroquinone up to 0.8 mM with high reproducibility and less than 5 % error.The electrode response for hydroquinone concentration of 0.25 mM was stable with repeated use for at least 800 assays without significant loss of activity.     

Alternative pathway activities in aged potato tuber slices measured by three methods

To find a simple and reliable oxygen electrode-based method to estimate the values of alternative pathway activity (V _alt) and its contribution to total respiration V _alt/V _t) in aged potato (Solanum tuberosum L.) tuber slices, we compared conventional hydroxamate-inhibiting method, improved hydroxamate-inhibiting method with 2,6-dichlorophenol indophenol (DCPIP), and the oxygen isotope discrimination (OID) method. The values of V _alt and V _alt/V _t obtained with an improved hydroxamate-inhibiting method with DCPIP in 12-h- and 24-h-aged slices were about twice higher than those with the conventional hydroxamate-inhibiting method. Only a relatively small difference in the values of V _alt and V _alt/V _t obtained by the OID method and the improved hydroxamate-inhibiting method with DCPIP in 12-h and 24-h-aged slices was observed. These results indicated that the improved hydroxamate-inhibiting method with DCPIP could be considered as a new, simple, and reliable technique for the noninvasive assay of the AP activity.From Fiziologiya Rastenii, Vol. 52, No. 2, 2005, pp. 311–315.Original English Text Copyright © 2005 by Hou, Zhou, Kong, Liang, Zhang.This article was submitted by the authors in English.This revised version was published online in April 2005 with a corrected cover date.     

Recording the oxygen production of a single Acetabularia cell for a prolonged period

A method for recording the O₂ evolution of an individual Acetabularia cell or cell fragment over a period of weeks is described. The method is based on the polarographic O₂ determination by means of a platinum electrode in a flow-through system. The mean O₂ evolution of a full-grown cell under constant conditions (2 5001m/m², 20 °C) was 3–6 μ1 O₂ per cell perh. Under these conditions the O₂ evolution exhibited a pronounced circadian rhythm with an average period of about 23 h and an amplitude of about 2.3 μ1 O₂ per cell per h. No significant differences were found between nucleate and anucleate cells.     

Eine Elektrode zur kontinuierlichen Messung des Gelöstsauerstoffs (pO2) in Fermenterkulturen

Zusammenfassung Die Arbeit beschreibt Aufbau und Eigenschaften einer Elektrode zur kontinuierlichen Messung des Sauerstoffpartialdruckes in Fermenterkulturen. Die nach dem polarographischen Prinzip arbeitende Elektrode leitet sich von der Clarkschen Meßanordnung ab. Ihr wesentliches Merkmal ist die Auftrennung in zwei selbständige Bauelemente, einen Membran-tragenden Glasmantel und den Elektroden-tragenden Meßgeber. Der Elektrodenmantel wird gemeinsam mit dem Fermentergefäß autoklaviert, der thermolabile Meßfühler dagegen erst nachträglich eingesetzt. Die Eichung der Elektrode kann vor dem Autoklavieren in einem Spezialgefäß oder danach im Fermentergefäß vorgenommen werden. Der sterilisierbare, in die Kulturlösung hineinragende Elektrodenmantel wird von der Lösung durch eine O₂-permeable, 250 starke Siliconglasgewebemembran abgeschlossen. Zwischen Siliconschicht und der blanken Platinoberfläche der Kathode befindet sich eine weitere, stabilisierende Membran von 12 Stärke (Cellophanfolie). Die Empfindlichkeit der Elektrode beträgt 10^–9 A/mm Hg pO₂ bei 37°C, die Einstellzeit auf 95% des Endwertes 50 sec bei 37°C. Die Abhängigkeit der Messungen von der Turbulenz des Mediums ist zwischen 400 und 1200 UpM Rührergeschwindigkeit zu vernachlässigen. Wiederholtes Autoklavieren der Siliconmembran hat keinen Einfluß auf die Einstellzeiten.

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An electrode for continuous measurement of oxygen tension in fermenter cultures

Summary The paper describes the construction and properties of an electrode for continuous measurement of oxygen tension in fermenter cultures. The equipment is a modified Clark electrode with a cathode of 99,99% pure Pt and a silver anode. It works polarographically. Its most outstanding feature is the subdivision into two separate construction elements: an outer glass-envelope containing a membrane, and a membrane-carrying electrode. The glass-envelope is autoclavable together with the fermenter vessel, whereas the thermolabile electrode must be inserted into the sterilized glass-envelope. The electrode can be calibrated either in a special vessel before autoclaving the glass envelope, or after sterilization in the air-saturated sterile culture medium, before inoculation.The steam resistent glass-envelope, which is immersed in the culture medium, is covered on the lower part with a silicone membrane of 250 thickness which is stabilized by glass fibers. The silicone membrane is separated from the polished Pt cathode by a second stabilizing membrane (cellophane) of 12 thickness, which is part of the electrode. The sensitivity of the electrode is 10^–9 A/mm Hg pO₂ at 37° C. When the oxygen tension is changed from 0 to 100%, the electrode follows this change up to 95% within 50 sec at 37° C. The reading is nearly independent of the agitation of the medium between 400 and 1,200 rpm of the impeller turbine in the culture vessel. Repeated steam-sterilization has no influence on the response time of the electrode.

     

Impedance of the electrogenic Cl− pump inAcetabularia: Electrical frequency entrainements,voltage-sensitivity,and reaction kinetic interpretation

Summary Reaction kinetic analysis of the electrical properties of the electrogenic Cl^– pump inAcetabularia has been extended from steady-state to nonsteady-state conditions: electrical frequency responses of theAcetabularia membrane have been measured over the range from 1 Hz to 10 kHz at transmembrane potential differences across the plasmalemma (V _m ) between –70 and –240 mV using voltage-clamp techniques. The results are well described by an electrical equivalent circuit with three parallel limbs: a conventional membrane capacitancec _m , a steadystate conductanceg _o (predominantly of the pump pathway plus a minor passive ion conductance) and a conductanceg _s in series with a capacitancec _p which are peculiar to the temporal behavior of the pump. The absolute values and voltage sensitivities of these four elements have been determined:c _m of about 8 mF m^–2 turned out to be voltage insensitive; it is considered to be normal.g _o is voltage sensitive and displays a peak of about 80 S m^–2 around –180 mV. Voltage sensitivity ofg _s could not be documented due to large scatter ofg _s (around 80 S m^–2).c _p behaved voltage sensitive with a notch of about 20 mF m^–2 around –180 mV, a peak of about 40 mF m^–2 at –120 mV and vanishing at –70 mV. When these data are compared with the predictions of nonsteady-state electrical properties of charge transport systems (U.-P. Hansen, J. Tittor, D. Gradmann, 1983,J. Membrane Biol. in press), model A (redistribution of states within the reaction cycle) consistently provides magnitude and voltage sensitivity of the elementsg _o ,g _s andc _p of the equivalent circuit, when known kinetic parameters of the pump are used for the calculations. This analysis results in a density of pump elements in theAcetabularia plasmalemma of about 50 nmol m^–2. The dominating rate constants for the redistribution of the individual states of the pump in the electric field turn out to be in the range of 500 sec^–1, under normal conditions.     

Bioprocess engineering characteristics of the horizontal stirred tank

Bioreactor performance studies of the recently developed horizontal stirred tank with a volume of 421 have been carried out for fermentation with Trichosporon cutaneum. Quantification on the basis of measured oxygen transfer capacity and power consumption is presented and compared with data for a conventional vertical tank bioreactor.During the experiments it has been observed that two different forms of morphology of Trichosporon, i.e. the normal yeast-form (Y) with single cells and a mycelium-form (M) with filamentous cells, are present in the horizontal stirred tank when working with the original strain (DSM 70698). After separation both forms were characterized and later on used for bioreactor performance studies in the horizontal and vertical stirred tank. Results of oxygen efficiency show the drastic effect of the morphology change on bioreactor performance. Finally different bioreactors are quantitatively compared on the basis of oxygen transfer, power consumption and productivity using the reference fermentation system Trichosporon cutaneum.List of Symbols F m³/h flow rate (volumetric) - k _La1/h volumetric transfer coefficient of OTR - M Nm torque - n 1/s rotational speed - P Nm/s power - V m³ volume - V _G1/min gas flow rate - x kg/m³ biomass concentration - ^* morphology index - ^* engineering (specific) viscosity - _app Ns/m² apparent viscosity - ₀ N/m² yield stress (Casson law) - t _1/e h measured time acc. to momentum method [17] - tEh characteristic time of electrode response - t _Gh mean residence time of gas phase Abbreviations CFR completely filled reactor - CRR cyclic ring reactor (torus) - JLR jet loop reactor - HSTR horizontal stirred tank reactor - M mycelium-form of Trichosporon cutaneum - O₂-eff O₂-efficiency - OUR O₂-uptake rate - OTR O₂-transfer rate - STR stirred tank reactor - ThLR thin layer reactor - VSTR vertical stirred tank reactor - Y yeast-form of Trichosporon cutaneum The work presented in this paper was supported by an Austrian Research Grant (FFWF, Project no. 4496)     

Oxygen equilibria and structural characteristics of the tetrameric and polymeric intracellular hemoglobins from the bivalve molluscBarbatia reeveana

Summary The arcid bivalveBarbatia reeveana contains within its erythrocytes two hemoglobins with remarkably different structures and oxygen equilibrium properties. A tetrameric hemoglobin (M _r about 60,000) with non-identical subunits (₂₂) constitutes about 60% of the erythrocytic heme protein. This hemoglobin has a relatively low oxygen affinity (P ₅₀=19 Torr at 20°C, pH 7.2), shows cooperativityn _H=2.2, shows no Bohr effect between pH 6.8 and 7.6 and a heat of oxygenation (H) of –5.4 kcal/mole between 15 and 35°C. Its oxygen affinity appears to be insensitive to ATP.B. reeveana erythrocytes also contain another hemoglobin withM _r=430,000, the largest intracellular hemoglobin known in any organism. The subunit of this hemoglobin is unusual, having aM _r of 32–34,000 and two heme oxygen binding sites per polypeptide chain. The large hemoglobin has a very low oxygen affinity (P ₅₀=33 Torr at 20°C, pH 7.2), shows slight cooperativity,n _H=1.8, and no Bohr effect (Grinich and Terwilliger 1980). The H at pH 7.2 equals –2.9 kcal/mole, a low value for most hemoglobins, and its O₂ affinity appears to be insensitive to ATP. The two hemoglobins ofB. reeveana, so different in their structure, are also different in their functional properties.     

Oxygen transfer in an airlift reactor with multiple net draft tubes

A pilot scale airlift reactor with multiple net draft tubes was developed to improve oxygen transfer in the reactor. The reactor was 0.29 m in diameter and 2 m height. A steadystate sulfite oxidation method was applied to determine an overall volumetric mass transfer coefficient. Oxygen transfer of the proposed airlift reactor can be 60–100% higher than that of bubble columns under the same operating conditions.List of Symbols C ^* mol·dm^–3 saturated concentration of dissolved oxygen - C _L mol·dm^–3 bulk concentration of dissolved oxygen - G mol/min nitrogen flow rate - k _L a hr^–1 the volumetric gas-liquid mass transfer coefficient - Mo ₂ g/mol molecular weight of oxygen - OTR g/min the oxygen transfer rate - U _g cm/s superficial air velocity - V _L dm³ volume of the liquid phase - _in oxygen mole ratio in the inlet gas - _out oxygen mole ratio in the outlet gas     

A simple dynamic method for on-line and off-line determination of kLa during cultivation of animal cells

Summary A new, fast method is described to determine k_La either off-line, or on-line during animal-cell cultivation. Since it does not need the equilibrium concentration of oxygen in the liquid phase (C*), it is not required to await a new steady state. Furthermore, the results do not depend on the calibration value of the dissolved-oxygen probe. The method yielded accurate values for k_La, both for an oxygen-consuming and a non-consuming system.Nomenclature C _L Dissolved-oxygen concentration [mol·m^-3] - C ^* C _L in equilibrium with the oxygen concentration in the gas phase [mol·m^-3] - C _L, Equilibrium oxygen concentration at stationary conditions [mol·m^-3] - k_La Volumetric oxygen transfer coefficient [s^-1] - r Specific oxygen consumption of biomass [mol·cell^-1·s^-1] - X Cell concentration [cells·m^-3] - t Time [s] - Noise of dissolved-oxygen probe [mol·m^-3] - Absolute error of k_La-measurement [s^-1]