Protein mass transfer studies on a spray column using the PEG-Reppal PES 100 aqueous two-phase system

The characterization of Bovine Serum Albumin mass transfer mechanisms in a spray column using an aqueous two-phase system composed of poly(ethylene glycol) and a modified starch-Reppal PES 100-is done. The poly(ethylene glycol) rich phase is used as the dispersed phase and protein transfer takes place from the dispersed phase to the continuous phase. The effect of dispersed phase superficial velocity, system composition, continuous phase height and distribution system design on either overall protein mass transfer coefficient or column hold-up is described. It is shown that continuous phase superficial velocity and phase composition are the main controlling factors for protein transfer. It is also observed that, with the tested system, only at very low dispersed phase superficial velocities is it possible to operate the spray column as an extraction column. In this system the upper operating limit of the dispersed phase velocity is ten times smaller than in other aqueous two-phase systems.List of Symbols ATPS Aqueous Two-Phase System - BSA Bovine Serum Albumin - C _i kg m^–3 inlet dispersed phase protein concentration - C ₀ kg m^–3 outlet dispersed phase protein concentration - C _d kg m^–3 dispersed phase protein concentration - C _c kg m^–3 continuous phase protein concentration - D m column internal diameter - H hold-up - h, h _d m dispersion height - h ₀ m initial dispersion height (initial continuous phase height) - k _da s^–1 overall mass transfer coefficient - m protein partition coefficient - n number of holes of distribution system - PEG Poly(ethylene glycol) - Q m³ s^–1 dispersed phase volumetric flow rate - S m² column internal area - V m³ dispersion volume A. Venâncio was supported by a JNICT (Junta Nacional de Investigaçäo Científica e Tecnológica) grant.     

Viscous media in tower bioreactors: Hydrodynamic characteristics and mass transfer properties

Studies in tower reactors with viscous liquids on flow regime, effective shear rate, liquid mixing, gas holdup and gas/ liquid mass transfer (k _La) are reviewed. Additional new data are reported for solutions of glycerol, CMC, PAA, and xanthan in bubble columns with diameters of 0.06, 0.14 and 0.30 m diameter. The wide variation of the flow behaviour index (1 to 0.18) allows to evaluate the effective shear rate due to the gas flow. New dimensionless correlations are developed based on the own and literature data, applied to predict k _La in fermentation broths, and compared to other reactor types.List of Symbols a(a) m^–1 specific interfacial area referred to reactor (liquid) volume - Bo Bond number (g D _c ² _L/) - c _L(c _L ^* ) kmol m^–3 (equilibrium) liquid phase oxygen concentration - C coefficient characterising the velocity profile in liquid slugs - C _s m^–1 coefficient in Eq. (2) - d _B(d_vs) m bubble diameter (Sauter mean of d _B) - d ₀ m diameter of the openings in the gas distributor plate - D _c m column diameter - D _L m²s^–1 diffusivity - E _L(E_W) m² s^–1 dispersion coefficient (in water) - E ² square relative error - Fr Froude number (u _G/(g D_c)^0.5) - g m s^–2 gravity acceleration - Ga Gallilei number (g D _c ³ _L ² / _eff ² ) - h m height above the gas distributor the gas holdup is characteristic for - k Pasⁿ fluid consistency index (Eq. 1) - k _L m s^–1 liquid side mass transfer coefficient - k _La(k_La) s^–1 volumetric mass transfer coefficient referred to reactor (liquid) volume - L m dispersion height - n flow behaviour index (Eq. 1) - P W power input - Re liquid slug Reynolds number ( _L(u _G +u _L) D _c/_eff) - Sc Schmidt number ( _eff/( _L D _L )) - Sh Sherwood number (k _La D _c ² /D_L) - t s time - u _B(u_sw) m s^–1 bubble (swarm) rise velocity - u _G(u_L) m s^–1 superficial gas (liquid) velocity - V(V_L) m³ reactor (liquid) volume Greec Symbols W m^–2 K^–1 heat transfer coefficient - y(y _eff) s^–1 (effective) shear rate - _G relative gas holdup - s relaxation time of viscoelastic liquid - _L(_eff) Pa s (effective) liquid viscosity (Eq. 1) - _L kg m^–3 liquid density - N/m surface tension     

Residence time distribution of liquid phase in an external-loop airlift bioreactor

The residence time distribution analysis was used to investigated the flow behaviour in an external-loop airlift bioreactor regarded as a single unit and discriminating its different sections. The experimental results were fitted according to plug flow with superimposed axial dispersion and tank-in-series models, which have proved that it is reasonable to assume plug flow with axial dispersion in the overall reactor, in riser and downcomer sections, as well, while the gas separator should be considered as a perfectly mixed zone. Also, the whole reactor could be replaced with 105-30 zones with perfect mixing in series, while its separate zones, that is the riser with 104-27, the downcomer with 115-35 and the gas separator with 25-5 perfectly mixed zones in series, respectively, depending on gas superficial velocity, A_D/A_R ratio and the liquid feed rate.List of Symbols A _D cross sectional area of downcomer (m²) - A _R cross sectional area of riser (m²) - A ₁ A ₂ length of connecting pipes (m) - Bo Bodenstein number (Bo=vL·L/D _ax (-) - C concentration (kg m^–3) - C residence time distribution function - C ₀ coefficientEquation (12) - C _r dimensionless concentration - D _D diameter of downcomer (m) - D _R diameter of riser column (m) - D _ax axial dispersion coefficient (m²s^–1) - H _d height of gas-liquid dispersion (m) - H _L height of clear liquid (m) - i number of complete circulations - L length of path (m) - m order of moments - N _eq number of perfectly mixed zones in series - n _c circulating number - Q _c recirculating liquid flow rate (m³ s^–1) - q _F liquid feed flow rate (m³s^–1) - Q _G gas flow rate (m³s^–1) - Q _T total liquid flow rate (m³s^–1) - r recycle factor - s exponent inEquation (12) regarded as logarithmic decrement of the oscillating part of RTD curve - t time (s) - t _C circulation time (s) - t _s mean residence time (s) - t ₉₉ time necessary to remove 99% of the tracer concentration (s) - V _A volume of connecting pipes (m³) - V _D volume of downcomer (m³) - V _L liquid volume in reactor (m³) - V _R volume of riser (m³) - V _LD linear liquid velocity in downcomer (m s^–1) - V _LR linear liquid velocity in riser (m s^–1) - V _SLD superficial liquid velocity in downcomer (m s^–1) - V _SLR superficial liquid velocity in riser (m s^–1) - x independent variable inEquation (1) - ¯x mean value of x - z axial coordinate - _GR gas holdup in riser - _m(x) central moment of m order - ² variance - dimensionless time     

Adsorption studies for the separation ofl-tryptophan froml-serine and indole in a bioconversion medium

l-tryptophan was produced froml-serine and indole by immobilized Escherichia coli cells in organic-aqueous systems. Selective adsorption was the method chosen to enable both product separation andl-serine reutilization. Amongst various adsorbents tested activated carbons and neutral polymeric resins (XAD-4 and XAD-7) showed good performance. The neutral resins could selectively concentrate thel-tryptophan from dilute aqueous solutions and adsorbed only 5% of the unconvertedl-serine. High separation factors (l-tryptophan/l-serine and indole/l-tryptophan) were obtained with these adsorbents. Despite a lower capacity, the XAD-7 resin had the advantage of desorbingl-tryptophan with basic or acidic solutions, while organic solvents were required to desorb, at the same concentration levels, this compound from XAD-4.In a packed bed column filled with XAD-4 resin or activated carbon, totall-tryptophan adsorption and recovery were achieved at linear velocities up to 5.0 cm/min and 3.2 cm/min respectively. Successive sorbent reutilization, following continuous sorption and elution steps, was carried out in packed bed columns with the neutral resins and activated carbon.Thel-form of tryptophan, after crystallization, was identified by HPTLC.List of Symbols HPLC High Performance Liquid Chromatography - HPTLC High Performance Thin Layer Chromatography - Trp tryptophan - Ser Serine - A amount of sorbent(g) - c equilibrium solute concentration in the aqueous phase (g/dm³) - c _i initial (before adding the sorbent) liquid phase concentration (g/dm³) - C _T tryptophan concentration in the inlet solution (g/dm³) - C _To tryptophan concentration in the outlet solution (g/dm³) - E _z axial dispersion coefficient (m²/s) - k experimental constant (Eq. 1, 2 and 3) - K ₁ rate constant of adsorption (min^–1) - L column length(m) - n experimental constant (eq. 1, 2 and 3) - q equilibrium solid phase concentration (g solute/g sorbent) - q _max maximum capacity of sorbent (g solute/g sorbent) - t time(s) - v liquid velocity (m/s) - V volume of liquid phase(dm³) - V _e eluted volume(dm³) - V _r volume needed to saturate the column (dm³)     

Hydrodynamics and mass transfer in an airlift loop fermentor

Summary The hydrodynamics and mass transfer behaviour of an airlift fermentor with an external loop (height 10m) has been investigated by measuring gas and liquid velocities, gas hold-up, liquid mixing and oxygen transfer coefficients. Liquid phase properties, i.e., ionic strength, viscosity and surface tension have been varied by altering the fermentation media. Results are compared with those from bubble column experiments performed in the same unit. It is shown, that more uniform two-phase flow in the airlift leads to advantages in scale-up and operation.Nomenclature a Specific interfacial area per volume of dispersion (m²/m³) - c Local concentration of tracer (kmol/m³) - c Concentration of tracer at infinite time (kmol/m³) - C_L Concentration of oxygen in the liquid bulk (kmol/m³) - C_L ^* Concentration of oxygen in the interface (kmol/m³) - D_ax Axial dispersion coefficient (cm²/s) - I Ionic strength (kmol/m³) - i Inhomogeneity [defined in Eq. (2)] - Rate of oxygen transfer (kmol/s) - t_c Circulation time (s) - t_M Mixing time (s) - V_R Volume of gas-liquid dispersion (m³) - V_SG Superficial gas velocity in up-flow column (m/s) Greek letter symbols _L Oxygen transfer coefficient (m/s) - Dynamic viscosity (m Pa s) - Surface tension (m N/m) Presented at the First European Congress on Biotechnology, Interlaken, September 25–29, 1978     

Mathematical models for mixing in deep-jet bioreactors: Calculation of parameters

The macroscopic mathematical model based on compartments with ideal mixing zones and tanks-in series was evaluated. Based on the experimental data obtained in a 300 dm³ pilot reactor and the dependence of mixing time on the volume of liquid phase, we have found mathematical relations between the ratio of vessel diameter to liquid level, adjustable parameters of model and the mixing time.List of Symbols V dm³ total volume of bioreactor - V _g dm³ total volume of liquid - V ₁ dm³ volume of ideally mixed zone in the vessel - V ₂ dm³ volume of macromixer in inner circulation flows - V ₃ dm³ volume of liquid phase in the pump - V ₄ dm³ volume of liquid phase in the pipe between the vessel and the pump - V ₅ dm³ volume of liquid phase in the pipe between the pump and air input system included falling jet - V _LT dm³ volume of liquid in the tank - V _LC dm³ volume of liquid in the circulation system - F _E dm³/s inner volumetric circulation flow rate across the macromixers - F _cir dm³/s external volumetric circulation flow rate, pumping capacity - t _A s time interval of the pulse application - t _AA s time point of the pulse application related to the free choosen starting point of the experiment - t _m s mixing time - t _c s circulation time - t _end s end time of simulation - C _*,* kg/m³ concentration of tracer in the indicated compartment - C ₀ kg/m³ concentration of the tracer before the injection - C _t kg/m³ concentration of the tracer at the indicated time - C kg/m³ theoretical concentration of the full mixed tracer - C _sim kg/m³ calculated concentration of tracer during numerical integration method - i index of an arbitrary tank - D _T m diameter of bioreactor - D 1/s dilution rate - H _L m level of liquid in the unaerated vessel - vector of inhomogenities     

Cell recovery by continuous flotation

Summary Cell recovery by means of continuous flotation of the Hansenula polymorpha cultivation medium without additives was investigated as a function of the cultivation conditions as well as of the flotation equipment construction and flotation operational parameters. The cell enrichment and separation is improved at high liquid residence times, high aeration rates, small bubble sizes, increasing height of the aerated column, and diameter of the foam column. Increasing cell age and cultivation with nitrogen limitation reduce the cell separation.Symbols C_P cell mass concentration in medium g·l^–1 - C_R cell mass concentration in residue g·l^–1 - C_S cell mass concentration in foam liquid g·l^–1 - V equilibrium foam volume cm³ - V gas flow rate through the aerated liquid column cm³·s^–1 - V_F feed rate to the flotation column ml/min - ¹ V _S/V foaminess s - mean liquid residence time in the column s     

Penicillin recovery from aqueous solutions by continuous foam flotation

Summary Penicillin G recovery is investigated in a continuous flotation column in the presence of different collectors which form a complex with penicillin. The performance of the penicillin recovery was investigated as a function of the mole ratio () of collector-to-penicillin and the aliphatic chain length of the collector. At =1 and low penicillin concentrations (e.g., 20 mg·1^-1), high foam liquid concentrations (680 mg·l^-1), low residue concentrations (12 mg·l^-1) and high penicillin separation (56) can be attained. At =4 the separation increases to 150, and 95% of the penicillin can be recovered.Symbols C_p penicillin concentration in feed (mg·l^-1) - C_R penicillin concentration in outlet liquid (mg·l^-1) - C_S penicillin concentration in foam liquid (mg·l^-1) - C_S/C_P penicillin enrichment (-) - C_S/C_R penicillin separation (-) - % Pen in S penicillin yield in foam liquid (%) - VV}_S foam liquid volume flow (ml·min^-1) - VV}_P feed (ml·min^-1) - VV_{N 2} nitrogen flow rate (ml·s^-1) - temperature     

Model for the production of L-tryptophan from L-serine and indole by immobilized cells in a three-phase liquid-impelled loop reactor

Production of L-tryptophan from L-serine and indole catalyzed by Escherichia coli, immobilized in k-carrageenan gel beads, is technically feasible in the liquidimpelled loop reactor (LLR), using an organic solvent, e.g. n-dodecane.With L-serine in large excess intrinsic reaction kinetics is approximately first order with respect to indole, with a reaction constant of 8.5×10^–5 m³ kg _dw ^–1 s^–1.The overall process kinetics is jointly controlled by intrinsic kinetics and by intraparticle mass transfer resistance, which can be quantified using an effectiveness factor.Mass transfer of indole from the organic to the aqueous phase and from the aqueous to the gel phase are relatively fast and thus have negligible influence in the overall process kinetics, under the operational conditions tested. However, they may become important if the process is intensified by increasing the cell concentration in the gel and/or the gel hold-up in the reactor.A simple model which includes indole mass balances over the aqueous and organic phases, mass transfer and reaction kinetics, with parameters experimentally determined in independent experiments, was successful in simulating L-tryptophan production in the LLR.List of Symbols a, b, c coefficients of the equilibrium curve for indole between organic and aqueous phases - A, B, C, D, E, F auxiliary variables used in liquid-liquid mass transfer studies - a _x specific interfacial area referred to the volume of the aqueous phase (m^–1) - A _x interfacial area (m²) - a _Y specific interfacial area referred to the volume of the organic phase (m^–1) - A _Y interfacial area (m²) - C _b substrate concentration in the bulk of the aqueous phase (kg m^–3) - C _e substrate concentration in exit stream (kg m^–3) - C _E biocatalyst concentration referred to the aqueous phase (kg m^–3) - C _{E s} biocatalyst concentration referred to the volume of gel (kg m^–3) - C _s substrate concentration at the gel surface (kgm^–3) - d, e, f coefficients of the equilibrium curve for indole between aqueous and organic phases - dp particle diameter (m) - K ₂ kinetic constant (s^–1) - K ₁ kinetic constant K₂/K_M (kg^–1 m³ s^–1) - K _M Michaälis-Menten constant (kgm^–3) - K _X mass transfer coefficient referred to the aqueous phase (ms^–1) - K _Xa_X volumetric mass transfer coefficient based on the volume of the aqueous phase (s^–1) - k _Y mass transfer coefficient referred to the organic phase (ms^–1) - K _Ya_Y volumetric mass transfer coefficient based on the volume of the organic phase (s^–1) - N _X mass flux of indole from organic to aqueous Phase (kg m^–2s^–1) - N _Y mass flux of indole from aqueous to organic phase (kg m^–2s^–1) - Q _e volumetric flow rate in exit stream (m³s^–1) - Q _f volumetric flow rate in feed stream (m³s^–1) - _obs observed reaction rate (kg s^–1 m^–3) - intrinsic reaction rate (kg s^–1 m^–3) - Re Reynolds number - Sc Schmidt number - Sh Sherwood number - t time (s) - u superficial velocity (m s^–1) - V _max maximum reaction rate (kg s^–1m^–3) - V _S volume of the support (m³) - V _X volume of aqueous phase (m³) - V _Y volume of the organic phase (m³) - X indole concentration in the aqueous phase (kgm^–3) - Y indole concentration in the organic phase (kg m^–3 Greek Letters overall effectiveness factor - _e external effectiveness factor - _i internal effectiveness factor - Thiele module A fellowship awarded to one of us (D.M.R.)by INICT is gratefuly acknowledged.     

Measurement of the evaporation rate of liquid in a shaking flask

The evaporation rate (N_H2O) of liquid in a shaking flask was measured under various shaking conditions: temperature, humidity, flask shape, liquid volume in the flask (V_L), length of the stopper in the flask neck (L_C), rotational speed of the shaker (N), and wind velocity (V_W). The rate was significantly affected by these factors, and the existence of a distribution of water vapor pressure was suggested inside and outside the flask. To predict the evaporation rate, the following empirical equation was derived by the least squares method: N_H2O=1.26×10⁻²S_F^1.18L_F^−1.3 (p_s–p)^1.24V_L^0.11N^0.05V_W^0.26L_C^−0.37 where, S_F is the cross-sectional area of flask neck, L_F is the length of flask neck, p_s is the saturated partial pressure of water vapor at the temperature of the air surrounding the flask, and p is the partial pressure of water vapor in the flask.     

Enhanced oxygen transfer rates in fermentation using soybean oil-in-water dispersions

Summary The effect of soybean oil on the volumetric oxygen transfer coefficient during the cultivation ofAerobacter aerogenes cells is presented. For our aeration-agitation conditions (0.278 vvm and 500 rpm), it has been demonstrated that the use 19% (v/v) of soybean oil enabled a 1.85-fold increase of thek _l a coefficient (calculated on a per liter aqueous phase basis). For smaller volumetric oil fractions,k _L a increased linearly with the oil loading. Because of the oxygen-vector properties of soybean oil, this oil is able to significantly increase thek _L a of a bioreactor.Nomenclature C^*, C saturation and actual dissolved oxygen concentrations respectively (g/m³) - K_La volumetric oxygen transfer coefficient (h^–1) - K_La_initial k _La measured before the oil addition (h^–1) - M_O2 molar mass of oxygen (dalton) - N oxygen transfer rate (g/m³. h) - P_O2. P_N2 partial pressures ofO ₂ andN ₂ in the gas (atm) - P_H2OT partial pressure of water in air at the temperatureT (atm) - P_T total pressure (atm) - Q₀ volumetric flow rate of outlet air before seeding (m³/h) - Sp spreading coefficient (dynes/cm) - T absolute temperature of outlet gas (K) - V_i volume of the liquidi in the fermentor (m³) - V_M molar volume at 273 K and 1 atm (m³/mole) - _ij interfacial tension betweeni andj componants (dynes/cm) - _v volumetric fraction of the oil (v/v) - G gas - O oil - W water - i inlet - o outlet     

Mass transfer and liquid mixing in an airlift reactor with a net draft tube

Mass transfer and liquid mixing in an airlift reactor with a net draft tube were experimentally investigated. Four different column diameters were considered. The mass transfer was measured using the volumetric gas-liquid mass transfer coefficient which was determined by the dynamic method. The mass transfer coefficients in the airlift reactors with different column diameters were not always higher than those in the bubble columns. The liquid mixing was measured using mixing time which was determined by a pulse technique. Under the same superficial gas velocity, the mixing times of the airlift reactors with a net draft tube were always less than those of the bubble columns.List of Symbols C mol·dm^–3 bulk concentration of dissolved oxygen - C ₀ mol·dm^–3 initial concentration of dissolved oxygen - C _e mol·dm^–3 saturated concentration of dissolved oxygen - ¯C dimensionless dissolved oxygen concentration - D _c cm diameter of column - D _N cm diameter of the nozzle hole - D _T cm diameter of the net draft tube - H _L cm static liquid height - H _T cm height of the net draft tube - k _L a hr^–1 volumetric mass transfer coefficient - L _T cm length of the net draft tube - t _M sec mixing time of the liquid phase - t ₀ sec mixing time of the liquid phase in a bubble column - V _L dm³ volume of the liquid phase - U _g cm/s superficial air velocity     

To what extent is a constant volume design worse than a minimum volume design for a series of CSTR's?

The balance equations for substrate in a cascade of CSTR's undergoing an enzyme-catalyzed reaction following Michaelis-Menten kinetics are developed in dimensionless form. Analytical expressions relating the intermediate concentrations are independently obtained for the cases of minimum overall volume and constant volume. The fractional deviations between the overall volumes following these two design criteria are calculated and presented for several values of the relevant parameters. For situations of practical interest, the fractional deviation is below 10%. Increasing values of the Michaelis-Menten parameter, K _m(or decreasing values of the number of reactors in the cascade, N) lead to lower values of the maximum deviation; this maximum deviation is attained at lower conversions of substrate when K _mis increased or N decreased.List of Symbols C _{S, i}mol.m^–3 concentration of substrate at the outlet of the i-th reactor - C _* ^{S, i} normalized concentration of substrate at the outlet of the i-th reactor - C _* ^{S, i, eq} normalized concentration of substrate at the outlet of the i-th reactor using the design criterion of constant volume - C _* ^{S, i, opt} normalized concentration of substrate at the outlet of the i-th reactor using the design criterion of minimum overall volume - C _{S, 0} mol.m^–3 concentration of substrate at the inlet to the first reactor - Da _i Damköhler number for the i-th reactor - Da _eq constant Damköhler number for each reactor of the cascade - Da _{tot, eq} overall Damköhler number for the cascade assuming equal-sized reactors - Da _{tot, min} minimum overall Damköhler number for the cascade - Er fractional deviation between the overall volumes using the two different design criteria - K _mmol. m^–3 Michaelis-Menten constant - K _* ^M dimensionless Michaelis-Menten constant - N number of reactors of the cascade - Q m³. s^–1 volumetric flow rate - V _im³ volume of the i-th reactor - v _max mol. m^–3. s^–1 reaction rate under saturation conditions of the enzyme with substrate - V _{tot, opt} m³ minimum overall volume of the cascade - V _{tot, eq} m³ overall volume of the cascade assuming equal-sized reactors     

Gas holdup and mass transfer characteristics of carboxymethyl cellulose solutions in a bubble column with a radial gas sparger

The gas phase holdup and mass transfer characteristics of carboxymethyl cellulose (CMC) solutions in a bubble column having a radial gas sparger have been determined and a new flow regime map has been proposed. The gas holdup increases with gas velocity in the bubbly flow regime, decreases in the churn-turbulent flow regime, and increases again in the slug flow regime. The volumetric mass transfer coefficient (k _La) significantly decreases with increasing liquid viscosity. The gas holdup and k _La values in the present bubble column of CMC solutions are found to be much higher than those in bubble columns or external-loop airlift columns with a plate-type sparger. The obtained gas phase holdup ( _g) and k _La data have been correlated with pertinent dimensionless groups in both the bubbly and the churn-turbulent flow regimes.List of Symbols a m^–1 specific gas-liquid interfacial area per total volume - A _d m² cross-sectional area of downcomer - A _r m² cross-sectional area of riser - d _b m individual bubble diameter - d _vs m Sauter mean bubble diameter - D _c m column diameter - D _L m²/s oxygen diffusivity in the liquid - Fr Froude number, U _g/(g D_c)^1/2 - g m/s² gravitational acceleration - G _a Galileo number, gD _c ^{3 2}/² _app - H _a m aerated liquid height - H _c m unaerated liquid height - K Pa · sⁿ fluid consistency index - k _L a s^–1 volumetric mass transfer coefficient - n flow behavior index - N _i number of bubbles having diameter d _bi - Sc Schmidt number, _app/( D _L) - Sh Sherwood number, k _L a D _c ² /D_L - U _sg m/s superficial gas velocity - U _gr m/s superficial riser gas velocity - V _a m³ aerated liquid volume - V _c m³ unaerated liquid volume - N/m surface tension of the liquid phase - _g gas holdup - _app Pa · s effective viscosity of non-Newtonian liquid - kg/m³ liquid density - ý s^–1 shear rate - Pa shear stress     

Protein recovery by continuous flotation

Summary Bovine serum albumin (BSA) was recovered from aqueous solutions by foam flotation. The protein concentrations in foam liquid C _S, in feed C _Pand in residue C _Rwere determined. The protein enrichment C _S/C_Pand the separation C _S/C_Ras well as the protein fraction in the foam liquid % BSA and foam liquid volume flow were determined as functions of the medium properties, operational conditions, and equipment parameters as well as concentrations of solid particles. At low protein concentrations in feed (e.g., C _P=40 mg · l^-1), and at 40° C, high performance was attained (C _X=2,000 mg · l^-1, C _R=4.4 mg · l^-1, C _S/C_P=50, C _S/C_R=450, 90% BSA. Continuous foam flotation is an efficient procedure for the recovery of low concentrations of proteins from liquid cultures.Abbreviations BSA bovine serum albumine - C _P protein concentration in feed (mg · l^-1) - C _R protein concentration in residue (mg · l^-1) - C _S protein concentration in foam liquid (mg · l^-1) - C _S/C_R protein separation (-) - C _S/C_P protein enrichment (-) - V _P feed rate (ml · min^-1) - V _R residue flow rate (ml · min^-1) - V _S foam liquid volume flow (ml · min^-1) - N number of theoretical stages in an ideal cascade (-) - temperature (° C) - mean residence time (min)     

Separation of supercoiled and open-circular plasmid DNA by liquid-liquid counter-current chromatography

We report for the first time the use of liquid-liquid counter-current chromatography (CCC) for the preparative scale fractionation of plasmid DNA. Almost complete fractionation of supercoiled and open circular plasmid DNA (6.9 kb) could be achieved using a phase system comprising 12.5% (w/w) PEG 600 and 18% (w/w) K₂HPO₄. Experiments were carried out on a Brunel J-type CCC machine (100 ml PTFE coil) at a mobile phase flow rate of 0.5 ml min^{– 1} and a rotational speed of 600 rpm. Compared to conventional HPLC techniques the capacity of CCC is not limited by the surface area of resin available for adsorption. Symbols: C _b, Concentration of plasmid in lower phase (g ml^–1); C _t, Concentration of plasmid in upper phase (g ml^–1); CV, Total volume of mobile phase present in the coil and connecting leads (ml); K, Equilibrium solute partition coefficient (K=C _t/C _b); OC, Open circular plasmid; SC, Supercoiled plasmid; S _f, Percentage stationary phase retention (S _f=V _s/V _c); t _s, Time for phase separation (s); V _b, Volume of bottom phase (ml); V _c, Coil volume (ml); V _m, Volume of mobile phase present in coil at equilibrium (ml); V _r, Volume ratio of two phases (V _r=V _t/V _b); V _s, Volume stationary phase present in coil at equilibrium (ml); V _t, Volume of top phase (ml); V _tot, Total volume of phase system (ml).     

Improved scale-up strategies of bioreactors

Effective scale-up is essential for successful bioprocessing. While it is desirable to keep as many operating parameters constant as possible during the scale-up, the number of constant parameters realizable is limited by the degrees of freedom in designing the large-scale operation. Scale-up of aerobic fermentations is often carried out on the basis of a constant oxygen transfer coefficient, k _L a, to ensure the same oxygen supply rate to support normal growth and metabolism of the desired high cell populations. In this paper, it is proposed to replace the scale-up criterion of constant k _L by a more direct and meaningful criterion of equal oxygen transfer rate at a predetermined value of dissolved oxygen concentration. This can be achieved by using different oxygen partial pressures in the influent gas streams for different scales of operation. One more degree of freedom, i.e., gas-phase oxygen partial pressure, is thus added to the process of scale-up. Accordingly, one more operating factor can be maintained constant during scale-up. It can be used to regulate the power consumption in large-scale fermentors for economical considerations or to describe the fluid mixing more precisely. Examples are given to show that the results of optimization achieved in the bench-scale study can be translated to the production-scale fermentor more successfully with only a small change in the gas-phase oxygen partial pressure employed in the bench-scale operation.List of Symbols a m²/m³ Specific gas/liquid interfacial area - C _L mole/m³ Dissolved oxygen concentration in bulk liquid phase - C ^* mole/m³ Equilibrium oxygen concentration at gas/liquid interface - D _i m Impeller diameter - D _T m Bioreactor diameter - H _L mole/m³ · atm Henry's-law constant - k _L m/s Liquid-phase mass transfer coefficient - N 1/s Impeller agitation speed - N _i Number of impellers - OTR mole/s · m³ Oxygen transfer rate per unit volume of the medium - P _g kW Power input in aerated fermentation - P _o kW Power input in non-gassed fermentation - p _g atm Gas-phase oxygen partial pressure - Q m³/s Volumetric gas flow rate - Re _i Impeller Reynolds number - T _Q Joule Torque applied to the mixer shaft - V m³ Liquid volume - v _s m/s Superficial gas velocity - kg/m · s Liquid viscosity - kg/m³ Liquid density     

Study of the Solute Flows of Multicomponent and Heterogeneous Non-Ionic Solutions in Double-Membrane System

The solute flows were studied in a double-membrane osmotic-diffusive cell, in which two membranes mounted in horizontal planes separate three compartments (l,m,r) containing the non-homogeneous, non-electrolytic binary and ternary solutions. The volume of inter-membrane compartment (m), which is the infinitesimally layer of solution, and volume of external compartments (l and r) fulfill the conditions V _m 0 and V _l =V _r , respectively. In an initial moment, the solution concentrations satisfy the condition (C ^o _s ) _l < (C ^o _s ) _m >(C ^o _s ) _r. The double-membrane osmotic-diffusive cell is composed of two complexes: boundary layer/membrane/boundary layer, mounted in horizontal planes. In the cell, solute flux was measured as a function of concentration and gravitational configuration. The linear dependencies of the solute flux on concentration difference in binary solutions and nonlinear – in ternary solutions were obtained. It was shown that the double-membrane osmotic-diffusive cell has rectifying and amplifying properties of solute flows.     

镍离子对尖叶莴苣氮素吸收和生长生理的影响

以‘全年油麦菜’尖叶莴苣为试验材料,采用水培方式,研究3个浓度(0 mg·L^-1、0.1 mg·L^-1、1 mg·L^-1)Ni²⁺在22.4 mg·L^-1 N处理下对尖叶莴苣氮素吸收的生长及生理影响。结果显示：(1)尖叶莴苣根系和地上部生物量随处理时间的增加呈上升趋势。与对照T₁(0 mg·L^-1 Ni²⁺、112 mg·L^-1 N)相比,T₂处理(0 mg·L^-1 Ni²⁺、22.4 mg·L^-1 N)对尖叶莴苣根系及叶片生长具有一定抑制作用,植株鲜重、干重、根冠比、根系长度、平均直径、表面积、体积、根尖数、分根数、叶片表面积和体积在T3处理(0.1 mg·L^-1 Ni²⁺、22.4 mg·L^-1 N)下显著高于对照,T₄处理(1 mg·L^-1 Ni²⁺、22.4 mg·L^-1 N)对尖叶莴苣根系及其叶片生长具有一定促进作用,但对其根尖数和分根数表现出一定抑制性。(2)随着Ni²⁺浓度的增加,尖叶莴苣叶片叶绿素a、叶绿素b和总叶绿素含量呈先升后降的变化规律,且均在T₃处理下显著提高。(3)随着处理时间的增加,尖叶莴苣叶片的净光合速率(P_n)、气孔导度(G_s)和蒸腾速率(T_r)逐渐上升,胞间CO₂浓度(C_i)逐渐下降,且T₃处理叶片的G_s显著高于对照,其C_i最低,P_n最大。(4)施加Ni²⁺对尖叶莴苣有机酸、可溶性蛋白和可溶性糖含量以及SOD和POD活性有显著影响,在T₃处理下有机酸含量降低,可溶性糖和可溶性蛋白含量显著增加,SOD和POD活性显著提高。(5)T₃处理尖叶莴苣根系中N及叶片中B和Ca含量较高;根系中Ni含量高于叶片,T₃处理叶片中的Ni含量较低,Mg含量较高;植株体内Cu含量随Ni²⁺浓度增加而下降。研究表明,外源Ni²⁺处理能影响低氮条件下(22.4 mg·L^-1 N)尖叶莴苣幼苗生长及生理状况,适宜浓度(0.1 mg·L^-1)Ni²⁺可有效提高尖叶莴苣根系对氮素的吸收利用效率,减少氮素施用量,促进尖叶莴苣根系和地上部叶片生长,增加光合色素含量,并提高净光合速率,进而改善植株的产量和营养品质。     

Synthesis,reactions, spectral and magnetic studies of bimetallic alkoxides of cobalt(II) with zirconium(IV)

A bimetallic isopropoxide of cobalt(II) with the formula Co[Zr₂(OPrⁱ)₉]₂, prepared by the reaction of COCl₂ with K[Zr₂(OPrⁱ)₉] in 1:2 molar ratio, has been shown to undergo alcoholysis reactions with graded alcohols (primary, secondary and tertiary) to afford products of the types, Co[Zr₂(OR)₉]₂ (where R = Me, Et, Prⁿ, Buⁿ, Buⁱ and Bu^s) Co[Zr₂(OPrⁱ)₃(OEt)₆]₂, Co[Zr₂(OPrⁱ)₆(OBu^s)₃]₂, CO[Zr₂(OPrⁱ)₃(OBu^s)₆]₂, Co[Zr₂(OPrⁱ)₆(OBu^t)₃]² and Co[Zr₂(OPrⁱ)₃(OR)₆]₂ (where R = Am^t or Bu^t). These derivatives have been characterized by elemental analyses and molecular weight determinations. Infrared, electronic (visible) spectral and magnetic susceptibility measurements suggest a distorted octahedral geometry for these derivatives.