Air slugs entrapped cross-flow filtration of bacterial suspensions

A novel cross-flow technique for membrane filtration of bacterial cell suspensions was established. This is an air slugs entrapped cross-flow method in which air slugs were generated by introducing air into the cross-flow stream. As air slugs moved along with cross-flow, the disturbance of cell sublayer formation on membrane surface was enhanced. As a consequence, filtration flux was improved and stabilized. The effect of air slugs on improving filtration flux was more pronounced in filtering gram-negative Escherichia coli cell than grampositive Brevibacterium flavum cell. Moreover, air slug was about 50% more effective on reducing filtration resistance using ultrafiltration (UF) membrane of 300,000 molecular weight cutoff (MWCO) than microfiltration (MF) membrane of 0.2 mum. (c)1993 John Wiley & Sons, Inc.     

Pressure effects in cross-flow microfiltration of suspensions of whole bacterial cells

The effect of Trans-Membrane Pressure (TMP) on permeate flux during cross-flow microfiltration of bacterial cell suspensions in tubular ceramic membranes is studied experimentally. Continuous filtration experiments with suspensions of whole bacterial cells (Mycobacterium M156) show a dramatic permeate flux decline with increasing TMP. During the very early stages of the filtration process, a linear relationship between permeate flux and TMP is observed, suggesting an initial surface sorption of cells on the membrane surface. At longer times, the permeate flux vs. TMP data exhibit a critical pressure beyond which the permeate flux declines with increasing trans-membrane pressure. This is interpreted in terms of the formation of a compressible cake, whose permeability can be described through the Carman-Kozeny equation.     

Dewatering of algal suspension using microfiltration with cross flow in the presence of magnetite as a filter aid

Dewatering algal suspensions is an important step in the extraction of oil and other useful substances from algae. In this study, spherical Nannochloropsis sp. and ellipsoidal Monoraphidium sp. suspensions were dewatered in the presence of different amounts of 350-nm magnetite particles using a microfiltration membrane with 360-nm pores in cross-flow mode. Magnetite functions as a filter aid by reducing the deformation of the cake of filtered algae on the membrane and providing paths for water to flow through the filtration cake of algae. In the case of Nannochloropsis sp., the highest dewatering rate was obtained when the number ratio, defined based on the size and ideal density, between Nannochloropsis sp. and magnetite was 1:12.5, but the addition of magnetite had no observable effect on the filtration of ellipsoidal Monoraphidium sp. suspensions through the membrane. After dewatering, magnetite was effectively separated from the concentrated algal suspension by the application of a magnetic field in an open flow system. Magnetite has the potential to enhance dewatering performance using a cross-flow membrane system.     

Microfiltration of recombinant yeast cells using a rotating disk dynamic filtration system

To develop a highly efficient cell harvest step under time constraint, a novel rotating disk dynamic filtration system was studied on the laboratory scale (0.147-ft.(2) nylon membrane) for concentrating recombinant yeast cells containing an intracellular product. The existing cross-flow microfiltration method yielded pseudo-steady state flux values below 25 LMH (L/m(2). h) even at low membrane loadings (10 L/ft.(2)). By creating high shear rates (up to 120,000(-1)) on the membrane surface using a rotating solid disk, this dynamic filter has demonstrated dramatically improved performance, presumably due to minimal cake buildup and reduced membrane fouling. Among the many factors investigated, disk rotating speed, which determines shear rates and flow patterns, was found to be the most important adjustable parameter. Our experimental results have shown that the flux increases with disk rotating speed, increases with transmembrane pressure at higher cell concentrations, and can be sustained at high levels under constant flux mode. At a certain membrane loading level, there was a critical speed below which it behaved similarly to a flat sheet system with equivalent shear. Average flux greater than 200 LMH has been demonstrated at 37-L/ft.(2) loading at maximum speed to complete sixfold concentration and 15-volume diafiltration for less than 100 min. An order of magnitude improvement over the crossflow microfiltration control was projected for large scale production. This superior performance, however, would be achieved at the expense of additional power input and heat dissipation, especially when cell concentration reaches above 80 g dry cell weight (DCW)/L. Although a positive linear relationship between power input and dynamic flux at a certain concentration factor has been established, high cell density associated with high viscosity impacted adversely on effective average shear rates and, eventually, severe membrane fouling, rather than cake formation, would limit the performance of this novel system. (c) 1995 John Wiley & Sons, Inc.     

Preferential deposition of smaller cells during cross-flow microfiltration of a yeast suspension

Summary By examining the size distribution of cells that do not deposit during cross-flow microfiltration of a yeast suspension, we demonstrate that the smaller cells in the suspension are preferentially deposited on the membrane. The extent to which the deposition process favours smaller cells was found to be unaffected by cell concentration or membrane type over the range of concentrations examined.     

Pilot-scale separation of lysozyme from hen egg white by integrating aqueous two-phase partitioning and membrane separation processes

A novel, cost-effective method of lysozyme separation from hen egg white was studied. This method integrates aqueous two-phase partitioning in the system EO₅₀PO₅₀/phosphates with membrane separation processes. The experiments were carried out in a pilot-scale on crude hen egg white.Initially, by forming an aqueous two-phase system, lysozyme was selectively extracted to the upper, polymer-rich phase while the other egg white proteins partitioned to the lower, phosphate-rich phase. Then, in order to recover lysozyme, thermoseparation of polymer-rich phase was applied. A novel approach for the simultaneous thermoseparation of the polymer-rich phase as well as for the recovery of the lysozyme was proposed, using a cross-flow microfiltration. Additionally, recovery of proteins by ultrafiltration from lower, phosphate-rich phase was also investigated.Lysozyme could be obtained after the thermal phase separation by means of microfiltration at a total recovery over the extraction steps of 47.5 and the purification factor of 10.5. The specific activity of lysozyme preparations was 34 188 U/mg of protein. Using cross-flow membrane techniques, it was found that the recovery of the polymer by microfiltration from the top phase was 83.9.     

Microfiltration conditions modify Lactobacillus bulgaricus cryotolerance in response to physiological changes

This work aimed at analyzing the effect of microfiltration conditions (cross-flow velocity and transmembrane pressure) on the quality of frozen Lactobacillus bulgaricus CFL1 starters produced on pilot scale. Microfiltered cells were less resistant during the concentration process than centrifuged cells. In contrast, bacterial cryotolerance during freezing was improved after microfiltration, in a range of 28–88%, depending on the microfiltration conditions. During frozen storage, cell resistance was also affected by microfiltration conditions, either positively or negatively, compared to centrifugation. The best cryotolerance was obtained for cells microfiltered at a cross-flow velocity of 2 m/s and a transmembrane pressure of 0.15 MPa. This improvement was explained by considering membrane fatty acid composition of Lb. bulgaricus CFL1. This condition increased unsaturated to saturated and cyclic to saturated fatty acid ratios, which enhanced membrane fluidity, thus helping the cells to better resist freezing and frozen storage.     

Mechanistic modeling of the loss of protein sieving due to internal and external fouling of microfilters

Fed‐batch and perfusion cell culture processes used to produce therapeutic proteins can use microfilters for product harvest. In this study, new explicit mathematical models of sieving loss due to internal membrane fouling, external membrane fouling, or a combination of the two were generated. The models accounted for membrane and cake structures and hindered solute transport. Internal membrane fouling was assumed to occur due to the accumulation of foulant on either membrane pore walls (pore‐retention model) or membrane fibers (fiber‐retention model). External cake fouling was assumed to occur either by the growth of a single incompressible cake layer (cake‐growth) or by the accumulation of a number of independent cake layers (cake‐series). The pore‐retention model was combined with either the cake‐series or cake‐growth models to obtain models that describe internal and external fouling occurring either simultaneously or sequentially. The models were tested using well‐documented sieving decline data available in the literature. The sequential pore‐retention followed by cake‐growth model provided a good fit of sieving decline data during beer microfiltration. The cake‐series and cake‐growth models provided good fits of sieving decline data during the microfiltration of a perfusion cell culture. The new models provide insights into the mechanisms of fouling that result in the loss of product sieving. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1323–1333, 2017     

Effect of membrane morphology on system capacity during normal flow microfiltration

Understanding the effects of membrane fouling on system capacity is critical for the successful design and scale-up of microfiltration systems. The underlying morphology and structure of the microfiltration membrane can have a significant effect on system capacity by altering the rate and extent of fouling. Experimental data were obtained for system capacity during protein microfiltration using several model membranes with both homogeneous and composite structures. Data were compared with predictions of a new model that can account for both pore blockage and cake formation, and also includes the effects of membrane morphology on internal flow profiles within the membrane. Membranes with highly interconnected pores have a significantly higher capacity due to the reduction in flux decline arising from the fluid flow under and around any surface blockage. The model calculations are in good agreement with the flux decline data, allowing far more accurate predictions of system capacity than for the commonly used V(max) analysis.     

Membrane fouling in a membrane bioreactor (MBR): sludge cake formation and fouling characteristics

A submerged membrane bioreactor (MBR) with a working volume of 1.4 L and a hollow fiber microfiltration membrane was used to treat a contaminated raw water supply at a short hydraulic retention time (HRT) of approximately 1 h. Filtration flux tests were conducted regularly on the membrane to determine various fouling resistances, and confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were employed to characterize the biofouling development and sludge cake formation on the membrane. The experimental results demonstrate that the MBR is highly effective in drinking water treatment for the removal of organic pollutants, ammonia, and UV absorbance. During the MBR operation, the fouling materials were not uniformly distributed on the entire surface of all of the membrane fibers. The membrane was covered partially by a static sludge cake that could not be removed by the shear force of aeration, and partially by a thin sludge film that was frequently washed away by aeration turbulence. The filtration resistance coefficients were 308.4 x 10(11) m(-1) on average for the sludge cake, 32.5 x 10(11) m(-1) on average for the dynamic sludge film, and increased from 10.5 x 10(11) to 59.7 x 10(11) m(-1) for the membrane pore fouling after 10 weeks of MBR operation at a filtration flux of 0.5 m3/m2 x d. Polysaccharides and other biopolymers were found to accumulate on the membrane, and hence decreased membrane permeability. More important, the adsorption of biopolymers on the membrane modified its surface property and led to easier biomass attachment and tighter sludge cake deposition, which resulted in a progressive sludge cake growth and serious membrane fouling. The sludge cake coverage on the membrane can be minimized by the separation, with adequate space, of the membrane filters, to which sufficient aeration turbulence can then be applied.     

Membrane fouling by cell-protein mixtures: in situ characterisation using multi-photon microscopy

Fouling of the membrane by cell and protein mixtures can result in severe flux declines, leading to the eventual need to clean or replace the membrane. In this study multi-photon microscopy, a fluorescence-based technique is used to 3-D image in situ the fouling of microfiltration membranes by suspensions containing combinations of washed yeast, bovine serum albumin (BSA) and ovalbumin. Appropriate fluorescent labelling allows the three foulant species to be clearly identified. Images correlate well with filtration data and clearly show the cake of yeast cells capturing protein aggregates. The proteins exhibited very different filtration behaviour. When filtering washed yeast together with ovalbumin and/or a 50:50 mixture by mass of BSA and ovalbumin, the ovalbumin fouling dominates the system. Capture of aggregates by the cake did not reduce fouling of the membrane by the protein and increased the resistance of the cake. For mixtures of BSA and washed yeast, the presence of a cake of yeast cells did reduce fouling of the membrane by the protein, however, the extra resistance due to the cake resulted in a flux lower than that when filtering BSA alone.     

Microfiltration of yeast suspensions with self-cleaning spiral vortices: possibilities for a new membrane module design

A novel method of producing controlled vortices was used to reduce both concentration polarization and membrane fouling during microfiltration of Saccharomyces cerevisiae broth suspensions. The method involves flow around a curved channel at a sufficient rate so as to produce centrifugal instabilities (called Dean vortices). These vortices depolarize the build-up of suspended particles such as yeast cells at the membrane-solution interface and allow for increased membrane permeation rates. Various operating conditions under which such vortices effectively reduced cake build-up of suspended particles such as yeast cells at the membrane-solution interface and allow for increased membrane permeation rates. Various operating conditions under which such vortices effectively reduced cake build-up during microfiltration of 0 to 0.55 dry wt% yeast broth were investigated. Flux improvements of over 60% for 0.25 dry wt% yeast broth for flow with over that without Dean vortices were observed. This beneficial effect increased with increasing retentate flow rate and increasing transmembrane pressure and decreased with increasing concentration of suspended matter. Similar behavior was observed whether the cells were viable of killed. the improvement in flux in the presence over that in the absence of vortices correlated well with centrifugal force or azimuthal velocity squared. The relative cake resistances increased with reservoir yeast concentration. These values with vortices increased from 62% to 75% of that without vortices with increasing yeast concentration. The ratio of the cake thicknesses in the limiting case (at high feed concentration) was 3.25. These results suggest that self-cleaning spiral vortices could be effective in maintaining good and steady microfiltration performance with cell suspensions other than those tested. (c) 1995 John Wiley & Sons, Inc.     

Transient and stationary operating conditions on performance of lactic acid bacteria crossflow microfiltration

A filtration rig equipped with a tubular alumina membrane was used to study the performance of crossflow microfiltration of Lactobacillus helveticus. Experiments were performed at constant permeation flux. High cell concentrations and fast transient conditions to the stationary J adversely affected permeability. Membrane fouling was due to a fast irreversible layer formation and to a reversible cell cake. This microbial deposit characteristics were dependent on the ratio permeation flux/wall shear stress, J/tau(w). Fouling was faster and more severe when J/tau(w) was greater than a critical value of 1.15 L(-1) . h(-1) . m(-2) . Pa(-1). The disordered structure of this cell cake seemed to lead to a macromolecule deposit between the cells which adversely affected the membrane permeability. (c) 1996 John Wiley & Sons, Inc.     

Cross-flow membrane microfiltration of a bacteriol fermentation broth

Although cross-flow membrane filtration is a very attractive option for harvesting cells and recovering enzymes from cell homogenates, the process is not without its problems. Foremost of these is the deposit of dissolved and suspended solutes onto the membrane surface during operation. The formation of these dense and sometimes compressive sublayers (often called cakes) offers additional resistance to axial and permeate flows and often affects the retention characteristics of the process. In view of the complex nature of the sublayer formation process and its sensitivity to cross-flow velocity, this investigation was undertaken to determine the main factors responsible for the decline in performance during the harvesting of B. polymyxa broth by membrane microfiltration. System parameters varied include axial flow rate, concentration of cells, proteins and other components in the feed, membrane materials (ceramic, polypropylene, and stainless steel), and cleaning methods. To help explain the observed results, a new mass transport model-the solids flux model-based on the assumptions that back migration of particles from the sublayer or membrane surface is negligible and that particles that reach the solid-solution interface attach (stick) completely, is tested. Using a variety of diagnostic methods, magnesium ammonium phosphate precipitate is formed during steam sterilization of the medium and is implicated as the major foulant in this study.     

Purification during crossflow electromicrofiltration of fermentation broth

The present study was to investigate the purification of a fermentation broth by an electromicrofiltration membrane. Microfiltration runs with a crude and a centrifuged broth, with solution of particles recovered from centrifugation and with permeates from microfiltration experiments were thus compared.Microfiltration performances were governed by colloids and small particles that induced sharp initial flux declines. For these results, the evolution of the overall membrane resistance was increased by 80% in comparison with the electromicrofiltration membrane. The main focus of this study was set on the enhancement of the filtrate flux by an electric field. This pressure electrofiltration leads to a drastic improvement of the filtration by 100% and the filtration time was thereby reduced. Pressure electrofiltration serves as an interesting alternative to the cross-flow filtration and it effectively separates advantageous constituents such as amino acids and biopolymers from a fermentation broth. They were equally maintained during the microelectrofiltration, although they were significantly reduced by 45% by the microfiltration without the application of an electric field. Accordingly, since the electrofiltration membrane was provided more permeability, this study experimentally demonstrates that the permeability inside a membrane can be controlled using an electric field.     

Microfiltration of streptococcal fermentation broths: study of the factors affecting the concentration effect

Streptokinase (SK) recovery from streptococcal fermentation broth by cross-flow microfiltration has been studied. Recovery of SK in the filtrate, independent of the volumetric concentration factor, is approximately two-fold lower than the initial SK activity in the fermentation broth; moreover, the SK activity in the retentate increase during the process, reaching a concentration factor of 2.73. These results show that the membrane works more as an ultrafiltration membrane, with rejection of S = 0.6, than as a microfiltration membrane. Under filtration conditions, the membrane permeation rate decreased with time. This decreased could be explained by deposition and interaction of material onto/with the membrane resulting in the concentration of permeable products. Studies of the individual concentration factors for the main streptococcal exocellular proteins, indicate clearly that the concentration of the proteins during the microfiltration process is independent of the size of the proteins, suggesting that other factors, such as charge and hydrophobicity, along with concentration-polarization, should be taken also into account for the understanding of this phenomenon. (c) 1994 John Wiley & Sons, Inc.     

藻源污染物特性对高藻水微滤处理过程中膜污染的影响

采用不同生长时期的藻细胞、藻源型有机物(AOM)及原藻液进行过滤实验,研究不同生长时期的藻源污染物对膜污染的影响特性及机制。利用UMFI法分析不同生长时期的藻细胞、AOM及原藻液的污染程度;采用CRITIC分析法定量分析了不同生长时期的AOM和藻细胞在混合过滤过程中对膜污染的贡献率,同时采用混合污染堵塞模型分析了不同生长时期的原藻液不同过滤阶段的主要污染堵塞类型。结果表明, 3个生长时期的藻细胞及AOM的膜污染程度均为对数期最轻;值得注意的是,在原藻液过滤过程中藻细胞及AOM的膜污染贡献率随着生长时期的不同而有所变化,其中AOM的污染权重随着生长时期的延长不断减小,而藻细胞的污染权重随着生长时期的延长不断增大。不同生长时期的原藻液过滤过程中均呈现两段式污染堵塞类型,并且后段均为滤饼堵塞。研究不仅阐明了藻源型污染物特性对微滤处理高藻水膜污染的影响机制,同时也为改善膜污染的技术开发提供参考。     

A novel approach to evaluate the permeability of cake layer during cross-flow filtration in the flocculants added membrane bioreactors

In order to obtain a better understanding of the cake layer formation mechanism in the flocculants added MBRs, a model was developed on the basis of particle packing model considering cake collapse effect and a frictional force balance equation to predict the porosity and permeability of the cake layers. The important characteristic parameters of the flocs (e.g., floc size, fractal dimensions) and operating parameters of MBRs (e.g., transmembrane pressure, cross-flow velocity) are considered in this model. With this new model, the calculated results of porosities and specific cake resistances under different MBR operational conditions agree fairly well with the experimental data.     

Combined in-fermenter extraction and cross-flow microfiltration for improved inclusion body processing

In this study we demonstrate a new in-fermenter chemical extraction procedure that degrades the cell wall of Escherichia coli and releases inclusion bodies (IBs) into the fermentation medium. We then prove that cross-flow microfiltration can be used to remove 91% of soluble contaminants from the released IBs. The extraction protocol, based on a combination of Triton X-100, EDTA, and intracellular T7 lysozyme, effectively released most of the intracellular soluble content without solubilising the IBs. Cross-flow microfiltration using a 0.2 microm ceramic membrane successfully recovered the granulocyte macrophage-colony stimulating factor (GM-CSF) IBs with removal of 91% of the soluble contaminants and virtually no loss of IBs to the permeate. The filtration efficiency, in terms of both flux and transmission, was significantly enhanced by in-fermenter Benzonase digestion of nucleic acids following chemical extraction. Both the extraction and filtration methods exerted their efficacy directly on a crude fermentation broth, eliminating the need for cell recovery and resuspension in buffer. The processes demonstrated here can all be performed using just a fermenter and a single cross-flow filtration unit, demonstrating a high level of process intensification. Furthermore, there is considerable scope to also use the microfiltration system to subsequently solubilise the IBs, to separate the denatured protein from cell debris, and to refold the protein using diafiltration. In this way refolded protein can potentially be obtained, in a relatively pure state, using only two unit operations.     

A study on the effect of operating parameters on the cross-flow microfiltration of yeasts

The effects of pump speed, cumulative permeate volume and concentration of feed (yeast cells) on the permeate flux have been studied on a batch cross-flow microfiltration process. The experiments were conducted for two different cellulose acetate membrane modules of 0.2 m and 0.45 m pore size. A three factor experiment was designed for this purpose and the effect of the operating parameters on the filtration rate was studied by the analysis of variance (ANOVA). It is concluded from the analysis of the experimental data that pump speed has the maximum bearing upon the permeate rate within the operating range of parameters. Fouling conditions were examined in the light of colloids deposition on membranes due to surface interactions. However this paper looks into the relationship and sensitivity of the operating parameters in a cross-flow microfiltration unit rather than exploring the theoretical principles behind the observed phenomena.This revised version was published online in October 2005 with corrections to the Cover Date.