Analysis of process integration and intensification of enzymatic cellulose hydrolysis in a membrane bioreactor

Enzymatic cellulose hydrolysis has been studied for many years, generating rich literatures and knowledge in respect to the underlying reaction mechanism, reaction kinetics, and bioreactor systems. This paper attempts to offer some additional information and new understanding of how reaction kinetics and reactor productivity can be improved in a process involving simultaneous reaction and product separation using a purpose‐built membrane reactor with a single combined reaction zone and separation zone. Different operating strategies of batch, fed batch and continuous cellulose hydrolysis were investigated with intermittent or simultaneous removal of products (reducing sugars) to reduce enzyme inhibition and improve reactor productivity. The effect of continuous and selective product removal, reduced enzyme inhibition and higher enzyme concentration in retention were examined for the potential benefit in process integration and intensification in order to lower the high process cost of the enzymatic hydrolysis process, mainly due to slow reaction kinetics and expensive enzymes. A mathematical model was offered to account for the effect of selective product (reducing sugars) separation, permeate flux, reduced cellulase inhibition, dynamic structural change of the solid substrate and possible shear deactivation of the enzyme. Computer analysis was also carried out to analyse the quasi‐steady state of the reaction intermediates in order to gain an insight into the reaction mechanism in simultaneous reaction and separation systems. Some original analysis and simulation of the effect of membrane separation parameters on the overall reactor performance is offered, including the effect of membrane selectivity (rejection coefficient) and flux. Copyright © 2005 Society of Chemical Industry     

酶解反应与膜分离耦合连续制备酪蛋白磷酸肽

采用酶解反应与膜分离耦合新工艺连续水解全酪蛋白制备酪蛋白磷酸肽(CPPs)。考察了超滤膜对胰蛋白酶及底物溶液的截留效果;研究了初始底物质量浓度、初始酶质量浓度、反应体积、膜渗透通量等参数对反应器性能和反应转化率的影响规律;利用高效凝胶排阻色谱系统(HPSEC)对酶解产物进行检测分析;建立了酶膜反应器连续水解动力学模型,并对间歇与连续酶解过程进行比较分析,证明反应-分离耦合技术可使酶解效率及蛋白酶利用率大幅提高,并使产物得到调控与富集,为CPPs的酶法制备提供了一种更为有效的方法。     

Membranes in the enzymatic synthesis of biotensides from renewable sources

A novel integrated process of enzymatic synthesis of sugar fatty acid esters from renewable sources was proposed for the system oleic acid/-methylglucoside focussing on the application of different membrane techniques. The operational parameters were studied and optimised carrying out the reaction in an enzymatic membrane reactor (EMR) where the catalyst remained retained by means of ultrafiltration. A pervaporation unit coupled to the EMR was applied for by-product removal (water). A proper product separation and isolation was achieved applying combined techniques including filtration, evaporation, extraction and alternatively stepwise elution chromatography or dialysis.     

米曲霉菌体光学拆分N-乙酰-DL-苯甘氨酸的动力学研究

用液相培养基培养的米曲霉菌体直接拆分N 乙酰 DL 苯甘氨酸,并对该酶促反应进行了初步研究。考察了反应温度、pH值、金属离子、底物浓度、产物浓度等因素对反应的影响。结果表明,酶促反应的最适反应温度和pH值分别为52℃和7.0,当底物浓度小于200mmol·L-1时,反应符合Michaelis Menten方程,当底物浓度大于200mmol·L-1时,反应存在底物抑制现象。进一步研究发现水解反应也存在着产物抑制现象。     

Dynamics of penicillin G hydrolysis in an electro‐membrane reactor

Analysis of penicillin G hydrolysis in a membrane reactor with membrane‐entrapped penicillin G acylase is performed using a mathematical model of the reactor system. An electric field imposed to the reactor is considered to enhance transport rates of reaction components and reaction rate. Diffusion, electrophoretic migration and electro‐osmotic flux across the membrane are considered. The analysis focuses on possible effects of the principal operational parameters (electric field intensity, inlet substrate concentration, membrane thickness) on reactor performance. Multiplicities of steady states are frequently encountered. The membrane reactor performance can be easily targeted towards the required reaction regime by applying a constant or periodically varying electric field to the system. The periodic alternation of the polarity of the electric field substantially increases the effectiveness factor of penicillin hydrolysis compared with the steady state operation. Proper adjustments of electric field intensity may also compensate for the decay in enzyme activity. © 2001 Society of Chemical Industry     

Mathematical modelling and simulation of a recycle dialysis membrane reactor in a reversed micellar system

A mathematical framework was developed for the evaluation of a recycle dialysis membrane reactor (RDMR). The lipase-catalyzed hydrolysis of olive oil in an AOT-iso-octane reversed micellar system was employed as a model. Three specific operational strategies have been considered, namely batch, fed-batch, and fed-batch-bleed. Simulation shows the conversion of substrate to be strongly dependent on efficient use of the substrate, since the permeability coefficients of both substrate and product are quite similar. Sensitivity analyses were performed to assess the influences of various parameters (membrane area, substrate feed rate, solvent bleed rate and permeability) on the performance of the reactor in different modes of operation. The analyses presented are useful to assist the optimization of the operational strategy used for the RDMR system.     

Effect of enzyme inhibition on the performance of packed-bed biological reactor - a theoretical study

The performance characteristic of a packed bed reactor has been analyzed by considering diffusional resistance of the biofilm. Model equations were solved by the method of orthogonal collocation for various classical enzyme inhibition kinetics, including partially non-competitive, partially competitive, partially uncompetitive, partially mixed and fully mixed. For all considered modes of inhibition, an increase in the inlet substrate concentration decreases the steady state conversion in the reactor. However, an increase in the Peclet number has been found to improve the conversion. The effects of various other process variables of physical importance were also investigated parametrically.     

MASS TRANSFER-REACTION COUPLING PRODUCES MULTIPLE SUBSTRATE CONCENTRATIONS AT FLOC SURFACE: SUBSTRATE INHIBITED KINETICS

Simplistically, the whole cell can be modelled as a well mixed substrate sink, enveloped by a diffusive barrier (the cell membrane). In this communication we use this model in describing diffusion and reaction within a cell floe, subject to substrate inhibition. The cell floe is pictured as an array of well mixed environments between which substrate transfer occurs. When the observed rate of substrate uptake by the floe is equated to the substrate transport rate across the extra-floe stagnant liquid film, multiple interface concentrations are possible. Their number increases as intra-floe diffusion influences the observed reaction rate. More fundamentally, this is the first time that more than three steady states have been predicted using the particular graphical technique with substrate inhibited kinetics.     

MASS TRANSFER-REACTION COUPLING PRODUCES MULTIPLE SUBSTRATE CONCENTRATIONS AT FLOC SURFACE: SUBSTRATE INHIBITED KINETICS

Simplistically, the whole cell can be modelled as a well mixed substrate sink, enveloped by a diffusive barrier (the cell membrane). In this communication we use this model in describing diffusion and reaction within a cell floe, subject to substrate inhibition. The cell floe is pictured as an array of well mixed environments between which substrate transfer occurs. When the observed rate of substrate uptake by the floe is equated to the substrate transport rate across the extra-floe stagnant liquid film, multiple interface concentrations are possible. Their number increases as intra-floe diffusion influences the observed reaction rate. More fundamentally, this is the first time that more than three steady states have been predicted using the particular graphical technique with substrate inhibited kinetics.     

PRODUCTION OF ETHANOL WITH SACCHAROMYCES CEREVISIAE IN A CONTINUOUS REACTOR Note I: kinetics of immobilized yeast

Continuous production of ethanol by fermentation with immobilized yeasts gives a better yield and capacity and can be better controlled than conventional processes with batch reactors and a suspended biomass. This paper (the first of three) examines the operative conditions for immobilizing Saccharomyces Cerevisiae on beech wood cubes. After preliminary evaluation of the non-influence of mass transfer phenomena and the extent of alcoholic inhibition, an experimental investigation of the kinetics for a metabolic reaction in a continuous recycle reactor was carried out. The kinetic equation was found to be a Michaelis-Menten type with uncompetitive inhibition by substrate, and linear inhibition by product.     

Evaluation of UF and RO in a cellulose saccharification process

The applications of membrane separation within the bioconversion of lignocellulosic materials to ethanol are studied, and this paper reports on cellulolytic enzyme recovery, and reduction of the product inhibition. The conversion of cellulose and hemicellulose to hexoses and pentoses is carried out in a laboratory scale UF-membrane reactor, and the sugars, which have an inhibiting effect on the enzymes, are continuously removed with the permeate. By optimizing the space velocity, the enzyme inhibition is minimized and the rate of hydrolysis is significantly increased.The recovery of the enzymes has also been investigated with regard to their mechanical stability and adsorption on the substrate. Prior to enzymatic hydrolysis, the raw material is subjected to a pretreatment to make the cellulose more accessible to the enzymes, and the effect of the pretreatment can easily be evaluated in the UF-membrane reactor.As a result of the continuous removal of products formed in the hydrolysis, the permeate has a low content of fermentable sugars, and to give the optimal conditions for the fermentation a concentration step is needed. Different RO-membranes are tested in a laboratory scale batch cell to be able to choose a membrane material, fit for filtration in larger modules.     

Optimal design of continuously stirred membrane reactors in series using Michaelis-Menten kinetics with competitive product inhibition: theoretical analysis

Analytical expressions were derived for the optimal design (based on the minimum of the volume of the total number of reactors) of N continuously stirred membrane reactors (CSMRs) performing the enzyme-catalyzed reaction described by Michaelis-Menten kinetics with competitive product inhibition. The influence of membrane selectivity for both substrate and product on the total dimensionless residence time of the reactors (overall volume) was determined. The optimal design of N CSMRs (variable volume reactors) was compared with equal volume membrane reactors required to achieve the same degree of substrate conversion. The effect of kinetic and operating parameters on the performance of membrane reactors was determined. Optimization results show that membrane reactors are superior to continuously stirred tank reactors (CSTRs) in series at a high substrate rejection coefficient and low product rejection coefficient, high substrate conversion and using a small number of reactors. Also a high dimensionless Michaelis-Menten constant, high dimensionless inhibition constant and low substrate concentration in the feed to the first reactor improved the performance of the membrane reactors vs. CSTRs in series. The reduction in total volume of the optimal membrane reactors compared to CSTRs in series was up to 86% for the conditions in this work. A comparison between the optimum and equal volume design of membrane reactors in series showed no major difference in total volume between the two design criteria at a practical range of operating conditions. A volume reduction up to 16% was observed for the conditions in this work.     

反胶团酶反应过程开发研究进展

反胶团酶反应具有水相与有机相酶反应的共同优点 ,因而受到人们的重视。制约其实际应用的重要原因在于如何解决酶的重复利用问题。最初人们采用水溶液萃取法回收酶 ,但由于条件苛刻 ,酶活回收率较低。相变法利用温度的改变使体系分相以实现酶的回收 ,操作简便 ,但仍有较大的酶活损失。连续流两相离心反应器可以实现反胶团酶反应的连续操作且酶的损失很小 ,但该反应器传质速率较低因而转化率不高。膜反应器是反胶团酶反应中研究最多也最有应用前途的一种反应器形式。文中介绍了反胶团酶反应中酶重复利用的方法 ,对其过程开发的最新进展进行了综述。     

Stability characteristics of a biochemical reactor with predator-prey relationship. A substrate inhibition case

The present work characterizes static behaviour of a CSTBR with existing food chain relationship, for a case when base carbon and energy source exhibits toxic influence on enzymatic pathways of all microorganisms. Some bifurcation diagrams for representative process parameters are presented, together with characteristic phase plane plots. Conditions leading to induction of persistent oscillations of substrate and cell concentration are derived, and ways to avoid them are also given herein.     

有机相中固定化酶催化反应拆分手性化合物--间歇反应器内非定态动力学模拟

建立了有机相间歇反应器内球形多孔载体固定化酶催化反应拆分手性化合物的非定态动力学模型.采用正交配置法求解,讨论了对映体选择性E、内外传质阻力以及底物和水的抑制作用对酶促拆分手性化合物的拆分效果和速率的影响.结果表明:对映体选择性E是决定手性化合物能否拆分的关键参数;若一拆分过程对S-对映体优先反应,传质阻力可以忽略,E≥100,则R-和S-对映体几乎完全被拆分;如果存在传质阻力,则酶的对映体选择性降低,拆分效果不好,且拆分速率下降;底物和水的抑制作用对酶促拆分的效果影响不大,但拆分所需的时间相应增加.     

Periodic solutions in a porous catalyst pellet—homoclinic orbits

The analysis performed as well as extensive numerical simulations have revealed the possibility of the generation of homoclinic orbits as a result of homoclinic bifurcation in the model which describes transport phenomena and chemical reaction in a porous catalyst pellet. A method has been proposed for the development of a special type of diagrams—the so-called bifurcation diagrams. These diagrams comprise the locus of homoclinic orbits together with the lines of limit points bounding the region of multiple steady states as well as the locus of the points of Hopf bifurcation. Thus, they define a set of parameters for which homoclinic bifurcation can take place. They also make it possible to determine conditions under which homoclinic orbits are generated.Two kinds of homoclinic orbits have been observed, namely semistable and unstable orbits. It is found that the character of the homoclinic orbit depends on the stability features of the limit cycle which is linked with the saddle point.Very interesting dynamic phenomena are associated with the two kinds of homoclinic orbits; these phenomena have been illustrated in the solution diagrams and phase diagrams.     

Mass transfer and biochemical reaction in enzyme membrane reactor systems—I. Single enzyme reactions

The effective use of an enzyme as a biological catalyst for process scale conversion is greatly enhanced by immobilizing the enzyme within the matrix of a membrane whose walls are permeable to the substrate and product. The enzyme is thus stabilized for reuse, but the apparent activity is reduced compared to that obtained with the free enzyme because of transport resistances in the bulk fluid near the membrane and within the membrane itself. To account for this reduction, solutions to the differential equations describing the mass transport-kinetic models can be obtained for certain important cases. The reaction is considered to be first-order and isothermal, and to occur in one of two modes of operation: fixed-bed or batch (limited volume). Irreversible and reversible reactions and plane sheet and spherical geometries of the membrane are considered. An example of the application of these solutions to actual data obtained with a laboratory scale immobilized enzyme reactor is presented.     

Polarization Enhanced Production in a Membrane Reactor

Abstract

The membrane-based catalytic reactor with “free” catalyst has been modeled simply and studied by the authors on a theoretical basis. The hypothesis supporting an advantage of the membrane reactor is twofold: a) perfect rejection by the membrane and small reject fraction concentrates the catalyst highly in a steady-state continuous flow reactor (SSCFR) and b) the membrane reactor produces a filtered, high quality product. Of further advantage is the potential for concentrating the catalyst highly near the membrane in a thin diffusion dependent zone wherein the reacting substrate is also concentrated. The conjunction of concentrated catalyst and substrate leads to less inhibition of the reaction. The model involves conventional diffusional theory, simple membrane characterization of uniform flux, and Michaelis-Menten kinetics.     

Effect of Polymer Growth Rate and Diffusion Resistance on the Behavior of Industrial Polyethylene Fluidized Bed Reactor

The two‐phase model developed for the UNIPOL polyethylene process is improved by introducing polymer diffusion resistance, this means modelling of polyethylene fluidized bed reactors has been examined on two levels, at small scale of individual polymer particle, and macroscale of the whole reactor. The model utilizes the multigrain model that accounts for the reaction rate at catalyst surface to explore the static and dynamic bifurcation behavior of the fluidized bed catalytic reactor. Detailed bifurcation diagrams are developed and analyzed for the effect of polymer growth factor and Thiele modulus (the significance of the porous medium transport resistance is characterized by Thiele modulus) on reactor dense phase monomer concentration and reactor temperature as well as polyethylene production rate and reactor single pass conversion for the safe temperature region. The observations reveal that significant diffusion resistance to monomer transport exists, and this can mask the intrinsic rate constants of the catalyst. The investigation of polymer growth factor indicates that, the nascent stage of polymerization is highly gas phase diffusion influenced. Intraparticle temperature gradients would appear to be negligible under most normal operating conditions.     

反应器操作方式对酶动力学拆分立体选择性的影响

描述了在批式反应器和连续流搅拌反应器（ＣＳＴＲ）中酶动力学拆分对映异构体的不同之处，从宏观反应器平衡角度，推导出了在ＣＳＴＲ反应器中不同于在批式反应器中的一定酶立体选择性（Ｅ）下，底物或产物的对映体过量值与反应的转化率之间关系的定量关系式。并通过商品脂肪酶及芽胞杆菌Ｅ－５３脂肪酶催化的萘普生甲酯的不对称水解反应得到了证实。分别在批式反应器和ＣＳＴＲ反应器中进行萘普生的酶法拆分，在一定转化率下，批式