Enzyme Membrane Reactor Research Articles

Exploiting UPO versatility to transform rutin in more soluble and bioactive products

The discovery of unspecific peroxygenases (UPOs) completely changed the paradigm of enzyme-based oxyfunctionalization reactions, as these enzymes can transform a wide variety of substrates with a relatively simple reaction mechanism. The fact that UPO can exert both peroxygenative and peroxidative activity in either aromatic or aliphatic carbons, represents a great potential in the production of high value-added products from natural antioxidants. In this work, the flavonoid rutin has been considered as possible substrate for UPO from Agrocybe aegerita, and its peroxygenation or its peroxidation and successive oligomerization have been studied. Different experiments were performed in order to reduce the range of process variables involved and gaining insight on the behavior of this enzyme, leading to a multivariable optimization of UPO-based rutin modification. While trying to preserve enzyme activity this optimization aimed for maximizing the production of more soluble antioxidants. Reusability of the enzyme was evaluated recovering UPO using an enzymatic membrane reactor, revealing challenges in enzyme stability due to inactivation during the filtration stages. The influence of the radical scavenger ascorbic acid on product formation was investigated, revealing its role in directing the reaction towards hydroxylated rutin derivatives, hence indicating a shift towards more soluble and bioactive products.

Modeling and simulation of bi‐continuous jammed emulsion membrane reactors for enhanced biphasic enzymatic reactions

AbstractBi‐continuous jammed emulsion (bijel) membrane reactors, integrating simultaneous reaction and separation, offer a promising avenue for enhancing membrane reactor processes. In this study, we present a comprehensive macroscopic‐scale physicochemical model for tubular bijel membrane reactors and a numerical solution strategy for solving the governing partial differential equations. The model captures the co‐continuous network of two immiscible phases stabilized by nanoparticles at the liquid–liquid interface. We present the derivation of model equations and an efficient numerical solution strategy. The model is validated with experimental results from a conventional enzymatic biphasic membrane reactor for oleuropein hydrolysis, already reported in the literature. Simulation results indicate accurate prediction of reactor behavior, highlighting the potential superiority of bijel membrane reactors over current technologies. This research contributes a valuable tool for scale‐up, design, and optimization of bijel membrane reactors, filling a critical gap in this emerging field.

Opting for polyamines with specific structural traits as a strategy to boost performance of enzymatic membrane reactors

Enzymatic membrane reactor (EMR) is a type of continuous-flow bioreactor offering easy separation and reuse of biocatalyst while giving opportunities to improve its performance. However, simultaneous enhancement of activity and stability of enzyme while retaining high membrane permeability poses a challenge for a single reactor. Herein, we propose a new method to optimize EMR system − by employing cationic polyelectrolytes with selected molecular weight, backbone, and amino group type for modification of membrane surface; after we have evaluated the effect of polyamine chemistry and membrane properties on each aspect of EMR performance. Polydopamine (PDA)-assisted co-deposition of polyamines (polyethyleneimine (PEI), PEI-graft-poly(ethylene glycol) (PEI-g-PEG), poly(allylamine hydrochloride) (PAH), and poly(diallyldimethylammonium chloride) (PDADMAC)) was used as the modification method; the membranes were characterized by water permeability, water contact angle (WCA), zeta potential (ZP), FTIR spectra, and SEM/EDX; and EMRs were assessed by biocatalytic activity at various pH, flux, immobilization yield and efficiency (activity recovery), enzyme leakage, pH/storage and thermal stability, and reusability. In this work, EMRs with immobilized alcohol dehydrogenase (ADH) showed activities up to 8.9 mU/cm2, retained up to 86 % of initial activity in three conversion cycles without any reduction in flux, and allowed to increase non-optimal pH activity (pH 10) by up to 81 % and improve storage stability by up to 53 % compared to the free enzyme. We discovered statistically significant (p < 0.01) relationships between: (1) polyamine backbone type (linear vs branched) and membrane permeability; (2) membrane isoelectric point and immobilization yield; and (3) membrane WCA and immobilization efficiency. To our knowledge, this is the first systematic study of the effect of polyelectrolyte chemistry on performance of immobilized enzyme − which showed the method’s strong potential for customizing properties and maximizing the productivity of EMRs.

Production of Antioxidant Peptides from Snakehead Fish Using Batch and Continuous Enzymatic Hydrolysis

Bioactive peptides are promising functional ingredients. Due to its high protein content, snakehead fish (Channa striata) extract (SHFE) is one of the suitable parent proteins for bioactive peptides. This study aimed to investigate the production of SHFE-based antioxidative peptides in a conventional batch and continuous system facilitated by an enzymatic membrane reactor (EMR). The effects of different proteases (Alcalase, Neutrase), substrate concentrations, and enzyme-to-substrate ratios were investigated in the batch process. Continuous hydrolysis was then performed under the optimum conditions obtained from the batch process. The optimum conditions based on the antioxidant capacity measured by DPPH and FRAP assays were employing Alcalase with a substrate concentration [S] of 3% (w/v) and an enzyme-to-substrate ratio [E]/[S] of 10% (w/w). Continuous operation was shown to have been performed over a prolonged period, based on the calculated fouling rate. Furthermore, filtration of the resulting permeate with a smaller membrane pore size (2-kDa) increased the antioxidant capacity. This study is expected to increase the production of functional ingredients in snakehead fish.

Stabilizing enzymatic membrane reactor for precise production of oligodextran with tailored molecular weight

Enzyme membrane reactor (EMR) provides an effective strategy for controlling molecular weight (Mw) of oligodextrans. However, the synergistic effect between dextran hydrolysis and product separation as well as continuous operation stability remains still unclear. We observed that covalently immobilized dextranase exhibited more obvious exo-hydrolysis behavior compared to free enzymes probably due to enzyme conformational alterations. Using non-covalently immobilized enzyme, suitable ultrafiltration membrane, optimal substrate/enzyme ratio and matched permeate flux, the yield of the target product reached 62.8%. However, the non-covalently immobilized dextranase exhibited reduced hydrolytic activity and more obvious membrane fouling (caused by high-Mw dextran). When free dextranase with high activity was used together with suitable permeate flux and high shear rate, the yield increased to 72.0%. The obtained EMR maintained stable for 8 h with slight fouling formation because enzymatic hydrolysis and membrane separation were well matched. The polydispersity index of the target product reached 1.3 (analytical grade) following diafiltration purification.

Assessment of the efficiency and stability of enzymatic membrane reaction utilizing lipase covalently immobilized on a functionalized hybrid membrane

Enzyme immobilization in membrane bioreactors has been considered as a practical approach to enhance the stability, reusability, and efficiency of enzymes. In this particular study, a new type of hybrid membrane reactor was created through the phase inversion method, utilizing hybrid of graphene oxide nanosheets (GON) and polyether sulfone (PES) in order to covalently immobilize the Candida rugosa lipase (CRL). The surface of hybrid membrane was initially modified by (3-Aminopropyl) triethoxysilane (APTES), before the use of glutaraldehyde (GLU), as a linker, through the imine bonds. The resulted enzymatic hybrid membrane reactors (EHMRs) were then thoroughly analyzed by using field-emission scanning electron microscopy (FE-SEM), contact angle goniometry, surface free energy analysis, X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, attenuated total reflection (ATR), and energy-dispersive X-ray (EDX) spectroscopy. The study also looked into the impact of factors such as initial CRL concentration, storage conditions, and immobilization time on the EHMR's performance and activity, which were subsequently optimized. The results demonstrated that the CRLs covalently immobilized on the EHMRs displayed enhanced pH and thermal stability compared to those physically immobilized or free. These covalently immobilized CRLs could maintain over 60% of their activity even after 6 reaction cycles spanning 50 days. EHMRs are valuable biocatalysts in developing various industrial, environmental, and analytical processes.

Production of Galacto‐Oligosaccharides by Enzymatic Membrane Reactors Using Free β‐Galactosidase

AbstractPrebiotic galacto‐oligosaccharides (GOSs) are synthesized by transgalactosylation of lactose using β‐galactosidases. Well‐established, large‐scale production of GOS is performed by stirred‐tank reactors (STR) operating in batch fashion. Main manufacturing expenses are attributed to enzyme purchase costs and the downstream operations required for subsequent enzyme inactivation and removal. In the last decades, several studies and patents have appeared on free enzyme membrane reactor (FEMR) as an economic alternative to STR due to enhanced enzyme recovery and reuse. In this review, we attempt to summarize the key design aspects, the catalytic performance and efficiency of FEMR compared to STR technology, and the main concerns related to stable product quality over long‐term operation.

Novel membrane coating methods involving use of graphene oxide and polyelectrolytes for development of sustainable energy production: Pressure Retarded Osmosis (PRO) and Enzymatic Membrane Reactor (EMR)

This study compares pressure retarded osmosis (PRO) and the enzymatic membrane reactor (EMR) for the production of green energy in the form of power density and biomethanol, respectively. A systematic design of the biocatalytic membrane reactor and the PRO membrane system was carried out where we combined physical adsorption of polyelectrolyte (PE) and the graphene oxide (GO) layer-by-layer (LbL) assembly system. The hybrid LbL structure is proposed as a strategy to simultaneously advance the operational stability of the enzymes in the EMR and to increase hydrophilicity and power density in the PRO approach. Using polydopamine (PDA), poly(diallyldimethylammonium chloride) (PDADMAC) and GO allowed functionalization of polysulfone (PSF) membranes for subsequent Alcohol Dehydrogenase (ADH) immobilization in the EMR and functionalization of polyamide (PA) membranes for PRO. Tailoring membrane surface chemistry allowed an increase in enzyme conversion rate in comparison to the pristine, unmodified membrane (99.6% vs 2%, respectively) without significantly compromising water permeability. Moreover, power density increased from 2.10 to 2.64 W/m2 for pristine and modified membrane, respectively. Energy production in kJ/m2·h was compared and the most efficient technology was chosen.

Mechanically stirred enzymatic membrane reactor containing HRP immobilized on Tau-SiO2@Fe3O4-GO nanocomposite for removal of tetracycline in synthetically concocted wastewater.

The process of mechanically stirred membrane reactor containing the suspension of horseradish peroxidase (HRP) immobilized on synthesized nanocomposite (Tau-SiO2@Fe3O4-GO) was designed for continuous degradation of tetracycline. The immobilized HRP was characterized in terms of kinetic parameters and catalytic activities as these parameters were improved highly through immobilization. The stability indices including pH and temperature were investigated in parallel. The immobilized HRP was more tolerable to pH changes as compared to free HRP and the optimum temperature obtained at 40°C. The reusability of HRP was promoted by immobilization as far as 70% of initial activity after ten cycles. The enzymatic degradation of optimum concentration of tetracycline was carried out in batch condition and 100% of tetracycline removed after 30min. The results also showed that higher concentration of H2O2 exhibited more oxidation of tetracycline. The optimal ratio of HRP/H2O2 was also obtained at 0.005. The simultaneous process including separation and the biocatalytic degradation established in the membrane stirrer reactor concluded that no amount of tetracycline was observed in the permeate stream coming from the membrane after 30min of operation.

Bioactive Peptides from Velvet Bean Tempe: Neutrase-Catalyzed Production in Membrane Reactor

Velvet beans are potential sources of parent proteins for bioactive peptide production. In this study, a combination of fermentation and neutrase-catalyzed continuous hydrolysis in an enzymatic membrane reactor was performed to produce antioxidative and angiotensin I-converting enzyme inhibitory (ACEi) peptides. The optimum operating conditions were τ = 6 h and [E]/[S] = 7.5%. The resulting permeate, which was a&lt;10-kDa fraction, exhibited antioxidant activity at 0.38 mg ascorbic acid equivalent antioxidant capacity (AEAC)/mL (2,2-diphenyl-1-picrylhydrazyl, DPPH inhibition) and 0.26 mg AEAC/mL (ferric reducing antioxidant power, FRAP), and ACEi activity of 81.02%. Further fractionation of the permeate increased the ACEi activity in which 2-kDa fraction showed the most potent activity (IC50 = 0.23 µg protein/mL). The IC50 value of the outcome was comparable to those reported in the literature for velvet bean-based peptides. Furthermore, this study suggests that neutrase is a good catalyst candidate for the synthesis of bioactive peptides from velvet beans.

Toward Renewable-Based Prebiotics from Woody Biomass: Potential of Tailored Xylo-Oligosaccharides Obtained by Enzymatic Hydrolysis of Beechwood Xylan as a Prebiotic Feed Supplement for Young Broilers.

Although antibiotic resistance emerges naturally, this process has been accelerated by the worldwide overuse and misuse of antibiotics. It is essential to find effective alternatives in the broiler industry to improve poultry health while maintaining production efficiency and product safety. In this study, we aimed to evaluate a potential alternative: wood-derived xylo-oligosaccharides (XOS). The objective of this research was to investigate the potential of XOS prepared using enzymatic hydrolysis of beechwood xylan as a prebiotic feed supplement for broilers. A pilot study was conducted to explore the optimal XOS fraction profile by in vitro fermentation. Subsequently, a semi-continuous enzyme membrane reactor was used, allowing for the production of tailored XOS in large quantities. Given the strong bidirectional relationship between intestinal health, nutrition, and intestinal microbiota composition in broilers, an in vivo experiment was performed to explore the potential of XOS as a prebiotic feed supplement by investigating growth performance, feed conversion ratio, caecal short and medium chain fatty acid (SCFA and MCFA) concentration, and microbiological composition of the caecal content. Results from the pilot study indicated that higher enzyme concentrations in the hydrolysis process yield a product that leads to a higher total SCFA and MCFA- and butyric acid production during in vitro fermentation by caecal bacteria. Supplementation of the tailored XOS to the broiler diet (day 1 (d1)-d8 0.13% wt/wt XOS, d9-d15 0.32% XOS) resulted in higher Bifidobacterium counts, beneficial to the health of birds, on d11 and d15.

Design of a novel multi-layer enzyme membrane reactor for low-fouling, tailored production of oligodextran

Enzymatic conversion processes face challenges in controlling oligosaccharide molecular weight (Mw). Enzymatic membrane reactors (EMRs) with immobilized enzymes address this, but direct enzyme immobilization on the membrane surface can lead to deactivation and reduced hydrolysis efficiency. This study proposes a novel EMR configuration: a three-layer structure. An electrospun porous fibrous layer, modified with PDA, TA, and APTES, serves as a mechanical support layer. A commercial separation membrane is positioned below. This configuration enhances enzyme activity and selectivity. Using a “fouling-induced” technique, immobilized activity of the enzyme (i.e. dextranase) significantly increased to 11.5 µmol-isomaltose/min, surpassing incubation-immobilized dextranase (0.075 µmol-isomaltose/min). The additional layer preserves catalytic patterns, reducing fouling and ensuring high selectivity. The EMR configuration excels in producing low Mw oligosaccharides. The catalytic layer achieves 11.3 µmol-isomaltose/min, while the membrane exhibits exceptional selectivity and stability. The hydrophilic RC10 membrane with small pores performs best. This study highlights the potential of the EMR system for efficient production of stable low Mw oligosaccharides. Insights into optimizing enzyme immobilization strategies and membrane selection benefit enzymatic conversion processes.

Enhancement of biocatalytic activity in enzymatic membrane reactors: Controlled modification with novel PAH/PDA composites as a tool to optimize reactor performance

Enzymatic membrane (EM) is a tool allowing to simultaneously upgrade enzymatic catalysis and membrane separation processes. However, their commercial use is often restricted by a trade-off between catalytic and mass transfer properties of the system. Herein, we report a new approach to the design of enzymatic membrane reactors (EMRs) - aimed to find the optimal balance between biocatalytic activity, water permeability, and enzyme loading; where we used co-deposited polyallylamine hydrochloride/polydopamine (PAH/PDA) membrane coatings as a tool to control the overall reactor performance. First, we demonstrate that tailoring the PAH/PDA coating chemistry can be more effective for promotion of total biocatalytic activity of EM with immobilized alcohol dehydrogenase (ADH) than increasing the amount of immobilized enzyme on a membrane support. Then, we show how the PAH/PDA ratio (0, 1:10, 1:2, 5:2), modification time (0.25, 1, 4, 16 h) and ionic strength (0.1–1.0 M NaCl) affect morphology of the membrane coating and alter the mechanisms of ADH-support interactions. Lastly, we illustrate the versatility of the PAH/PDA-coated membranes for application in other enzymatic systems on the example of glycerol dehydrogenase (GDH) and evaluate the performance of two EMRs as a function of applied pressure (1–3 bar). The final values of biocatalytic activities 25.4 ± 4.0 and 3.5 ± 0.3 mU/cm2 (which correspond to 140 ± 22 and 67 ± 5% immobilization efficiencies) for immobilized ADH and GDH, respectively, demonstrate that PAH/PDA-coated membranes not only can be used for simultaneous reaction and recovery of the biocatalyst, but also for enhancement of enzymatic activity - due to opportunity of reaction rate control in the membrane reactor.

Detoxification and decolorization of complex textile effluent in an enzyme membrane reactor: batch and continuous studies.

There is an urgent need to look for bio-based technologies to address the pollution related to textile dyes in waterbodies. The aim of this study was to evaluate an engineered laccase variant, LCC1-62 of Cyathus bulleri, expressed in recombinant Pichia pastoris, for the decolorization and detoxification of real textile effluent. The partially purified laccase effectively (~60-100%) decolorized combined effluent from different dyeing units at a laccase concentration of 500 U/L at a 50-mL level. Decolorization and detoxification of the combined effluents, from a local textile mill, were evaluated at 0.3 L volumetric level in a ray-flow membrane reactor in batch and continuous modes of operation. In batch studies, maximum decolorization of 97% and detoxification of 96% occurred at a hydraulic retention time (HRT) of 6 h without any additional laccase requirement. In continuous studies, the reactor was operated at an HRT of 6 h with a lower enzyme dosage (~120 U/L of the effluent). Decolorization was accompanied by a loss in laccase activity which was restored to ~120 U/L by the addition of laccase in two regimes. The addition of laccase, when the residual laccase activity decreased to 40% (~50 U/L), resulted in high decolorization (~5 ppm residual dye concentration) and low variance (σ2) of 2.77, while laccase addition, when the residual dye concentration decreased to ~8% (~10 U/L), resulted in an average dye concentration of 13 ppm with a high variance of 62.08. The first regime was implemented, and the continuous reactor was operated for over 80 h at an HRT of 3 and 6 h, with the latter resulting in ~95% decolorization and 96% reduction in the mutagenicity of the effluent. Less than 10% membrane fouling was observed over long operations of the reactor. The findings strongly suggest the feasibility of using LCC1-62 in an enzyme membrane reactor for large-scale treatment of textile effluents.

Synthesis of butyl cinnamate using enzymatic membrane reactor in a free solvent system

Cinnamic acid is a phenolic compound that has the potential to act as a natural antioxidant. Cinnamic acid esterification can be performed by adding alcohol as an alkyl group donor. This esterification is carried out to increase the antioxidant capacity of cinnamic acid. Esterification of cinnamic acid with butanol as the alkyl group donor was the best condition to modify cinnamic acid. In this study, the synthesis of butyl cinnamate using a enzymatic membrane reactor (EMR) is carried out continuously with Lipozyme TL IM as the catalyst. This study aimed to determine the optimal conditions for the synthesis of butyl cinnamate using various concentrations of cinnamic acid, biocatalyst, residence time, and the presence of molecular sieves. The synthesis of butyl cinnamate at 40°C was optimum when molecular sieves were present with 0.01 M cinnamic acid concentration, 1% (w/v) biocatalyst, and 12 h of residence time.

Continuous production of velvet bean-based bioactive peptides in membrane reactor with dual enzyme system

One of the main challenges hindering the commercialization of bioactive peptides is the lack of scalable and consistent production methods. To overcome this obstacle, an automated enzyme membrane reactor was used to continuously produce bioactive peptides from velvet bean (Mucuna pruriens). The optimum operating conditions were [E]/[S] = 5%, pH = 7.5, and τ = 12 h. The long-term continuous operation of the EMR system demonstrated its ability to maintain steady-state conditions. To minimize membrane fouling, an industrially viable strategy was employed, which combines operation at threshold flux and performing regular membrane cleaning. Further fractionation of the hydrolysates with a 2-kDa PES membrane resulted in the highest bioactivity. The IC50 values for antioxidant and ACE inhibition were 17.85 and 4.58 µg protein/mL, respectively. To map the overall bioactivities of the hydrolysates, LC-MS analysis coupled with BIOPEP-UWM database was performed and obtained DPP-4 and ACE inhibitors as the predominant bioactive activities.

Experimental study on the continuous production of velvet bean-based bioactive peptides in a membrane reactor and bioactivity mapping

This study investigates the continuous production of bioactive peptides from fermented velvet bean (Mucuna pruriens) using a cost-effective enzymatic membrane reactor. The optimal operating conditions for continuous hydrolysis were residence time of 9 h, enzyme-to-substrate ratio of 10%, and the use of a 5-kDa PES membrane for separation. The resulting permeate exhibited notable antioxidant activity with DPPH and FRAP assay values of 0.28 and 0.12 mg AEAC/mL, respectively, and an angiotensin-converting enzyme (ACE) inhibition of 83.28%. Further fractionation of the permeate with a 2-kDa membrane led to increased antioxidant activity and ACE inhibition, with resulting half maximal inhibitory concentration (IC50) values of 7.6 and 0.6 μg protein/mL, respectively. Liquid chromatography-mass spectrometry was used to identify peptide sequences from the <2-kDa fraction, which were matched with the BIOPEP-UWM™ database. The results revealed that the fermented velvet bean peptides were predominantly dipeptidyl peptidase (DPP)-4 inhibitors and ACE inhibitors. Industrial relevanceThe combination of a stirred tank reactor and membrane separation has the potential to enable the continuous production of bioactive peptides from velvet bean through the use of a freely suspended enzyme system. This reactor design offers an alternative solution to the existing delay in commercializing bioactive peptides, by (i) simplifying the industrial process development for continuous bioactive peptide production, (ii) eliminating the need for enzyme immobilization, and (iii) eliminating batch-to-batch variability in product quality and unproductive time associated with batch-wise production.

Production of Modified Nucleosides in a Continuous Enzyme Membrane Reactor

Nucleoside analogues are important compounds for the treatment of viral infections or cancers. While (chemo-)enzymatic synthesis is a valuable alternative to traditional chemical methods, the feasibility of such processes is lowered by the high production cost of the biocatalyst. As continuous enzyme membrane reactors (EMR) allow the use of biocatalysts until their full inactivation, they offer a valuable alternative to batch enzymatic reactions with freely dissolved enzymes. In EMRs, the enzymes are retained in the reactor by a suitable membrane. Immobilization on carrier materials, and the associated losses in enzyme activity, can thus be avoided. Therefore, we validated the applicability of EMRs for the synthesis of natural and dihalogenated nucleosides, using one-pot transglycosylation reactions. Over a period of 55 days, 2'-deoxyadenosine was produced continuously, with a product yield >90%. The dihalogenated nucleoside analogues 2,6-dichloropurine-2'-deoxyribonucleoside and 6-chloro-2-fluoro-2'-deoxyribonucleoside were also produced, with high conversion, but for shorter operation times, of 14 and 5.5 days, respectively. The EMR performed with specific productivities comparable to batch reactions. However, in the EMR, 220, 40, and 9 times more product per enzymatic unit was produced, for 2'-deoxyadenosine, 2,6-dichloropurine-2'-deoxyribonucleoside, and 6-chloro-2-fluoro-2'-deoxyribonucleoside, respectively. The application of the EMR using freely dissolved enzymes, facilitates a continuous process with integrated biocatalyst separation, which reduces the overall cost of the biocatalyst and enhances the downstream processing of nucleoside production.

Continuous synthesis of cinnamate ester using membrane reactor in a solvent-free system

Cinnamic acid is a phenolic compound with the potential to act as a natural antioxidant. Cinnamic acid esterification can be performed by adding a long-chain alcohol to one of its hydroxyl groups to improve its antioxidant activity. In this study, cinnamic esters were synthesized in an enzymatic membrane reactor (EMR) and were carried out continuously. This study aimed to determine the optimum conditions for the synthesis of cinnamic esters using various types of alcohols (e.g., butanol, hexanol, and octanol) as alkyl group donors and the impact of the esterification on the antioxidant activity. The continuous synthesis of cinnamic esters with TL IM lipozyme as the biocatalyst had the highest conversion (20.91%) with 1-butanol as the alkyl donor, in a synthesis strategy of a 12-h batch followed by continuous operation with a residence time of 9 h. The resulting ester increased the antioxidant activities by 2.18 and 3.85-fold, respectively based on the DPPH and FRAP assays.

Potential of Integrated Semi‐Continuous Enzymatic Synthesis and Filtration Processes for Efficiency Enhancement

AbstractGalacto‐oligosaccharides (GOS), increasingly popular prebiotics, are synthesized by enzymatic conversion of lactose. Among others, the total production costs are significantly influenced by the costly enzyme. Therefore, it was investigated if the reuse and full recovery of the enzyme is feasible, followed by the development of a semi‐continuous process in order to maintain a consistent high GOS yield. As a preliminary step, the enzyme‐catalyzed reaction was recorded within the permissible operating parameters. It was successfully shown that steady high GOS yields can be synthesized semi‐continuously within a filtration plant functioning as an enzymatic membrane reactor.