Free Enzyme System Research Articles

Dual-enhanced enzyme cascade hybrid hydrogel for the construction of optical biosensor

The biomimetic enzyme cascade system plays a key role in biosensing as a sophisticated signal transduction and amplification strategy. However, constructing a regulated enzyme cascade sensing system remains challenging due to the mismatch of multiple enzyme activities and poor stability. Herein, we design an efficient dual-enhanced enzyme cascade hybrid system (UFD-DEC) containing DNA-controlled nanozymes (Fe-cdDNA) and enzyme (urease) via combining the electrostatic contact effect with the hydrogel-directed confinement effect. Precise modulation of Fe-cdDNA nanozyme by DNA offers a means to control its catalytic efficiency. This regulated UFD-DEC system accelerates the reaction rate and provides remarkable stability compared with the free enzyme system. Benefiting from the plasticity properties of hydrogels, a “lab-in-a-tube” platform was constructed by encapsulating UFD-DEC in a microcentrifuge tube. Such a UFD-DEC-based hydrogel tube exhibits sufficient adaptability to profile urea when used in conjunction with a smartphone-assisted image processing algorithm, which on-site delivers urea information with a detection limit of 0.12 mmol L−1. This customizable and inexpensive miniaturized biosensor platform for monitoring urea may facilitate point-of-care testing applications.

Multi-point immobilization of GH 11 endo-β-1,4-xylanase on magnetic MOF composites for higher yield of xylo-oligosaccharides

GH 11 endo-β-1,4-xylanase (Xy) was a crucial enzyme for xylooligosaccharides (XOS) production. The lower reusability and higher cost of purification has limited the industrial application of Xy. Addressing these challenges, our study utilized various immobilization techniques, different supports and forces for Xy immobilization. This study presents a new method in the development of Fe3O4@PDA@MOF-Xy which is immobilized via multi-point interaction forces, demonstrating a significant advancement in protein loading capacity (80.67 mg/g), and exhibiting remarkable tolerance to acidic and alkaline conditions. This method significantly improved Xy reusability and efficiency for industrial applications, maintaining 60 % activity over 10 cycles. Approximately 23 % XOS production was achieved by Fe3O4@PDA@MOF-Xy. Moreover, the yield of XOS from cobcorn xylan using this system was 1.15 times higher than that of the free enzyme system. These results provide a theoretical and applicative basis for enzyme immobilization and XOS industrial production.

In-CdS@ZIF-8&FDH photo-enzyme nanosystem with high NADH regeneration ability via indium doping for enhanced CO2 reduction to formic acid

Replacement of fossil fuel by renewable resources is a central resource and energy efficiency strategy, where CO2 is the key molecule to proceed effectively in this direction. Here, we synthesized In-doped CdS and constructed a new photo-enzyme nanosystem 10 %In-CdS@ZIF-8&FDH, which can catalyze CO2 to formic acid (HCOOH) via formate dehydrogenase immobilized on ZIF-8 while photocatalytic regenerating NADH via indium-doped CdS. Indium doping greatly enhances the photocatalytic performance of CdS. The NADH regeneration rate reaches 94.10 %. The nanosystem can produce 267.18 μM HCOOH, which is 4.27 times that of the free enzyme system. And 85.98 % of the residual activity can be retained after reusing 8 times. This study will inspire more research efforts for light-driven enzymatic nanosystems in CO2 fixation and lay the foundation for realizing renewable energy-driven CO2 conversion to produce chemical products.

Advancements in wearable technology for monitoring lactate levels using lactate oxidase enzyme and free enzyme as analytical approaches: A review

Lactate is a metabolite that holds significant importance in human healthcare, biotechnology, and the food industry. The need for lactate monitoring has led to the development of various devices for measuring lactate concentration. Traditional laboratory methods, which involve extracting blood samples through invasive techniques such as needles, are costly, time-consuming, and require in-person sampling. To overcome these limitations, new technologies for lactate monitoring have emerged. Wearable biosensors are a promising approach that offers non-invasiveness, low cost, and short response times. They can be easily attached to the skin and provide continuous monitoring. In this review, we evaluate different types of wearable biosensors for lactate monitoring using lactate oxidase enzyme as biological recognition element and free enzyme systems.

Dendrobium nobile Lindl. Polysaccharides protect fibroblasts against UVA-induced photoaging via JNK/c-Jun/MMPs pathway

Ethnopharmacological relevanceDendrobium nobile Lindl. is an orchid species that is found throughout Asia, including Thailand, Laos, Vietnam, and China. It has been used to treat tumors, hyperglycemia, hyperlipidemia, and neurological disorders caused by aging in recent decades. Aim of the studyTo investigate the antagonistic effect of Dendrobium nobile Lindl. Polysaccharides (DNLP) on UVA-induced photoaging of Human foreskin fibroblasts (HFF-1) and explore its possible anti-aging mechanisms. Materials and methodsAn in vitro photoaging model of dermal fibroblasts was established with multiple UVA irradiations. Fibroblasts were treated with 0.06 mg/ml, 0.18 mg/ml, 0.54 mg/ml of DNLP one day before photodamage induction. The levels of reactive oxygen species (ROS), Malondialdehyde (MDA), cell viability and longevity, Superoxide Dismutase (SOD), Catalase (CAT), and Glutathione peroxidase (GSH-Px) enzymatic activities were determined. We examined how DNLP ameliorates the effects of photoaging, the JNK/c-Fos/c-Jun pathway, senescence-associated β-galactosidase (SA-β-Gal), and MMP expression levels were measured. ResultsUVA irradiation reduced the viability, lifespan, and proliferation of HFF-1 cells, increased ROS and lipid peroxidation and decreased the activities of free radical scavenging enzyme systems SOD, CAT, and GSH-Px. DNLP treatment can reverse UVA damage, reduce SA-β-Gal expression, reduce phosphorylation activation of the JNK/c-Fos/c-Jun pathway and inhibit MMP-1, MMP-2 MMP-3, and MMP-9 protein expression. ConclusionsDNLP can effectively inhibit UVA damage to HFF-1 and prevent cell senescence. Its mechanism of action may increase antioxidant enzyme activity while inhibiting JNK pathway activation and MMPs expression.

Hierarchically porous metal organic framework immobilized formate dehydrogenase for enzyme electrocatalytic CO2 reduction

The utilization of CO2 has become an important issue aiming to the achievement of carbon neutrality. The enzyme electrocatalytic CO2 reduction to produce carbonaceous fuels has been increasingly appealing. While the low CO2 aqueous solubility severely limits the CO2 reduction efficiency, and the expensive cofactor restricts the application potential. In this study, microporous UiO-66-NH2 was firstly converted into hierarchically porous structure (HP-UiO-66-NH2) containing both micropores and mesopores to simultaneously enhance the enrichment of CO2 and the immobilization of formate dehydrogenase by optimizing the pore structure. The relative contents of the protein secondary structures were similar after immobilization. The mechanism of enzyme electrocatalytic CO2 reduction was proposed. The enzyme stability was highly enhanced after immobilization and the CO2 reduction was enhanced by the enzyme electrocatalytic system through the combination of CO2 enrichment and electrocatalytic NADH regeneration. The optimized catalytic system can obtain formate yield of 1.826 mM in 3 h (5.57 times higher than the free enzyme system) with a generation rate of 6086.7 μmol gcat‑1h‑1.

GOx/Hb Cascade Oxidized Crosslinking of Silk Fibroin for Tissue-Responsive Wound Repair.

Promising wound dressings can achieve rapid soft-tissue filling while refactoring the biochemical and biophysical microenvironment to recruit endogenous cells, facilitating tissue healing, integration, and regeneration. In this study, a tissue biomolecule-responsive hydrogel matrix, employing natural silk fibroin (SF) as a functional biopolymer and haemoglobin (Hb) as a peroxidase-like biocatalyst, was fabricated through cascade enzymatic crosslinking. The hydrogels possessed mechanical tunability and displayed adjustable gelation times. A tyrosine unit on SF stabilised the structure of Hb during the cascade oxidation process; thus, the immobilized Hb in SF hydrogels exhibited higher biocatalytic efficiency than the free enzyme system, which provided a continuously antioxidative system. The regulation of the dual enzyme ratio endowed the hydrogels with favourable biocompatibility, biodegradability, and adhesion strength. These multifunctional hydrogels provided a three-dimensional porous extracellular matrix-like microenvironment for promoting cell adhesion and proliferation. A rat model with a full-thickness skin defect revealed accelerated wound regeneration via collagen deposition, re-epithelialisation and revascularisation. Enzyme-loaded hydrogels are an attractive and high-safety biofilling material with the potential for wound healing, tissue regeneration, and haemostasis.

Potential and Scale-Up of Pore-Through-Flow Membrane Reactors for the Production of Prebiotic Galacto-Oligosaccharides with Immobilized β-Galactosidase

The production of prebiotics like galacto-oligosaccharides (GOS) on industrial scale is becoming more important due to increased demand. GOS are synthesized in batch reactors from bovine lactose using the cost intensive enzyme β-galactosidase (β-gal). Thus, the development of sustainable and more efficient production strategies, like enzyme immobilization in membrane reactors are a promising option. Activated methacrylatic monoliths were characterized as support for covalent immobilized β-gal to produce GOS. The macroporous monoliths act as immobilized pore-through-flow membrane reactors (PTFR) and reduce the influence of mass-transfer limitations by a dominating convective pore flow. Monolithic designs in the form of disks (0.34 mL) and for scale-up cylindric columns (1, 8 and 80 mL) in three different reactor operation configurations (semi-continuous, continuous and continuous with recirculation) were studied experimentally and compared to the free enzyme system. Kinetic data, immobilization efficiency, space-time-yield and long-term stability were determined for the immobilized enzyme. Furthermore, simulation studies were conducted to identify optimal operation conditions for further scale-up. Thus, the GOS yield could be increased by up to 60% in the immobilized PTFRs in semi-continuous operation compared to the free enzyme system. The enzyme activity and long-time stability was studied for more than nine months of intensive use.

Effects of Bacillus velezensis zk1 on the physiology and metabolism of peaches

SummaryBacillus velezensis zk1 is the dominant bacterium that causing rot in peaches. However, the mechanisms through that this bacterium causes rot have not been elucidated. Here, we explored the mechanisms of peach decay caused by B. velezensis zk1. The invasion of B. velezensis zk1 in peaches resulted in an increase in glucose and arabinose contents in fruit tissues. Moreover, the relative conductivity of the fruit reached 84% after 4 days of culture with bacterial invasion. With the destruction of cells, the malondialdehyde content increased, whereas the vitamin C, dialdehyde, flavonoid and total phenol contents decreased. Polyphenol oxidase, superoxide dismutase, catalase, peroxidase, ascorbate peroxidase and ammonia lyase activities also decreased. Overall, these findings demonstrated that B. velezensis zk1 infection damaged peach chloroplasts, mitochondria, respiratory chain activity and related free radical scavenging enzyme systems, thereby disrupting the normal physiological metabolism of peaches and causing rot.

Au-Au/IrO2@Cu(PABA) Reactor with Tandem Enzyme-Mimicking Catalytic Activity for Organic Dye Degradation and Antibacterial Application.

Herein, a Au-Au/IrO2 nanocomposite with tandem enzyme-mimicking activity was innovatively synthesized, which can show outstanding glucose oxidase (GOx)-like activity and peroxidase-like activity simultaneously under neutral conditions. Moreover, a Au-Au/IrO2@Cu(PABA) reactor was prepared via encapsulation of the Au-Au/IrO2 nanocomposite in a Cu(PABA) metal organic framework. The reactor not only exhibits excellent organic solvent stability, acid resistance, and reusability but also displays better cascade reaction catalytic efficiency (kcat/Km = 148.86 min-1 mM-1) than the natural free enzyme system (GOx/HRP) (kcat/Km = 98.20 min-1 mM-1) and Au-Au/IrO2 nanocomposite (kcat/Km = 135.24 min-1 mM-1). In addition, it is found that the reactor can catalyze glucose or dissolved oxygen to produce active oxygen species (ROS) including HO, 1O2, and O2-· through its enzyme-mimicking activity. Finally, the novel reactor was successfully used in organic dye degradation and antibacterial application. The results show that it can effectively degrade methyl orange, methylene blue, and rhodamine B, which all can reach a degradation rate of nearly 100% after interacting with Au-Au/IrO2@Cu (PABA) for 3.5 h. Furthermore, the reactor also exhibits excellent antibacterial activity, so as to achieve a complete bactericidal effect to Staphylococcus aureus and Escherichia coli at a concentration of 12.5 μg mL-1.

Co-immobilization of an Enzyme System on a Metal-Organic Framework to Produce a More Effective Biocatalyst

In many respects, enzymes offer advantages over traditional chemical processes due to their decreased energy requirements for function and inherent greener processing. However, significant barriers exist for the utilization of enzymes in industrial processes due to their limited stabilities and inability to operate over larger temperature and pH ranges. Immobilization of enzymes onto solid supports has gained attention as an alternative to traditional chemical processes due to enhanced enzymatic performance and stability. This study demonstrates the co-immobilization of glucose oxidase (GOx) and horseradish peroxidase (HRP) as an enzyme system on Metal-Organic Frameworks (MOFs), UiO-66 and UiO-66-NH2, that produces a more effective biocatalyst as shown by the oxidation of pyrogallol. The two MOFs utilized as solid supports for immobilization were chosen to investigate how modifications of the MOF linker affect stability at the enzyme/MOF interface and subsequent activity of the enzyme system. The enzymes work in concert with activation of HRP through the addition of glucose as a substrate for GOx. Enzyme immobilization and leaching studies showed HRP/GOx@UiO-66-NH2 immobilized 6% more than HRP/GOx@UiO-66, and leached only 36% of the immobilized enzymes over three days in the solution. The enzyme/MOF composites also showed increased enzyme activity in comparison with the free enzyme system: the composite HRP/GOx@UiO-66-NH2 displayed 189 U/mg activity and HRP/GOx@UiO-66 showed 143 U/mg while the free enzyme showed 100 U/mg enzyme activity. This increase in stability and activity is due to the amine group of the MOF linker in HRP/GOx@UiO-66-NH2 enhancing electrostatic interactions at the enzyme/MOF interface, thereby producing the most stable biocatalyst material in solution. The HRP/GOx@UiO-66-NH2 also showed long-term stability in the solid state for over a month at room temperature.

Effect of Xylanase Immobilisation Conditions by Combination of Entrapment and Covalent Binding on Alginate Beads

The immobilisation of enzymes offer improvement in enzyme stability and characteristics as well as overcome the limitations of free enzyme systems for commercial purposes. In the current study, xylanase was immobilised using a combination technique of entrapment and covalent binding within and onto calcium alginate beads. The sodium alginate and calcium chloride (CaCl2) concentration used for the preparation of alginate beads which is the support matrix for xylanase immobilisation were fixed at 3% (w/v) and 0.3 M, respectively. The effect of immobilisation conditions (agitation rate, enzyme loading, and glutaraldehyde concentration) were studied using One-Factor-At-a-Time (OFAT) approach. The best condition for optimum immobilisation yield (83.93%) was found to be made up of the following parameter combination: agitation rate, 200 rpm; xylanase loading, 200 U; and glutaraldehyde concentration, 12% (w/w). The study shows the immobilisation conditions play a significant role towards the immobilisation yield of xylanase.

Impact of Murraya koenigii (L.) Sprengel (Rutaceae) extracts on lifespan and oxidative stress markers in Drosophila melanogaster Meigen, 1830 (Diptera)

The free radicals (ROS and RNS) damage to proteins, DNA, lipids of the cell. These free radicals creates the imbalance in physiological functions and acts as a prevalent cause of various diseases such as cancer, diabetes, cardiovascular diseases, aging, oxidative stress and metabolic syndrome by dysfunction of antioxidant enzyme system of cell. Using the fruit fly Drosophila melanogaster Meigen, 1830 (Diptera) as a model we examined the antioxidant properties of Murraya koenigii (L.) Sprengel (Rutaceae) on the life history parameters. We demonstrate a novel physiological interaction between free radicals, oxidative stress and antioxidant enzyme system by using extracts of M. koenigii in standard diet of the fly. This study describes how this interaction impacts a very early cellular defect associated with ageing and ageing associate diseases. We also describe progressive deficits in flies expressing the superoxide dismutase gene, catalase and lipid peroxidation. Collectively, our work demonstrates that Drosophila can be used to study the cellular, physiological and behavioral basis of human ageing related diseases.

Combined cross-linked enzyme aggregates of glycerol dehydrogenase and NADH oxidase for high efficiency in situ NAD+ regeneration

Cofactor regeneration is an important method to avoid the consumption of large quantities of oxidized cofactor NAD+ in enzyme-catalyzed reactions. Herein, glycerol dehydrogenase (GDH) and NADH oxidase preparations by aggregating enzymes with ammonium sulphate followed by cross-linking formed aggregates for effective regeneration of NAD+. After optimization, the activity of combi-CLEAs and separate CLEAs mixtures were 950 and 580 U/g, respectively. And the catalytic stability of combi-CLEAs against pH and temperature was superior to the free enzyme mixture. After ten cycles of reuse, the catalytic efficiency could still retain 63.3% of its initial activity, indicating that the constructed combi-CLEAs system had excellent reusability. Also, the conversion of glycerol to 1,3-dihydroxyacetone (DHA) was improved by the constructed NAD+ regeneration system, resulting in 4.6%, which was 2.5 times of the free enzyme system. Thus, wide applications of this co-immobilization method in the production of various chiral chemicals could be expected in the industry for its high efficiency at a low cost.

Construction of ordered multienzyme systems using multifunctional polymer brush-grafted magnetic nanoparticles as scaffolds

A block copolymer brush based on functional monomers (N-succinimidyl acrylate and N-adamantylacrylamide) successfully grew on the surface of Fe3O4@SiO2 nanoparticles by surface-initiated reversible addition-fragmentation chain transfer polymerization (SI-RAFT). According to the multiple recognition mechanisms of this material, horseradish peroxidase (HRP) and β-cyclodextrin-modified glucose oxidase (GOx-CD) were constructed into a multienzyme system with an ordered distribution for the cascade reaction, where multifunctional polymer brush-grafted magnetic nanoparticles were used as the scaffolds. The catalytic activity of GOx-CD was enhanced compared with free GOx. In addition to being easily recycled, this multienzyme system exhibited an outstanding conditional stability compared with the free enzyme system. After being reused 8 times, the activity of multienzyme system retained 67.21% of the initial catalytic activity. This strategy provides a novel idea for reusable ordered enzyme immobilization.

Bifunctional Hybrid Enzyme-Catalytic Metal Organic Framework Reactors for α-Glucosidase Inhibitor Screening.

The screening strategy based on α-glucosidase inhibition has been widely employed for the discovery of antidiabetic drugs, but it still faces some challenges in practical applications, such as poor stability of enzyme, high consumption of test compounds, low sensitivity of screening methods and so on. In this work, a bifunctional hybrid enzyme-catalytic metal organic framework reactor (GAA@GOx@Cu-MOF) with a flower-shaped globular structure was innovatively prepared via self-assembling of α-glucosidase (GAA), glucose oxidase (GOx), Cu2+, and 4,4'-bipyridine. It was found that GAA@GOx@Cu-MOF not only enjoyed merits of high stability, selectivity, and sensitivity but also possessed the character of assembly line work, with about 4.58 times enhanced enzyme activity compared with the free enzyme system. Based on the above characteristics, a highly sensitive screening of GAA inhibitors could be achieved with the detection limit of 7.05 nM for acarbose. Furthermore, the proposed method was successfully applied to the screening of oleanolic acid derivatives as potential antidiabetic drugs. Therefore, it was expected that this work could provide new insights and inspirations for the screening of clinical antidiabetic drugs and for further exploration of functional MOF composites.

Comparison of immobilized and free enzyme systems in industrial production of short‐chain fructooligosaccharides from sucrose using a techno‐economic approach

AbstractShort‐chain fructooligosaccharides (scFOS) are nutraceuticals with numerous applications in the food and pharmaceutical industries. The production of scFOS using immobilized biocatalysts offers some functional and technical advantages over free enzyme counterparts. To investigate the economic potential of the immobilized enzyme system relative to the free enzyme system, a techno‐economic comparison was conducted on three methods of scFOS production (powder and syrup forms) at a capacity of 2000 t per annum (tpa) by enzymatic synthesis from sucrose: the free enzyme (FE), calcium alginate immobilized enzyme (CAIE), and amberlite IRA 900 immobilized enzyme (AIE) systems. These processes were simulated in Aspen Plus to obtain the mass and energy balances and to estimate the operating and capital costs, followed by economic evaluation and sensitivity analysis. Profitability analysis showed that all three systems are economically viable as their associated minimum selling prices (MSP) were well below the scFOS market price of 5 $ kg−1. However, the FE system was the most profitable with the lowest MSP of 2.61 $ kg−1 because the savings on cost as a result of enzyme immobilization could not offset the additional costs associated with immobilization. Sensitivity analysis demonstrated that total operating cost, fixed capital investment, and internal rate of return (% IRR) have the greatest effects on the MSP. Furthermore, the syrup form of scFOS production leads to 29% less MSP, compared to powder form. In addition, the studied plant capacities of 5000 and 1000 tpa showed 10% and 16% reductions on MSP respectively. © 2019 Society of Chemical Industry and John Wiley & Sons, Ltd

Bioconversion of xylose to xylonic acid via co-immobilized dehydrogenases for conjunct cofactor regeneration

Enzymatic cofactor-dependent conversion of monosaccharides can be used in the bioproduction of value-added compounds. In this study, we demonstrate co-immobilization of xylose dehydrogenase (XDH, EC 1.1.1.175) and alcohol dehydrogenase (ADH, EC 1.1.1.1) using magnetite-silica core-shell particles for simultaneous conversion of xylose into xylonic acid (XA) and in situ cofactor regeneration. The reaction conditions were optimized by factorial design, and were found to be: XDH:ADH ratio 2:1, temperature 25 °C, pH 7, and process duration 60 min. Under these conditions enzymatic production of xylonic acid exceeded 4.1 mM and was more than 25% higher than in the case of a free enzymes system. Moreover, the pH and temperature tolerance as well as the thermo- and storage stability of the co-immobilized enzymes were significantly enhanced. Co-immobilized XDH and ADH make it possible to obtain higher xylonic acid concentration over broad ranges of pH (6–8) and temperature (15–35 °C) as compared to free enzymes, and retained over 60% of their initial activity after 20 days of storage. In addition, the half-life of the co-immobilized system was 4.5 times longer, and the inactivation constant (kD = 0.0141 1/min) four times smaller, than those of the free biocatalysts (kD = 0.0046 1/min). Furthermore, after five reaction cycles, immobilized XDH and ADH retained over 65% of their initial properties, with a final biocatalytic productivity of 1.65 mM of xylonic acid per 1 U of co-immobilized XDH. The results demonstrate the advantages of the use of co-immobilized enzymes over a free enzyme system in terms of enhanced activity and stability.

Enzymatic CO2 reduction to formate by formate dehydrogenase from Candida boidinii coupling with direct electrochemical regeneration of NADH

Enzymatic conversion of CO2 to formate was carried out in the cathodic cell of a two-chamber electrochemical apparatus where NAD+ was reduced on the surface of a Copper foam electrode. Formate dehydrogenase (FDH) was used as the biocatalyst in both free form and immobilized on the modified electrospun polystyrene nanofibers (EPSNF). The fabricated EPSNF were modified by a multistage procedure including acid treatment, silanization followed by activation with glutaraldehyde. The effects of regenerated NADH concentration and time of enzymatic reaction on the formate production in the both systems were studied. The results indicated that the EPSNF immobilized FDH had a desirable activity, long-term storage stability (41% after 20 days) and reusability after eight cycles of successive reactions (53% of the initial activity). Moreover, it was revealed that the increase of cofactor concentration at the early times of reaction was favorable to the formate production. However, an inhibitory effect was observed at higher concentrations of NADH, and the optimum values of 0.45 mM and 0.51 mM were obtained for the maximum enzyme activity by the free and immobilized enzymes respectively. The produced formate at the optimum cofactor concentration after 300 min was 0.61 mM and 0.31 mM for the free and immobilized enzyme systems. Finally, it can be concluded that the presented process is a promising approach to the enzymatic conversion of CO2.

Assembly of Synthetic Functional Cellulosomal Structures onto the Cell Surface of Lactobacillus plantarum, a Potent Member of the Gut Microbiome.

Heterologous display of enzymes on microbial cell surfaces is an extremely desirable approach, since it enables the engineered microbe to interact directly with the plant wall extracellular polysaccharide matrix. In recent years, attempts have been made to endow noncellulolytic microbes with genetically engineered cellulolytic capabilities for improved hydrolysis of lignocellulosic biomass and for advanced probiotics. Thus far, however, owing to the hurdles encountered in secreting and assembling large, intricate complexes on the bacterial cell wall, only free cellulases or relatively simple cellulosome assemblies have been introduced into live bacteria. Here, we employed the "adaptor scaffoldin" strategy to compensate for the low levels of protein displayed on the bacterial cell surface. That strategy mimics natural elaborated cellulosome architectures, thus exploiting the exponential features of their Lego-like combinatorics. Using this approach, we produced several bacterial consortia of Lactobacillus plantarum, a potent gut microbe which provides a very robust genetic framework for lignocellulosic degradation. We successfully engineered surface display of large, fully active self-assembling cellulosomal complexes containing an unprecedented number of catalytic subunits all produced in vivo by the cell consortia. Our results demonstrate that the enzyme stability and performance of the cellulosomal machinery, which are superior to those seen with the equivalent secreted free enzyme system, and the high cellulase-to-xylanase ratios proved beneficial for efficient degradation of wheat straw.IMPORTANCE The multiple benefits of lactic acid bacteria are well established in health and industry. Here we present an approach designed to extensively increase the cell surface display of proteins via successive assembly of interactive components. Our findings present a stepping stone toward proficient engineering of Lactobacillus plantarum, a widespread, environmentally important bacterium and potent microbiome member, for improved degradation of lignocellulosic biomass and advanced probiotics.