Modification Of Protein Structure Research Articles

Effect of processing and formulation factors on Catalase activity in tablets

The manufacturing of tablets containing biologics exposes the biologics to thermal and shear stresses, which are likely to induce structural changes (e.g., aggregation and denaturation), leading to the loss of their activity. Saccharides often act as stabilizers of proteins in formulations, yet their stabilizing ability throughout solid oral dosage processing, such as tableting, has been barely studied. This work aimed to investigate the effects of formulation and process (tableting and spray-drying) variables on catalase tablets containing dextran, mannitol, and trehalose as potential stabilizers. Non-spray-dried and spray-dried formulations were prepared and tableted (100, 200, and 400 MPa). The enzymatic activity, number of aggregates, reflecting protein aggregation and structure modifications were studied. A principal component analysis was performed to reveal underlying correlations. It was found that tableting and spray-drying had a notable negative effect on the activity and number of aggregates formed in catalase formulations. Overall, dextran and mannitol failed to preserve the catalase activity in any unit operation studied. On the other hand, trehalose was found to preserve the activity during spray-drying but not necessarily during tableting. The study demonstrated that formulation and process variables must be considered and optimized together to preserve the characteristics of catalase throughout processing.

How pulse electric field treatment affects anti-nutritional factors and plant protein digestibility: A concise review

Consumption of foods rich in anti-nutritional factors (ANFs) can lead to inadequate absorption of vitamins and minerals. Pulsed electric field (PEF) treatment is a non-thermal food processing technology that uses short bursts of high voltage electric fields to induce structural modifications in biomolecules, including proteins, through electrochemical reactions and polarization of structural moieties. PEF offers potential benefits in enhancing the nutritional quality of food products by altering the structure and function of ANFs. However, the effects of PEF on ANFs in food have not been well established. Additionally, PEF-treated samples may undergo protein structural orientation modification due to electroporation and free radical species, potentially unfolding the native structure and influencing protein digestibility. Moreover, by increasing the mass transfer rate, electroporation can enhance the extraction of the initially trapped fractions into the solvent medium, which may include target compounds that affect protein functionality and digestibility. However, the protein digestibility of PEF-treated food products is yet to be thoroughly explored. Therefore, this review objectively investigates the possible mechanisms involved and the effectiveness of PEF in reducing different ANFs and modifying plant protein digestibility. This review also provides in-depth information about the possible applications of PEF-treated food items in the food industry.

Molecular Docking, Quantum Mechanics and Molecular Dynamics Simulation of Anti-CAD Drugs Against High-Risk Xanthine Dehydrogenase Variants Associated with Oxidative Stress Pathways

Xanthine dehydrogenase (XDH) contributes significantly to generating reactive oxygen species in coronary artery disease (CAD). XDH has been proposed as a therapeutic target, but its genetic variants could affect protein structure and drug response. We aimed to assess protein structure modification occur due to genetic variants and to screen 215 CAD drugs for their utility in personalized CAD treatment against the XDH variants. A series of computational methods were implemented to identify pathogenic variants that cause XDH structure instability localized at the conserved regions contributing to functional significance. Then, the XDH structures with the pathogenic variants were modeled using two different approaches to select the best models for docking with the CAD drugs. Finally, the stability of the docked complexes and their ability to transfer electrons were evaluated using molecular dynamics (MD) simulations and quantum mechanics/molecular mechanics (QM/MM) calculation. Among 751 variants examined; R149C and Q919R showed high pathogenicity, localized in conserved regions could alter protein structure and function. Further, docking of CAD drugs against XDH (native, R149C and Q919R) showed vericiguat with higher affinity, ranging from -7.95 kcal/mol to -10.41 kcal/mol, than the well-known XDH inhibitor (febuxostat, -5.73 kcal/mol to -8.35 kcal/mol). This indicates that vericiguat will be effective in CAD treatment, regardless of the XDH variants. Additionally, MD simulation and QM/MM confirmed vericiguat stability and electron transfer ability to form hydrogen bonds with the XDH protein. In conclusion, vericiguat will be beneficial for the personalized treatment of CAD by inhibiting XDH variants. Additional clinical studies are necessary to confirm our findings.

The role of membrane physiology in sHSP Lo18-lipid interaction and lipochaperone activity

To cope with environmental stresses, organisms, including lactic acid bacteria such as O. oeni, produce stress proteins called HSPs. In wine, O. oeni is constantly confronted by stress affecting its membrane fluidity. To survive through in these deleterious conditions, O. oeni synthesizes Lo18, a unique, small HSP which acts as a molecular chaperone and a lipochaperone. The molecular mechanism underlying its lipochaperone activity, particularly regarding membrane lipid composition, remains poorly understood. In this context, Lo18 lipochaperone activity and the associated modification in protein structure were studied during interaction with different liposomes from O. oeni cultures representing unstressed, stressed and stressed-adapted physiological states. The results showed that the presence of the membrane (whatever its nature) induces a modification of Lo18’s structure. Also, the presence of oleic acid and/or phosphatidylglycerol is important to favor Lo18-membrane interaction, allowing lipochaperone activity. This research enhances understanding of sHSP-membrane interactions in bacterial systems.

Biomimetic Spider Silk by Crosslinking and Functionalization with Multiarm Polyethylene Glycol

AbstractSpider silk is renowned for its exceptional mechanical properties, surpassing those of other natural and many synthetic fibers. Yet, replicating its remarkable properties through synthetic production remains a challenge. The variability in the mechanical properties of synthetic spider silks lacking protective coatings, exacerbated by factors such as spinning conditions and humidity levels, poses an additional challenge, impacting their application potential. Bioconjugation offers a versatile synthetic method to modify protein structures, enhancing their pharmacokinetics, solubility, stability, and immune response. In particular, polyethylene glycol (PEG)‐ylation has emerged as a successful strategy with numerous marketed PEG–protein conjugates. This study introduces synthetic spider silk—multiarm PEG bioconjugates, facilitating spidroin crosslinking, and chemical functionalization while retaining a biomimetic spinning approach. Two different examples demonstrate the potential of this approach to improve the fiber's tensile strength and extensibility, respectively, both leading to an increased toughness modulus. Furthermore, the approach could allow the tuning of fiber mechanical properties without developing a new mini‐spidroin construct and fiber coating with lipids attached to multiarm PEG, potentially mitigating the impact of environmental conditions on synthetic spider silk fibers.

Dissecting the interaction between Graphene Oxide and Human Transferrin via biophysical, biochemical and in-silico approaches: Implications for nanomaterial anti-fibrillation properties and protein structure

Human Transferrin (HTF) plays a pivotal role in numerous neurological therapies by carrying drugs and nanomaterials through the challenging blood-brain barrier. Nonetheless, the binding of HTF with highly relevant nanomaterials is not fully comprehended. Here, the nanomaterial graphene oxide (GO) has been obtained and their interaction with HTF in the native and fibrillar form has been investigated, by employing multi-spectroscopic, modeling, and microscopic techniques. The fluorescence results reveal that GO quenches the HTF intrinsic fluorescence by following a mix of static and dynamic mechanisms, and the GO-HTF complex formation has been confirmed by UV-Vis spectroscopy. Additionally, HTF attaches firmly to GO (KS≈ 106M−1) through mainly by hydrophobic and hydrogen bonds (according to determined ΔH=-23.77±1.03 kJ.mol−1 and ΔS= +39.53 ± 1.69 J.mol−1.K−1). Further, the in-silico investigation has evidenced the residues and chemical groups involved in HTF-GO interaction. Furthermore, SEM images reveal the formation and inhibition of HTF aggregates by GO, and spectroscopic methods reveal the related protein structural modification involved in the fibrillation process. In summary, all the results open questions concerning GO use for many neurological therapies.

Discovery of a novel homocysteine thiolactone hydrolase and the catalytic activity of its natural variants.

Homocysteine thiolactone (HTL), a toxic metabolite of homocysteine (Hcy) in hyperhomocysteinemia (HHcy), is known to modify protein structure and function, leading to protein damage through formation of N-Hcy-protein. HTL has been highly linked to HHcy-associated cardiovascular and neurodegenerative diseases. The protective role of HTL hydrolases against HTL-associated vascular toxicity and neurotoxicity have been reported. Although several endogeneous enzymes capable of hydrolyzing HTL have been identified, the primary enzyme responsible for its metabolism remains unclear. In this study, three human carboxylesterases were screened to explore new HTL hydrolase and human carboxylesterase 1 (hCES1) demonstrates the highest catalytic activity against HTL. Given the abundance of hCES1 in the liver and the clinical significance of its single-nucleotide polymorphisms (SNPs), six common hCES1 nonsynonymous coding SNP (nsSNPs) variants were examined and characterized for their kinetic parameters. Variants E220G and G143E displayed 7.3-fold and 13.2-fold lower catalytic activities than its wild-type counterpart. In addition, the detailed catalytic mechanism of hCES1 for HTL hydrolysis was computational investigated and elucidated by Quantum mechanics/molecular mechanics (QM/MM) molecular dynamics (MD) method. The function of residues E220 and G143 in sustaining its hydrolytic activity of hCES1 was analyzed, and the calculated energy difference aligns well with experimental-derived results, supporting the validity of our computational insights. These findings provide insights into the potential protective role of hCES1 against HTL-associated toxicity, and warrant future studies on the possible association between specific genetic variants of hCES1 with impaired catalytic function and clinical susceptibility of HTL-associated cardiovascular and neurodegenerative diseases.

Impact of non‐thermal plasma generated using air and nitrogen on functional properties of milk protein dispersions

AbstractThis study focuses on improving the functionalities of milk protein concentrate (MPC) using a nonthermal plasma jet and plasma‐activated water (PAW) with atmospheric air and nitrogen as source gases. The 5% and 10% MPC dispersions were directly treated with a plasma jet, and PAW was used to make the MPC dispersions. The dispersions were analyzed for changes in protein structure and functional properties. The treatment altered the secondary structure of MPC protein structure by increasing β‐components and changing the order of random coils. The solubility of the PAW‐treated 5% protein dispersions doubled due to plasma‐induced modification of hydrophilic and covalent bonds of the protein, but this increase was not significant for the 10% dispersions due to less hydration. Emulsifying capacity increases by around 7% for plasma jet and PAW with air, owing to hydrophobicity on the particle surface. The gelation capacity and heat coagulation time rise by almost twice; however, foaming capacity decreases, indicating protein structural modifications and aggregations caused by plasma exposure. The viscosity of the 5% dispersions decreased due to high solubility, while that of the 10% dispersions increased due to less hydration. Principle component analysis was used to correlate the change in functionality with different operating parameters. In conclusion, this study illustrates that the functional properties of MPC can be significantly modified through plasma treatment. The observed changes depend on several factors, including the mode of plasma exposure, the source gas used to generate the plasma, and the concentration of the protein solution.Practical applicationsMilk protein concentrate (MPC) is a promising ingredient in food products such as cheese, cultured dairy items, nutritional goods, infant milk formulas, ice cream, dairy‐based drinks, sports beverages, and a variety of health‐focused products. However, the application is profoundly hindered due to its poor solubility and solubility‐related functional properties. MPC powders lose functionalities during manufacturing and gradually lose them again during processing and storage. This study explores the potential of non‐thermal plasma to improve the functional properties of proteins. This study explores the potential of two different discharges: plasma jet and plasma‐activated water (PAW), using atmospheric air and nitrogen as source gases. The findings of this study would also help to understand the types of plasma discharge (jet vs PAW) and types of sources of gases that can be used in industrial applications. Some optimization parameters of this study can be used for scaling up the plasma processing of milk or dairy products. In addition, PAW is being very well studied to improve food safety, and this study would provide information on the physicochemical properties of MPC. Therefore, it would also help to understand how to apply non‐thermal plasma to achieve both microbial and physical effects.

Effect of supercritical carbon dioxide on protein structure modification and antimicrobial peptides production of Mongolian cheese and its in vitro digestion

The aim of this research was to investigate the effects of supercritical carbon dioxide (SC-CO2) treatment on protein structure in Mongolian cheese. The peptides during the digestive process of the SC-CO2 treated cheese were also studied. SC-CO2 technology was utilized to treat Mongolian cheese at three temperatures (45, 55 and 65 °C) and three pressures (7.5, 12.5 and 17.5 MPa). The results of fluorescence, ultraviolet–visible, Fourier transform infrared spectroscopy and free sulfhydryl groups showed that SC-CO2, particularly at 65 °C and 17.5 MPa, modified the protein structure in Mongolian cheese effectively. The data of LC-MS/MS-based peptidomics showed that the content of antimicrobial peptides found in the SC-CO2 treated Mongolian cheese was 1.55 times that of the untreated Mongolian cheese; the content of unique antimicrobial peptides in the digested SC-CO2 treated Mongolian cheese was 1.46 times that of the digested untreated Mongolian cheese, which proved that SC-CO2 could help produce antimicrobial peptides in cheese not only during the process of SC-CO2 treatment but during subsequent simulated gastrointestinal digestion as well. In conclusion, SC-CO2 could be considered a promising method to develop cheese products with potential health benefits.

Effect of moderate heating during alkaline extraction on composition and functional properties of brewer's spent grain protein concentrates

Brewer's spent grain (BSG) is a low-cost protein-rich residue and a valuable source of plant proteins. Extraction conditions and parameters can influence the composition and functionality of the resultant BSG protein concentrates. In this study, alkaline extraction at room temperature and 50 °C was investigated to explore the alterations induced by moderate heating in the composition, fatty acid and amino acid profiles, emulsion stability, and foaming properties of BSG protein concentrates. The protein content of the sample extracted at 50 °C decreased by 6%. An increase of 29.4% in the carbohydrate content and 2.5% in the lipid content was observed due to extraction at 50 °C. Hydrophobic amino acid content and saturated/unsaturated fatty acid ratio decreased by 20.9% and 4.8% respectively, in the protein concentrate extracted at 50 °C. Fourier transform infrared spectroscopy and gel electrophoresis showed some degree of protein structural modification and aggregation in the sample extracted at 50 °C. Although the heat-induced compositional alterations in the protein concentrate were not in favour of functionality, its emulsion stability and foaming capacity were significantly improved. These findings demonstrate that the heat-induced improvements in the surface hydrophobicity and conformational flexibility of proteins play a dominant role in the functionality of BSG protein concentrates. It can provide insights into modulating the performance of these valuable side-stream proteins for various food, pharmaceutical, and cosmetics products.

Alkyl anchor–modified artificial viral capsid budding outside-to-inside and inside-to-outside giant vesicles

ABSTRACT The budding of human immunodeficiency virus from an infected host cell is induced by the modification of structural proteins bearing long-chain fatty acids, followed by their anchoring to the cell membrane. Although many model budding systems using giant unilamellar vesicles (GUVs) induced by various stimuli have been developed, constructing an artificial viral budding system of GUVs using only synthesized molecules remains challenging. Herein, we report the construction of an artificial viral capsid budding system from a lipid bilayer of GUV. The C-terminus of the β-annulus peptide was modified using an octyl chain as an alkyl anchor via a disulfide bond. The self-assembly of the β-annulus peptide with an octyl chain formed an artificial viral capsid aggregate. The fluorescence imaging and transmission electron microscopy observations revealed that the addition of the tetramethylrhodamine (TMR)-labeled octyl chain–bearing β-annulus peptide to the outer aqueous phase of GUV induced the budding of the capsid-encapsulated daughter vesicle outside-to-inside the mother GUV. Conversely, the encapsulation of the TMR-labeled octyl chain–bearing β-annulus peptide in the inner aqueous phase of GUV induced the budding of the capsid-encapsulated daughter vesicle inside-to-outside the mother GUV. Contrarily, the addition of the TMR-labeled β-annulus peptide to GUV barely induced budding. It was demonstrated that the higher the membrane fluidity of GUV, the more likely budding would be induced by the addition of the alkyl anchor–modified artificial viral capsid. The simple virus-mimicking material developed in this study, which buds off through membrane anchoring, can provide physicochemical insights into the mechanisms of natural viral budding from cells.

From Bulk to Single Molecules: Surface-Enhanced Raman Scattering of Cytochrome C Using Plasmonic DNA Origami Nanoantennas.

Cytochrome C, an evolutionarily conserved protein, plays pivotal roles in cellular respiration and apoptosis. Understanding its molecular intricacies is essential for both academic inquiry and potential biomedical applications. This study introduces an advanced single-molecule surface-enhanced Raman scattering (SM-SERS) system based on DNA origami nanoantennas (DONAs), optimized to provide unparalleled insights into protein structure and interactions. Our system effectively detects shifts in the Amide III band, thereby elucidating protein dynamics and conformational changes. Additionally, the system permits concurrent observations of oxidation processes and Amide bands, offering an integrated view of protein structural and chemical modifications. Notably, our approach diverges from traditional SM-SERS techniques by de-emphasizing resonance conditions for SERS excitation, aiming to mitigate challenges like peak oversaturation. Our findings underscore the capability of our DONAs to illuminate single-molecule behaviors, even within aggregate systems, providing clarity on molecular interactions and behaviors.

Effects of two types of corn starches on physicochemical and structural properties of red bean protein isolate acid-induced cold gels

A mixture of red bean protein isolate (RBPI) with common starch (CS) or waxy starch (WS) from corn was heated, and induced by glucono-δ-lactone (GDL) to form a RBPI-starch complex cold gel. The physicochemical and structural properties of these gels were also investigated. Compared with the RBPI gels, the complex gels with CS had higher gel strength, water retention, viscoelasticity, and deformation resistance, forming a denser and more continuous microstructure with a high degree of crosslinking and small pores. Compared with the RBPI-CS complex gels, the RBPI-WS complex gels exhibited higher water retention and stronger transformation ability of free water to immobilized water; however, they exhibited poor mechanical properties and microstructures. The difference in the physicochemical properties of the gels was mainly related to the change in the hydrophobic interactions between the two gel systems. In addition, protein structure modification affected the gelation properties of the RBPI-CS and RBPI-WS complexes. The RBPI-CS complex gel exhibited strong X-ray peaks and fluorescence quenching ability. The addition of CS promoted the flexibility of the secondary structure and tightness of the tertiary conformation of the protein in the complex gel, forming a stronger and more compact non-covalent gel structure. However, the protein in the RBPI-WS complex gel showed an ordered secondary structure and a loose tertiary conformation, which weakened the gel network.

Food-Oral Processing: Current Progress, Future Directions, and Challenges.

Oral processing refers to the series of physical, chemical, and biological processes inside the oral cavity when we consume food. This process affects the taste, quality, and nutrient absorption of the body. In the human diet, oral processing plays a crucial role because it impacts not only the food flavor and texture but also the absorption and utilization of nutrients. With the progress of science and technology and the increasing demand for food, the study of oral processing has become increasingly important. This paper reviews the history and definition of oral processing, its current state of research, and its applications in food science and technology, focusing on personalized taste customization, protein structure modification, food intake and nutrition, and bionic devices. It also analyzes the impact of oral processing on different types of food products and explores its potential in the food industry and science research.

Genetic variation in the CYP1A gene caused by laboratory exposure of benzo (a) pyrene to common carp

Polycyclic aromatic hydrocarbons (PAHs) are pervasive environmental contaminants, with benzo(a)pyrene (B(a)P) standing out as a prototypical example. B(a)P is a known mutagen and carcinogen that, upon metabolic activation stimulated by cytochromes P450 (CYP1A) with microsomal epoxide hydrolase, reacts with DNA. In this study, common carp (Cyprinus carpio) were subjected to three different doses of B(a)P (1μg/kg and 10μg/kg) to investigate the mutagenic effects on B(a)P-derived DNA in vivo.Blood samples were collected, followed by thorough DNA extraction and polymerase chain reaction analysis. The findings revealed a pronounced mutational impact of B(a)P on the amplified segment of the CYP1A gene. Specifically, at a concentration of 1μg/kg, B(a)P induced 8 mutations, 3 amino acid alterations, and the emergence of 5 new haplotype patterns. In contrast, at a concentration of 10μg/kg, B(a)P resulted in 21 mutations, 10 changes in amino acids, and the generation of 7 new haplotype patterns. Additionally, alterations in the three-dimensional protein composition were observed in both dosage groups.This study underscores the significant mutagenic potential of B(a)P, shedding light on its capacity to induce genetic changes and protein structure modifications, thereby emphasizing the importance of monitoring and addressing the environmental presence of PAHs for both ecological and human health considerations

In-Silico Method for Predicting Pathogenic Missense Variants Using Online Tools: AURKA Gene as a Model.

In-silico analysis provides a fast, simple, and cost-free method for identifying potentially pathogenic single nucleotide variants. To propose a simple and relatively fast method for the prediction of variant pathogenicity using free online in-silico (IS) tools with AURKA gene as a model. We aim to propose a methodology to predict variants with high pathogenic potential using computational analysis, using AURKA gene as model. We predicted a protein model and analyzed 209 out of 64,369 AURKA variants obtained from Ensembl database. We used bioinformatic tools to predict pathogenicity. The results were compared through the VarSome website, which includes its own pathogenicity score and the American College of Medical Genetics (ACMG) classification. Out of the 209 analyzed variants, 16 were considered pathogenic, and 13 were located in the catalytic domain. The most frequent protein changes were size and hydrophobicity modifications of amino acids. Proline and Glycine amino acid substitutions were the most frequent changes predicted as pathogenic. These bioinformatic tools predicted functional changes, such as protein up or down-regulation, gain or loss of molecule interactions, and structural protein modifications. When compared to the ACMG classification, 10 out of 16 variants were considered likely pathogenic, with 7 out of 10 changes at Proline/Glycine substitutions. This method allows quick and cost-free bulk variant screening to identify variants with pathogenic potential for further association and/or functional studies.

Effects of myofibril-palatinose conjugate as a phosphate substitute on meat emulsion quality

The objective of this study was to investigate a replacement for phosphate in meat products. Protein structural modification was employed in this study, and grafted myofibrillar protein (MP) with palatinose was added to meat emulsion without phosphate. Here, 0.15% of sodium polyphosphate (SPP) was replaced by the same (0.15%) concentration and double (0.3%) the concentration of grafted MP. Although the thermal stability was decreased, the addition of transglutaminase could increase stability. The rheological properties and pH also increased with the addition of grafted MP and transglutaminase. The addition of grafted protein could be perceived by the naked eye by observing a color difference before cooking, but it was not easy to detect after cooking. The cooking loss, emulsion stability, water holding capacity, lipid oxidation, and textural properties improved with the addition of grafted MP. However, the excessive addition of grafted MP and transglutaminase was not recommended to produce a high quality of phosphate replaced meat emulsion, and 0.15% was identified as a suitable addition ratio of grafted MP.

Protein Post-Translational Modifications Based on Proteomics: A Potential Regulatory Role in Animal Science.

Genomic studies in animal breeding have provided a wide range of references; however, it is important to note that genes and mRNA alone do not fully capture the complexity of living organisms. Protein post-translational modification, which involves covalent modifications regulated by genetic and environmental factors, serves as a fundamental epigenetic mechanism that modulates protein structure, activity, and function. In this review, we comprehensively summarize various phosphorylation and acylation modifications on metabolic enzymes relevant to energy metabolism in animals, including acetylation, succinylation, crotonylation, β-hydroxybutylation, acetoacetylation, and lactylation. It is worth noting that research on animal energy metabolism and modification regulation lags behind the demands for growth and development in animal breeding compared to human studies. Therefore, this review provides a novel research perspective by exploring unreported types of modifications in livestock based on relevant findings from human or animal models.

Unravelling the impact of ohmic heating on commercial pea protein structure

Pea protein (PP) has attracted the attention of the food industry and the scientific community as a substitute for more established plant proteins such as soy proteins. However, commercial PP often displays poor solubility which limits its functionality and further use by the food industry. To overcome this challenging issue, plant protein modification techniques for improved functionality have been investigated. Ohmic heating (OH) is an emergent thermo-electrical processing technology which has demonstrated potential in modifying protein structure and therefore its functionality. In this work, the potential use of OH has a physical technique for protein modification was investigated. Therefore, a commercial PP was submitted to OH application, at treatment temperatures ranging from 90 to 150 °C and the potential changes in PP solubility upon OH application were evaluated. An increase in protein solubility of ca. 240 % was observed after OH application at 150 °C. Protein structural characterization was performed, suggesting PP structural rearrangement, rather than unfolding, upon OH application. Furthermore, OH application presumably promoted the dissociation of the insoluble aggregates already present in the commercial PP sample. Moreover, higher treatment temperatures (particularly from 110 °C onwards) may have induced protein hydrolytic cleavage of the peptide bond to some extent, leading to the formation of mainly small soluble aggregates and peptide fragments. These results open novel perspectives for the use of OH as a promising tool for improving commercial PP solubility and, therefore, enabling its tailored application in the food industry.

Behind the Scenes of PluriZyme Designs

Protein engineering is the design and modification of protein structures to optimize their functions or create novel functionalities for applications in biotechnology, medicine or industry. It represents an essential scientific solution for many of the environmental and societal challenges ahead of us, such as polymer degradation. Unlike traditional chemical methods, enzyme-mediated degradation is selective and environmentally friendly and requires milder conditions. Computational methods will play a critical role in developing such solutions by enabling more efficient bioprospecting of natural polymer-degrading enzymes. They provide structural information, generate mechanistic studies, and formulate new hypotheses, facilitating the modeling and modification of these biocatalysts through enzyme engineering. The recent development of pluriZymes constitutes an example, providing a rational mechanism to integrate different biochemical processes into one single enzyme. In this review, we summarize our recent efforts in this line and introduce our early work towards polymer degradation using a pluriZyme-like technology, including our latest development in PET nanoparticle degradation. Moreover, we provide a comprehensive recipe for developing one’s own pluriZyme so that different laboratories can experiment with them and establish new limits. With modest computational resources and with help from this review, your first pluriZyme is one step closer.