Enzyme Stability Research Articles

Stabilization of galactose oxidase by high hydrostatic pressure: Insights on the role of cavities size.

High hydrostatic pressure stabilized galactose oxidase (GaOx) at 70.0-80.0°C against thermal inactivation. The pseudo-first-order rate constant of inactivation kinact decreased by a factor of 8 at 80°C and by a factor of 44 at 72.5°C. The most pronounced effect of pressure was at the lowest studied temperature of 70.0°C with an activation volume of inactivation ΔV‡ of 78.8 cm3 mol-1. The optimal pressure against thermal inactivation was between 200 and 300 MPa. Unlike other enzymes, as temperature increased the ΔV‡ of inactivation decreased, and as pressure increased the activation energy of inactivation Eai increased. Combining the results for GaOx with earlier research on the pressure-induced stabilization of other enzymes suggests that ΔV‡ of inactivation correlates with the total molar volume of cavities larger than ~100 Å3 in enzyme monomers for enzymes near the optimal pH and whose thermal unfolding is not accompanied by oligomer dissociation.

The multiple enhancements of deep eutectic solvent on cellulase significantly improve the saccharification efficiency of lignocellulosic biomass

The introduction of specific choline chloride (ChCl)- and betaine (Bet)-based deep eutectic solvents (DESs) into the conventional acetic acid–sodium acetate solution led to a significant increase in cellulase activity and stability. In particular, a higher enhancement in β-glucosidase activity was obtained, which plays a key role in improving cellulase function. The DESs prolonged the half-life of cellulase. The addition of 10 % (v/v) ChCl/1,3-propanediol (1:2) and 5 % (v/v) Bet/glycerol (1:2) resulted in the most significant improvement, with the half-life of cellulase reaching three times the initial half-life value. The above both DESs allowed the cellulase to maintain an excellent level of activity over time, even after 72 h. The DESs tightened the structure of the enzyme by changing its secondary structure, which had a stabilizing effect on the overall conformation. The density functional theory calculations of DESs revealed that the DES with a more stable structure more effectively protected the active conformation. The molecular dynamics simulation on β-glucosidase indicated that the hydrogen bond donor end of the DES primarily formed hydrogen bonds with the enzyme, which induced a spatially compact conformation by increasing the α-helix content while reducing the β-sheet content. It also restricted the movement of amino acids, particularly stabilizing the active site, and increased the hydrophilic region, thereby enhancing enzyme activity and stability. In addition, the DES formed a protective layer that prevented the unproductive adsorption of lignin on the enzymes during the hydrolysis of lignocellulosic feedstocks. These multiple improvements provided by DESs significantly increased the enzymatic saccharification efficiency of corn stover and poplar.

A Critical Review on Immobilized Sucrose Isomerase and Cells for Producing Isomaltulose.

Isomaltulose is a novel sweetener and is considered healthier than the common sugars, such as sucrose or glucose. It has been internationally recognized as a safe food product and holds vast potential in pharmaceutical and food industries. Sucrose isomerase is commonly used to produce isomaltulose from the substrate sucrose in vitro and in vivo. However, free cells/enzymes were often mixed with the product, making recycling difficult and leading to a significant increase in production costs. Immobilized cells/enzymes have the following advantages including easy separation from products, high stability, and reusability, which can significantly reduce production costs. They are more suitable than free ones for industrial production. Recently, immobilized cells/enzymes have been encapsulated using composite materials to enhance their mechanical strength and reusability and reduce leakage. This review summarizes the advancements made in immobilized cells/enzymes for isomaltulose production in terms of refining traditional approaches and innovating in materials and methods. Moreover, innovations in immobilized enzyme methods include cross-linked enzyme aggregates, nanoflowers, inclusion bodies, and directed affinity immobilization. Material innovations involve nanomaterials, graphene oxide, and so on. These innovations circumvent challenges like the utilization of toxic cross-linking agents and enzyme leakage encountered in traditional methods, thus contributing to enhanced enzyme stability.

Influences of Polyphenols on the Properties of Crosslinked Acellular Fish Swim Bladders: Experiments and Molecular Dynamic Simulations.

Acellular fish swim bladders (AFSBs) are a promising biomaterial in tissue engineering, however, they may suffer from rapid degradation due to enzyme invasion. In this work, natural polyphenols, including epigallocatechin gallate (EGCG), proanthocyanidin (PC), tannic acid (TA) and protocatechuic acid (PCA), were utilized to improve the properties of AFSBs through crosslinking modifications. Fourier transform infrared (FTIR) results indicate that the triple helix of the collagen in AFSBs is well preserved after crosslinking. The differential scanning calorimetry (DSC), water contact angle (WCA) and in vitro degradation tests indicate that the polyphenol-crosslinked AFSBs exhibit improved thermal stability, enzymatic stability, hydrophilicity and mechanical properties. Among them, EGCG with multiple phenolic hydroxyl groups and low potential resistance is more favorable for the improvement of the mechanical properties and enzymatic stability of AFSBs, as well as their biocompatibility and integrity with the collagen triple helix. Moreover, the crosslinking mechanism was demonstrated to be due to the hydrogen bonds between polyphenols and AFSBs, and was affected by the molecular size, molecular weight and the hydroxyl groups activity of polyphenol molecules, as clarified by molecular dynamic (MD) simulations. The approach presented in this work paves a path for improving the properties of collagen materials.

Photobleaching FRET-FLIM-ICS for quaternary structure quantification on cells. Theory and simulations

The oligomerization of proteins is an important biological control mechanism and has several functions in activity and stability of enzymes, structural proteins, ion channels and transcription factors. The determination of the relevant oligomeric states in terms of geometry (spatial extent), oligomer size (monomer or dimer or oligomer) and affinity (amounts of monomer, dimer and oligomer) is a challenging biophysical problem. Förster resonance energy transfer and fluorescence fluctuation spectroscopy are powerful tools that are sensitive to proximity and oligomerization respectively. Here it is proposed to combine image-based lifetime-detected Forster resonance energy transfer with image correlation spectroscopy and photobleaching to determine distances, oligomer sizes and oligomer distributions. Simulations for simple oligomeric forms illustrate the potential to improve the discrimination between different quaternary states in the cellular milieu.

The Application of CRISPR Gene Editing Technology in PET Biodegradation

Plastics used in people's daily life bring convenience on the one hand, and on the other hand, a large amount of plastic waste also brings great pollution to the environment. Polyethylene terephthalate (PET) is a polymer polymerization material, which has become one of the most widely used plastics in the world because of its durability and easy processing. The stability of the ecological environment around it is seriously threatened by the large amount of PET used and its difficult degradation.PET even endangered human health. The traditional methods of PET treatment are physical and chemical methods such as incineration, landfill, and heat cracking, etc. Comoared with the first two generations of gene-editing system, CRISPR/Cas9 system has the advantage of easy design, low cost and high efficiency. Currently, the most favorable method of treating PET to protect the environment is biodegradation. This paper introduces the nature of PET and the research progress of PET degradation enzymes, seeking to improve the activity and thermal stability of degradation enzymes by CRISPR/Cas9 gene editing technology, this paper explores the construction of mutants with high thermal stability and high degradation activity of the fusion dual enzyme of PETase and MHETase by CRISPR/Cas9 gene editing technology.

Methods for Determining and Predicting Shelf Life of Food Products

The length of shelf life can be determined using storage tests that evaluate the physical, chemical, enzymatic, and microbiological stability of the food. The design of the tests as well as the interpretation of the results require microbiological and technical knowledge and skills. Fresh foods can become microbiologically risky before they noticeably spoil. These products will require additional studies such as predictive microbiological models or challenge tests in addition to direct ones. Deterioration of shelf-stable or frozen foods at the end of their shelf life affects quality and consumer acceptance without impacting health and safety. In such a case, accelerated shelf-life testing is recommended, and the Arrhenius model is commonly used to determine the relationship between chemical reaction rate and temperature change.

Construction, characterization, antioxidant activity and effects on properties in vitro digestion of selenium nanoparticles decorated with Cyperus esculentus polysaccharides

In this study, in order to obtain selenium complex with high antioxidant activity and low release rate, selenium nanoparticles (SeNPs) were decorated by chemical reduction with Cyperus esculentus polysaccharides (CEP) as stabilizers (CEP-SeNPs). The characterization, construction mechanism, stability, and effects on properties in vitro digestion of CEP-SeNPs were investigated. CEP-SeNPs were orange-red in color and uniformly distributed in a spherical shape with an average diameter of 138.33 ± 0.57 nm, exhibiting good stability which could be stored at 4 °C. The selenium element in CEP-SeNPs was selenium in a zero-valent amorphous state. The O-H and C=O bonds in CEP were bound to selenium by an inductive effect to construct CEP-SeNPs for infrared analysis, and no new covalent bonds were formed in this process. CEP as stabilizer on selenium nanoparticles were physical combination. At the same time, the synergistic effects at antioxidant activity of polysaccharides and selenium was found which made CEP-SeNPs had a higher antioxidant activity than CEP. Above all, CEP-SeNPs showed better pH and enzyme stability in simulated digestion. The selenium release rate of CEP-SeNPs were only 18.52 % ± 0.17 % after intestinal digestion. The antioxidant activity of CEP-SeNPs were higher than that of CEP in the simulated digestion. It provides a theoretical basis for the development and utilization of rich selenium products.

Evaluation of free and immobilized cellulase on chitosan-modified magnetic nanoparticles for saccharification of sorghum residue.

Enzymatic hydrolysis plays a pivotal role in transforming lignocellulosic biomass. Addressing alternate techniques to optimize the utilization of cellulolytic enzymes is one strategy to improve its efficiency and lower process costs. Cellulases are highly specific and environmentally benign biocatalysts that break down intricate polysaccharides intosimple formsofsugars. In contrast to the most difficult and time-consuming enzyme immobilization processes, in this research, we studied simple, mild, and successful techniques for immobilization of pure cellulase on magnetic nanocomposites using glutaraldehyde as a linker and used in the application of sorghum residue biomass. Fe3O4 nanoparticles were coated with chitosan from the co-precipitation method, which served as an enzyme carrier. The nanoparticles were observed under XRD, Zeta Potential, FESEM, VSM, and FTIR. The size morphology results presented that the Cs@Fe3O4 have 42.2nm, while bare nanoparticles (Fe3O4) have 31.2nm in size. The pure cellulase reaches to 98.07% of loading efficiency and 71.67% of recovery activity at optimal conditions. Moreover, immobilized enzyme's pH stability, thermostability, and temperature tolerance were investigated at suitable conditions. The kinetic parameters of free and immobilized enzyme were estimated as Vmax; 29 ± 1.51 and 27.03 ± 2.02µmolmin-1mg-1, Km; 4.7 ± 0.49mM and 2.569 ± 0.522mM and Kcat; 0.13s-1, and 0.89s-1. Sorghum residue was subjected to 2% NaOH pre-treatment at 50℃. Pre-treated biomass contains cellulose of 64.8%, used as a raw material to evaluate the efficiency of reducing sugar during hydrolysis and saccharification of free and immobilized cellulase, which found maximum concentration of glucose 5.42g/L and 5.12g/L on 72h. Thus, our study verifies the use of immobilized pure cellulase to successfully hydrolyze raw material, which is a significant advancement in lignocellulosic biorefineries and the reusability of enzymes.

Implications of Natural 3′–5′– Linkages in the Loop Region of Isomeric 2′–5′–Linked Thrombin–Binding Aptamer

AbstractThe backbone modification of the thrombin‐binding aptamer (TBA) in the TT and TGT loop regions by isomeric 2′–5′–linkages was found to impose additive destabilizing effects on the thermal stability of the G‐quadruplex structure. In contrast, the thermal stability of isomeric 2′–5′–linked TBA, i. e., isoTBA, was significantly improved by isomeric 3′–5′–phosphodiester linkages. The isoTBA, when modified with 3′–5′–linkages in both lateral TT loops (isoTBA202), exhibited higher thermal stability and enzymatic stability in comparison to other oligomers in the present study, and TBA202 showed higher antithrombin activity than other loop‐modified TBA oligomers.

Urease stabilization in urea–urease–H+ system and its influence on the clock reaction dynamics

AbstractThe inherent autocatalytic kinetics of the urea–urease–H+ system positions it as a promising candidate for the design of dynamic materials with time‐domain programmable functions. Nevertheless, the stability of the enzyme can markedly influence the temporal evolution dynamics of the system and curtail its widespread applicability. This work employs several kinds of enzyme stabilization methods, including chemical cross‐linking, physical coating, solvent stabilization, and solvent‐physical coating co‐modification, to systematically explore the impact of enzyme stabilization on clock reaction dynamics. Extensive experimental tests and analysis indicate that solvent and chemical cross‐linking stabilization methods can better preserve clock dynamics with sensitive switching ability. Nevertheless, due to significant pH changes in the reacting system, the reusability of the enzyme is better retained in the physical coating and solvent‐physical coating co‐modification methods.

Recent Progress on Conversion of Lignocellulosic Biomass by MOF-Immobilized Enzyme.

The enzyme catalysis conversion of lignocellulosic biomass into valuable chemicals and fuels showed a bright outlook for replacing fossil resources. However, the high cost and easy deactivation of free enzymes restrict the conversion process. Immobilization of enzymes in metal-organic frameworks (MOFs) is one of the most promising strategies due to MOF materials' tunable building units, multiple pore structures, and excellent biocompatibility. Also, MOFs are ideal support materials and could enhance the stability and reusability of enzymes. In this paper, recent progress on the conversion of cellulose, hemicellulose, and lignin by MOF-immobilized enzymes is extensively reviewed. This paper focuses on the immobilized enzyme performances and enzymatic mechanism. Finally, the challenges of the conversion of lignocellulosic biomass by MOF-immobilized enzyme are discussed.

Dual enzyme self-assembly cluster for high efficient asymmetric reduction of ethyl 6-oxo-8-chlorooctanoate

(R)-ECHO is synthesized through the asymmetric reduction of ethyl 6-oxo-8-chlorooctanoate (ECOO) using carbonyl reductase(CR2) as a catalyst and co-enzyme NADPH as a hydrogen donor with addition of glucose dehydrogenase (GDH) for NADPH regeneration, thereby enhancing biotransformation efficiency. To improve the reaction efficiency further, a dual enzyme self-assembly cluster EutM-SC-ST-CR2(GDH) was developed specifically for this purpose. This co-immobilised system demonstrated a 50% increase in the conversion of ECOO compared to when SpyTag-CR2(ST-CR2) and SpyTag-GDH(ST-GDH) were co-catalysts. The optimal conditions for this increase were observed to be at a pH of 8.0, 30°C, when concentrations of 250 mM ECOO, 270 mM glucose and 0.1 mM NADPH, and a molar ratio of ST-CR2 to ST-GDH to EutM-SpyCatcher(EutM-SC) of 1:1:2 (molecular concentrations of ST-CR2, ST-GDH and EutM-SC were 3.5 mM, 3.5 mM and 7 mM). Under these conditions, the conversion of ECOO reached up to 95% and the enantiomeric excess (e.e.) of (R)-ECHO achieved 99.9%. Moreover, the EutM-SC-ST-CR2(GDH) construct displayed enhanced enzyme activity, stability, and kinetics, as well as improvements in electrostatic and ionic strengths. In conclusion, this study presents the development of a novel self-assembled protein scaffold, EutM-SC, for the co-immobilisation of ST-CR2 and ST-GDH, thus optimizing the biological reaction conditions and offering potential for the industrial production of (R)-α-lipoic acid.

Boosted Enzyme Activity via Encapsulation within Metal-Organic Frameworks with Pores Matching Enzyme Size and Shape.

A novel and versatile approach called "physical imprinting" is introduced to modulate enzyme conformation using mesoporous materials, addressing challenges in achieving improved enzyme activity and stability. Metal-organic frameworks with tailored mesopores, precisely matching enzyme size and shape, are synthesized. Remarkably, enzymes encapsulated within these customized mesopores exhibit over 1670% relative activity compared to free enzymes, maintaining outstanding efficiency even under harsh conditions such as heat, exposure to organic solvents, wide-ranging pH extremes from acidic to alkaline, and exposure to a digestion cocktail. After 18 consecutive cycles of use, the immobilized enzymes retain 80% of their initial activity. Additionally, the encapsulated enzymes exhibit a substantial increase in catalytic efficiency, with a 14.1-fold enhancement in kcat/KM compared to native enzymes. This enhancement is among the highest reported for immobilized enzymes. The improved enzyme activity and stability are corroborated by solid-state UV-vis, electron paramagnetic resonance, Fourier-transform infrared spectroscopy, and solid-state NMR spectroscopy. The findings not only offer valuable insights into the crucial role of size and shape complementarity within confined microenvironments but also establish a new pathway for developing solid carriers capable of enhancing enzyme activity and stability.

Reconstructing dynamics correlation network to simultaneously improve activity and stability of 2,3-butanediol dehydrogenase by design of distal interchain disulfide bonds

The complete enzyme catalytic cycle includes substrate binding, chemical reaction and product release, in which different dynamic conformations are adopted. Due to the complex relationship among enzyme activity, stability and dynamics, the directed evolution of enzymes for improved activity or stability commonly leads to a trade-off in stability or activity. It hence remains a challenge to engineer an enzyme to have both enhanced activity and stability. Here, we have attempted to reconstruct the dynamics correlation network involved with active center to improve both activity and stability of a 2,3-butanediol dehydrogenase (2,3-BDH) by introducing inter-chain disulfide bonds. A computational strategy was first applied to evaluate the effect of introducing inter-chain disulfide bond on activity and stability of three 2,3-BDHs, and the N258C mutation of 2,3-BDH from Corynebacterium glutamicum (CgBDH) was proved to be effective in improving both activity and stability. In the results, CgBDH-N258C showed a different unfolding curve from the wild type, with two melting temperatures (Tm) of 68.3 °C and 50.8 °C, 19.7 °C and 2 °C higher than 48.6 °C of the wild type. Its half-life was also improved by 14.8-fold compared to the wild type. Catalytic efficiency (kcat/Km) of the mutant was increased by 7.9-fold toward native substrate diacetyl and 8.8-fold toward non-native substrate 2,5-hexanedione compared to the wild type. Molecular dynamics simulations revealed that an interaction network formed by Cys258, Arg162, Ala144 and the catalytic residues was reconstructed in the mutant and the dynamics change caused by the disulfide bond could be propagated through the interactions network. This improved the enzyme stability and activity by decreasing the flexibility and locking more “reactive” pose, respectively. Further construction of mutations including A144G showing a 44-fold improvement in catalytic efficiency toward meso-2,3-BD confirmed the role of modifying dynamics correlation network in tunning enzyme activity and selectivity. This study provided important insights into the relationship among dynamics, enzyme catalysis and stability, and will be useful in the designing new enzymes with co-evolution of stability, activity and selectivity.

MODIFICATION OF HYALURONIDASE MICROENVIRONMENT: INNOVATORY APPROACHES FOR DEVELOPMENT OF BIOCATALYTIC MEDICAL PREPARATIONS

The computational study of 3D-model hyaluronidase interaction with shortchain glycosaminoglycan ligands had performed demonstrating the diversity and significance of their reaction on enzyme structure. The purpose of this review was evolution of limiting enzyme functioning interactions (impact on stability, biocatalyst activity) with grounding of recommendations for experimental modification of hyaluronidase for obtaining of its derivative of medicine destination (according the results of theoretical researches). The analysis was performed on databases of PubMed, Web of Science, MedLine, E-library in frames last 15 years. The binding of chondroitin trimers (on centers cn6, cn3, cn1) to hyaluronidase molecular surface increased the enzyme stability, binding of chondroitin sulfate trimers (on centers cs2, cs4, cs7, cs8 or cs1, cs2, cs4, cs7, cs8) decreased the inhibition of enzyme by tetramer heparin. It should be noted the importance of ligand binding for regulation of enzyme functioning and existence of multiform and multicomponent microenvironment of enzyme. The sequence of preferable coupling of ligands with hyaluronidase is elicited in our study and with its help was evaluate reality of experimental selective modification of enzyme (possibly noncovalent or covalently, for instance, with chondroitin sulfate trimers on centers cs7, cs1, cs5) for experimental obtaining of stabilized enzyme forms. The perspective approaches for this aim may be the noncovalent reaction on hyaluronidase by chondroitin or chondroitin sulfate trimers as well covalent modification of biocatalyst by chondroitin sulfate trimers.

Biomineralized synthesis of luminescent protease-(NH4)2Y3F11•H2O hybrid nanospheres and their applications as a stable and reusable enzyme reactor

Proteases, such as trypsin, are essential for extracting collagen in various industrial applications. The potential applications of rare earth nanomaterials, specifically yttrium nanoparticles, have attracted significant interest across various fields due to their distinctive characteristics, including high dielectric constant and thermal stability. Biomineralization has emerged as a promising approach to synthesize protein-inorganic nanomaterials with hierarchical structures and desired functions. In the present investigation, a novel protease-templated biomineralization strategy was developed for synthesizing protease-(NH4)2Y3F11•H2O hybrid nanomaterials using a one-pot method under very mild conditions. For modifying the morphologies of (NH4)2Y3F11•H2O throughout biomineralization, protease has been demonstrated to be a highly promising biotemplate. Protease was utilized as a template for morphological control in the biomineralization procedure, which resulted in a gradual transformation of the initially formed (NH4)2Y3F11•H2O octahedral structures into uniform nanospheres. The applicability of this approach was supported by successfully utilizing various proteases to synthesize protease-(NH4)2Y3F11•H2O hybrid nanospheres. In addition to a strong and desirable luminescent signal, these hybrid nanospheres demonstrated extensive recycling because of their high enzymatic activity, stability and durability. The protease-mediated biomineralization approach offers an easy and robust approach to develop innovative protease-inorganic composites. Its moderate reaction conditions and simple operation render it a viable tool for developing stable and reusable enzyme reactors in various industrial applications.Graphical

N-terminal domain truncation yielded a unique dimer of polysaccharide hydrolase with enhanced enzymatic activity, stability and calcium ion independence

Domain engineering, including domain truncation, fusion, or swapping, has become a common strategy to improve properties of enzymes, especially glycosyl hydrolases. However, there are few reports explaining the mechanism of increased activity from a protein structure perspective. Amy703 is an alkaline amylase with a unique N-terminal domain. Prior studies have shown that N-Amy, a mutant without an N-terminal domain, exhibits improved activity, stability, and calcium ion independence. In this study, we have used X-ray crystallography to determine the crystal structure of N-Amy and used AlphaFold2 to model the Amy703 structure, respectively. We further used size exclusion chromatography to show that Amy703 existed as a monomer, whereas N-Amy formed a unique dimer. It was found that the N-terminus of one monomer of N-Amy was inserted into the catalytic domain of its symmetrical subunit, resulting in the expansion of the catalytic pocket. This also significantly increased the pKa of the hydrogen donor Glu350, thereby enhancing substrate binding affinity and contributing to increased N-Amy activity. Meanwhile, two calcium ions were found to bind to N-Amy at different binding sites, which also contributed to the stability of protein. Therefore, this study provided new structural insights into the mechanisms of various glycosyl hydrolases.

Evaluation of Heavy Metal Concentrations in Enzyme Embedded Poly Lactic Acid (EPLA) by ICP-OES: Lipase from Candida Rugosa.

Abstract: Enzymes like lipases are essential for catalyzing important reactions in the fields of biotechnology and industrial operations. For quality control and safety evaluations, however, the quantification of these contaminants is crucial because the presence of heavy metals in enzyme preparations can negatively affect their activity and stability. Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES), a key component of this investigation, is used to measure the levels of heavy metals in lipase enzyme preparations embedded in polylactic acid (PLA). The lipase enzyme being studied comes from Candida rugosa, which is well-known for its considerable industrial applications. According to preliminary findings, the lipase enzyme is successfully protected from external heavy metal pollutants by the encapsulating technique, maintaining its enzymatic activity and stability. The lipase-PLA composite had only minimal quantities of heavy metals, according to the ICP-OES study. The research's conclusions show great potential for the creation of reliable and contaminant-free lipase enzyme preparations, which will increase their suitability for usage in a variety of industrial processes and encourage the use of biodegradable polymers, supporting environmental sustainability. By highlighting the significance of quality control and safety assessment through the determination of heavy metal concentrations, this research contributes to the larger field of enzyme biotechnology. It also highlights the potential of Candida Rugosa lipase implanted in PLA matrices for eco-friendly and sustainable biocatalytic applications in sectors ranging from food and pharmaceuticals to biofuel generation and other biodegradable polymers.

Design of a Novel Lysine Isopeptide 1018KI11 with Potent Antimicrobial Activity as a Safe and Stable Food Preservative Candidate.

Antimicrobial peptides are potent food additive candidates, but most of them are sensitive to proteases, which limits their application. Therefore, we substituted arginine for lysine and introduced a lysine isopeptide bond to peptide IDR-1018 in order to improve its enzymatic stability. Subsequently, the protease stability and antimicrobial/antibiofilm activity of the novel peptides (1018K2-1018KI11) were investigated. The data revealed that the antienzymatic potential of 1018KI11 to bromelain and papain increased by 2-8 folds and 16 folds, respectively. The minimum inhibitory concentration (MIC) of 1018KI11 against methicillin-resistant Staphylococcus aureus (MRSA) ATCC43300 and Escherichia coli (E. coli) ATCC25922 was reduced 2-fold compared to 1018K11. Mechanism exploration suggested that 1018KI11 was more effective than 1018K11 in disrupting the cell barrier and damaging genomic DNA. Additionally, 1018KI11 at certain concentration conditions (2-64 μg/mL) reduced biofilm development of MRSA ATCC43300 by 4.9-85.9%. These data indicated that novel peptide 1018KI11 is a potential food preservative candidate.