Increasing Enzyme Concentration Research Articles

Cooperative DNA Binding and Protein/DNA Fiber Formation Increases the Activity of the Dnmt3a DNA Methyltransferase

The Dnmt3a DNA methyltransferase has been shown to bind cooperatively to DNA and to form large multimeric protein/DNA fibers. However, it has also been reported to methylate DNA in a processive manner, a property that is incompatible with protein/DNA fiber formation. We show here that the DNA methylation rate of Dnmt3a increases more than linearly with increasing enzyme concentration on a long DNA substrate, but not on a short 30-mer oligonucleotide substrate. We also show that addition of a catalytically inactive Dnmt3a mutant, which carries an amino acid exchange in the catalytic center, increases the DNA methylation rate by wild type Dnmt3a on the long substrate but not on the short one. In agreement with this finding, preincubation experiments indicate that stable protein/DNA fibers are formed on the long, but not on the short substrate. In addition, methylation experiments with substrates containing one or two CpG sites did not provide evidence for a processive mechanism over a wide range of enzyme concentrations. These data clearly indicate that Dnmt3a binds to DNA in a cooperative reaction and that the formation of stable protein/DNA fibers increases the DNA methylation rate. Fiber formation occurs at low μm concentrations of Dnmt3a, which are in the range of Dnmt3a concentrations in the nucleus of embryonic stem cells. Understanding the mechanism of Dnmt3a is of vital importance because Dnmt3a is a hotspot of somatic cancer mutations one of which has been implicated in changing Dnmt3a processivity.

Chain and pore-blocking effects on matrix degradation in protein-loaded microgels.

Factors affecting matrix degradation in protein-loaded microgels were investigated for dextran-based microgels, the sugar-binding protein Concanavalin A (ConA), and the dextran-degrading enzyme Dextranase. For this system, effects of enzyme, protein, and glucose concentrations, as well as pH, were considered. Microgel network degradation was monitored by micromanipulator-assisted light microscopy, whereas enzyme and protein distributions were monitored by confocal microscopy. Results show that Dextranase-mediated microgel degradation increased with increasing enzyme concentration, whereas an increased ConA loading in the dextran microgels caused a concentration-dependent decrease in microgel degradation. In the presence of glucose, competitive release of microgel-bound ConA restored the microgel degradation observed in the absence of ConA. To clarify effects of mass transport limitations, microgel degradation was compared to that of non-cross-linked dextran, demonstrating that ConA limits enzyme substrate access in dextran microgels primarily through pore blocking and induction of pore shrinkage. The experimentally observed effects were qualitatively captured by a modified Michaelis-Menten approach for spherical symmetry, in which network blocking by ConA was included. Taken together, the results demonstrate that matrix degradation of protein-loaded microgels depends sensitively on a number of factors, which need to be considered in the use of microgels in biomedical applications.

β-Galactosidase Immobilization on Chitosan-Hydroxyapatite Complex: Effects of Immobilization Conditions

In this study, ?-galactosidase from Kluyvermoyces lactis was immobilized onto chiosan-hydroxyapatite. Hydroxyapatite (HAP) is a nontoxic ceramic biomaterial generally that has been used for dental applications, drug delivery, immobilization of metals, enzymes and recently some yeast cells. Effects of immobilization parameters including medium temperature and pH, enzyme concentration and coupling time was evaluated to obtain the best reaction conditions. The optimal immobilization pH was determined as pH 7.5 with the highest enzyme activity of 1337.2 U g-1 chitosan-HAP beads and highest immobilization efficiency of 81.4%. Furthermore, the immobilized protein amount showed increase with increasing enzyme concentration as expected. However, highest specific activity (365.2 mg-1 protein) was achieved at low enzyme concentrations. Characterization of immobilized enzyme was tested using o-nitrophenol-?-D-galactopyranoside (ONPG) as a substrate. Immobilized enzyme exhibited better thermostability and higher stability at lower (pH 5.0) and higher (pH 9.0) pH values. Immobilization also resulted in a shift of 0.5 units in pH optimum for maximum enzyme activity. In contrast to optimum pH value, after immobilization no shift occurred in optimum temperature. Furthermore enzyme retained 81.8% of its initial activity during consecutive eight cycles. After 15 days of storage, immobilized enzyme obtained 82.6% of its original activity, whereas free enzyme showed a significant decrease.

UDP-glucose Dehydrogenase Activity and Optimal Downstream Cellular Function Require Dynamic Reorganization at the Dimer-Dimer Subunit Interfaces

UDP-glucose dehydrogenase (UGDH) provides precursors for steroid elimination, hyaluronan production, and glycosaminoglycan synthesis. The wild-type UGDH enzyme purifies in a hexamer-dimer equilibrium and transiently undergoes dynamic motion that exposes the dimer-dimer interface during catalysis. In the current study we created and characterized point mutations that yielded exclusively dimeric species (obligate dimer, T325D), dimeric species that could be induced to form hexamers in the ternary complex with substrate and cofactor (T325A), and a previously described exclusively hexameric species (UGDHΔ132) to investigate the role of quaternary structure in regulation of the enzyme. Characterization of the purified enzymes revealed a significant decrease in the enzymatic activity of the obligate dimer and hexamer mutants. Kinetic analysis of wild-type UGDH and the inducible hexamer, T325A, showed that upon increasing enzyme concentration, which favors the hexameric species, activity was modestly decreased and exhibited cooperativity. In contrast, cooperative kinetic behavior was not observed in the obligate dimer, T325D. These observations suggest that the regulation of the quaternary assembly of the enzyme is essential for optimal activity and allosteric regulation. Comparison of kinetic and thermal stability parameters revealed structurally dependent properties consistent with a role for controlled assembly and disassembly of the hexamer in the regulation of UGDH. Finally, both T325A and T325D mutants were significantly less efficient in promoting downstream hyaluronan production by HEK293 cells. These data support a model that requires an operational dimer-hexamer equilibrium to function efficiently and preserve regulated activity in the cell.

Chemical and biophysical properties of gelatins extracted from alkali-pretreated skin of cuttlefish (Sepia officinalis) using pepsin

Gelatins from alkali-pretreated skin of cuttlefish (Sepia officinalis) were extracted with different concentrations of pepsin at pH2.0 and characterized. The increase of enzyme concentration increased the yield of gelatin extraction. The yields of gelatins G5, G10 and G15, extracted, respectively, with 5, 10, and 15U of pepsin/g of alkali-treated skin of cuttlefish, were 6.59%, 7.69% and 9.22%, respectively, while in the absence of pepsin (G0) the yield was only 2.68%. Nevertheless, the extraction of gelatin from alkali-pretreated skin of cuttlefish with pepsin at pH2.0 affected the molecular weight, biophysical and functional properties of resulting gelatins. Gelatin extracted without pepsin showed the higher gel strength, and transition, gelling and melting temperatures, which are slightly higher than G5. All of these values decreased with increasing enzyme concentration. In addition, foam expansion (FE) was affected by enzymatic treatment, and the values decreased as pepsin increased. Nevertheless, the partially hydrolyzed gelatins showed the same emulsifying properties.

Enzymatic Hydrolysis of Gelatin Layers of X-Ray Films and Release of Silver Particles Using Keratinolytic Serine Proteases from Purpureocillium lilacinum LPS # 876

Enzymatic decomposition of gelatin layers on used X-ray films and repeated utilization of the enzyme for potential application in silver recovery were investigated using keratinolytic serine proteases from Purpureocillium lilacinum LPS # 876. At pH 9.0, the enzymatic reaction was enhanced by the increase of enzyme concentration or by the increase of the temperature up to 60℃. Under the conditions of 6.9 U/ml, 60℃, and pH 9.0, hydrolysis of the gelatin layers and the resulting release of silver particles were achieved within 6 min. The protective effect of polyols against thermal denaturation was investigated. The presence of glycerol and propylene glycol increased enzyme stability. When the reusability of the enzyme for gelatin hydrolysis was tested, it could be seen that it could be effectively reused for more cycles when glycerol was added, compared with the enzyme without protective agents. The results of these repeated treatments suggested that a continuous process of recycling silver from used X-ray is feasible. Keeping in mind that recycling is (at the present time) needed and imperative, it can be remarked that, in this research, three wastes were successfully used: hair waste in order to produce serine proteases; glycerol in order to enhance enzyme thermal stability; and used Xray films in order to recover silver and PET films.

Effect of an Error in the Forward Difference based tabular values of the Rate of Enzymatic Reaction based Mathematical Functions

In this research paper, we explained the effect of an error in the tabular values of substrate concentration, enzyme concentration and temperature with the rate of enzymatic reaction. All values are in interval. These values are obtained by mathematical functions of the rate of enzymatic reaction. Mathematical functions show that the concentration of substrate is the limiting factor, as the substrate concentration increases, the Enzyme reaction rate increases. Assuming a sufficient concentration of substrate is available, increasing Enzyme concentration will increase the rate of enzymatic reaction. Temperature, concentration of substrate and concentration of enzyme are increased the rate of enzymatic reaction at a limit which is called optimum limit. We take these values in tabular form with forward difference and added an error in the tabular values of substrate concentration, enzyme concentration and temperature with the rate of enzymatic reaction. Tabular values show the erroneous values. There are changes in tabular values. The effect of an error goes increasing with the succession. The coefficients of ? are the binomial coefficients with alternating signs. In various difference columns of the difference table, the algebraic sum of the error is zero. The maximum error occurs in the same line as the erroneous tabular value. This is show the effect of tabular values of substrate concentration, enzyme concentration and temperature with the rate of enzymatic reaction.

Two cold-induced family 19 glycosyl hydrolases from cherimoya (Annona cherimola) fruit: An antifungal chitinase and a cold-adapted chitinase

Two cold-induced chitinases were isolated and purified from the mesocarp cherimoyas (Annona cherimola Mill.) and they were characterised as acidic endochitinases with a Mr of 24.79 and 47.77kDa (AChi24 and AChi48, respectively), both family 19 glycosyl hydrolases. These purified chitinases differed significantly in their biochemical and biophysical properties. While both enzymes had similar optimal acidic pH values, AChi24 was enzymatically active and stable at alkaline pH values, as well as displaying an optimal temperature of 45°C and moderate thermostability. Kinetic studies revealed a great catalytic efficiency of AChi24 for oligomeric and polymeric substrates. Conversely, AChi48 hydrolysis showed positive co-operativity that was associated to a mixture of different functional oligomeric states through weak transient protein interactions. The rise in the AChi48 kcat at increasing enzyme concentrations provided evidence of its oligomerisation. AChi48 chitinase was active and stable in a broad acidic pH range, and while it was relatively labile as temperatures increased, with an optimal temperature of 35°C, it retained about 50% of its maximal activity from 5 to 50°C. Thermodynamic characterisation reflected the high kcat of AChi48 and the remarkably lower ΔH(‡), ΔS(‡) and ΔG(‡) values at 5°C compared to AChi24, indicating that the hydrolytic activity of AChi48 was less thermodependent. In vitro functional studies revealed that AChi24 had a strong antifungal defence potential against Botrytis cinerea, whereas they displayed no cryoprotective or antifreeze activity. Hence, based on biochemical, thermodynamic and functional data, this study demonstrates that two acidic endochitinases are induced at low temperatures in a subtropical fruit, and that one of them acts in an oligomeric cold-adapted manner.

Electrochemical RNase A Detection Using an Electrode with Immobilized Ferrocenyl Deoxyribooligonucleotide Containing Cytidine Residue

AbstractA ferrocenyl deoxyribooligonucleotide (FcODN(rC)) with contiguous cytosine bases and a single ribonucleotide, cytidine, was immobilized on a gold electrode, and this electrode was used to detect RNase A. RNase A activity in a solution was assessed using cyclic voltammetry, and it was found that the current response of the sensor electrode decreased with increasing enzyme concentration. An extremely low detection limit of 1.0×10−11 g mL−1 RNase A was observed, with 15–90 % changes in the current signal. RNase activity can be an indicator of a number of diseases; therefore, this probe has great potential for applications in medical diagnostics.

Degradation of barnacle nauplii: implications to chitin regulation in the marine environment

The exoskeleton of most invertebrate larval forms is made of chitin, which is a linear polysaccharide of β (1→4)-linked N-acetylglucosamine (GlcNAc) residues. These larval forms offer extensive body surface for bacterial attachment and colonization. In nature, degradation of chitin involves a cascade of processes brought about by chitinases produced by specific bacteria in the marine environment. Microbial decomposition of larval carcasses serves as an alternate mechanism for nutrient regeneration, elemental cycling and microbial production. The present study was undertaken to assess the influence of chitinase enzyme on the degradation of the nauplii of barnacle, Balanus amphitrite. The survival and abundance of bacteria during the degradation process under different experimental conditions was monitored. To the best of our knowledge, no such study is conducted to understand the degradation of larval exoskeleton using chitinase and its influence on bacteria. An increase in the chitinase activity with increase in temperature was observed. Scanning electron micrographs of chitinase treated nauplii showed scars on the surface of the barnacle nauplii initially and further disruption of the exoskeleton was observed with the increase in the treatment time. Bacterial abundance of the chitinase treated nauplii increased with the increase in enzyme concentration. Pathogenic bacteria such as Vibrio cholerae, V. alginolyticus, V. parahaemolyticus which were initially associated with the exoskeleton were absent after chitinase treatment, however, Bacillus spp. dominated subsequent to chitinase treatment and this might have important implications to marine ecosystem functioning.

Mathematical functions of the rate of enzymatic reaction with Temperature, concentration of substrate and concentration of enzyme are proved that the divide differences are symmetrical in all their arguments

In this research paper, I used mathematical functions of the rate of enzymatic reaction for proving the divide differences are symmetrical in all their argument. The concentration of substrate is the limiting factor, as the substrate concentration increases, the Enzyme reaction rate increases. Assuming a sufficient concentration of substrate is available, increasing Enzyme concentration will increase the enzymatic reaction rate. The rise in Temperature accelerates an Enzyme reaction but at the same time causes inactivation of the protein. At certain Temperature known as the optimum Temperature the activity is maximum. Temperature, concentration of substrate and concentration of enzyme are increased the rate of enzymatic reaction at a limit which is called optimum limit. I take their values in mathematical form where “n” is optimum limit.

Aldol addition of dihydroxyacetone to N-Cbz-3-aminopropanal catalyzed by two aldolases variants in microreactors

Aldol addition of dihydroxyacetone to N-Cbz-3-aminopropanal catalyzed by two d-fructose-6-phosphate aldolase variants, FSA A129S and FSA A129S/A165G, overexpressed in Escherichia coli was studied in microreactors. The presence of organic solvent was necessary due to poor solubility of N-Cbz-3-aminopropanal in water. Hence, three co-solvents were evaluated: ethyl acetate, acetonitrile and dimethylformamide (DMF). The influence of these solvents and their concentration on the enzyme activity was independently tested and it was found that all solvents significantly reduce the activity of FSA depending on their concentration. The reaction was carried out in three different microreactors; two without and one with micromixers. By increasing enzyme concentration, it was possible to achieve higher substrate conversion at lower residence time. Enzyme activity measured at the outlet flow of the microreactor at different residence time revealed that enzymes are more stable at lower residence times due to shorter time of exposure to organic solvent. The reaction in the batch reactor was compared with the results in microreactor with micromixers. Volume productivity was more than three fold higher in microreactor with micromixers than in the batch reactor for both aldolases. It was found to be 0.88Md−1 and 0.80Md−1 for FSA A129S and FSA A129S/A165G, respectively.

A novel process to efficiently form transglutaminase-set soy protein isolate-stabilized emulsion gels

A novel process to efficiently form soy protein isolate (SPI)-stabilized emulsion gels, induced by microbial transglutaminase (MTGase) was reported, with the enzyme added before the emulsification (instead of after the emulsification in the conventional case). The rheological behavior and microstructure of the formed emulsion gels (at a constant protein concentration of 6 g/100 mL and oil fractions (ø) of 0.2–0.6) were characterized using dynamic oscillatory measurement and confocal laser scanning microscope. The gel stiffness of the formed gels with the novel process progressively increased with increasing enzyme concentration (1.0–5.0 U/g), or with the ø increasing from 0.2 to 0.6. Interestingly, the gel stiffness at a given ø with the novel process was considerably greater than that with the conventional process, though the enzyme concentration was less (e.g., 1.0 vs 20 U/g). Furthermore, a pre-incubation (15–60 min) of the proteins with the enzyme (before the emulsification) greatly improved the gelation. The analyses of droplet size and amount of entrapped proteins within the gel matrix indicated that during the initial gel network, the bridging flocculation of oil droplets played a prominent role, but the full development of the gel stiffness mainly depended on the inter-droplet covalent cross-linking.

Competition between Anion Binding and Dimerization Modulates Staphylococcus aureus Phosphatidylinositol-specific Phospholipase C Enzymatic Activity

Bacterial phosphatidylinositol-specific phospholipase C targets PI and glycosylphosphatidylinositol-linked proteins of eukaryotic cells. Functional relevance of a homodimeric S. aureus PI-PLC crystal structure is supported by enzyme kinetics and mutagenesis. Nonsubstrate phosphatidylcholine increases activity by facilitating enzyme dimerization. Activating transient dimerization is antagonized by anions binding to a discrete site. Interplay of protein oligomerization and anion binding controls enzyme activity. Staphylococcus aureus phosphatidylinositol-specific phospholipase C (PI-PLC) is a secreted virulence factor for this pathogenic bacterium. A novel crystal structure shows that this PI-PLC can form a dimer via helix B, a structural feature present in all secreted, bacterial PI-PLCs that is important for membrane binding. Despite the small size of this interface, it is critical for optimal enzyme activity. Kinetic evidence, increased enzyme specific activity with increasing enzyme concentration, supports a mechanism where the PI-PLC dimerization is enhanced in membranes containing phosphatidylcholine (PC). Mutagenesis of key residues confirm that the zwitterionic phospholipid acts not by specific binding to the protein, but rather by reducing anionic lipid interactions with a cationic pocket on the surface of the S. aureus enzyme that stabilizes monomeric protein. Despite its structural and sequence similarity to PI-PLCs from other Gram-positive pathogenic bacteria, S. aureus PI-PLC appears to have a unique mechanism where enzyme activity is modulated by competition between binding of soluble anions or anionic lipids to the cationic sensor and transient dimerization on the membrane.

Structural Basis for Morpheein-type Allosteric Regulation of Escherichia coli Glucosamine-6-phosphate Synthase: EQUILIBRIUM BETWEEN INACTIVE HEXAMER AND ACTIVE DIMER

The amino-terminal cysteine of glucosamine-6-phosphate synthase (GlmS) acts as a nucleophile to release and transfer ammonia from glutamine to fructose 6-phosphate through a channel. The crystal structure of the C1A mutant of Escherichia coli GlmS, solved at 2.5 Å resolution, is organized as a hexamer, where the glutaminase domains adopt an inactive conformation. Although the wild-type enzyme is active as a dimer, size exclusion chromatography, dynamic and quasi-elastic light scattering, native polyacrylamide gel electrophoresis, and ultracentrifugation data show that the dimer is in equilibrium with a hexameric state, in vitro and in cellulo. The previously determined structures of the wild-type enzyme, alone or in complex with glucosamine 6-phosphate, are also consistent with a hexameric assembly that is catalytically inactive because the ammonia channel is not formed. The shift of the equilibrium toward the hexameric form in the presence of cyclic glucosamine 6-phosphate, together with the decrease of the specific activity with increasing enzyme concentration, strongly supports product inhibition through hexamer stabilization. Altogether, our data allow us to propose a morpheein model, in which the active dimer can rearrange into a transiently stable form, which has the propensity to form an inactive hexamer. This would account for a physiologically relevant allosteric regulation of E. coli GlmS. Finally, in addition to cyclic glucose 6-phosphate bound at the active site, the hexameric organization of E. coli GlmS enables the binding of another linear sugar molecule. Targeting this sugar-binding site to stabilize the inactive hexameric state is therefore suggested for the development of specific antibacterial inhibitors.

Effects of Enzymatic Hydrolysis of Protein on the Pasting Properties of Different Types of Wheat Flour

As one of the most effective methods to modify proteins, enzymatic hydrolysis is used widely in the preparation of wheat products in the food industry. During the same process, starch pasting occurs frequently. The effects of wheat protein hydrolysis with papain, pepsin, and trypsin on the pasting properties of 3 different kinds of flour were investigated in 5 concentrations. Results showed that the peak viscosity, trough, final, and integral area of pasting curve of these flours decreased with increasing enzymatic hydrolysis of protein, and decreased significantly with the increasing enzyme concentrations. Medium-gluten flour was the least sensitive to enzymatic activity and weak-gluten the most sensitive. Downtrends appeared with increasing papain and trypsin concentrations in the form of breakdown. Enzymes had no significant different effect on the peak times of strong- and medium-gluten flour, but prolonged peak time slightly in weak-gluten flour. The pasting time and temperature of strong- and medium-gluten flour were significantly increased in a concentration-dependent manner. However, there were no significant effects on the pasting times of weak-gluten flour. These results could supply a basis for utilization of enzymatic hydrolysis of wheat protein in food industry and for further studies into the interactions between hydrolyzed protein and starch in food or processing industries.

Conjugated polyelectrolyte complexes with single-walled carbon nanotubes for amperometric detection of glucose with inherent anti-interference properties

Supramolecular complexes between single-walled carbon nanotubes and the conjugated polyelectrolytes poly[3-(3-N,N-diethylaminopropoxy)thiophene] (PDAOT), poly[3-(3′-thienyloxy)propanesulfonate] (P3TOPS), and poly[2,5-bis(3-sulfonatopropoxy)-1,4-ethynylphenylene-alt-1,4-ethynylphenylene] (aPPE) were prepared in homogeneous aqueous solution, and mixed with the enzyme glucose oxidase. The resulting solutions were drop cast onto the surface of glassy carbon electrodes, producing uniform coatings that contained high concentrations of carbon nanotubes and the enzyme. It was found that electrochemical cells comprising the coated electrodes were responsive to the addition of glucose, resulting in biosensors having a detection limit of 5.0 μM and a linear dynamic range of 0.5 to 5.0 mM glucose. It was shown that sensitivity increased with increasing nanotube concentration up to 4 g L−1, and increasing enzyme concentration up to 20 g L−1. The high sensitivity of the nanotube-based sensors results from an effective increase in the surface area of the electrode as a result of contact with a percolation network of carbon nanotubes that is in close contact with glucose oxidase throughout the coating. By using the anionic conjugated polyelectrolyte aPPE in the coating, it was found that charge repulsion of anionic interferents such as uric acid and ascorbic acid could be achieved without the need for a Nafion overcoat. These results pave the way for development of new sensors to a variety of other analytes.

Effect of Cellulase Hydrolysis on Dyeing Properties of Linen

This paper investigated the effect of cellulase on dyeing properties of linen fabric. Acid cellulase of different concentrations was used for treating linen fabric with and without simulated mechanical agitation. After the cellulase treatment, linen fabrics were dyed to different depths, with two reactive dyes (one was bifunctional and the other was monofunctional). Results revealed that colour yield was reduced as shown by K/S Sum value in relation to the increased enzyme concentration and mechanical action. The X-ray diffraction analysis revealed that crystallinity ratio was increased with increase in amount of enzyme used in the treatment. Colourfastness to crocking and laundering were improved after enzyme treatment.

Enzymatic activation of cellulose acetate membrane for reducing of protein fouling

In this study, the surface of cellulose acetate (CA) ultrafiltration membrane was activated with serine protease (Savinase) enzyme to reduce protein fouling. Enzyme molecules were covalently immobilized with glutaraldehyde (cross-linking agent) onto the surface of CA membranes. The membrane activation was verified using filtration experiments and morphological analysis. Scanning electron microscopy (SEM) images and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy of the activated membrane when compared with raw membrane were confirmed that the enzyme was immobilized onto the membrane surface. The immobilization efficiencies changed from 13.2 to 41.2% according to the enzyme ratios from 2.5 to 10.0mg/mL. However, the permeability values decreased from 232±6 to 121±4L/m2hbar with increasing enzyme concentration from 2.5 to 10.0mg/mL. In fouling experiments, bovine serum albumin (BSA) was used as the protein model solution and activated sludge was used as the model biological sludge. Enzyme-activated membranes exhibited good filtration performances and protein rejection efficiencies were compared with raw CA membrane. Also the relative flux reduction (RFR) ratios of membranes were calculated as 97% and 88% for raw CA and enzyme-activated membranes (5mg/mL savinase), respectively. The membrane activated with Savinase enzyme could be proposed as a surface treatment method before filtration to mitigate protein fouling.

Recycling of Cellulosic Fibers by Enzymatic Process

In this research, enzymatic treatment as an environmental friendly process has been used for recycling process of old cellulosic wastes such as cotton, viscose, and lyocell. Cellulase hydrolyses cellulosic chains and shortens cellulosic fibers. This study investigates to detect the optimum enzyme concentration and time of treatments for suitable changes of length and weight loss. The main purposes of this article are shortening of cellulosic fibers and evaluating of enzymatic treatment in different kind of cellulosic fibers. According to the data of experiments, with the increase of enzyme concentration and the treatment time, the length and weight loss percentage of the cellulosic fibers has been decreased. The length and weight loss percentage of treated viscose is more than that of lyocell and cotton fibers. Optimized condition, reaction time, and enzyme concentration have been determined by mean length of treated cellulosic samples. Suitable longitudinal distribution of fiber for papermaking industries is in the range of 0 to 4 mm. Optimum enzyme concentration and treatment time for recycling cotton, lyocell, and viscose fibers are 2% and 48 h for cotton and lyocell and 0.5% and 48 h for viscose, respectively. According to the data of experiment, the length of treated fibers is appropriate for its usage as a raw material in papermaking industries.