Sephadex G-100 Column Chromatography Research Articles

Extraction Optimization, Composition Analysis, and Antioxidation Evaluation of Polysaccharides from White Jelly Mushroom, Tremella fuciformis (Tremellomycetes).

We investigated the sonication-assisted extraction (SAE) conditions of Tremella fuciformis polysaccharides. Single-factor experiments showed that the optimal sonication intensity was 6 W/cm2; the other SAE conditions were further optimized using response surface experimental design. A Box-Behnken experiment exhibited that the optimal SAE conditions were temperature, 85°C; ratio of liquid to solid, 46 mL/g; and sonication time, 2 hours. The maximal polysaccharide yield (8.95 g/100 g) was obtained under the optimal extraction conditions. Polysaccharides extracted by sonication or traditional hot water extraction (TWE) were further separated and purified using DEAE-Sepharose and Sephadex G-100 column chromatography. Polysaccharide fractions (SAE-1, SAE-2, TWE-1, and TWE-2) were obtained; their molecular weights were 9.85 × 104, 6.64 × 104, 4.75 × 105, and 2.81 × 105 Da, respectively. The total sugar content of SAE-1 and SAE-2 was lower and the protein and nucleic acid contents were higher than those of TWE-1 and TWE-2. In addition, SAE-1 and SAE-2 showed stronger Fe2+-chelating activities and superoxide radical scavenging activities than TWE-1 and TWE-2. This suggests that polysaccharides extracted by sonication possess high antioxidant activities.

Structural characterization and biological activities of a novel polysaccharide from Phyllanthus emblica.

A novel water-soluble polysaccharide named PEPW80-1, with a molecular mass of 4.7 kDa, was isolated from the pulp tissues of Phyllanthus emblica, and purified by sephadex G-100 column and sephacryl S-300 HR chromatography. The structural features of PEPW80-1 were investigated by a combination of acid hydrolysis, periodate oxidation-Smith degradation, methylation analysis, gas chromatography-mass spectrometry, scanning electron microscope, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectroscopy. The results showed that PEPW80-1 had a specific optical rotation of [α]25/D = +113° (c = 0.5 mg/mL) and its backbone composed of (1,3)-linked-β-L-rhamnose and (1,3,6)-linkage-β-D-galactose, with two branch chains of (1,4)-linked-α-D-galactose and (1,6)-linked-β-D-galactose and terminated with 1-α-L-arabinose. The antioxidant assays showed that PEPW80-1 possess 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity and hydroxyl radical-scavenging activity, enhancing reductive power. The results of immunomodulatory assays in vitro showed that PEPW80-1 could promote the proliferation of mouse splenocytes. Those proposed that PEPW80-1 might be developed as a potential value-added product with the activities of immunomodulator and free-radical inhibitors.

Preparation, characterisation, antioxidant and antiglycation activities of the novel polysaccharides from the pileus of Dictyophora rubrovolvata

SummaryThe polysaccharides from the pileus of Dictyophora rubrovolvata (DPP) were extracted by ultrasonic‐assisted extraction (UAE) and the operating parameters were optimised by orthogonal experiment. The polysaccharides were separated and purified by DEAE‐52 and Sephadex G‐100 column chromatography to obtain two fractions of DPP‐1s and DPP‐2s. The average of molecular weights, monosaccharide compositions, antioxidant and antiglycation activities of DPP‐1s and DPP‐2s were investigated. The results showed DPP‐1s and DPP‐2s had high inhibitory effects on oxidation and glycation, and DPP‐2s showed stronger bioactivity than DPP‐1s, which might be due to their differences in the molecular weight and uronic acid content. Furthermore, the UV and FTIR spectra were used to evaluate the characterisations of DPP‐2s, which demonstrated that DPP‐2s had no protein and nuclear acid with the pyranose ring structure. This study indicated that DPP‐2s could be used as a source of natural antioxidant and antiglycation components.

Structural elucidation and immunostimulatory activity of polysaccharide isolated by subcritical water extraction from Cordyceps militaris

Water-soluble polysaccharides were obtained from Cordyceps militaris (C. militaris) (CMP) by subcritical water extraction (SWE). Two polysaccharides fractions, CMP-W1 and CMP-S1, were isolated from CMP using DEAE-52 cellulose and Sephadex G-150 column chromatography. The structural characteristics of CMP-W1 and CMP-S1 were investigated. The results showed that the molecular weight of CMP-W1 and CMP-S1 are 3.66×105Da and 4.60×105Da, respectively, and both of them were heteropolysaccharides composed of d-mannose, d-glucose, d-galactose with the molar ratios of 2.84:1:1.29 and 2.05:1:1.09, respectively. FT-IR spectra analysis suggested that CMP-W1 and CMP-S1 belonged to pyranose form sugar and protein free. For immunostimulatory activity assay in vitro, CMP-W1 and CMP-S1 significantly promoted lymphatic spleen cell proliferation of mice. Therefore, the polysaccharides obtained from C. militaris by SWE can be used as potential natural immunostimulant in functional foods or medicine.

Purification, partial characterization and bioactivity of sulfated polysaccharides from Grateloupia livida

Purification, preliminary characterization and bioactivity of polysaccharides from Grateloupia livida (GL) were investigated. Three water-soluble sulfated polysaccharide fractions (GLP-1, GLP-2 and GLP-3) were isolated and purified from the edible and medicinal red seaweed, Grateloupia livida (Harv.) Yamada by DEAE Sepharose CL–6B and Sephadex G-100 column chromatography, and chemical characterization was performed by HPGPC, GC–MS, FT-IR and SEM. In addition, anticoagulant activities were determined by activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) using normal human plasma in vitro. The antioxidant activities against DPPH and ABTS+ radicals were evaluated and compared. The molecular weights of GLP-1, GLP-2 and GLP-3 were 39.5, 60.4 and 3.36kDa, respectively. Monosaccharide analysis revealed that three polysaccharide fractions were homopolysaccharides and comprised of galactose only. Anticoagulant assays indicated that crude GLP, and purified GLP-1 and GLP-2 effectively prolonged APTT and TT, but not PT. All polysaccharide fractions exhibited significant in vitro antioxidant activities in a dose-dependent manner. GLP-2 showed consistently better anticoagulant and antioxidant activities compared with GLP, GLP-1 and GLP-3. These results demonstrate that sulfated polysaccharides isolated from Grateloupia livida can serve as readily available alternative natural sources of anticoagulant and antioxidant agents.

In Vitro Antioxidant activities and anti-diabetic effect of a polysaccharide from Schisandra sphenanthera in rats with type 2 diabetes

A water soluble polysaccharide (SSPW1), with an average molecular weight of 191.18KD, was isolated and purified from the water extract of Schisandra sphenanthera by DEAE-52 and Sephadex G-100 column chromatography. SSPW1 contained 48.92% neutral sugar, 5.56% proteins and 42.83% uronic acid, and were composed of rhamnose, arabinose, mannose, galactose, glucose at a molar ratio of 13.52, 5.69, 3.92, 41.28 and 35.59. In vitro experiment showed that SSPW1 have a satisfactory scavenging effect on superoxide anion free radical, hydroxyl radical and DPPH free radical. In vivo experiment showed that SSPW1 could increase the body weight, improve the glucose tolerance, reduce FBG, and elevate the levels of FINS and the value of ISI in the diabetic rats. In addition, SSPW1 could reduce the MDA content, and increase GSH-PX, CAT and SOD activities. These results suggest that SSPW1 has both in vitro and in vivo antioxidant effect, which may be closely related to its antidiabetic effect.

Purification, characterization and in vitro antioxidant activities of polysaccharide from Chaenomeles speciosa

A water-soluble polysaccharide named as CSP-2 was isolated and purified from Chaenomeles speciosa by DEAE-Sepharose and Sephadex G-100 column chromatography. CSP-2 was composed of galactose (Gal), rhamnose (Rha), glucose (Glc) and xylose (Xyl) with a relative molar ratio of 3.8:1.6:1.2:0.4. The average molecular weight of CSP-2 was estimated to be about 4.6×104Da. The in vitro antioxidant assay showed that CSP-2 exhibited remarkable antioxidant activities to scavenge the DPPH radical, superoxide radical, hydroxyl radical and ABTS radical and significant cytoprotective effects on H2O2-induced PC12 cells injury in a concentration-dependent manner. Overall, CSP-2 may be useful as a naturally potential antioxidant agent for application in food and medicinal fields.

Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus

Boletus snicus (BS) is one of the commercially important mushroom species. Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. The average of molecular weight of BSP-1b and BSP-2b were 59.21kDa and 128.74kDa. BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. The antiglycation activities of BSP-1b and BSP-2b were investigated. The results showed BSP-1b and BSP-2b had high inhibitory effects on glycation and exhibited dose-dependent responses. BSP-2b showed stronger antiglycation activity than BSP-1b. This study indicated that the BSP-2b could effectively inhibit the formation of advanced glycation end-products.

Purification and characterization of low molecular weight alkaline xylanase from Neosartorya tatenoi KKU-CLB-3-2-4-1

A low molecular weight alkaline xylanase from Neosartorya tatenoi KKU-CLB-3-2-4-1 was purified to homogeneity by ammonium sulfate precipitation, Sephadex G-100 and DEAE-cellulose column chromatography. The xylanase was purified 32.09 fold, with the specific activity of 16.07 U mg−1 protein and a recovery yield of 2.67%. The purified xylanase was estimated by SDS-PAGE with a molecular weight of 20 kDa. The purified xylanase was active at pH 10.0 and retained over 75% of the original activity in pH range 7.0–11.0 after incubation at 4 °C for 24 h. The optimum temperature of the purified xylanase was 45 °C. As for thermo-stability, more than 80% of original xylanase activity was retained after heating at 45 °C for 120 min. Xylanase activity was stimulated by Ag+ and inhibited by Hg2+. The purified xylanase was specific to beechwood xylan, birchwood xylan and oat spelt xylan. The Km and Vmax values for beechwood xylan were 10.34 mg/mL and 45.66 μmol/min/mg, respectively. In addition, the xylanase hydrolyzed beechwood xylan yielded mainly xylotriose and xylotetraose as end products, suggesting it was an endo-xylanase. Thus, its properties may make it attractive for application in the pulp and paper industry.

Isolation and anti-Verticillium dahliae activity from Bacillus axarquiensis TUBP1 protein

Anti-Verticillium dahliae proteins were purified by applying (NH4)2SO4 (70%) precipitation, DEAE Sepharose Fast Flow (DEAE FF), and Sephadex G-100 column chromatography to analyze the anti-V. dahliae proteins produced by Bacillus axarquiensis TUBP1. The anti-V. dahliae fraction was identified by using LC–MS/MS. The relative molecular mass and isoelectric point (pI) of purified anti-V. dahliae protein were 6.22kDa and 4.82, respectively. We obtained the amino acid sequences of four peptide segments, among them, two novels, whereas the other two were similar to peptidase T(tr154685475). Our results showed that the anti-V. dahliae protein from B. axarquiensis TUBP1 possessed good thermal stability, and that an alkaline condition was conducive for the anti-V. dahliae activity of purified proteins. Scanning electron microscopy (SEM) experiments showed that protein-treated V. dahliae exhibited anomalies in the morphology of the fungal hyphae and spores. Transmission electron microscopy (TEM) experiments suggested that the plasma membrane of V. dahliae may be main target of the antifungal protein from B. axarquiensis TUBP1. The strong anti-V. dahliae activity of B. axarquiensis TUBP1 implied that its components might provide an alternative source of the biocontrol of Verticillium wilt.

Purification and characterization of a surfactant-compatible lipase from Aspergillus tamarii JGIF06 exhibiting energy-efficient removal of oil stains from polycotton fabric.

An extracellular lipase with 23,666.66 U/ml/min activity was produced by Aspergillus tamarii JGIF06 under submerged fermentation in mineral salt medium containing coconut oil (2.5 % v/v), tryptone (2 % w/v) and ammonium chloride (2 % w/v), with initial pH of 5 ± 0.2, incubated at 25 °C for 7 days on a rotary shaker at 120 rpm. A 7.9-fold increase in lipase-specific activity was recorded after purification by DEAE Sepharose ion exchange and Sephadex G200 column chromatography. The apparent molecular mass of this enzyme was revealed as 50 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The optimal lipase activity was recorded at pH 4 and 37 °C. The enzyme revealed broad specificity towards different vegetable oils. The K m and V max of the lipase on olive oil was found to be 330.4 mg and 53,690 U/ml/min, respectively. The lipase activity was stable in the presence of surfactants such as cetrimonium bromide, sodium dodecyl sulphate and Tween 80, and metal ions and reagents such as Ca2+, Ba2+ and 2-mercaptoethanol. However, the activity was greatly reduced in the presence of organic solvents such as chloroform. The stain removal potential of the crude lipase was determined on polycotton fabric pieces stained with peanut oil. Lipase added to cold water alone significantly enhanced the removal of stain by 152 %. The addition of lipase also improved the stain removal efficiency of a commercially available detergent in the presence of either cold (25 ± 2 °C) or hot (65 ± 2 °C) water. The current findings suggest the potentiality of this enzyme for energy-efficient biocatalytic application.

Purification and characterization of a melanin biodegradation enzyme from Geotrichum sp.

Melanin is a black or brown phenolic polymer present mainly in skin and hair. Although melanin can be degraded by some microbial species, the melanin degradation capacity of Geotrichum sp. is unknown. The aim of this study was to characterize a melanin biodegradation enzyme from Geotrichum sp. In this study, we assessed the melanin degradation activity of Geotrichum sp. in comparison with the major melanin-degrading enzymes, manganese-dependent peroxidase (MnP), manganese-independent peroxidase, lignin peroxidase and laccase. Furthermore, the effect of several carbohydrates on melanin degradation by Geotrichum sp. was determined. The MnP enzyme was purified using ammonium sulphate precipitation and Sephadex G-200 column chromatography, and then the conditions for optimal enzymatic activity were determined by adjusting the pH, temperature and Tween-80 concentration. Compared with extracellular ligninolytic enzymes of Geotrichum sp., MnP had the highest ligninolytic enzyme activity; and the highest enzymatic activity was observed in the presence of glucose. The final purified MnP enzyme exhibited 6 U mL-1 activity and had a molecular weight of 54.2 kDa. The enzymatic activity was highest at pH 4.5 and 25-35°C in the absence of Tween-80. These results indicate the potential of MnP purified from Geotrichum sp. as a skin-lightening agent in the cosmetic industry.

Extraction of polysaccharides and its antitumor activity on Magnolia kwangsiensis Figlar & Noot.

The crude polysaccharides from the leaves of Magnolia kwangsiensis Figlar & Noot. were extracted by hot water extraction, the yield was 5.09%. Two polysaccharide fractions (P-2 and P-3) were isolated by DEAE-52 cellulose chromatography and Sephadex G-100 column chromatography in order, respectively. P-2 and P-3 were characterized by IR, HPGPC and GC–MS. P-2 was comprised of only glucose. Its molecular weight was 11.2×103Da and the formula was C414H690O345. P-3 was comprised of xylose and rhamnose in the ratio of 1:4. Its molecular weight was 7.8×103Da, and the formula was C319H528O220. The antitumor activities of P-2 and P-3 on the growth of human lung cancer (A549) cells and human gastric carcinoma (SGC7901) cells in vitro were evaluated. The results indicated that P-3 exhibited marked antitumor activities with IC50 value of 8.48 and 5.66μg/mL.

Ultrasound-assisted extraction of polysaccharides from Artemisia selengensis Turcz and its antioxidant and anticancer activities.

Artemisia selengensis Turcz (AST) is a perennial herb with therapeutic and economic applications in China. The effects of ultrasound-assisted extraction (UAE) parameters upon extraction yield (EY%), antioxidant and antitumor activities of the polysaccharides extracts were studied by using a factorial design and response surface methodology. The optimal conditions determined were as: ultrasonic power 146W, extraction time 14.5min. and extraction temperature 60°C. The average molecular weights of two homogeneous polysaccharides (APS1 and APS2) purified by DEAE cellulose-52 and Sephadex G-100 column chromatography were 125.4 and 184.1kDa, respectively. Monosaccharide analysis showed that APS1 and APS2 were composed of five common monomers i.e., galactose, mannose, arabinose, xylose and rhamnose and one different monomer glucose and galacturonic acid respectively, with a most abundant part in molar % of APS1 and APS2 were glucose (83.01%) and galacturonic acid (48.87%) while least were xylose (0.80%) and mannose (1.73%) respectively. The antioxidant properties were determined by evaluating DPPH, hydroxyl radical scavenging activity and reducing power which indicated both APS1 and APS2 showed strong scavenging activities and anticancer activities on HT-29, BGC823 and antitumor activity on HepG-2. As UAE improved the polysaccharides yield than CSE, meanwhile, no significant difference of polysaccharides chemical compositions. Therefore, the present study suggests that the consumption of AST leaves may beneficial for the treatment of many diseases.

Purification and characterization of a novel thermo stable L-methioninase from Streptomyces sp. DMMMH4 and its evaluation for anticancer activity

L-methioninase has been purified 2.55-fold from the crude extract of Streptomyces sp. DMMMH4. The purification procedure was carried out by heat treatment and gel filtration on Sephadex G-200 column chromatography. SDS-PAGE electrophoresis showed a migrating protein band molecular mass of 48 kDa. The kinetic properties determined for the purified enzyme displayed optimum activity at 70OC and thermal stability was 70OC for 30 min. The enzyme activity showed maximum activity at pH 6 using acetate buffer 0.05M and was relatively stable across a broad range of pH values (5.5-8 pH). The enzyme strongly inhibited by Cr+2, Fe+2, Ni+2, Cd+2, PMSF, β-mercaptoethanol and SDS while Hg+2,Cu+2 and iodoacetate completely inhibited the enzyme activity at a final concentration of 10mM. The purified enzyme exhibited a Km of 0.7, 0.15 and 0.25 mM for L-methionine, DL-ethionine and L-cystine respectively. Cytotoxicity test demonstrate that enzyme was active against liver HepG2, breast MCF-7, lung A549, prostate PC3 and colon HCT116 cancer cell lines and has negligible toxicity toward a normal melanocyte cell line HFB4 .

Ultrasonic extraction, antioxidant and anticancer activities of novel polysaccharides from Chuanxiong rhizome

Ultrasonic-assisted extraction technology was employed to prepare Ligusticum chuanxiong Hort polysaccharide. Single factor test and orthogonal experimental design were used to optimize the extraction conditions. The results showed that the optimal extraction conditions consisted of ultrasonic temperature of 80°C, ultrasonic time of 40min and water to raw material ratio of 30mL/g. Three novel polysaccharides fractions, LCX0, LCX1 and LCX2, were isolated and purified from the crude polysaccharides using DEAE-52 cellulose and Sephadex G-100 column chromatography. The molecular weight and monosaccharide composition of three LCX polysaccharides fractions were analyzed with gel permeation chromatography (GPC) and HPLC analysis, respectively. Furthermore, the antioxidant and in vitro anticancer activities of the polysaccharides were investigated. Compared with LCX0, LCX2 and LCX1 showed relative higher antioxidant activity and inhibitory activity to the growth of HepG2, SMMC7721, A549 and HCT-116 cells. It is suggested that the novel polysaccharides from rhizome of L. chuanxiong could be promising bioactive macromolecules for biomedical use.

Novel functional polysaccharides from Radix Polygoni Multiflori water extracted residue: Preliminary characterization and immunomodulatory activity

The alkali-extractable polysaccharides (APMPs) were isolated from the water extracted residues of Radix Polygoni Multiflori, and further purified by DEAE-52 cellulose and Sephadex G-100 column chromatography to obtain a homogeneous polysaccharide (APMP-2) with molecular weights of 7724.8Da. HPLC chromatography analysis identified that APMP-2 was a heteropolysaccharides and mainly composed of Galactose and Xylose with a molar ratio of 4.31: 1.06. It was shown that both APMP and APMP-2 were of activation effects on splenocytes and peritoneal macrophages, and also significantly restore the proliferation rate, phagocytic index and cytokine (IL-2 and TNF-α) production level of 5-FU-treated splenocytes/peritoneal macrophages in a dosage-dependent manner. The results suggested that polysaccharides presented in Radix Polygoni Multiflori water-extracted residues possessed immunomodulatory activity and could be used as potential immunomodulators, and this finding could be a reference for the utilization of Radix Polygoni Multiflori water extracted residues.

Decolorization of biomethanated distillery effluent by immobilized enzymes

The massive quantity of colored distillery wastewater called molasses spent wash (MSW), if disposed untreated can cause considerable stress on the water sources leading to widespread damage to aquatic life. Microbial decolourization is an environment-friendly technique for removing this color.Recent research has focused on the development of enzymatic processes for the treatment of wastewaters. Enzymes like peroxidases, oxid oreductases, cellulolytic enzymes cyanidase, proteases, amylases, etc.from variety of different plants and microorganisms have been reported to play an important role in waste treatment. Practical problems, such as the high-cost isolation, purification, instability in organic media and high temperatures, restrict their use. In present work, the possibility of using immobilized enzyme to enhance the biodegradability of the distillery-spent wash has been investigated. Four bacterial strains showing decolorization of 30% Biomethanated wastewater were isolated from the sites where distillery effluent being discharged. The extracellular enzyme responsible for decolorization was purified, using a series of purification steps that included ammonium sulphate precipitation and gel permeation using sephadex G-100 column chromatography. The purified enzymes from each isolate were with a molecular mass of 68.7 kDa and were capable of 61.8%decolorization of 60% effluent. Alginate immobilization of these enzymes led to 62.97% decolorization after 120 hours. Re usability of the immobilized enzyme showed that it could be used with similar efficiency for 3 cycles and thereby it decreased. This data can be referred to in developing a technology for the removal of a specific colorant from effluent in future.

Extraction, purification, characterization and antioxidant activity of polysaccharides from Piteguo fruit

Piteguo fruit (PTG) picked from the trees growing at a high altitude of 2100–2400m in Gansu Province, China, is rich in a variety of nutrients, especially the polysaccharide. The optimum conditions for ultrasonic-assisted extraction Piteguo polysaccharide (PTGP) were investigated by response surface methodology (RSM), including ratio of water to raw material, extraction time, extraction temperature and ultrasonic power. The result was shown as follow, ratio of water to raw material at 13:1mL/g, extraction time of 66min, extraction temperature at 70°C, ultrasonic power at 230W, and the yield was 5.16%±1.81%. While the predicted yield of PTGP was 5.25% under the optimum conditions. Two homogenous polysaccharide (PTGP1 and PTGP2) were obtained by DEAE-cellulose-52 and Sephadex G-100 column chromatography. PTGP1 and PTGP2 were confirmed to be composed with three and two different kinds of polymers by HPSEC-LLS, respectively. The monosaccharide composition detected by GC–MS indicated that PTGP1 was a heteropolysaccharide consisting of rhamnose, arabinose, xylose, mannose, glucose and galactose with the molar ratio of 3.06:2.32:1.56:4.87:3.28:2.18, however mannose has not been detected in PTGP2. Evaluation of the antioxidant activity in vitro suggested that PTGP1 and PTGP2 had significant scavenging activity of hydroxyl, superoxide anion and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. Overall, the polysaccharides from Piteguo Fruit may have potential applications in the medical and food industries.

Characterization of a Solvent-Tolerant Manganese Peroxidase (MnP) from G anoderma Lucidum and Its Application in Fruit Juice Clarification

Manganese peroxidase (MnP) is necessary in fruit juice clarification for the degradation of phenolic compounds. In the present study, MnP was produced using basal medium supplemented with tamarind shell (1% w/v), ethanol (2% v/v) and gallic acid (1 mM) to a maximum activity of 65 U/mL. A 43.0 kDa MnP was purified by ammonium sulfate fractionation, Sephadex G-100 and DEAE-cellulose column chromatography with 6.5-fold and 13% yield. The optimal activity of the purified MnP was achieved at pH 5.5 and temperature of 45C with 2,6-dimethoxyphenol as substrate. The inhibitors sodium azide, cyanide and EDTA inhibited the MnP activity up to 100, 93 and 84%, respectively. MnP showed high activity and stability in the presence of different metal ions, surfactants and organic solvents. Clarification of different fruit juices with MnP showed its potential application in the food and beverage industry. Practical Applications Manganese peroxidase (MnP) is extensively used in various applications, such as pulp delignification, dye decoloration, biotransformation, biosensor applications and fruit juice clarification. Ganoderma lucidum-derived purified MnP is a solvent-tolerant metalloenzyme that showed its potential for industrial application in clarification of various fruit juices. The stability of MnP in the presence of various divalent cations, surfactants and solvents suggests that it could be used in industrial effluent treatments as well as the food and beverage industry.