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1.
This paper reports the first isolation from cultures of two endoxylanases secreted by Fusarium graminearum Schwabe [teleomorph Gibberella zeae (Schweinitz) Petch]. When F. graminearum is grown on wheat bran hydrated with a modified synthetic medium, high xylanase activity can be extracted. The two endoxylanases were identified by LC-MS/MS as the products of genes FGSG_6445 (Genbank gene id 2788192 ) (xylanase 1) and FGSG_3624 (GenBank accession no. AJ863566 ) (xylanase 2) with 61 and 51% sequence coverage, respectively. Both enzymes showed a pH optimum at pH 6, with xylanase 1 exhibiting a wider active pH range (5.5-9) than xlylanase 2 (5.5-7.5). Their temperature dependences were similar, >60% between 35 and 60 °C, with optimal temperatures of 45 °C for xylanase 1 and 50 °C for xylanase 2. Kinetic studies found that both enzymes had a lower K(m) for linear beachwood xylan than arabinoxylan. For xylanase 2, the V(max) increased with arabinoxylan, but decreased for xylanase 1.  相似文献   

2.
On the basis of glycine-dimethyl phosphite synthesis of glyphosate, a new synthesis approach of glyphosate, without using triethylamine and the establishment of triethylamine recovery equipment, was designed in the laboratory. The environmental pollutants were reduced. The influences of reactant amount (mol), reaction temperature (°C), and reaction time (min) on the glyphosate yield and purity were investigated. The results showed that the glyphosate yield and purity could be 80.12 and 86.31 wt %, respectively, under the optimum scheme for glyphosate yield (glycine consumption, 0.1 mol; dimethyl phosphite consumption, 0.12 mol; condensation reaction temperature, 50 °C; hydrochloric acid consumption in hydrolysis reaction, 0.35 mol; temperature of acidification with hydrochloric acid, 10 °C; adjusting hydrolysis product pH value, 1.0; time of dropping esterifying liquid into hydrochloric acid in hydrolysis reaction, 80 min; hydrolysis reaction temperature, 120 °C; and vacuum distillation time, 90 min), and the glyphosate yield and purity could be 77.92 and 94.94 wt %, respectively, under the optimum scheme for glyphosate purity (glycine consumption, 0.1 mol; dimethyl phosphite consumption, 0.1 mol; condensation reaction temperature, 50 °C; hydrochloric acid consumption in hydrolysis reaction, 0.35 mol; temperature of acidification with hydrochloric acid, 10 °C; adjusting hydrolysis product pH value, 1.5; time of dropping esterifying liquid into hydrochloric acid in hydrolysis reaction, 60 min; hydrolysis reaction temperature, 110 °C; and vacuum distillation time, 90 min). The product structures under the two schemes were confirmed by means of FTIR (Fourier transform infrared spectroscopy) and (1)H NMR ((1)H nuclear magnetic resonance spectroscopy).  相似文献   

3.
Polyphenol oxidase (PPO) of cauliflower was purified to 282-fold with a recovery rate of 8.1%, using phloroglucinol as a substrate. The enzyme appeared as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The estimated molecular weight of the enzyme was 60 and 54 kDa by SDS-PAGE and gel filtration, respectively. The purified enzyme, called phloroglucinol oxidase (PhO), oxidized phloroglucinol (K(m) = 3.3 mM) and phloroglucinolcarboxylic acid. The enzyme also had peroxidase (POD) activity. At the final step, the activity of purified cauliflower POD was 110-fold with a recovery rate of 3.2%. The PhO and POD showed the highest activity at pH 8.0 and 4.0 and were stable in the pH range of 3.0-11.0 and 5.0-8.0 at 5 °C for 20 h, respectively. The optimum temperature was 55 °C for PhO and 20 °C for POD. The most effective inhibitor for PhO was sodium diethyldithiocarbamate at 10 mM (IC(50) = 0.64 and K(i) = 0.15 mM), and the most effective inhibitor for POD was potassium cyanide at 1.0 mM (IC(50) = 0.03 and K(i) = 29 μM).  相似文献   

4.
Bombyx mori silk fibroin is a protein-based macromolecular biopolymer with remarkable biocompatibility. Silk fiber was degummed and subjected to a series of treatments, including dissolution and dialysis, to yield an aqueous solution of silk fibroin, which was introduced rapidly into excess acetone to produce crystalline silk fibroin nanoparticles (SFNs). The SFNs were conjugated covalently with a neutral protease (NP) using glutaraldehyde as the cross-linking reagent. The objective of this study was to determine the optimal conditions for biosynthesis of the SFN-NP bioconjugates. First, SFN-NP was obtained by covalent cross-linking of SFN and NP at an SFN/NP ratio of 5-8 mg:1 IU with 0.75% glutaraldehyde for 6 h at 25 °C. When adding 50 IU of the enzyme, the residual activity of biological conjugates was increased to 31.45%. Studies on the enzyme activity of SFN-NP and its kinetics showed that the stability of SFN-NP bioconjugates was greater than that of the free enzyme, the optimum reactive temperature range was increased by 5-10 °C, and the optimum pH value range was increased to 6.5-8.0. Furthermore, the thermal stability was improved to some extent. A controlled hydrolysis test using the poorly water-soluble protein sericin as a substrate and SFN-NP as the enzyme showed that the longer the reaction time (within 1 h), the smaller the molecular mass (<30 kDa) is of the sericin peptide produced. The SFN-NP bioconjugate is easily recovered by centrifugation and can be used repeatedly. The highly efficient processing technology and the use of SFN as a novel vector for a protease has great potential for research and the development of food processing.  相似文献   

5.
Temperature and pH influence the conformation of the whey protein β-lactoglobulin (β-Lg) monomer, dimer, and octamer formation, its denaturation, and solubility. Most hydrolyses have been reported at trypsin (EC 3.4.21.4) optimum conditions (pH 7.8 and 37 °C), while the hydrolysate mass spectrometry was largely limited to peptides with <4 kDa. There are few reports on trypsin peptide release patterns away from optimum. This work investigated the influence of alkaline (8.65 and 9.5) and optimum (7.8) pH at different temperatures (25, 37.5, and 50 °C) on β-Lg (7.5%, w/v) hydrolysis. Sample aliquots were drawn out before the addition of trypsin (blank sample) and at various time intervals (15 s to 10 min) thereafter. Matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS/MS) was used to monitor peptide evolution over time with the use of two matrixes: α-cyano-4-hydroxycinnamic acid (HCCA) and 2.5-dihydroxyacetophenone (DHAP). Mass analysis showed that the N- and C-terminals (Lys(8)-Gly(9), Lys(100)-Lys(101), Arg(124)-Thr(125), Lys(141)-Ala(142), and Arg(148)-Leu(149)) of β-Lg were cleaved early (15 s) implying the ease of trypsinolysis at the exposed terminals. Hydrolyses at 25 °C and pH 7.8 as well as at 50 °C and pH 9.5 were slowed down and ordered. Nonspecific chymotrypsin-like behavior occurred more at higher temperatures (50 °C) than at lower ones (25 and 37.5 °C). In addition to our earlier work in the acid pH region, it can be concluded that there is potential for controlled hydrolysis outside the trypsin optimum, where different target peptides with predictable biofunctionalities could be produced.  相似文献   

6.
An enzyme having activity toward n-hexanol was purified from apple, and its biochemical characteristics were analyzed. The purification steps consisted of sedimentation with ammonium sulfate, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephadex G-100 column. The obtained enzyme had a yield of 16.00% with a specific activity of 18879.20 U/mg protein and overall purification of 142.77-fold. The enzyme showed activity to isoamylol, 1-propanol, n-hexanol, and isobutanol but not toward methanol and ethanol. With n-hexanol as a substrate, the optimum conditions were pH 4.0 and 30 °C for enzyme activity and pH 3.0-4.0 and temperatures below 40 °C for enzyme stability. The enzyme activity was increased significantly by adding l-cysteine and Fe(2+) at all tested concentrations and slightly by Zn(2+) at a high concentration but decreased by additions of EDTA, Ga(2+), K(+), Mg(2+), sodium dodecyl sulfate (SDS), sodium aluminum sulfate (SAS), dithiothreitol (DTT), and glutathione (GSH). The enzyme activities toward n-hexanol and n-hexanal were increased by NADH but decreased by NAD(+), in contrast to a decrease toward n-hexane by addition of both NAD(+) and NADH.  相似文献   

7.
Subtilisin was immobilized on polycaprolactam and used for food packaging applications to reduce the transference of microorganisms from the packaging material to the packaged food material. The optimized conditions for subtilisin immobilization was as follows: pH, 8; temperature, 4 °C; glutaraldehyde, 0.5%; incubation time, 25 h; and subtilisin concentration, 600 μL. The formation of -CH═N- at 1576 cm(-1) in the Fourier transform infrared (FTIR) spectrum confirmed the immobilization. Subtilisin-immobilized polycaprolactam (SIP) exhibited the highest residual activity of 106.67 ± 4.41% and 104.67 ± 0.88% at 40 °C and pH 8 and retained residual activity of 94% at the end of 56 days when compared to 21.33 ± 4.10% in the case of free subtilisin. SIP significantly (p < 0.05) lowered the colony forming units (CFU), dry weight, and protein and carbohydrate contents in bacterial and fungal biofilm. Practical application of the SIP on ham steaks at 4 and 20 °C showed a 2-3 times reduction of Staphylococcus aureus as well as Escherichia coli cells in the range of p < 0.05.  相似文献   

8.
1,3-Dioleoyl-2-palmitoylglycerol, an important triacylglycerol in infant formulas, was effectively enriched by a two-step process: (a) dry fractionation of leaf lard and (b) enzymatic acidolysis of the fractionated leaf lard. In step a, the 1,3-dioleoyl-2-palmitoylglycerol content was increased from 16.77 to 30.73% after programmed temperature treatment of the leaf lard at 60 °C for 20 min followed by 34 °C for 10 h. In step b, 43.72% of the 1,3-dioleoyl-2-palmitoylglycerol content was obtained at the optimal conditions of enzymatic acidolysis: a substrate molar ratio of 1:4 (the fractionated leaf lard/camellia oil fatty acids), 6% (w/w) of enzyme loading, and 6 h of reaction time at 45 °C. On the basis of gas chromatography determination and "deducting score" principle, a model was properly established for characterizing the quality of triacylglycerols enriched with 1,3-dioleoyl-2-palmitoylglycerol. This approach would be a valuable contribution in structured lipids industries because only gas chromatography determination was involved.  相似文献   

9.
Soil from a field under long-term grass was treated with 0.02m sodium periodate for various periods up to 1176 h, followed by 0.1 d sodium tetraborate for 6 h. This destroyed an increasing proportion of microaggregates >45 μm and carbohydrate. After periodate treatment for 6 h about 70% of the soil sugars remained in the residue as measured by reducing sugar content and about 67% as individual sugars measured by gas-liquid chromatography. After 48 h the reducing sugar content was about 45%. An inverse linear relationship was established between the proportion by weight of microaggregates >45 μm and residual carbohydrate. The residual carbohydrate showed an enrichment in sugars commonly found in plant materials; glucose, arabinose and xylose, suggesting that the microbial carbohydrate had been preferentially destroyed. When the concentration of the periodate was increased to 0.05 m the residue contained about 50% of the original carbohydrate after 6 h treatment, and 25% after 48 h and an additional increase of about 10% in the proportion by weight of particles in the <45 μm range. These results throw doubt on the validity of assumptions made in a number of studies about the limited extent to which soil polysaccharide is involved in aggregation.  相似文献   

10.
The purification and partial enzymology characteristics of polyphenol oxidase (PPO) from rape flower were studied. After preliminary treatments, the crude enzyme solution was in turn purified with ammonium sulfate, dialysis, and Sephadex G-75 gel chromatography. The optimal conditions and stability of PPO were examined at different pH values and temperatures. Subsequently, PPO was also characterized by substrate (catechol) concentrations, inhibitors, kinetic parameters, and molecular weight. Results showed that the optimal pH for PPO activity was 5.5 in the presence of catechol and that PPO was relatively stable at pH 3.5-5.5. PPO was moderately stable at temperatures from 60 to 70 °C, whereas it was easily denatured at 80-90 °C. Ethylenediaminetetraacetic acid, sodium chloride, and calcium chloride had little inhibitive effects on PPO, whereas citric acid, sodium sulfite, and ascorbic acid had strongly inhibitive effects. The Michaelis-Menten constant (K(m)) and maximal reaction velocity (V(max)) of PPO were 0.767 mol/L and 0.519 Ab/min/mL of the crude PPO solution, respectively. PPO was finally purified to homogeneity with a purification factor of 4.41-fold and a recovery of 12.41%. Its molecular weight was 60.4 kDa, indicating that the PPO is a dimer. The data obtained in this research may help to prevent the enzymatic browning of rape flower during its storage and processing.  相似文献   

11.
The noncharacterized protein ACL75304 encoded by the gene Ccel_0941 from Clostridium cellulolyticum H10 (ATCC 35319), previously proposed as the xylose isomerase domain protein TIM barrel, was cloned and expressed in Escherichia coli . The expressed enzyme was purified by nickel-affinity chromatography with electrophoretic homogeneity and then characterized as d-psicose 3-epimerase. The enzyme was strictly metal-dependent and showed a maximal activity in the presence of Co(2+). The optimum pH and temperature for enzyme activity were 55 °C and pH 8.0. The half-lives for the enzyme at 60 °C were 6.8 h and 10 min when incubated with and without Co(2+), respectively, suggesting that this enzyme was extremely thermostable in the presence of Co(2+) but readily inactivated without metal ion. The Michaelis-Menten constant (K(m)), turnover number (k(cat)), and catalytic efficiency (k(cat)/K(m)) values of the enzyme for substrate d-psicose were estimated to be 17.4 mM, 3243.4 min(-1), and 186.4 mM min(-1), respectively. The enzyme carried out the epimerization of d-fructose to d-psicose with a conversion yield of 32% under optimal conditions, suggesting that the enzyme is a potential d-psicose producer.  相似文献   

12.
Wine lactone (i.e., 3a,4,5,7a-tetrahydro-3,6-dimethylbenzofuran-2(3H)-one, 1a/1b) was formed hydrolytically at wine pH from both racemic (E)-2,6-dimethyl-6-hydroxyocta-2,7-dienoic acid (3) and the corresponding glucose ester 2a at 45 °C but at room temperature was only formed from the acid 3. The glucose ester does not appear to be a significant precursor for the formation of wine lactone in wine. The slow formation of wine lactone from the free acid 3 indicates that the acid is not likely to be an important precursor to wine lactone in young wines unless present in high concentration (? 1 mg/L), but could be a significant precursor to wine lactone in wine that is several years old. The wine lactone formed in hydrolysates of the (6R)-enantiomer of 3 was partially enriched in the (3S,3aS,7aR)-enantiomer 1a when the hydrolysis was conducted at pH 3.2 and 100 °C in a closed vessel or under simultaneous distillation-extraction (SDE) conditions, and the enantiomeric excess (ee) varied from 5 to 22%. Hydrolysis of (6R)-3 in sealed ampules at 45 °C and at pH 3.0, 3.2, or 3.4 gave near-racemic wine lactone, but when the hydrolyses were conducted at room temperature, the product was enriched in the (3S,3aS,7aR)-enantiomer 1a and the ee was greater at higher pH (up to 60% at pH 3.4).  相似文献   

13.
Production, purification, and characterization of a novel β-1,3-1,4-glucanase (lichenase) from thermophilic Rhizomucor miehei CAU432 were investigated. High-level extracellular β-1,3-1,4-glucanase production of 6230 U/mL was obtained when oat flour (3%, w/v) was used as a carbon source at 50 °C. The crude enzyme was purified to homogeneity with a specific activity of 28818 U/mg. The molecular weight of purified enzyme was estimated to be 35.4 kDa and 33.7 kDa by SDS-PAGE and gel filtration, respectively. The optimal pH and temperature of the enzyme were pH 5.5 and 60 °C, respectively. The K(m) values of purified β-1,3-1,4-glucanase for barley β-glucan and lichenan were 2.0 mM and 1.4 mM, respectively. Furthermore, the gene (RmLic16A) encoding the β-1,3-1,4-glucanase was cloned and its deduced amino acid sequence showed the highest identity (50%) to characterized β-1,3-1,4-glucanase from Paecilomyces thermophila. The high-level production and biochemical properties of the enzyme enable its potential industrial applications.  相似文献   

14.
Resistant starches (RS) were prepared by phosphorylation of wheat, waxy wheat, corn, waxy corn, high‐amylose corn, oat, rice, tapioca, mung bean, banana, and potato starches in aqueous slurry (≈33% starch solids, w/w) with 1–19% (starch basis) of a 99:1 (w/w) mixture of sodium trimetaphosphate (STMP) and sodium tripolyphosphate (STPP) at pH 10.5–12.3 and 25–70°C for 0.5–24 hr with sodium sulfate or sodium chloride at 0–20% (starch basis). The RS4 products contain ≤100% dietary fiber when assayed with the total dietary fiber method of the Association of Official Analytical Chemists (AOAC). In vitro digestion of four RS4 wheat starches showed they contained 13–22% slowly digestible starch (SDS) and 36–66% RS. However after gelatinization, RS levels fell by 7–25% of ungelatinized levels, while SDS levels remained nearly the same. The cross‐linked RS4 starches were distinguished from native starches by elevated phosphorus levels, low swelling powers (≈3g/g) at 95°C, insolubilities (<1%) in 1M potassium hydroxide or 95% dimethyl sulfoxide, and increased temperatures and decreased enthalpies of gelatinization measured by differential scanning calorimetry.  相似文献   

15.
An esterase from rice ( Oryza sativa ) bran was identified on two-dimensional gel using 4-methylumbelliferyl butyrate as a substrate. The esterase cDNA (870 bp) encoded a 289 amino acid protein (designated OsEST-b) and was expressed in Escherichia coli . The molecular weight of recombinant OsEST-b (rOsEST-b) was 27 kDa, as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochemical characterization demonstrated that rOsEST-b was active over a broad temperature range (optimum at 60 °C) and preferred alkaline conditions (optimum at pH 9.0). The rOsEST-b showed maximum activity toward p-nitrophenyl butyrate (C(4)) among various p-nitrophenyl esters (C(4)-C(18)), indicating that rOsEST-b is an esterase for short-chain fatty acids. The kinetic parameters under optimal conditions were K(m) = 27.03 μM, k(cat) = 49 s(-1), and k(cat)/K(m) = 1.81 s(-1) μM(-1). The activity of rOsEST-b was not influenced by ethylenediaminetetraacetic acid, suggesting that it is not a metalloenzyme. The amino acid sequence analysis revealed that OsEST-b had a conserved pentapeptide esterase/lipase motif but that the essential active site serine (GXSXG) was replaced by cysteine (C). These results suggest that OsEST-b is distinct from traditional esterases/lipases and is a novel lipolytic enzyme in rice bran.  相似文献   

16.
Locust bean gum (LBG) was employed to screen mannanase-producing bacteria. The bacterium with highest mannanase ability was identified as Paenibacillus cookii. It revealed highest activity (6.67 U/mL) when cultivated in 0.1% LBG with 1.5% soytone and 0.5% tryptone after 4 days incubation at 27 °C. Its mannanase was purified to electrophoretical homogeneity after DEAE-Sepharose and Sephacryl S-100 separation. The purified mannanase, with an N-terminus of GLFGINAY, had pH and temperature optimum at 5.0 and 50 °C, respectively, and was stable at pH 5.0-7.0, ≤ 50 °C. It was strongly activated by β-mercaptoethanol, dithiothreitol, cysteine, and glutathione, but inhibited by Hg(2+), Cu(2+), Zn(2+), Fe(3+), PMSF, iodoacetic acid, and EDTA. According to substrate specificity study, the purified mannanase had high specificity to LBG and konjac.  相似文献   

17.
For the first time, alkaline hydrogen peroxide (AHP) extraction conditions were used to isolate hemicellulose (arabinoxylan) from destarched corn fiber. Yields of the water-soluble hemicellulose B ranged from 35% (24 hr extraction at 25°C) to 42% (2 hr extraction at 60°C). The hemicellulose B resulting from the 2 hr extraction (pH 11.5) was off-white in color, and a very low proportion (1.7%) of water-insoluble hemicellulose A was extracted. AHP treatment caused delignification and facilitated the alkaline extraction of hemicellulose from the lignocellulosic fiber matrix. In the absence of H2O2, yields were reduced by more than one-third when using otherwise identical extraction conditions of time, temperature and pH. In the standard protocol, corn fiber, NaOH solution, and H2O2 were mixed in a 1:25:0.25 (w/v/w) ratio. Extractions were conducted at pH 11.5 at 25°C or 60°C. The pH was adjusted to 11.5 by addition of NaOH at ambient and elevated temperatures. The optimum hemicellulose yield (51.3%; dry, starch-free basis) was obtained when the pH was increased to 12.5 for the final one-half of the extraction period. Products obtained after extraction at pH values greater than 11.5 were tan in color, however, and the goal of the research has been to isolate white hemicellulose B and then evaluate its properties. Under most conditions, the yields ofhemicellulose B, potentially the most useful form for food and industrial applications, exceeded those of hemicellulose A by more than 10-fold. The hemicellulose B products were lighter in color than those obtained using traditional alkaline extraction conditions of refluxing with calcium or sodium hydroxide. Steps prior to extractions with alkaline H2O2, such as grinding to 20 mesh and extracting with azeotropic toluene-ethanol, were found to be unnecessary.  相似文献   

18.
Prebiotic galacto-oligosaccharides (GOS) were effectively synthesized from lactose in organic-aqueous biphasic media by a novel metagenome-derived β-galactosidase BgaP412. A maximum GOS yield of 46.6% (w/w) was achieved with 75.4% lactose conversion rate in the cyclohexane/buffer system [95:5 (v/v) cyclohexane/buffer] under the optimum reaction conditions (initial lactose concentration = 30% (w/v), T = 50 °C, pH 7.0, and t = 8 h). The corresponding productivity of GOS was approximately 17.5 g L(-1) h(-1). The GOS mixture consisted of tri-, tetra-, and pentasaccharides. Trisaccharides were the chief component of reaction products. These experimental results showed that a low water content, a high initial lactose concentration, and an elevated reaction temperature could significantly promote the transgalactosylation activity of β-galactosidase BgaP412; at the same time, the enhanced GOS yield in an organic-aqueous biphasic system is because of the fact that thermodynamic equilibrium can be shifted to the synthetic direction by reversing the normal hydrolysis.  相似文献   

19.
Protein solubility (PS) values of different soy protein isolate (SPI) films were determined in water, 0.01 N HCl, 0.01 N NaOH, 4 M urea, and 0.2 M 2-mercaptoethanol. Tensile and color (L, a, and b values) properties of films also were determined. Control films were cast from heated (70 degrees C for 20 min), alkaline (pH 10) aqueous solutions of SPI (5 g/100 mL of water) and glycerin (50% w/w of SPI). Additional films were cast after incorporation of dialdehyde starch (DAS) at 10% w/w of SPI or small amounts of formaldehyde in the film-forming solutions. Also, control film samples were subjected to heat curing (90 degrees C for 24 h), UV radiation (51.8 J/m(2)), or adsorption of formaldehyde vapors. PS of control films was highest (P < 0.05) in 2-mercaptoethanol, confirming the importance of disulfide bonds in SPI film formation. All treatments were effective in reducing (P < 0.05) film PS in all solvents. Both DAS and adsorbed formaldehyde rendered the protein in films practically insoluble in all solvents. Adsorption of formaldehyde vapors and heat curing also substantially increased (P < 0.05) film tensile strength from 8.2 to 15.8 or 14.7 MPa, respectively. However, heat curing decreased (P < 0.05) film elongation at break from 30 to 6%. Most treatments had small but significant (P < 0.05) effects on b color values, with DAS-containing films having the greatest (P < 0. 05) mean b value (most yellowish). Also, DAS-containing, heat-cured, and UV-irradiated films were darker, as evidenced by their lower (P < 0.05) L values, than control films. It was demonstrated that PS of SPI films can be notably modified through chemical or physical treatments prior to or after casting.  相似文献   

20.
Pectinase was covalently immobilized onto the macroporous polyacrylamide (PAM) microspheres synthesized via an inverse suspension polymerization approach, resulting in 81.7% immobilization yield. The stability of the macroporous PAM support, which has a large surface area, is not impeded by the adsorbed proteins despite the fact that up to 296.3 mg of enzyme is immobilized per gram of the carrier particles. The immobilized enzyme retained more than 75% of its initial activity over 30 days, and the optimum temperature/pH also increased to the range of 50-60 °C/3.0-5.0. The immobilized enzyme also exhibited great operational stability, and more than 75% residual activity was observed after 10 batch reactions. The kinetics of a model reaction catalyzed by the immobilized pectinase was finally investigated. Moreover, the immobilized pectinase could be recovered by centrifuging and showed durable activity at the process of recycle.  相似文献   

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