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1.
The membrane-bound hydrogenase from autotrophically cultured Rhizobium japonicum cells has been purified and partially characterized. During the purification process it was necessary to exclude O_2, add reducing agents and include the non-ionic detergent Genapol in buffers in order to maintain activity and prevent the enzyme frjom precipitation. The purified enzyme retained 78% of its activity after storage under H_2 at —80℃ for six months. The most highly purified fraction with a specific activity of 142 μmol methylene blue reduced·min~(-1)·mg protein~(-1), showed two major bands on SDS polyacrylamide electrophoresis gels with molecular weights of about 60,000 and 30,000. There was no positive correlation between hydrogenase specific activity and cytochrome b content of fractions obtained from the different steps in the purification process. No cytochrome b could be detected in the fraction with the highest specific activity. Some other characteristics of the hydrogenase from autotrophically cultu 相似文献
2.
Studies on Manganese Peroxidase Immobilized in Gelatin-containing Microemulsion-based Gels 总被引:4,自引:0,他引:4
SONGShao-fang LUANYu-xia SUXiu-rong 《高等学校化学研究》2005,21(4):423-425
The immobilized technique of manganese peroxidase (MnP) in gelatin-containing microemulsion-based gels and the effects of storage time and reuse times on its catalytic activity were studied. The results show that the MnP immobilized together with Mn^2 and H2O2 could effectively oxidize syringaldazine in n-heptane. The immobilized MnP still had a high catalytic activity after one-month storage under a freezing condition. The reuse times have a relation to the amount of the immobilized H2O2. When the amount of the immobilized H2O2 is sufficient, the microemulsion-based gels containing MnP could be used many times. 相似文献
3.
Nanopolystyrene was used as a solid support for the covalent immobilization of Candida antarctica lipase B(Cal B) using the photoreactive reagent 1‐fluoro‐2‐nitro‐4‐azido benzene(FNAB) as a cou‐pling reagent. The obtained derivative was then used as a biocatalyst in a microwave assisted ester‐ification experiment. Factors such as contact time, pH, and enzyme concentration were investigated during immobilization. The hydrolytic activity, thermal, and operational stability of immobi‐lized‐Cal B were determined. The maximum immobilized yield(218 μg/mg support) obtained at pH 6.8 exhibited optimum hydrolytic activity(4.42 × 103 mU p‐nitrophenol/min). The thermal stability of Cal B improved significantly when it was immobilized at pH 10, however, the immobilized yield was very low(93.6 μg/mg support). The immobilized‐Cal B prepared at pH 6.8 and pH 10 retained 50% of its initial activity after incubation periods of 14 and 16 h, respectively, at 60 °C. The opera‐tional stability was investigated for the microwave assisted esterification of oleic acid with metha‐nol. Immobilized‐Cal B retained 50% of its initial activity after 15 batch cycles in the micro‐wave‐assisted esterification. The esterification time was notably reduced under microwave irradia‐tion. The combined use of a biocatalyst and microwave heating is thus an alternative total green synthesis process. 相似文献
4.
RENLing-ling HEJing EvansD.G. DUANXue 《高等学校化学研究》2003,19(3):324-329
A hydrotalcite-like Mg2 /Al3 layered double hydroxide (LDH) material was prepared by means of amodified coprecipitation method involving a rapid mixing step followed by a separate aging process. LDH calcined at 500℃ , denoted as CLDH, was characterized by XRD, IR and BET surface area measurements.CLDH has a poor crystalline MgO-like structure with a high surface area and porosity. CLDH was used as asupport for the immobilization of penicillin G acylase(PGA). The effect of varying the immobilization conditions, such as pH, contact time and the ratio of enzyme to support, on the activity of the immobilized enzymein the hydrolysis of penicillin G has been studied. It was found that the activity of the immobilized enzyme decreased slightly with decreasing pH and reached a maximum after a contact time of 24 h. The activity of theimmobilized enzyme increased with increasing the ratio of enzyme to support. It was found that the adsorption of PGA inhibited the expected reaction of CLDH with an aqueous medium to regenerate a LDH phase. Itsoriginal activity(36%) after 15 cycles of reuse of the immobilized enzyme was retained, but no further loss in the activity was observed. 相似文献
5.
SiO2 monoliths with double-pore structure were synthesized with tetraethyl orthosilicate, polyethylene glycol, starch, ethanol and ammonia as the main raw materials via a sol-gel route. The effect of different content of starch of the samples was studied on the mesopore structure, macropore appearance, hydrothermal stability and enzyme activity to immobilize gluczyme. The results indicate that porous blocks with mesopores of 10~12 nm and macropores of 10~30 μm can be formed by the removal of polyethylene glycol and starch after heating at 600 ℃. The minimal density of the obtained samples was 0.34 g·cm-3 and the maximal porosity was 76%. After being immersed in water at 80 ℃ for 7 d, the density, porosity and shape of N2 adsorption-desorption curves of the samples were almost unchanged, showing that the sample has a good hydrothermal stability. The initial enzyme activity of sample P2KD30 was 11 190 U, and definite enzyme activity could be maintained after five cycles. 相似文献
6.
7.
Reported here is a protocol to fabricate a biocatalyst with high enzyme loading and activity retention, from the conjugation of electrospun nanofibrous membrane having biomimetic phospholipid moiety and lipase. To improve the catalytic efficiency and activity of the immobilized enzyme, poly(acrylonitrile-co-2-methacryloyloxyethyl phosphorylcholine)s(PANCMPCs) were, respectively, electrospun into nanofibrous membranes with a mean diameter of 90 nm, as a support for enzyme immobilization. Lipase from Candida rugosa was immobilized on these nanofibrous membranes by adsorption. Properties of immobilized lipase on PANCMPC nanofibrous membranes were compared with those of the lipase immobilized on the polyacrylonitrile(PAN) nanofibrous and sheet membranes, respectively. Effective enzyme loading on the nanofibrous membranes was achieved up to 22.0 mg/g, which was over 10 times that on the sheet membrane. The activity retention of immobilized lipase increased from 56.4% to 76.8% with an increase in phospholipid moiety from 0 to 9.6%(molar fraction) in the nanofibrous membrane. Kinetic parameter Km was also determined for free and immobilized lipase. The Km value of the immobilized lipase on the nanofibrous membrane was obviously lower than that on the sheet membrane. The optimum pH was 7.7 for free lipase, but shifted to 8.3-8.5 for immobilized lipases. The optimum temperature was determined to be 35 ℃ for the free enzyme, but 42-44℃ for the immobilized ones, respectively. In addition, the thermal stability, reusability, and storage stability of the immobilized lipase were obviously improved compared to the free one. 相似文献
8.
Two peptide chains A_1 and A_2 of the Lys active fragment, linked via a couple ofinter-disulfide bonds, could be separated from each other after reduction with dithiothreitoland gel filtration on Sephadex G-25. Reoxidation of the reduced peptide chain A_1 resultedin recovering the inhibitory activity with 25% yield, based on the original activity of theLys fragment. The A_1 active fragment was further purified by affinity chromatographywith immobilized trypsin. Sephadex G-25 gel filtration produced two forms of the A_1 activefragment, the major fraction being a monomer and the minor one being a dimeer with loweractivity. The results obtained offered evidence of the evolution of mung bean inhibitorfrom an ancestral single-headed inhibitor by fused gene duplication with A_2 as a connectingpeptide. The CD spectra of the Lys fragment and the reoxidized peptide chain A_1 werealso compared. 相似文献
9.
ZOU XiaoChuan CHEN ShaoCheng REN YanRong SHI KaiYun LI Jun & FU XiangKai 《中国科学:化学(英文版)》2012,(11):2397-2407
Chiral Mn Ⅲ (salen) (Jacobsen’s catalyst) was axially immobilized onto a new type of organic polymer-inorganic hybrid materialzirconium poly(styrene-isopropenyl phosphonate)-phosphate(ZPS-IPPA) with different linkage lengths and evaluated as catalysts for the epoxidation of unfunctionalized olefins. The results demonstrated that the prepared catalysts exhibited moderate to good activity and enantioselectivity in the asymmetric epoxidation of unfunctionalized olefins. Furthermore, the immobilized catalysts were relatively stable and could be conveniently separated from the reaction system by simple precipitation in hexane. Moreover, higher enantioselectivity was obtained with catalyst 2c than that of homogeneous counterpart catalyzed even after eight times. The excellent recycling of the catalyst was attributed to its structure feature of ZPS-IPPA which is different from either pure polystyrene or pure zirconium phosphates. 相似文献
10.
Phosphofructokinase-2 was purified to homogeneity from chicken livers by homogeniza-tion, polyethylene glycol fractionation and column chromatography on DEAE-Sephadex A-50 and Blue-Sepharose 4B. Some properties of the enzyme were as follows: (i) The saturation curve of the enzyme for fructose 6-phosphate showed hyperbolic and the Km of fructose 6-phosphate was affected by inorganic phosphate while Vmax was not; (ii) the binding of ATP to the enzyme was of negative cooperativity with a Hill coefficient of 0.56; (iii) the activity of the enzyme was completely lost in the presence of EDTA. The enzyme was activated by Mg2+ at low concentrations, but inhibited by Mg2+ at high concentrations; (iv) the enzyme was stable below 30℃ and easily lost its activity when the temperature was above 40℃; (v) the activity of the enzyme was stable at the range of pH 7-9, increased at pH 9.0-9.5 and decreased when pH was over 9.5; (vi) the enzyme was sensitive to trypsin and ATP protected the enzyme against the proteolysi 相似文献
11.
An indigenous and industrially important manganese peroxidase (MnP) isolated from solid state culture of Pleurotus ostreatus IBL-02 was 2.1-fold purified and immobilized using sol-gel matrix of trimethoxysilane and proplytetramethoxysilane. Lower KM, higher Vmax, hyper-activation, enhanced acidic and thermal stability up to 70℃, were the improved/enhanced catalytic features of the presently sol-gel immobilized MnP. A maximum of 100% decolorization was achieved within a short time period for the different real dyes containing textile industry effluents i.e. Nishat textile (NIT), K&N textile (KIT), Sitara textile (SIT), and Crescent textile (CRT). 相似文献
12.
Chairattanamanokorn P Imai T Kondo R Ukita M Prasertsan P 《Applied biochemistry and biotechnology》2006,128(3):195-204
To select a thermotolerant fungal strain for decolorization of wastewaters, ligninolytic enzyme production (lignin peroxidase,
manganese peroxidase [MnP], and laccase), decolorization, and removal of total phenol and chemical oxygen demand (COD) were
detected. Thirty-eight fungal strains were studied for enzyme production at 35 and 43°C on modified Kirk agar medium including
2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and MnCl2. Thirteen strains grew on manganese-containing agar and provided green color on ABTS-containing agar plates under culture
at 43°C. Decolorization of wastewater from alcohol distillery (WAD) by these strains was compared under static culture at
43°C, and Pycnoporus coccineus FPF 97091303 showed the highest potential. Thereafter, immobilized mycelia were compared with free mycelia for WAD decolorization
under culture conditions of 43°C and 100 rpm. The immobilized mycelia on polyurethane foam enhanced the ligninolytic enzyme
production as well as total phenol and color removal. At about the same COD removal, MnP and laccase produced by immobilized
mycelia were 2 and 19 times higher than by free mycelia; the simultaneous total phenol and color removal were 3.1 and 1.5
times higher than the latter. Moreover, decolorization of synthesis dye wastewater was carried out at 43°C and 100 rpm. More
than 80% of 300 mg/L of reactive blue-5 was decolorized by the immobilized mycelia within 1 to 2 d for four cycles. 相似文献
13.
The objective of this study was to exploit the decolorization potential of a newly isolated white-rot fungus Schizophyllum commune IBL-6 for the biodegradation of reactive textile dye Cibacron Red FN-2BL. In the initial decolorization study of 10 days, it was observed that S. commune IBL-6 was a better decolorizer of Cibacron Red FN-2BL. Various process parameters like composition of basal nutrient medium, pH, temperature, additional carbon and nitrogen sources, and initial dyestuff concentration were optimized to develop an economic decolorization process. The optimum dye decolorization was achieved in basal nutrient medium II containing 0.1% Cibacron Red FN-2BL and supplemented with 1% glucose after 3 days incubation at pH 4.5 and 30 degrees C. All the additional carbon sources were found to enhance decolorization process, whereas most of the nitrogen supplements caused fungal-growth inhibition. The pattern of enzymes involved in the biodegradation of this dye was studied, and manganese peroxidase was found to be the major peroxidase with minor lignin peroxidase and laccase activities. 相似文献
14.
Mahreen Matto Rukhsana Satar Qayyum Husain 《Applied biochemistry and biotechnology》2009,158(3):512-523
Calcium alginate–starch entrapped bitter gourd peroxidase has been employed for the treatment of a textile industrial effluent
in batch as well as in continuous reactor. The textile effluent was recalcitrant to decolorization by bitter gourd peroxidase;
thus, its decolorization was examined in the presence of a redox mediator, 1.0 mM 1-hydroxybenzotriazole. Immobilized enzyme
exhibited same pH and temperature optima for effluent decolorization as attained by soluble enzyme. Immobilized enzyme could
effectively remove more than 70% of effluent color in a stirred batch process after 3 h of incubation. Entrapped bitter gourd
peroxidase retained 59% effluent decolorization reusability even after its tenth repeated use. The two-reactor system containing
calcium alginate–starch entrapped enzyme retained more than 50% textile effluent decolorization efficiency even after 2 months
of its operation. The absorption spectra of the treated effluent exhibited a marked difference in the absorption at various
wavelengths as compared to untreated effluent. The use of a two-reactor system containing immobilized enzyme and an adsorbent
will be significantly successful for treating industrial effluents at large scale, and it will help in getting water free
from aromatic pollutants. 相似文献
15.
Wastewater emissions from textile factories cause serious environmental problems. Manganese peroxidase (MnP) is an oxidoreductase with ligninolytic activity and is a promising biocatalyst for the biodegradation of hazardous environmental contaminants, and especially for dye wastewater decolorization. This article first summarizes the origin, crystal structure, and catalytic cycle of MnP, and then reviews the recent literature on its application to dye wastewater decolorization. In addition, the application of new technologies such as enzyme immobilization and genetic engineering that could improve the stability, durability, adaptability, and operating costs of the enzyme are highlighted. Finally, we discuss and propose future strategies to improve the performance of MnP-assisted dye decolorization in industrial applications. 相似文献
16.
Anthony C. Grabski Jerald K. Rasmussen Patrick L. Coleman Richard R. Burgess 《Applied biochemistry and biotechnology》1996,60(1):1-17
Manganese peroxidase (MnP) is secreted by white-rot fungi and participates in the degradation of lignin by these organisms.
MnP uses H2O2 as an oxidant to oxidize MnII to MnIII as the manganic ion Mn3+. The Mn3+ stabilized by chelation, is a highly reactive nonspecific oxidant capable of oxidizing a variety of toxic organic compounds.
Previous attempts at immobilization of MnP, purified fromLentinula edodes through reactive amino groups, have been hindered by the protein’s low lysine content of only 1% and its instability above
pH 6.0. As an alternative to amine coupling, the enzyme has now been covalently immobilized through its carboxyl groups, using
an azlactonefunctional copolymer derivatized with ethylenediamine and 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ)
as a coupling reagent. The immobilization reaction was performed under acidic (pH 5.25) conditions, and 90% coupling efficiency
was achieved within 2 h. Net immobilization efficiencies, expressed as the product of protein coupling efficiency and enzyme
activity, have been measured at > 95% within 4 h. The MnP-NH-polymer and the free soluble protein were characterized and compared
for their pH, temperature, and storage stabilities, as well as their H2O2 dependence and kinetics. The tethered MnP, employed in an immobilized enzyme bioreactor for generation of chelated Mn3+ may have industrial applications as a nonspecific oxidant of organopollutants. 相似文献
17.
Anthony C. .Grabski Patrick L. Coleman Gary J. Drtina Richard R. Burgess 《Applied biochemistry and biotechnology》1995,55(1):55-73
Manganese peroxidase (MnP) purified fromLentinula edodes was covalently immobilized on 3M’s azlactone-functional copolymer, 3M EmphazeTM AB1 Biosupport Medium. Tethered MnP is capable
of generating Mn3+ from Mn2+ and H2O2. Mn3+, properly chelated, can be used as a nonspecific oxidant of organopollutants. A variety of conditions designed to maximize
coupling efficiency while maintaining Mn3+ -generating catalytic activity were tested. Biochemical characteristics of the MnP enzyme, including amino acid composition,
pH and temperature stability, and concentration of its Mn2+ substrate, influenced chemical conditions necessary for the coupling reaction. The physical parameters of immobilization
reaction time, protein concentration, ionic conditions, pH, and temperature were examined. Results of these experiments indicated
maximum coupling efficiency and enzyme activity were achieved by immobilizing at MnP concentrations < 2 mg/mL for at least
2 h using pH 7.0 buffer containing 1.0M sodium sulfate and 1.0 mM Mn2+. Increasing coupling reaction temperature also improved coupling efficiency. A synthesis of these optimized immobilizations
yielded MnP coupling efficiencies of 40–50% with 35% of the coupled protein retaining enzymatic activity. Results of MnP immobilizations
on nonporous azlactone-functional dispersion polymers more hydrophobic than Emphaze are also reported, and coupling efficiencies
> 65% with 100% of the coupled enzyme active have been measured. 相似文献
18.
Sami Sayadi Fathi Zorgani Radhouane Ellouz 《Applied biochemistry and biotechnology》1996,56(3):265-276
This paper describes the decolorization and chemical oxygen demand (COD) removal of olive mill waste-waters (OMW) byPhanerochaete chrysosporium grown in agitated submerged cultures. WhenP. chrysosporium was cultivated in the form of pellet, no decolorization of crude OMW was observed. Decolorization occured only after removing by ultrafiltration, the high-mol-wt (HM) polyphenolic fraction (> 60 kDa). The use of high lignin peroxidase (LiP) producing medium yielded the highest levels of OMW decolorization and COD removal. In this case, extensive depolymerization and subsequent accumulation of phenolics with intermediates molecular weight were observed. Furthermore, increasing the concentration of the HM fraction decreased the color and COD removals. The decolorizing activity was lost when the concentration of the HM fraction reached 25% (v/v). Consequently, LiP activity was found to be completely inhibited in the presence of HM fraction, but not with the low-mol-wt (LM) polyphenolic fraction (<8 kDa). The use ofP. chrysosporium immobilized on polyurethane foam resulted in efficient decolorization of crude OMW. Moreover, the addition of an induction medium was shown to perform several repeated batch cultures for OMW decolorization and COD removal. 相似文献
19.
F. A. Rodríguez E. P. Rivero L. Lartundo-Rojas I. González 《Journal of Solid State Electrochemistry》2014,18(11):3153-3162
Textile industry is one of the major generators of wastewaters containing recalcitrant compounds such as dyes that jeopardize public health and environment. Electro-oxidation is an alternative method for treating recalcitrant compounds, and the key element for efficient degradation is the adequate use of dimensionally stable anode (DSA) electrodes to efficiently generate active chlorine, which degrades dyes contained in effluents into more environment-friendly compounds. This work is thereby aimed at preparing a novel DSA electrode for efficient generation of active chlorine. Two different dimensionally stable anodes (Ti/RuO2 and Sb2O5-doped Ti/RuO2-ZrO2) were prepared and then characterized by grazing incidence X-ray diffraction, X-ray photoelectron spectroscopy, and scanning electron microscopy, which corroborated the presence of RuO2, ZrO2, and Sb2O5. The comparison of electroactive areas, assessed by chronoamperometry, showed that Zr helps increase the area of the ternary electrode facilitating the formation of active chlorine. Active chlorine formation was further studied by cyclic voltammetry that revealed a reduction peak attributed to chlorine (product of chloride oxidation). Additionally, decolorization of model solutions that simulate textile effluents containing indigo carmine and reactive black 5 in media with and without chlorides was performed. In the chloride-containing medium, decolorization occurred at a faster rate than in the presence of sulfates. Decolorization of carmine indigo and reactive black 5 in the chloride-containing medium took 40 min and 2 h, respectively. In conclusion, the DSA electrode made of Sb2O5-doped Ti/RuO2-ZrO2 can efficiently generate the active chlorine for degradation of recalcitrant compounds. 相似文献
20.
Degradation of textile dyes mediated by plant peroxidases 总被引:3,自引:0,他引:3
The peroxidase enzyme from the plants Ipomea palmata (1.003 IU/g of leaf) and Saccharum spontaneum (3.6 IU/g of leaf) can be used as an alternative to the commercial source of horseradish and soybean peroxidase enzyme for
the decolorization of textile dyes, mainly azo dyes. Eight textiles dyes currently used by the industry and seven other dyes
were selected for decolorization studies at 25–200 mg/L levels using these plant enzymes. The enzymes were purified prior
to use by ammonium sulfate precipitation, and ion exchange and gel permeation chromatographic techniques. Peroxidase of S. spontaneum leaf (specific activity of 0.23 IU/mg) could completely degrade Supranol Green and Procion Green HE-4BD (100%) dyes within
1 h, whereas Direct Blue, Procion Brilliant Blue H-7G and Chrysoidine were degraded >70% in 1 h. Peroxidase of Ipomea (I. palmata leaf; specific activity of 0.827 U/mg) degraded 50 mg/L of the dyes Methyl Orange (26%), Crystal Violet (36%), and Supranol
Green (68%) in 2–4 h and Brilliant Green 54%), Direct Blue (15%), and Chrysoidine (44%) at the 25 mg/L level in 1 to 2 h of
treatment. The Saccharum peroxidase was immobilized on a hydrophobic matrix. Four textile dyes, Procion Navy Blue HER, Procion Brilliant Blue H-7G,
Procion Green HE-4BD, and Supranol Green, at an initial concentration of 50 mg/L were completely degraded within 8 h by the
enzyme immobilized on the modified polyethylene matrix. The immobilized enzyme was used in a batch reactor for the degradation
of Procion Green HE-4BD and the reusability was studied for 15 cycles, and the halflife was found to be 60 h. 相似文献