A novel knotted cotton-thread carrier: its potential use in achieving the biomass renewal of Phanerochaete chrysosporium in an immobilized growth system

In order to achieve an innovative strategy to renew the biomass of Phanerochaete chrysosporium in an immobilized growth system which can maintain white-rot fungi biomass, a novel knotted cotton-thread carrier was designed and made. By using a high-speed stirring apparatus under the conditions of 1400 r/min stirring speed for 6 min, mycelia immobilized on the knotted cotton-thread carriers were exfoliated completely and homogenized to a proper size. Furthermore, the homogenized mycelia from the immobilized mycelia can resume their growth on the knotted cotton-thread carriers in agitated flask cultures. The average regrowth biomass on the new carriers and the reused carriers was 0.0171 g/carrier and 0.0314 g/carrier, respectively. It proves that the knotted cotton-thread carrier performs perfectly in homogening the immobilized mycelia to achieve the biomass renewal of P. chrysosporium in an immobilized growth system. Supported by the National Natural Science Foundation of China (Grant No. 206770 33)     

High production of β-glucosidase from a marine Aspergillus niger immobilized on towel gourd vegetable sponges

To produce β-glucosidase by consecutive batch fermentation, a marine Aspergillus niger was immobilized on a natural carrier, towel gourd vegetable sponges. The immobilized mycelia were 0.15 g/g carrier with the immobilized biomass percentage of over 95%. The immobilized mycelia possessed the long durability(22.5 days). The maximum production occurred 1.5 day earlier by the immobilized mycelia than by the free mycelia. β-Glucosidase production of five consecutive batches was over 110 U/m L. At high salinity,the activity and stability of β-glucosidase from the marine A. niger increased remarkable. Immobilizing the marine A. niger on the novel natural carrier achieved the efficient production of β-glucosidase.     

Feasibility of treating swine manure in an anaerobic sequencing batch biofilm reactor with mechanical stirring

Anaerobic sequencing batch reactors containing granular or flocculent biomass have been employed successfully in the treatment of piggery wastewater. However, the studies in which these reactors were employed did not focus specifically on accelerating the hydrolysis step, even though the degradation of this chemical oxygen demand (COD) fraction is likely to be the limiting step in many investigations of this type of wastewater. The mechanically stirred anaerobic sequencing batch biofilm reactor offers an alternative for hastening the hydrolysis step, because mechanical agitation can help to speed up the reduction of particle sizes in the fraction of particulate organic matter. In the present study, a 4.5-L reactor was operated at 30°C, with biomass immobilized on cubic polyurethane foam matrices (1 cm of side) and mechanical stirring provided by three flat-blade turbines (6 cm) at agitation rates varying from 0 to 500 rpm. The reactor was operated to treat diluted swine waste, and mechanical stirring efficiently improved degradation of the suspended COD. The operational data indicate that the reactor remained stable during the testing period. After 2 h of operation at 500 rpm, the suspended COD decreased by about 65% (from 1500 to 380 mg/L). Apparent kinetic constants were also calculated by modified first-order expressions.     

A technique for mycelial development of ectomycorrhizal fungi on agar media

A technique was established to study ectomycorrhizal fungi on agar media. Petri dishes, 60 mm in diameter, containing 10 mL of culture medium covered with a cellophane disk were used for easy collection of the mycelium after growth. For analysis of fungal biomass production, a sterilized cellophane sheet was placed on the medium’s surface. Inoculation was achieved by placing a mycelial block onto the center of the cellophane sheet and then incubating at 25°C in the dark. Colony radial growth was measured and biomass dry wt was determined. Fresh mycelia were homogenized with 10 mL of acetate buffer (pH 5.5) for enzyme analysis. A crude extract was obtained by adding all culture medium to 90 mL of distilled water and homogenizing in a Potter. Reducing sugars, enzyme concentration, and pH were determined. Three fungal strains, Suillus collinitus, Pisosithus arrhizus, and Hebeloma cylindrosporum, were grown in different culture media (potato dextrose agar or Pintro’s medium). Parameters measured over time included glucose concentration, phosphatase activity, biomass, and pH.     

Screening thermotolerant white-rot fungi for decolorization of wastewaters

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.     

Entrapment of Lentinus sajor-caju into Ca-alginate gel beads for removal of Cd(II) ions from aqueous solution: preparation and biosorption kinetics analysis

A white rot fungus species Lentinus sajor-caju biomass was entrapped into alginate gel via a liquid curing method in the presence of Ca(II) ions. The biosorption of cadmium(II) by the entrapped live and dead fungal biomass has been studied in a batch system. The heat-treatment process enhanced the biosorption capacity of the immobilized fungal biomass. The effect of initial cadmium concentration, pH and temperature on cadmium removal has been investigated. The maximum experimental biosorption capacities for entrapped live and dead fungal mycelia of L. sajur-caju were found to be 104.8±2.7 mg Cd(II) g⁻¹ and 123.5±4.3 mg Cd(II) g⁻¹, respectively. The kinetics of cadmium biosorption was fast, approximately 85% of biosorption taking place within 30 min. The biosorption equilibrium was well described by Langmuir and Freundlich adsorption isotherms. The change in the biosorption capacity with time is found to fit pseudo-second-order equations. Cadmium binding properties of entrapped fungal preparations have been determined applying the Ruzic equations. Since the biosorption capacities are relatively high for both entrapped live and dead forms, they could be considered as suitable biosorbents for the removal of cadmium in wastewater treatment systems. The biosorbents were reused in three consecutive adsorption/desorption cycles without significant loss in the biosorption capacity.     

超高效液相色谱-串联质谱法同时测定鸡肝中残留的四环素类、磺胺类和喹诺酮类药物

采用超高效液相色谱-串联质谱(UPLC-MS/MS)在正离子模式下通过多反应监测(MRM)方式同时测定了鸡肝脏组织中3种四环素类药物、10种磺胺类药物以及8种喹诺酮类药物的残留。试样由McIlvaine缓冲液-乙腈(体积比为1:4)、乙腈提取,合并上清液并用氮气吹干,用0.05 mol/L磷酸三乙胺缓冲液-乙腈(体积比为85:15)溶解残余物,经正己烷脱脂后,采用UPLC-MS/MS进行定性、定量分析。该方法对测定的21种药物的检出限均为2 μg/kg,定量限均为5 μg/kg。在添加水平分别为5,10和50 μg/kg时,21种药物的加标回收率为66.8%～128.5%,日内测定的相对标准偏差(RSD)为0.8%～20.2%,日间测定的RSD为2.2%～15.3%。该方法可作为动物源性食品中这3类药物残留检测的确证方法。     

Enhanced Production of High-Quality Biomass, δ-Aminolevulinic Acid,Bilipigments, and Antioxidant Capacity of a Food Alga <Emphasis Type="Italic">Nostochopsis lobatus</Emphasis>

The growing interest in natural food has raised the global demand for nutraceuticals. We studied enhanced production of biomass, delta-aminolevulinic acid (delta-ALA), bili pigments and antioxidant capacity of a food alga Nostochopsis lobatus in a full-factorial (three level) design with supplemental Zn, glutamine, and Zn + glutamine in batch culture. Production of biomass, pigments, and antioxidant capacity all were higher under immobilized cell cultures in comparison to free cell cultures. Maximum biomass (2,390 mg dry wt l(-1)), delta-ALA (2.715 microg mg(-1) dry wt h(-1)), phycocyanin (98.50 mg g(-1) dry wt), phycoerythrin (158.0 mg g(-1) dry wt), and antioxidant capacity (140.50 mumoles ascorbic acid equivalent capacity g(-1) fresh wt) were recorded when Zn and glutamine were supplemented together in the growth medium at pH 7.8. These effects were found to be significantly related to the activities of glutamine synthetase (GS(max): 490.2 nmoles mg protein(-1) min(-1)), glutamate synthase (GOGAT(max): 27.0 nmoles mg protein(-1) min(-1)), and glutamate dehydrogenase (GDH(max): 159.9 nmoles mg protein(-1) min(-1)). This study shows that N. lobatus could be a promising bioresource for the production of nutritionally rich biomass, delta-ALA, bili pigments, and antioxidants. Use of immobilized cells in batch culture supplemented with Zn and glutamine could be an effective approach for scaling up production for commercial use.     

Biosorption of mercury by carboxymethylcellulose and immobilized Phanerochaete chrysosporium

Phanerochaete chrysosporium basidiospores immobilized onto carboxymethylcellulose were used for the removal of mercury ions from aqueous solutions. The biosorption of Hg(II) ions onto carboxymethylcellulose and both immobilized live and heat-inactivated fungal mycelia of Phanerochaete chrysosporium was studied using aqueous solutions in the concentration range 30-700 mg l⁻¹. The biosorption of Hg(II) ions by the carboxymethylcellulose and both live and heat-inactivated immobilized preparations increased as the initial concentration of mercury ions increased in the medium. Maximum biosorption capacity for immobilized live and heat-inactivated fungal mycelia of Phanerochaete chrysosporium was found to be 83.10 and 102.15 mg Hg(II) g⁻¹, respectively, whereas the amount of Hg(II) ions adsorbed onto the plain carboxymethylcellulose beads was 39.42 mg g⁻¹. Biosorption equilibria were established in approximately 1 h and the correlation regression coefficients show that the adsorption process can be well defined by a Langmuir equation. Temperature changes between 15 and 45 °C did not affect the biosorption capacity. The effect of pH was also investigated and the maximum adsorption of Hg(II) ions onto the carboxymethylcellulose and both live and heat-inactivated immobilized fungal mycelia was observed at pH 6.0. The carboxymethylcellulose-fungus beads could be regenerated using 10 mM HCl, with up to 95% recovery. The biosorbents were used in three biosorption-desorption cycles and no significant loss in the biosorption capacity was observed.     

A novel knotted cotton-thread carrier: its potential use in achieving the biomass renewal of Phanerochaete chrysosporium in an immobilized growth system

Science China Chemistry - In order to achieve an innovative strategy to renew the biomass of Phanerochaete chrysosporium in an immobilized growth system which can maintain white-rot fungi biomass,...     

Production and characterization ofPhanerochaete chrysosporium lignin peroxidases for pulp bleaching

The production of lignin peroxidase fromPhanerochaete chrysosporium was studied using immobilized mycelia in nylon-web cubes in semicontinuous fermentation using glucose pulses or ammonium tartrate pulses. Consistent enzyme production was achieved when glucose pulses were used, leading to an average activity of 253 U/L. The crude enzyme was added to eucalyptus kraft pulp before conventional and ECF bleaching sequences. Optimization of the enzymatic pretreatment led to the following operational conditions: enzyme load of 2 U/g of pulp, hydrogen peroxide addition rate of 10 ppm/h, and reaction time of 60 min. Pulp final characteristics were dependent on the chemical treatment sequence that followed enzymatic pretreatment. The chief advantage of enzymatic pretreatment was pulp viscosity preservation, which was observed in most of the experiments carried out with seven different chemical treatment sequences     

Preparation,Characterization and Optimization of Chitosan Nanoparticles as Carrier for Immobilization of Thermophilic Recombinant Esterase

Immobilization of biologically important molecules on myriad nano-sized materials has attracted great attention. Through this study, thermophilic esterase enzyme was obtained using recombinant DNA technology and purified applying one-step His-Select HF nickel affinity gel. The synthesis of chitosan was achieved from chitin by deacetylation process and degree of deacetylation was calculated as 89% by elemental analysis. Chitosan nanoparticles were prepared based on the ionic gelation of chitosan with tripolyphosphate anions. The physicochemical properties of the chitosan and chitosan nanoparticles were determined by several methods including SEM (Scanning Electron Microscopy), FT-IR (Fourier Transform Infrared Spectroscopy) and DLS (Dynamic Light Scattering). The morphology of chitosan nanoparticles was spherical and the nanospheres’ average diameter was 75.3 nm. The purified recombinant esterase was immobilized efficiently by physical adsorption onto chitosan nanoparticles and effects of various immobilization conditions were investigated in details to develope highly cost-effective esterase as a biocatalyst to be utilized in biotechnological purposes. The optimal conditions of immobilization were determined as follows; 1.0 mg/mL of recombinant esterase was immobilized on 1.5 mg chitosan nanoparticles for 30 min at 60°C, pH 7.0 under 100 rpm stirring speed. Under optimized conditions, immobilized recombinant esterase activity yield was 88.5%. The physicochemical characterization of enzyme immobilized chitosan nanoparticles was analyzed by SEM, FT-IR and AFM (Atomic Force Microscopy).     

Lipase Immobilized in Organic-Inorganic Matrices

Enzyme lipase was immobilized with ferrite powder and deposited in layers on glass slides from lipase to a solution of silicone alkoxides. The highest hydrolytical activity was observed with the magnetic lipase prepared by mixing the paste of ferrite powder and lipase with tetramethoxysilane, 3-aminopropyltriethoxysilane and propyltrimethoxysilane. In a mixed reactor, the particles of the magnetic lipase were desintegrated by mechanical stirring which caused loosing the lipase linked to magnetic material and resulted in a significant drop of activity after magnetic separation. Transparent layers were prepared by dip- or spin-coating from partially hydrolyzed tetraethoxysilane and solutions containing methyltriethoxysilane with 3-aminopropyltriethoxysilane or tetraethoxysilane with 3-mercaptopropyltriethoxysilane. The lipases immobilized in films with magnetic particles were active in tests with 4-nitrophenyl butyrate and were not inhibited by 0,0-dimethyl-0-(2,2-dichlor-vinyl)-phosphate.     

AC electrothermal enhancement of heterogeneous assays in microfluidics

AC-driven electrothermal flow is used to enhance the temporal performance of heterogeneous immuno-sensors in microfluidic systems by nearly an order of magnitude. AC electrokinetic forces are used to generate electrothermal flow, which in turn produces a circular stirring fluid motion that enhances the transport of diffusion-limited proteins. This provides more binding opportunities between suspended antigens and wall-immobilized antibodies. We investigate experimentally the effectiveness of electrothermal stirring, using a biotin-streptavidin heterogeneous assay, in which biotin is immobilized, and fluorescently-labeled streptavidin is suspended in a high conductivity buffer (sigma = 1.0 S m(-1)). Microfabricated electrodes were integrated within a microwell and driven at a frequency of f= 200 kHz and 10 V(rms). Fluorescent intensity measurements show that for a five minute assay, electrothermal stirring increases the binding rate by a factor of almost nine. Similar binding improvement was measured for longer assays, up to fifteen minutes. The electrothermal enhancement of this assay was modeled numerically and agrees with experimental binding rates. The measured fluid velocity of 22 +/- 2 microm s(-1) was significantly lower than that predicted by the numerical model, 1.1 mm s(-1), but nevertheless shows the same fourth power dependence on applied potential. The results demonstrate the ability for electrothermal stirring to reliably improve the response time and sensitivity within a given time limit for microfluidic diffusion-limited sensors.     

Immobilization of CdS nanoparticles formed in reverse micelles onto aluminosilicate supports and their photocatalytic properties

CdS nanoparticles, prepared in reverse micellar system, were immobilized onto thiol-modified aluminosilicate particles (ASSH) by a simple operation: addition of ASSH in the micellar solution and mild stirring. The resulting CdS nanoparticles-aluminosilicate composites (ASCdS) were used as photocatalysts for H2 generation from 2-propanol aqueous solution. The chemical properties of the aluminosilicate, such as affinity for water and other reactants, were found to affect the photocatalytic property of the CdS nanoparticles immobilized. Zeolite particles, having affinity for water and 2-propanol, gave a good ASCdS photocatalyst with respect to H2 generation.     

A Trefoil Knotted Polymer Produced through Ring Expansion

A synthetic strategy is reported for the production of a trefoil knotted polymer from a copper(I)‐templated helical knot precursor through ring expansion. The expected changes in the properties of the knotted polymer compared to a linear analogue, for example, reduced hydrodynamic radius and lower intrinsic viscosity, together with an atomic force microscopy (AFM) image of individual molecular knots, confirmed the formation of the resulting trefoil knotted polymer. The strategies employed here could be utilized to enrich the variety of available polymers with new architectures.     

Polypyrrole/hexagonally ordered silica nanocomposite as a novel fiber coating for solid-phase microextraction

A highly porous fiber coated polypyrrole/hexagonally ordered silica (PPy/SBA15) materials were prepared for solid-phase microextraction (SPME). The PPy/SBA15 nanocomposite was synthesized by an in situ polymerization technique. The resulting material was characterized by the scanning electron microscopy, thermogravimetric analysis and differential thermal analysis. The prepared nanomaterial was immobilized onto a stainless steel wire for fabrication of the SPME fiber. The fiber was evaluated for the extraction of some polycyclic aromatic hydrocarbons (PAHs) from aqueous sample solutions in combination with gas chromatography-mass spectrometry (GC-MS). A one at-the-time optimization strategy was applied for optimizing the important extraction parameters such as extraction temperature, extraction time, ionic strength, stirring rate, desorption time and desorption temperature. In optimum conditions (extraction temperature 70°C, extraction time 20 min, ionic strength 20% (WV(-1)), stirring rate 500 rpm, desorption temperature 270°C, desorption time 5 min) the repeatability for one fiber (n=3), expressed as relative standard deviation (R.S.D. %), was between 5.0% and 9.3% for the tested compounds. The quantitation limit for the studied compounds were between 13.3 and 66.6 pg mL(-1). The life span and stability of the PPy/SBA15 fiber are good, and it can be used more than 50 times at 260°C without any significant change in sorption properties. The developed method offers the advantage of being simple to use, with shorter analysis times, lower cost of equipment, thermal stability of fiber and high relative recovery in comparison to conventional methods of analysis.     

氧化铁磁性纳米粒子固定化酶*

纳米粒子作为酶固定化的载体,当其具有磁性时,制备的固定化酶易于从反应体系中分离和回收,操作简便;并且利用外部磁场可以控制磁性材料固定化酶的运动方式和方向,替代传统的机械搅拌方式,提高固定化酶的催化效率。在众多纳米材料中,氧化铁因其在磁性、催化等多方面的良好特性而倍受瞩目。本文对近年来各种氧化铁磁性纳米粒子固定化酶,尤其是固定化脂肪酶和蛋白酶的制备方法及其应用做了较为详细的阐述,对这些氧化铁磁性纳米粒子固定化酶的优缺点和发展前景进行了讨论。     

Preparation and characterization of pancreatic lipase immobilized in Eudragit-matrix

Pancreatic lipase (EC 3.1.1.3) was immobilized by entrapping in a commercial preparation of acrylic/methacrylic acid ester-based copolymer (Eudragit E 30 D). The activity of the immobilized lipase beads with a diameter of 1.5-2.0 mm was found to be lower than that of the free lipase. The optimum pH was shifted to the alkaline region and the thermal stability increased, whereas the optimum temperature level remained unchanged. The most important reason for the decreased activity was diffusion limitations. The diffusion of the substrate and products became more pronounced, and lipolytic activity increased upon addition of n-hexane into the reaction medium. The storage and operational stabilities of the immobilized lipase were investigated, and both characteristics were found to be increased when compared to the free enzyme. Furthermore, mechanical or magnetic stirring during the operation were found to have no influence on the carrier-matrix as determined by nephelometric measurements.     

AnSBBR Applied to a Personal Care Industry Wastewater Treatment: Effects of Fill Time,Volume Treated Per Cycle,and Organic Load

A study was performed regarding the effect of the relation between fill time, volume treated per cycle, and influent concentration at different applied organic loadings on the stability and efficiency of an anaerobic sequencing batch reactor containing immobilized biomass on polyurethane foam with recirculation of the liquid phase (AnSBBR) applied to the treatment of wastewater from a personal care industry. Total cycle length of the reactor was 8 h (480 min). Fill times were 10 min in the batch operation, 4 h in the fed-batch operation, and a 10-min batch followed by a 4-h fed batch in the mixed operation. Settling time was not necessary since the biomass was immobilized and decant time was 10 min. Volume of liquid medium in the reactor was 2.5 L, whereas volume treated per cycle ranged from 0.88 to 2.5 L in accordance with fill time. Influent concentration varied from 300 to 1,425 mg COD/L, resulting in an applied volumetric organic load of 0.9 and 1.5 g COD/L.d. Recirculation flow rate was 20 L/h, and the reactor was maintained at 30 °C. Values of organic matter removal efficiency of filtered effluent samples were below 71% in the batch operations and above 74% in the operations of fed batch followed by batch. Feeding wastewater during part of the operational cycle was beneficial to the system, as it resulted in indirect control over the conversion of substrate into intermediates that would negatively interfere with the biochemical reactions regarding the degradation of organic matter. As a result, the average substrate consumption increased, leading to higher organic removal efficiencies in the fed-batch operations.