Facile Preparation of Dopamine-Modified Magnetic Zinc Ferrite Immobilized Lipase for Highly Efficient Synthesis of OPO Functional Lipid

1,3-Dioleoyl-2-palmitoylglycerol(OPO)has been a hotspot of functional oils research in recent years,but due to the high cost of sn-1,3 specific lipase in enzymatic synthesis and the lack of biocatalyst stability,large-scale industrial application is difficult.In this study,the prepared magnetic ZnFe_(2)O_(4) was functionalized with dopamine to obtain ZnFe_(2)O_(4)@PDA,and the nano-biocatalyst ZnFe_(2)O_(4)@PDA@RML was prepared by immobilizing sn-1,3 specific lipase of Rhizomucor miehei lipase(RML)via a cross-linking method.The existence of RML on ZnFe_(2)O_(4)@PDA was confirmed by XRD,FTIR,SEM,and TEM.This strategy proved to be simple and effective because the lipase immobilized on magnetic nanoparticles could be quickly recovered using external magnets,enabling reuse of the lipase.The activity,adaptability to a high temperature,pH value,and operational stability of immobilized RML were superior to those of free RML.After optimizing the synthesis conditions,the OPO yield was 42.78%,and the proportion of PA at the sn-2 position(PA-Sn2)was 54.63%.After the first four cycles,the activity of ZnFe_(2)O_(4)@PDA@RML was not significantly affected.The magnetically immobilized lipase has good thermal stability,long-term storage stability,reusability,and high catalytic activity.It can be used as a green and efficient biocatalyst to synthesize the OPO functional lipid.

Effective desalination and anti-biofouling performance via surface immobilized MWCNTs on RO membrane

Desalination is considered a viable method to overcome the issue of water scarcity either from waste water or seawater. For this purpose, this study employed a facile approach to develop surface immobilized oxidized-MWCNTs(o-MWCNTs) onto crosslinked polyvinyl alcohol(PVA) membrane. Firstly, modified polysulphone substrate was synthesized on to which crosslinked PVA layer was spread onto it. PVA layer act as active layer for surface immobilization of o-MWCNTs in varying concentration. The functional group analysis, morphology and roughness of membranes surface was conducted out using FTIR, SEM and AFM respectively. The results showed that modified membranes, immobilized o-MWCNTs enhanced the salt rejection(Na_(2)SO_(4)) upto 99.8%. After contacting with Escherichia coli and Staphylococcus aureus for 2.5 h the bacteria mortalities of the fabricated membrane could reach 96.9%. Furthermore, the antibiofouling tests showed that OP-MWCNTs(1-5) modified membranes have higher anti-biofouling property than the control membrane.

Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose

Cytidine 5'-monophosphate(5'-CMP)is an essential nucleotide for additives.In this study,enhanced production of 5'-CMP was realized by the transformation of cytidine using co-immobilized di-enzymes,uridine-cytidine kinase(UCK)and acetate kinase(AcK).The immobilization yield of the enzyme had a clear correlation with the surface charges as zeta potential(ξ).Among them,ε-polylysinefunctionalized sepharose(SA-EPL,ξ=9.31 m V)showed high immobilization yield(78.8%),which was4.9-fold than that of nitrilotriacetic acid functionalized sepharose(SA-NTA,ξ=-12.6 m V).The residual activity of affinity co-immobilized enzyme(EPL-Ni/EPL@Ac K-UCK)was higher than 70.6%after recycled 10 times.Thus,this study provides an effective approach for the production of 5'-CMP with the advantages of low adenosine 5'-triphosphate(ATP)consumption,reduced side reactions,and improved reusability by co-immobilized UCK and Ac K on the functionalized Sepharose.

Preparation of Ionic Liquids Immobilized on FMIL-101 Catalysts for Conversion of CO_(2)to Propylene Carbonate

Metal-organic frameworks(MOFs)have attracted considerable research attention as a new type of porous material for catalytic applications.Herein,2,5-dihydroxyterephthalic acid was proposed to replace conventional terephthalic acid and reacted with chromic nitrate nonahydrate to synthesize a functional metal–organic framework(FMIL-101).This was then used to immobilize various compound ionic liquids to prepare three ionic liquids immobilized on FMIL-101 catalysts,namely,FMIL-101-[HeMIM]Cl/(ZnBr_(2))_(2),FMIL-101-[CeMIM]Cl/(ZnBr_(2))_(2),and FMIL-101-[AeMIM]Br/(ZnBr_(2))_(2).After characterization by Fourier-transform infrared spectroscopy,X-ray diffraction,ultraviolet spectroscopy,thermogravimetry,specific surface area analysis,and scanning electron microscopy,the catalysts were used to mediate cycloaddition reactions between carbon dioxide(CO_(2))and propylene oxide.The effects of reaction temperature,reaction pressure,reaction time,and catalyst dosage on the catalytic performance were investigated.The results revealed that the FMIL-101-supported CIL catalysts afforded the target product propylene carbonate with good catalytic performance and thermal stability.The optimal catalyst,FMIL-101-[CeMIM]Cl/(ZnBr_(2))_(2),displayed a propylene oxide conversion of 98.64%and a propylene carbonate selectivity of 96.63%at a reaction temperature of 110℃,a reaction pressure of 2.0 MPa,a catalyst dosage of 2.0%relative to propylene oxide,and a reaction time of 2.5 h.In addition,the conversion and selectivity of the catalyst decreased slightly after four cycles.Additionally,the catalyst decreased slightly in catalytic performance after being recycled four times.

Effect Research of Immobilized Algae-bacteria Removal Ammonia Nitrogen of Aquaculture Wastewater and Proposed Model

Applied Immobilized algae bacteria (ABI) to remove ammonia of freshwater aquaculture wastewater. Temperature (T),PH,light intensity (I),dissolved oxygen (DO) and filling rate five factors plays important role in the process of ammonia nitrogen removal ,related data between ammonia removal and five factors was received through multi-factor orthogonal test,and established relations model between the five factor and nitrogen removal. The results show that five-factors had significant effect on AR,and the best combinations for removing AR was temperature 30 ℃,pH=7.0,light intensity 6 000 lux,dissolved oxygen 5.0 mg/L and the fill rate 10%. According to the experimental data,equation model was proposed and coefficient of determination R2 =0.864 8,P<0.05. Samples T-test was done between the model predictions and the actual measured values.Test results showed that the significant difference of overall mean value sig. (2-tailed) was 0.978 (P>0.05),it Shows that had no significant difference between model predictions and the actual measured value,and model had a high degree of fitting.

Recent advances in immobilized enzymes on nanocarriers

Recent progress in nanotechnology has provided high-performance nanomaterials for enzyme immobilization.Nanobiocatalysts combining enzymes and nanocarriers are drawing increasing attention because of their high catalytic performance,enhanced stabilities,improved enzyme-substrate affinities,and reusabilities.Many studies have been performed to investigate the efficient use of cellulose nanocrystals,polydopamine-based nanomaterials,and synthetic polymer nanogels for enzyme immobilization.Various nanobiocatalysts are highlighted in this review,with the emphasis on the design,preparation,properties,and potential applications of nanoscale enzyme carriers and nanobiocatalysts.

Biodegradation of phenol by free and immobilized Acinetobacter sp.strain PD12

A new phenol-degrading bacterium with high biodegradation activity and high tolerance of phenol, strain PD 12, was isolated from the activated sludge of Tianjin Jizhuangzi Wastewater Treatment Facility in China. This strain was capable of removing 500 mg phenol/L in liquid minimal medium by 99.6% within 9 h and metabolizing phenol at concentrations up to 1100 mg/L. DNA sequencing and homologous analysis of 16S rRNA gene identified PD12 to be an Acinetobacter sp. Polyvinyl alcohol （PVA） was used as a gel matrix to immobilize Acinetobacter sp. strain PDI2 by repeated freezing and thawing. The factors affecting phenol degradation of immobilized cells were investigated, and the results showed that the immobilized cells could tolerate a high phenol level and protected the bacteria against changes in temperature and pH. Storage stability and reusability tests revealed that the phenol degradation functions of immobilized cells were stable after reuse for 50 times or storing at 4℃ for 50 d. These results indicate that immobilized Acinetobacter sp. strain PD 12 possesses a good application potential in the treatment of phenol-containing wastewater.

Comparison of Di-n-methyl Phthalate Biodegradation by Free and Immobilized Microbial Cells

To compare the biodegradation of di-n-methyl pathalate by free and immobilized microbial cells. Methods The enrichment and isolation technique was used to isolate the microorganism. The PAV-entrapment method was utilized to immobilize the microorganisms. The scanning electron microscophy (SEM) was used to observe the growth and distribution of microbial cells immobilized inside the PVA bead gels. The GC/MS method was used to identify the main intermediates of DMP degradation. Results The microbial cells could grow quite well in PVA gel. The metabolic pathway did not change before and after immobilization of the microbial cells. The degradation rate of immobilized cells was higher than that of free cells. Conclusion The immobilized microbial cells possess advantages than free cells when applied to the biodegradation of toxic organic pollutants.

Enzymatic Synthesis of Fatty Acid Methyl Esters from Crude Rice Bran Oil with Immobilized Candida sp. 99-125

The non-edible crude rice bran oil was extracted from white rice bran, and then was catalyzed by immobilized lipase for biodiesel production in this study. The effects of water content, oil/methanol molar ratio, temperature, enzyme amount, solvent,number of methanol added times and two-step methanolysis by using Candida sp. 99-125 as catalyst were investigated. The optimal conditions for processing 1 g rice bran oil were: 0.2 g immobilized lipase, 2 ml n-hexane as solvent, 20% water based on the rice bran oil mass, temperature of 40 °C and two-step addition of methanol. As a result, the fatty acid methyl esters yield was 87.4%. The immobilized lipase was proved to be stable when it was used repeatedly for 7 cycles.

Production of laccase by Coriolus versicolor and its application in decolorization of dyestuffs: (Ⅱ) Decolorization of dyes by laccase containing fermentation broth with or without self-immobilized mycelia

The capability of decolorization for commercial dyes by Coriolus versicolor fermentation broth containing laccase with or without immobilized mycelium was evaluated. With cell free fermentation broth containing laccase, high decolorization ratio was achieved for acid orange 7, but not for the other dyes concerned. The immobilized mycelium was proved to be more efficient than the cell free system. All the four dyestuffs studied were found being decolourized with certain extent by immobilized mycelium. The repeated batch decolorization was carried out with satisfactory results. The experimental data showed that the continuous decolorization of wastewater from a printing and dyeing industry was possible by using the self immobilized C. Versicolor.

Removal of Nitrogen, Phosphorus, and Organic Pollutants From Water Using Seeding Type Immobilized Microorganisms

Objective To study the possibility of removing nitrogen, phosphorus, and organic pollutants using seeding type immobilized microorganisms. Methods Lakes P and M in Wuhan were chosen as the objects to study the removal of nitrogen, phosphorus, and organic pollutants with the seeding type immobilized microorganisms. Correlations between the quantity of heterotrophic bacteria and the total nitrogen （TN）, total phosphorus （TP）, and total organic carbon （TOC） in the two lakes were studied. The dominant bacteria were detected, inoculated to the sludge and acclimated by increasing nitrogen, phosphorus and decreasing carbon source in an intermittent, time-controlled and fixed-quantity way. The bacteria were then used to prepare the seeding type immobilized microorganisms, selecting diatomite as the adsorbent cartier. The ability and influence factors of removing nitrogen, phosphorus, and organic pollutant from water samples by the seeding type immobilized microorganisms were studied. Results The coefficients of the heterotrophic bacterial quantity correlated with TOC, TP, and TN were 0.9143, 0.8229, 0.7954 in Lake P and 0.9168, 0.7187, 0.6022 in Lake M. Ten strains of dominant heterotrophic bacteria belonging to Pseudomonas, Coccus, Aeromonas, Bacillus, and Enterobateriaceae, separately, were isolated. The appropriate conditions for the seeding type immobilized microorgansims in purifying the water sample were exposure time=24 h, pH=7.0-8.0, and quantity of the immobilized microorganisms=0.75-1g/50 mL. The removal rates of TOC, TP, and TN under the above conditions were 80.2%, 81.6%, and 86.8%, respectively. Conclusion The amount of heterotrophic bacteria in the two lakes was correlated with TOC, TP, and TN. These bacteria could be acclimatized and prepared for the immobilized microorganisms which could effectively remove nitrogen, phosphorus, and mixed organic pollutants in the water sample.

Copper biosorption on immobilized seaweed biomass:Column breakthrough characteristics

The biosorption of copper by the brown seaweed Sargassum baccularia, immobilized onto polyvinyl alcohol （PVA） gel beads, was investigated with fixed-bed experiments. Laboratory-scale column tests were performed to determine breakthrough curves with varying flow rates and feed concentrations. A theoretical fixed-bed model, known as the Bohart-Adams equation, was evaluated in simulating the experimental breakthrough curves. The Bohart-Adams model qualitatively predicted the breakthrough trends. PVA- immobilized seaweed biomass beads were amenable to efficient regeneration with aqueous solution containing the chelating agent ethylenediaminetetraacetic acid （EDTA）. The biosorbent retained most of its original uptake capacity over three cycles of use. The excellent reusability of the biosorbent could lead to the development of a viable metal remediation technology.

Studies on Manganese Peroxidase Immobilized in Gelatin-containing Microemulsion-based Gels

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 H_2O_2 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 H_2O_2. When the amount of the immobilized H_2O_2 is sufficient, the microemulsion-based gels containing MnP could be used many times.

Repeated-Batch and Continuous Production of L-Lactic Acid by Rhizopus oryzae Immobilized in Calcium Alginat Beads:ReactorPerformance and Kinetic Model

Repeated-batch and continuous production of L-lactic acid by immobilized Rhizopusoryzae with calcium alginate entrapment method in a three-phase fluidized-bed bioreactor was stud-ied.The operation conditions were optimized.The productivity based on total reactor volume wasabout 3 times higher than that with free cells in a traditional stirred tank bioreactor.A mathemat-ical model was proposed and the model predictions were in good agreement with the experimentaldat.

Removal of Di-n-butyl Phthalate Using Immobilized Microbial Cells

The biodegradation of di-n-butyl phthalate （DBP） using immobilized microbial cells was carded out in an internal airlift loop reactor with ceramic honeycomb supports. A strain that is capable of degrading DBP was isolated from the activated sludge and identified as Bacillus sp. using 16S rDNA sequential analysis. Bacillus sp. could be rapidly attached onto the ceramic honeycomb supports. The immobilized cells could effectively degrade DBP in batch and continuous experiments. When the influent concentration of DBP was 50mg·L^-1, the effluent DBP reached less than lmg.L i with 6h hydraulic retention time （HRT） in continuous experiment. The immobilized microbial cells could grow and accumulate through the biodegradation of DBP, and the rate of degradation is accordingly increased. The possible pathway of DBP biodegradation using immobilized cells was tentatively proposed.

An efficient synthesis of ampicillin on magnetically separable immobilized penicillin G acylase

Penicillin G acylase(PGA) was immobilized on the magnetic hydrophilic polymer microspheres with average pore size of 17.1 nm,specific surface area of 128.2 m2/g and saturate magnetization of 6.4 emu/g.The 96.7%ampicillin yield with 1.60 of the synthesis/hydrolysis(S/H) ratio from 6-aminopenicillanic acid(6-APA) and D-(-)-alpha-phenylglycine methyl ester(D-PGME) can be achieved using the resultant magnetic biocatalyst in ethylene glycol,where only 82.1%yield with 1.40 of the S/H ratio was obtained using the free PGA under the identical reaction conditions.The immobilized PGA can be separated magnetically and recycled for five times without obvious loss of its catalytic activity.

Decolourization of Reactive Brilliant Blue KN-R by immobilized cells of Aspergillus ficuum

Aspergillus ficuum was immobilized with sodium alginate, and decolourization of Reactive Brilliant Blue KN-R was studied on immobilized and free Aspergillus ficuum. The optimal preparation condition of the strain immobilization was obtained by the orthogonal test, it is sodium alginate 3%, CaCl_2 5%, wet mycelia 30 g/L, calcific time 8 h. It was found that the immobilized cells could effectively decolourize Reactive Brilliant Blue KN-R, the optimum temperature and pH were 33℃ and 5.0, respectively. The kinetics study of decolourization of immobilized cells showed that the decolourization of Aspergillus ficuum immobilized conformed to zero-order reaction model. The decolourization efficiency of immobilized cell compared with that of free cell in different physical conditions. Results showed that the decolourization of immobilized cells with mycelia had the best efficiency. The immobilized cells could be reused after the first decolourization.

Lipase immobilized on HOOC-MCF:A highly enantioselective catalyst for transesterification resolution of (R,S)-1-phenylethanol

Pseudomonas cepacia lipase （PSL） immobilized on the carboxyl-functionalized meso-cellular foams （HOOC-MCF） was used for the transesterification resolution of （R,S）-l-phenylethanol in organic solvent. The results showed that the ee value of （S）-1- phenylethanol and （R）-1-phenylethyl acetate reached 99% with 50% conversion of 1-phenylethanol using toluene as solvent. Furthermore, it was found that PSL/HOOC-MCF exhibited high enantioselectivity in organic solvent with log P ≤ 2 such as toluene and hexane.

Hydrogen Production with High Evolution Rate and High Yield by Immobilized Cells of Hydrogen-producing Bacteria Strain B49 in a Column Reactor

To improve the hydrogen evolution rate in continuous hydrogen production of a fermentative hydrogen-producing bacteria strain B49 (AF481148 in EMBL), 4% immobilized cells by polyvinyl alcohol-boric acid method, with the addition of a small amount of calcium alginate in a column reactor obtain hydrogen yield of 2.31 mol H2/mol glucose and hydrogen evolution rate of 1435.4 ml/L culture&middoth respectively at medium retention time of 2 h with a medium containing l0 g glucose/L. As the cell density in gel beads is increased to 8%, hydrogen yield and hydrogen evolution rate for l0 g glucose/L are 2.34 mol H2/mol glucose and 2912.4 ml/L culture &middot h respectively at medium retention time of 1 h, and for molasses wastewater COD of 7505.9 mg/L hydrogen production potential of 205.6 ml/g COD and hydrogen evolution rate of 2057.7 ml/L culture&middoth at hydraulic retention time of 0.75 h are observed. In the continuous culture pH value keeps around 3.9 by self regulation.

Microfluidic reactor with immobilized enzyme-from construction to applications:A review

Microfluidic,as the systems for using microchannel(micron-or sub-micron scale)to process or manipulate microflow,is being widely applied in enzyme biotechnology and biocatalysis.Microfluidic immobilized enzyme reactor(MIER)is a tool with great value for the study of catalytic property and optimal reaction parameter in a flourishing and highly producing manner.In view of its advantages in efficiency,economy,and addressable recognition especially,MIER occupies an important position in the investigation of life science,including molecular biology,bioanalysis and biosensing,biocatalysis etc.Immobilization of enzymes can generally improve their stability,and upon most occasions,the immobilized enzyme is endowed with recyclability.In this review,the enzyme immobilization techniques applied in MIER will be discussed,followed by summarizing the novel developments in the field of MIER for biocatalysis,bioconversion and bioanalysis.The preponderances and deficiencies of the current state-of-the-art preparation ways of MIER are peculiarly discussed.In addition,the prospects of its future study are outlined.