Comparative analysis of hydrogen-producing bacteria and its immobilized cells for characteristics of hydrogen production

A strain of hydrogen producing bacteria was immobilized by polyvinyl alcohol-boric acid method, with the addition of a small amount of calcium alginate. The immobilized cells were insensitive to the presence of traces of O2. Moreover, the immobilized cells increased both the evolution rate and the yield of hydrogen production. Batch experiments with a medium containing 10 g/L glucose demonstrated the yields of hydrogen production by the immobilized and free cells were 2.14 mol/mol glucose and 1.69 mol/mol glucose, respectively. In continuous cultures at medium retention time of 2.0 h, the yield and the evolution rate of hydrogen production by the immobilized cells were 2.31 mol/mol glucose and 1 435.4 ml/（L·h） respectively. However, at medium retention time of 6.0 h, the yield and the evolution rate of hydrogen production by free cells were only 1.75 mol/mol glucose and 362.9 ml/（L·h）, respectively.

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.

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.

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 Agmatine by Immobilized Escherichia coli Cells with Arginine Decarboxylase Activity

A new method for the enzymatic synthesis of agmatine by immobilized Escherichia coli cells with arginine decarboxylase（ADC） activity was established and a series of optimal reaction conditions was set down. The arginine decarboxylase showed the maximum activity when the pyridoxal phosphate（PLP） concentration was 50 mmol/L, pH=7 and 45 °C. The arginine decarboxylase exhibited the maximum production efficiency when the substrate concentration was 100 mmol/L and the reaction time was 15 h. It was also observed that the appropriate concentration of Mg2＋, especially at 0.5 mmol/L promoted the arginine decarboxylase activity; Mn2＋ had little effect on the arginine decarboxylase activity. The inhibition of Cu2＋ and Zn2＋ to the arginine decarboxylase activity was significant. The immobilized cells were continuously used 6 times and the average conversion rate during the six-time usage was 55.6%. The immobilized cells exhibited favourable operational stability. After optimization, the maximally cumulative amount of agmatine could be up to 20 g/L. In addition, this method can also catalyze D,L-arginine to agmatine, leaving the pure optically D-arginine simultaneously. The method has a very important guiding significance to the enzymatic preparation of agmatine.

Asymmetric Synthesis of (-)-1-Trimethylsilyl-ethanol with Immobilized Saccharomyces Cerevisiae Cells in Water/Organic Solvent Diphasic System

Asymmetric synthesis of (-)-1-trimethylsilyl-ethanol with immobilized Saccharomyces cerevisiae cells in water/organic solvent biphasic system was studied. The effects of shake speed, hydrophobicity of organic solvent, volume ratio of water phase to organic phase, pH value of aqueous phase and reaction temperature on the initial reaction rate, maximum yield and enantiomeric excess (ee) of the product were systematically explored. All the above-mentioned factors had significant influence on the reaction. n-Hexane was found to be the best organic solvent for the reaction. The optimum shake speed, volume ratio of water phase to organic phase, pH value and reaction temperature were 150 r.min-1, 1/2, 8 and 30 ℃ respectively, under which the maximum yield and enantiomeric excess of the product were as high as 96.8% and 95.7%, which are 15% and 16% higher than those of the corresponding reaction performed in aqueous phase. To our best knowledge, this is the most satisfactory result obtained.

Asymmetric Reduction of 3,5-Bistrifluoromethyl Acetophenone with NADH Regeneration by Immobilized Cells of Saccharomyces rhodotorula in Aqueous-Organic Solvent Biphasic System

Asymmetric reduction of 3,5-bistrifluoromethyl acetophenone to produce(S)-3,5-bistrifluoromethylphenyl ethanol was successfully carried out with sodium alginate immobilized Saccharomyces rhodotorula cells in an aqueous-organic solvent biphasic system.The possible influential factors were examined thoroughly according to their effects on conversion rate and e.e of the product.Organic solvents were rated by their biocompatibility and conversion potential.The immobilized cells [125 mg/mL in 20 mmol/L Tris-HCl buffer and 5%(j) octane at pH 8] showed the best conversion with a substrate concentration of 1.42 g/L at 30℃ with glucose as co-substrate for cofactor regeneration.Sequential 8-batch process was carried out with immobilized cells with a slow decrease in conversion and e.e.The immobilized cells showed stable catalytic activity with 50% reserved activity and are superior especially in reusability in comparison with resting cells.

Production of D-p-Hydroxyphenylglycine from DL-5-p-Hydroxyphenyl Hydantoin by Immobilized Pseudomonas Putida Cells in a Hollow Fiber Membrane Bioreactor

An immobilized cell membrane bioreaction system was developed to promote cell stability. The hollow fiber membrane bioreactor with immobilized Pseudomonas putida cells, operating in continual repeated batch operation mode was used for producing D-p-hydroxyphenylglycine from DL-5-p-hydroxyphenyl hydantoin. The concentration of N-carbamyl-D-p-hydroxyphenylglycine and D-p-hydroxyphenylglycine in the efflux was analyzed by high performance liquid chromatography at different intervals.

Production of Indigo by Immobilization of E. coli BL21 （DE3） Cells in Calcium-Alginate Gel Capsules

The ability of catalyzing indole into indigo of gene engineering strain expressing P450 BM3 immobi- lized by entrapment in calcium-alginate gel capsules was examined,and various characteristics of immobilized cells were assessed.Optimum conditions for cells activity were not affected after immobilization,and pH and tempera- ture for both free and immobilized cells were found to be pH 7.5 and 35℃,respectively.The immobilized cells ex- hibited a markedly improved thermal stability than free cells.After five repeated experiments,the yield of indigo with the immobilized cells retained over 94%of their original activity,which indicated that the operational stability for recycling in batch processes was improved.

Rational design of Aspergillus flavus A5p1-immobilized cell system to enhance the decolorization of reactive blue 4(RB4)

Anthraquinone dyes are a class of typical carcinogenic and hard-biodegradable organic pollutants.This study aimed to enhance the decolorization of anthraquinone dye by rationally designing an expected immobilized system.Reactive blue 4(RB4) was used as a substrate model and a previous isolated dyedegrading strain Aspergillus flavus A5pl was purposefully immobilized.Considering the effects of cell attachment and mass transfer,the polyurethane foam(PUF) with open pore structure was selected as the immobilization carrier.Results showed that the RB4 decolorization efficiency was significant enhanced after immobilization.Compared to the free mycelium system,the decolorization time of200 mg·L^(-1)RB4 was shortened from 48 h to 28 h by the PUF-immobilized cell system.Moreover,the PUF-immobilized system could tolerate RB4 up to 2000 mg-L^(-1).In the packed bed bioreactor(PBBR),an average decolorization efficiency of 93.3% could be maintained by the PUF-immobilized system for26 days.The decolorization process of RB4 was well described by the logistic equation and the degradation pathway was discussed.It was found that the higher specific growth rate of the PUF-immobilized cells was one of reasons for the enhanced decolorization.The good performance of the PUFimmobilized cell system would make it have potential application value for RB4 bioremediation.

Batch CGTase Production with Free and Immobilized <i>Bacillus firmus</i>Strain 37 in Bovine Bone Charcoal

The present study aimed to study the batch production of CGTase (cyclomaltodextrin-glucanotransferase) with Bacillus firmus strain 37 free and immobilized in bovine bone charcoal in batch mode and in a fluidized bed batch reactor, respectively. The bovine bone charcoal is an innovative support material for the immobilization of microorganisms’ producers of enzymes and the use of this microbial support allows its reuse to a significant cost reduction of the process. The batch fermentation with free cells was investigated for 96 h and reached a CGTase activity equal to 0.77 U/mL. When the microorganism was immobilized on bovine bone charcoal (7 g) and cultivated in fluidized bed batch reactor with air supplementation (1 volume of air/volume of medium * minute), the same activity could be achieved in 24 h. The results of enzymatic activity achieved show the potential of CGTase production in a short time with Bacillus firmus strain 37 immobilized in bovine bone charcoal matrix and using air supplementation in the production medium.

Repeated-batch Cultivation of Encapsulated Monascus purpureus by Polyelectrolyte Complex for Natural Pigment Production

In general,productions of natural pigment in submerged microorganism culture were much less than that in solid-state fermentation,because the solid-state culture can provide a support carrier for the mycelium. To improve natural pigment production,the cultivation of Monascus purpureus in submerged encapsulated cell was investigated. Monascus purpureus immobilized in polyelectrolyte complex(PEC) microcapsules,which were pre-pared by sodium cellulose sulphate(NaCS) and poly-dimethyl-diallyl-ammonium chloride(PDMDAAC),was a good substitute for submerged cell culture because it mimicked the solid-state environment. The repeated-batch process with encapsulated cells was studied in flasks and a bubble column. The results indicated that the bubble column was more suitable for the encapsulation culture than the shaking flasks because of its good mass transfer performance and minor shear stress on cells. Owing to the protection of the microcapsule's membrane,Monascus purpureus in microcapsules increased approximately three times over that in free cell culture with negligible cell leakage to the medium. The pigment production in the bubble column finally reached 3.82(OD500) ,which was two times higher than in free cell culture. In addition,the duration of each batch was shortened to 15% of that in free cell culture.

Inhibiting Smooth Muscle Cell Proliferation via Immobilization of Heparin/Fibronectin Complexes on Titanium Surfaces

The aim of this study was to investigate the inhibitory effect of heparin/fibronectin （Hep/Fn） complexes on neointimal hyperplasia following endovascular intervention. Hep/Fn complexes were immobilized onto titanium （Ti） surfaces, with subsequent X-ray photoelectron spectroscopy （XPS）, Toluidine Blue 0 （TBO） and immunohistochemistry methods were used to characterize surface properties. Smooth muscle cell （SMC） cultures were used to evaluate the effect of Hep/Fn complexes on SMC proliferation. Results showed that Hep/Fn complexes successfully immobilized onto Ti surfaces and resulted in an inhibition of SMC proliferation. This study suggests that Hep/Fn surface-immobilized biomaterials develop as a new generation of biomaterials to prevent neointimal hyperplasia, particularly for use in cardiovascular implants.

Improved Method for Lacosamide Synthesis with Chemoenzymatic Method

Lacosamide was prepared by chemical method coupled with enzymatic method. N-Acetyl-D, L-3-methoxy-alanine, derived from D,L-3-methoxy-alanine, was used in the resolution process catalyzed by immobilized Escherichia coli cells with aminoacylase（EC3.5.1.14） activity. N-Acetyl-D-3-methoxy-alanine and L-3- methoxy-alanine were obtained from the resolution system. Lacosamide was synthesized by the amidation of N-acetyl-D-3-methoxy-alanine with benzylamine.

Adaptation of intestine-based microbial functions to bioethanol production

Animal intestine is a favorable habitat to microbes. It facilitates the evolution of dense and diversified microbial communities that are highly active and persistent throughout life span. Here, we stimulate this unique biosystem to develop high-efficient continuous bio-manufacturing processes. The pig small intestine was explored as a novel bioreactor with industrial Saccharornyces cerevisiae for biofuel production. Results showed that the small intestine was a beneficial material for cell adherence. The cells on the intestine exhibited the abilities of self- immobilization, self-duplication and self-repairing. Therefore the intestine-based S. cerevisiae could be continu- ously used for a long time at high metabolic activities. Both the fermentation speed and ethanol yield were im- proved. This study provides valuable insights into the functions of intestine-based biosystem and should inspire the development of bionic industrial processes. Future dissection of the interface mechanism and design of more bionic materials will make bioprocesses more economically favorable and environmentally sustainable.

Biodegradation of Crude Oil in Contaminated Soils by Free and Immobilized Microorganisms

The efficiencies of free and immobilized bacterial cultures of petroleum hydrocarbon degraders were evaluated and compared in this study. Hydrocarbon-degrading microbial communities with high tolerance to and high degrading ability of crude oil were obtained from the soil contaminated with crude oil in the Yellow River Delta. Then, the microbial cells were immobilized in sodium alginate （SA） beads and sodium Mginate-diatomite （SAD） beads. The biodegradation of crude oil in soil by immobilized cells was compared with that by free cells at three inoculation concentrations, 1× 104 colony forming units （cfu） kg-^（-1）（low concentration, L）, 5 × 104 cfu kg^（-1） （medium concentration, M）, and 1× 105 cfu kg^（-1） （high concentration, H）. At 20 d after inoculation, the maximum degradation rate in the immobilized systems reached 29.8% （SAD-M）, significantly higher （P 〈 0.05） than that of the free cells （21.1%）, and the SAD beads showed greater degradation than the SA beads. Moreover, both microbial populations and total microbial activity reached significantly higher level （P 〈 0.05） in the immobilized systems than free cell systems at a same initial inoculation amount. The scanning electronic microscope （SEM） images also confirmed the advantages of the immobilized microstructure of SAD beads. The enhanced degradation and bacterial growth in the SAD beads indicated the high potential of SAD beads as an effective option for bioremediation of crude oil-contaminated soils in the Yellow River Delta.

A new method of immobilizing HEK293 cells on the inner wall of a capillary column as stationary phase for drug screening

It is a new strategy to immobilize cells on the inner wall of a capillary column and use affinity capillary electrophoresis（ACE） to study receptor-ligand interactions or to screen natural products and compounds synthesized by combinatorial chemistry. In this paper, we developed a new method of immobilizing HEK293 cells on the inner wall of a capillary column. Four important experimental conditions were optimized, including cell injection density, PLL concentration, cell culturing time and sterile processing method. Immobilized cell-coated capillary columns prepared under the optimized experimental conditions exhibited good uniformity, stability and durability, which were suitable for capillary electrophoresis. The method could also be used to immobilize HEK293 cells over-expressing certain membrane receptors on the inner wall of a capillary. In this way, cell-coated capillary columns could be applied to ACE drug screening targeting certain membrane proteins.

An innovative approach to minimize excess sludge production in sewage treatment using integrated bioreactors

The present investigation deals with an application of integrated sequential oxic and anoxic bioreactor（SOABR） and fluidized immobilized cell carbon oxidation（FICCO） reactor for the treatment of domestic wastewater with minimum sludge generation. The performance of integrated SOABR-FICCO system was evaluated on treating the domestic wastewater at hydraulic retention time（HRT） of 3 hr and 6 hr for 120 days at organic loading rate（OLR）of 191 ± 31 mg/（L·hr）. The influent wastewater was characterized by chemical oxygen demand（COD） 573 ± 93 mg/L; biochemical oxygen demand（BOD5） 197 ± 35 mg/L and total suspended solids（TSS） 450 ± 136 mg/L. The integrated SOABR-FICCO reactors have established a significant removal of COD by 94% ± 1%, BOD5 by 95% ± 0.6% and TSS by 95% ± 4% with treated domestic wastewater characteristics COD 33 ± 5 mg/L; BOD59 ± 0.8 mg/L and TSS 17 ± 9 mg/L under continuous mode of operation for 120 days. The mass of dry sludge generated from SOABR-FICCO system was 22.9 g/m3. The sludge volume index of sludge formed in the SOABR reactor was 32 mL/g and in FICCO reactor it was 46 mL/g. The sludge formed in SOABR and FICCO reactor was characterized by TGA, DSC and SEM analysis. Overall, the results demonstrated that the integrated SOABR-FICCO reactors substantially removed the pollution parameters from domestic wastewater with minimum sludge production.

Preliminary study of groundwater denitrification using a composite membrane bioreactor

A composite membrane bioreactor(CMBR)integrating the immobilized cell technique and the membrane separation technology was developed for groundwater denitrification.The CMBR had two well mixed compartments with one filled with the nitratecontaining influent and the other with a dilute ethanol solution;the compartments were separated by the composite membrane consisting of a microporous membrane facing the influent and an immobilized cell membrane facing the ethanol solution.Nitrate and ethanol molecules diffused from the respective compartments into the immobilized cell membrane where nitrate was reduced to gaseous nitrogen by the denitrifying bacteria present there with ethanol as the carbon source.The microporous membrane was attached to one side of the immobilized cell membrane for retention of the disaggregated bacteria.Relative to the single dose of external ethanol,the twodose supplementation produced better treatment results as evidenced by the lower concentrations of NO_(3)^(-)-N and ethanol(as measured by total organic carbon)of the effluent.The batch treatment in CMBR removed most of the nitrate in the influent and attained a stable denitrification rate of 0.1 g·m^(-2)·h^(-1)for most of the 96-h cycles during the 30-cycle study.The effluent was essentially free of ethanol and nitrite nitrogen.