OBJECTIVETo use a new kind of fixing material, i.e. Sol-Gel organic-inorganic hybridized material to immobilize bacterium to detect Biochemical oxygen demand quickly.

METHODSThe biosensor was fabricated using a thin film in which Hansenula anomala was immobilized by sol-gel and an oxygen electrode. The optimum measurement for biochemical oxygen demand was at pH 7.0; 28 degrees C; response time 3 - 12 min. Pure organic compound, sewage and rate of recovery were detected with the biosensor.

RESULTSIt shows that the BOD biosensor can be used to detect many organic compounds such as amino acid, glucide. It is suitable to monitor sewage and industrial waste water which has low level alcohols and phenols. The microbial membrane can work 3 months and remain its 70% activity. It is measured that the rate of recovery of BOD is between 90% to 105% in sewage.

CONCLUSIONThe study confirmed the effectiveness and usefulness of BOD sensor, which is quick, convenient, low cost and reliable with little interference.

Bacteria ; Biosensing Techniques ; instrumentation ; Cells, Immobilized ; Gels ; Membranes, Artificial ; Nylons ; Oxygen ; analysis ; Sewage ; analysis ; microbiology

In order to carry out the process of the repeated intermittent L-lactic acid fermentation by self-immobilized Rhizopus oryzae, we investigated the effect of medium compositions on the morphology of Rhizopus oryzae. In submerged culture, fungi can be grown as broths of freely suspended mycelia and pellets or clumps, the pellets can be immobilized by themselves. The optimum medium composition for the first patch of fermentation was: 120 g/L glucose, 3 g/L NH4NO3, 0.14 g/L KH2PO4, 0.16 g/L NaH2PO4 (the concentration ratio of K+ and Na+ was 1:1). After 72 h fermentation, the physical form of Rhizopus oryzae was mostly uniform pellet with the diameter of 1.0-2.0 mm, the concentration of L-lactic acid was 100.8 g/L, and the conversion rate of glucose was 84%. During 16 batches of repeated fermentation, the L-lactic acid level was above 60.0 g/L and the conversion rate of glucose was 75%. The fermentation time of every batch was 24 h.
Cells, Immobilized ; metabolism ; Culture Media ; Fermentation ; Lactic Acid ; biosynthesis ; Rhizopus ; growth & development ; metabolism

Repeated batch fermentation with Rhizopus arrhizus immobilized by polyurethane was optimized in details including the time to replace, the volume of the replaced medium and the optimal composition of the medium to replace. Immobilized cells showed stability for repeated use. Immobilized cells could be used for 9 batches, lasting 140h in flask and 6 batches in 5L fermenter when the substitute culture medium consisted of (%): soybean flour 3.0, earthnut oil 0.5 and the liquid was replaced by 40% . The time for fermentation was reduced largely and the lipase productivity was 3 - 6 times as high as that in batch fermentation.
Biotechnology ; methods ; Cells, Immobilized ; metabolism ; Fermentation ; Lipase ; biosynthesis ; Plant Oils ; Rhizopus ; enzymology ; Soybeans

Lipase-mediated biodiesel production becomes increasingly important because of mild reaction conditions, pollution free during the process and easy product separation. Compared with traditional immobilized lipase, whole cell biocatalyst is promising for biodiesel production because it is easy to prepare and has higher enzyme activity recovery. Rhizopus oryzae IFO4697 can be used as the catalyst for biodiesel production. To further study the stability of the whole cell biocatalyst is extremely important for its further application on large scale. This paper focuses on the stability study of Rhizopus oryzae IFO4697 when used for the methanolysis of renewable oils for biodiesel production. The results showed that water content was important for achieving high catalytic activity and good stability of the biocatalyst. The optimum water content was found to be 5%-15%. Both particle size and desiccation methods showed no obvious effect on the stability of the biocatalyst. With GA cross-linking pretreatment, the stability of the biocatalyst could be improved significantly. When Rhizopus oryzae IFO4697 repeatedly used for next batch reaction, direct vacuum filtration was found to be a good way for the maintenance of good stability of the biocatalyst. Under the optimum reaction conditions, the methyl ester yield could keep over 80% during 20 repeated reaction batches.
Biocatalysis ; Bioelectric Energy Sources ; microbiology ; Biofuels ; Cells, Immobilized ; metabolism ; Lipase ; metabolism ; Rhizopus ; metabolism ; Soybean Oil ; metabolism

A diblock copolymer, poly(ethylene glycol) methacrylate-block-glycidyl methacrylate (PEGMA-GMA), was prepared on glass substrate by surface-initiated atom transfer radical polymerization (SI-ATRP), and endothelial specific peptide Arg-Glu-Asp-Val (REDV) was immobilized at the end of the PEGMA-GMA polymer brush by ring opening reaction through the rich epoxy groups in the GMA. The structure and hydrophilicity of the polymer brushes were characterized by static water contact angle, X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The results showed that the REDV modified copolymer brushes were successfully constructed on the glass substrates. The REDV peptide immobilized onto surface was quantitatively characterized by ultraviolet-visible spectroscopy (UV-VIS). The blood compatibility of the coating was characterized by recalcification time and platelet adhesion assay. The results showed that the polymer coating had good blood compatibility. The multifunctional active polymer coating with PEGMA and peptide produced an excellent prospect in surface construction with endothelial cells selectivity.
Biocompatible Materials ; Cells, Cultured ; Endothelial Cells ; Glass ; Humans ; Immobilized Proteins ; Methacrylates ; Oligopeptides ; Platelet Adhesiveness ; Polyethylene Glycols ; Polymers ; Surface Properties

OBJECTIVETo study the preparation of seeding type immobilized microorganisms and their degradation characteristics on di-n-butyl phthalate (DBP).

METHODSDiatomite, clinoptilolite, silk zeolite, and coal fly ash were chosen as reserved materials and modified. Their adsorption capacity and intensity in the bacteria were determined and the best carrier was picked out. The seeding type immobilized microorganisms were prepared by the best carrier and then it degraded DBP under different primary concentration, vibration rate, pH, temperature in the presence of metal compounds.

RESULTSThe adsorption capacity of the modified coal fly ash, silk zeolite, clinoptilolite and zeolite was 44.2%, 71.6%, 84.0%, and 94.4%, respectively, which was 1.66, 1.49, 1.37, and 1.16 times as high as that of their natural state. Their adsorption intensity was 72.1%, 90.5%, 90.1%, and 91.1% in turn. The modified diatomite was selected to prepare the seeding type immobilized microorganisms. When the primary DBP concentration was 100 to 500 mg/L, the DBP-degraded rate of the immobilized microorganisms could be above 80%. The degradation activity of both the dissociative and immobilized microorganisms was higher in vibration than in stillness. When pH was 6.0 to 9.0, the DBP-degraded rate of the immobilized microorganisms was above 82%, which was higher than the dissociative microorganisms. When the temperature was between 20 degrees C and 40 degrees C, the DBP-degraded rate could reach 84.5% in 24 h. The metal compounds could inhibit the degradation activity of both the dissociative and immobilized microorganisms. The degradation process of the immobilized microorganisms could be described by the first-order model.

CONCLUSIONThe adsorption capacity of the diatomite, clinoptilolite, silk zeolite and coal fly ash on DBP-degrading bacteria can be improved obviously after they are modified. The modified diatomite is best in terms of its adsorption capacity and intensity. Its seeding type immobilized microorganisms could degrade DBP effectively and is more adaptable to DBP load, temperature, pH than the dissociative microorganisms. The metal compounds could inhibit the activity of both the immobilized and dissociative microorganisms. The degradation reaction of the immobilized microorganisms on DBP is consistent with the first-order model.

Adsorption ; Bacteria ; metabolism ; Biodegradation, Environmental ; Biotechnology ; methods ; Cells, Immobilized ; Diatomaceous Earth ; Dibutyl Phthalate ; metabolism ; Hydrogen-Ion Concentration ; Kinetics ; Temperature ; Zeolites

The cis-epoxysuccinate hydrolase (CESH) from Rhizobium strain BK-20 is the key enzyme for L(+)-tartaric acid production. To establish a highly efficient and stable production process, we first optimized the enzyme production from Rhizobium strain BK-20, and then developed an immobilized cell-culture process for sustained production of L(+)-tartaric acid. The enzyme activity of free cells reached (3 498.0 +/- 142.6) U/g, and increased by 643% after optimization. The enzyme activity of immobilized cells reached (2 817.2 +/- 226.7) U/g, under the optimal condition with sodium alginate as carrier, cell concentration at 10% (W/V) and gel concentration at 1.5% (W/V). The immobilized cells preserved high enzyme activity and normal structure after 10 repeated batches. The conversion rate of the substrate was more than 98%, indicating its excellent production stability.
Alginates ; chemistry ; Cells, Immobilized ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Hydrolases ; metabolism ; Rhizobium ; enzymology ; metabolism ; Tartrates ; metabolism

Sugarcane bagasse modified by polyethylenimine (PEI) and glutaraldehyde (GA) was used as a carrier to immobilize Clostridium acetobutylicum XY16 in the process of butanol production. The effects of chemically modified sugarcane bagasse on batch and repeat-batch fermentations were investigated. Batch fermentation was conducted with an addition of 10 g/L modified sugarcane bagasse and 60 g/L glucose, resulting in a high solvent concentration of 21.67 g/L and productivity of 0.60 g/(L x h) with the treatment of 4 g/L PEI and 1 g/L GA. Compared to the fermentations by free cells and immobilized cells on unmodified sugarcane bagasse, the productivity increased 130.8% and 66.7%, respectively. The fibrous-bed bioreactor also maintained a stable butanol production during repeat-batch fermentations, achieving a maximum productivity of 0.83 g/(L x h) with a high yield of 0.42 g/g.
Batch Cell Culture Techniques ; Bioreactors ; Butanols ; metabolism ; Cells, Immobilized ; Cellulose ; metabolism ; Clostridium acetobutylicum ; metabolism ; Fermentation ; Saccharum ; chemistry

OBJECTIVETo investigate the nitrifying characteristics of both suspended- and attached-biomass in a hybrid bioreactor.

METHODSThe hybrid biological reactor was developed by introducing porous ceramic particles into the reactor to provide the surface for biomass attachment. Microorganisms immobilized on the ceramics were observed using scanning electron microscopy (SEM). All chemical analyses were performed in accordance with standard methods.

RESULTSThe suspended- and attached-biomass had approximately the same nitrification activity. The nitrifying kinetic was independent of the initial biomass concentration, and the attached-biomass had a stronger ability to resist the nitrification inhibitor.

CONCLUSIONThe attached biomass is superior to suspended-biomass for nitrifying wastewater, especially that containing toxic organic compounds. The hybrid biological reactor consisting of suspended- and attached-biomass is advantageous in such cases.

Biomass ; Bioreactors ; Cells, Immobilized ; Ceramics ; Microscopy, Electron, Scanning ; Nitrogen ; metabolism ; Phenol ; pharmacology ; Sewage ; Waste Disposal, Fluid ; instrumentation

Cell transplantation is a promising technology in cancer therapy, however, immunological rejection is the major problem of cell transplantation. Based on the permselective property of microcapsule membrane, encapsulated cells can be immuno-protected. The normal physiological state and function expression of cells can be maintained so as to realize allo- or xenotransplantation. The microencapsulated cells grow in three dimensions, giving a more biologically representative in vivo model, which hints difference in characters of growth and metabolism compared to the monolayer cells. Therefore, characterization of growth and metabolism of microencapsulated recombinant CHO cells is essential for further large-scale culture. In present study, the effect of concentration of glutamine on the growth, metabolism and endostatin production of microencapsulated cells was investigated. In the experimental range of initial glutamine concentrations from 2.69mmol/L to 9.05mmol/L in the culture of microencapsulated recombinant CHO cells, the maximum density of active cells and multiplication ratios almost kept constant. The specific consumption rate of glucose increased with lower initial glutamine concentration (2.69mmol/L). When initial glutamine concentration was much higher (7.91mmol/L to approximately 9.05mmol/L), the specific consumption rates of both glucose and glutamine increased while the efficiencies of glucose and glutamine decreased. The highest efficiencies of glucose and glutamine utilization were observed with initial glutamine concentration of 4.97mmol/L. It was also demonstrated that glutamine had significant effect on the accumulation of endostatin. The accumulative concentration of endostain reached its peak of 546.36 ng/mL with the initial glutamine concentration of 4.97mmol/L.
Animals ; CHO Cells ; Cell Culture Techniques ; Cell Proliferation ; drug effects ; Cells, Immobilized ; Cricetinae ; Cricetulus ; Culture Media ; Endostatins ; biosynthesis ; genetics ; Genetic Engineering ; Glucose ; metabolism ; Glutamine ; pharmacology ; Recombination, Genetic