Colonisation of polyurethane reticulated foam biomass support particle by methanogen species

Summary The ability of methanogen species to colonise reticulated polyurethane foam biomass support particles (BSP) in a continuous culture system using formate as carbon source was investigated. Scanning electron micrograph evidence and biomass measurements indicate that two methanogen species effectively colonised within the the matrix of the support particle.The freely suspended colonised BSP are resistant to washout, and a comparison of methane output of the immobilised culture and a liquid culture of the methanogens indicates the potential for process intensification of methane production.     

Sulphur and sulphate reduction with acetate and propionate in an aerobic process for sulphide removal

Summary Sulphide production rates of sulphur-and sulphate-reducing bacteria up to 50 mg per biomass support particle per day were observed in an aerated sulphide-removal reactor with polyurethane (PUR) foam as carrier material. The optimal pH and temperature for the sulphide-producing bacteria were 8.0 and 30°C respectively. Raschig rings and four types of cube-shaped PUR particles were tested as carrier materials. When using PUR particles, the sulphide production rate was always between 3% and 4% of the sulphide removal rate, dependent on the dimensions and pore size of the polyurethane support particles. With the Raschig rings this ratio was only 2% and for reactors in which no carrier materials were present it was even lower (0.6%). Media containing different mixtures of acetate, propionate, sulphur and sulphate inoculated under anoxic conditions with sludge from the aerated reactor showed the presence of acetate-degrading sulphur-reducing, but not of acetate-degrading sulphate-reducing, bacteria. With propionate as sole electron donor no degradation occurred in the presence of sulphur within 2 weeks, whereas sulphate-dependent propionate oxidation started after 5–6 days incubation. Bacteria were isolated and resembled Desulfuromonas acetoxidans and Desulfobulbus propionicus morphologically and physiologically.     

Tissue reactions to breast implants coated with polyurethane.

The histological features noted in the capsules from 7 polyurethane coated silicone breast prostheses are described. The polyurethane provoked a definite foreign body reaction and was slowly degraded, with some particles ejected from the capsule into the surrounding tissues. Separation of the polyurethane coating from the silicone prosthesis and the degradation of the polyurethane took about two years. Another much more resistant foreign material was found to occur in conjunction with the polyurethane in the capsules. It may be an adhesive or flakes off the silicone shell. Vacuolated spaces were noted in the inner layers of 3 capsules; it was assumed that they contained liquid silicone.     

Growth of Acinetobacter gerneri P7 on polyurethane and the purification and characterization of a polyurethanase enzyme

A soil microorganism, designated as P7, was characterized and investigated for its ability to degrade polyurethane (PU). This bacterial isolate was identified as Acinetobacter gerneri on the basis of 16?s rRNA sequencing and biochemical phenotype analysis. The ability of this organism to degrade polyurethane was characterized by the measurement of growth, SEM observation, measurement of electrophoretic mobility and the purification and characterization of a polyurethane degrading enzyme. The purified protein has a molecular weight of approximately 66?kDa as determined by SDS-PAGE. Substrate specificity was examined using p-nitrophenyl substrates with varying carbon lengths. The highest substrate specificity was observed using p-nitrophenyl-propanate with an activity of 37.58?±?0.21?U?mg(-1). Additionally, the enzyme is inhibited by phenylmethylsulfonylfluoride and by ethylenediamine-tetra acetic acid. When grown on Impranil DLN(?) YES medium, a lag phase was noted for the first 3?h which was followed by logarithmic growth for 5?h. For the linear portion of growth between 5 and 9?h, a μ value of 0.413?doublings?h(-1) was calculated. After 9?h of incubation the cell number dramatically decreased resulting in a chalky precipitate. Measurements of electrophoretic mobility indicated the formation of a complex between the PU and A. gerneri P7 cells. A hybrid zeta potential had been generated between the cells and polyurethane. Further evidence for a complex was provided by SEM observation where cells appeared to cluster along the surface of polyurethane particles and along edges of polyurethane films. Occasionally, the cells established an anchor-like structure that connected the cells to polyurethane particles.     

Citric acid production by Aspergillus niger immobilized on polyurethane foam

Summary Citric acid was produced using Aspergillus niger immobilized on polyurethane foam in a bubble column reactor. Most of the adsorbed cells remained on the support and, as a result, high oxygen tension was maintained during the reactor operation. However, uncontrolled growth of the pellets made continuous reactor operation difficult. The citric acid productivity obtained from 15 vol.% foam particles containing immobilized cells was 0.135 g/l per hour. This productivity of immobilized cells was almost the same as that of free cells. The oxygen level dropped to half saturation in 5 days in the immobilized cell culture in contrast to 2 days in the free cell culture.     

Operational optimization of a turbine blade reactor using macroporous support and its application to rice callus regeneration

Optimal operation condition was investigated for immobilized rice callus culture using a turbine blade reactor (TBR²) with polyurethane foam supports. By using polyurethane foam block as immobilization support, the inhibition of cell growth at a high agitation speed was avoided because the hydrodynamic stress against immobilized cell was probably reduced. Experimental results in each operational condition were assessed by means of rice callus growth, immobilization ratio in TBR and those regeneration frequencies in regeneration culture using solid medium. Concerning with pore size of polyurethane foam and support size, three-millimeter cube support of polyurethane foam with an average pore size of 1.3 mm was the most suitable support. The maximum immobilization ratio was 50% under 5% support volume by volume of growth medium. For improving the immobilization ratio of rice callus in the TBR, the optimum TBR operation and modification were investigated further. By repeating a periodic operation 3 times (agitating at 300 rpm for 5 min and then 50 rpm for 2 min, and then 200 rpm of constant agitation speed during the remaining time), almost all supports could entrap rice callus and homogeneous immobilization was attained. The immobilization ratio was improved as compared with that using a constant operation at 200 rpm. Next, the TBR was modified by setting an air sparger inside the stainless mesh cylinder. In the modified TBR, the floating support by air bubbles was reduced, and the immobilization ratio increased further and reached 86.3% when we increased the support volume to 15% under periodic operation on a daily basis. The regeneration frequency of immobilized callus was also slightly increased by periodic operation and modification of the TBR.     

Use of immobilized Bacillus subtilis for the stereospecific hydrolysis of d,l-menthyl acetate

Bacillus subtilis possessing a stereospecific menthyl esterase has been immobilized in a polyurethane foam and used for the hydrolysis of menthyl ester. The specific activity of cells immobilized in polyurethane foam decreased as the cell loading was increased. The aqueous content of the immobilized biocatalyst particles influenced the activity. The activity half-life of about 400 h observed for the immobilized biocatalyst is 66 × greater than that observed for free bacteria.     

Mathematical model of the oxidation of ferrous iron by a biofilm of Thiobacillus ferrooxidans

Microbial oxidation of ferrous iron may be a viable alternative method of producing ferric sulfate, which is a reagent used for removal of H(2)S from biogas. The paper introduces a kinetic study of the biological oxidation of ferrous iron by Thiobacillus ferrooxidans immobilized on biomass support particles (BSP) composed of polyurethane foam. On the basis of the data obtained, a mathematical model for the bioreactor was subsequently developed. In the model described here, the microorganisms adhere by reversible physical adsorption to the ferric precipitates that are formed on the BSP. The model can also be considered as an expression for the erosion of microorganisms immobilized due to the agitation of the medium by aeration.     

Factors affecting the immobilisation of plant cells on reticulated polyurethane foam particles

Summary Factors affecting the immobilisation and subsequent growth of plant cells in reticulated polyurethane foam particles have been studied using three plant species. Polyurethane foam from a number of commercial sources has been screened and a foam having a low phytotoxicity and good retention of plant cells selected for use. Particles (8×8×8 mm) of the material were seeded with plant cells from suspension culture and cells grown immobilised in particles until they occupied >80% of the available volume. For all species, foams containing small pores (60–80 ppi) were most effective in immobilising and retaining cells. For efficient use of the inoculum, high partial volumes of foam particles are required; with partial volumes above 40%, over 80% of the inoculum is taken up by the particles. While the initial immobilisation process presumably involves weak interactions between cells and the support material, factors such as inoculum size and the length of the loading period have been found to affect the immobilisation of cells and their subsequent growth within the matrix. A preliminary study of the requirements for the maintenance of viability of immobilised cultures at high cell densities has been made.     

Aspergillus niger inulinase immobilized in polyurethane foam and treated in pressurized LPG: A potential catalyst for enzymatic synthesis of fructooligosaccharides

Inulinase from Aspergillus niger was immobilized in polyurethane foam (PU). Immobilized catalyst was treated in pressurized liquefied petroleum gas (LPG) system. This biocatalyst was used in the fructooligosaccharide production using sucrose as substrate in aqueous system. The main objective of this study was to evaluate the reaction yield and productivity by using polyurethane foam as a low-cost support for enzyme immobilization in an alternative processes for fructooligosaccharide production in pressurized LPG system with potential for industrial application. The total FOS concentration obtained were 31% as a result of sucrose concentration reduction, and formation of FOS long chain (GF3 and GF4) from kestose (GF2). FOS concentrations of 5%, 22%, and 3% were obtained for GF2, GF3, and GF4, respectively. The methodology suggested in this research work, enzyme immobilization in a low-cost support, and treatment in LPG, showed potential technology for fructooligosaccharide synthesis.     

Enhanced activity of intracellular lipases from Rhizomucor miehei and Yarrowia lipolytica by immobilization on biomass support particles

The aim of this investigation was to obtain an efficiently immobilized intracellular lipase from Rhizomucor miehei and Yarrowia lipolytica. The activity of intracellular lipases from R. miehei and Y. lipolytica was enhanced by the addition of waste fats (beef tallow or poultry fat) to the medium and by cell immobilization on biomass support particles (BSPs, cubic particle of polypropylene or polyurethane foams). The highest intracellular activity of lipases was obtained after adding 20 and 50 BSPs to the medium of R. miehei (130.5 U) and Y. lipolytica (90.3 U), respectively. The best carrier for immobilizing intracellular lipases was polyurethane foam and the lipolytic activity of immobilized lipases was 2.1–4.3-times higher than the activity of lipases obtained from free biomass. The properties of the immobilized enzymes were very similar to the free enzymes but the immobilized intracellular lipases were more useful for the hydrolysis of waste fats. The highest reaction ratio (72%) and content of free fatty acids (68% (w/w)) in the reaction mixture was obtained after 72 h for beef tallow hydrolysis in a batch reaction with the immobilized lipases from R. miehei.     

Kinetic studies on urea hydrolysis by immobilized urease in a batch squeezer and flow reactor

The immobilization of urease on the reticulated polyurethane foam, and the kinetic phenomenon of urea hydrolysis by the resulting immobilized urease in both batch squeezer and circulated flow reactors were studied. Urease was immobilized with bovine serum albumin and glutaraldehyde on polyurethane foam support of 7 to 15 mum thickness. The residual apparent activity of urease after immobilization was about 50%. The good hydrodynamic property and flexibility of polyurethane foam were retained in solution after immobilization. A modified biofilm reactor model was used to describe the kinetic phenomenon of urea hydrolysis in both batch squeezer and circulated flow reactors. The characteristic parameters of the reactor model for both bioreactors were obtained by combining the Rosenbrock optimization method, the Rungs-Kutta method, and the Newton-Raphson method. The best-fit results were in good agreement with the experimental data. This study suggests another application of polyurethane foam in enzyme immobilization and immobilized enzyme reactors, which offers potential for practical applications in various bioreactors. (c) 1992 John Wiley & Sons, Inc.     

Immobilization ofPhanerochaete chrysosporium on porous polyurethane particles with application to biodegradation of 2-chlorophenol

Porous polyurethane particles were prepared and used for the immobilization of white rot fungusPhanerochaete chrysosporium. The immobilized cells were employed for the production of lignin peroxidase. Polyurethane immobilized spores, or mycelial pellets ofPhanerochaete chrysosporium as well as freely suspended mycelial pellets of fungus were used as biocatalyst for the degradation of 2-chlorophenol. The polyurethane carriers appear to be superior to the other carriers already used for the immobilization of fungus.     

Biosynthesis of pullulan using immobilized Aureobasidium pullulans cells

Immobilization of Aureobasidium pullulans by adsorption on solid supports and entrapment in open pore polyurethane foam were attempted. By adsorption, the highest cell loading of 0.012-0.018 g dry wt/cm(2) support was obtained in pH 2.0 medium. Under this acidic condition, the net surface charges (zeta potentials) of both the cells and supports were close to zero and no pullulan was synthesized. Cationic coatings of Cytodex and polyethylenimine were not efficient in enhancing the binding strength between the cells and the supports. Surface immobilized cells and polyurethane foam entrapped cells exhibited a similar fermentation characteristics resulting in ca. 18 g/L pullulan and ca. 5 g/L leaked cells. However, cells entrapped in the polyurethane foam were more shear resistant. The immobilized cells thus could be repeatedly used for pullulan biosynthesis.     

Liquid circulation velocity and overall gas holdup in a modified jet loop bioreactor with low density particles

Effect of low density particles on the apparent liquid circulation velocity and overall gas holdup was studied in a modified reversed flow jet loop bioreactor. Experiments were conducted using polyurethane beads, polystyrene particles which are comparable to bioparticles found in biological applications and glass beads. Influence of gas and liquid flow rates, draft tube to reactor diameter ratio and solids loading on these hydrodynamic properties were studied. The liquid circulation velocity was found to increase with an increase in liquid flow rate but decrease with an increase in gas flow rate or solids loading. The overall gas holdup increased with an increase in gas or liquid flow rate but decreased with an increase in solids loading. The range of optimum draft tube to reactor diameter ratio was found to be 04–0.5. The results obtained with low density particles were comparatively better than those with glass beads. Correlations were proposed to evaluate liquid circulation velocity and overall gas holdup in terms of operational and geometrical variables.     

Sorption isotherms and kinetics in the primary biodegradation of anionic surfactants by immobilized bacteria:: II. Comamonas terrigena N3H

Comamonas terrigena N3H was immobilized by covalent linking on silanized inorganic supports and by physical entrapment of cells within calcium alginate beads and reticulated polyurethane foam. Both entrapped cells were efficient for the primary biodegradation of the anionic surfactants dihexyl sulphosuccinate (DHSS) and dioctyl sulphosuccinate (DOSS), furthermore, exhibiting, in the case of polyurethane immobilized cells, a positive fractionating effect of the substrate by adsorption onto the polymer matrix. The overall kinetics for the surfactant removal from water were well-fitted to a biphasic process, a rapid passive sorption step of the surfactant onto the cell-loaded support and the intrinsic primary biodegradation slower step, both acting synergically.     

Ethanol production from sucrose by immobilizedZymomonas mobilis cells in polyurethane foam

Summary The possibility of using polyurethane foam as a support for the immobilization ofZymomonas mobilis cells to carry out sucrose conversion to ethanol was investigated. Sucrose hydrolysis efficiencies of 90% and higher, volumetric reactor productivity of 20 gL^–1h^–1 and final ethanol concentration of 6.3% (v/v) at a dilution rate of 0.4 h^–1 show the good performance of polyurethane foams for whole cell immobilization.     

Methanogenesis by methanogen species immobilised on reticulated foam biomass support particles. Inhibition of methane production at high substrate concentrations

Summary The rate of methanogenesis by a substrate depleted methanogenic culture immobilised on reticulated polyurethane foam particles became increasingly inhibited at –S>300 mM. (–S=Reactor substrate concentration). Inhibition was duplicated by the sequential addition of metabolically inert sorbitol to cultures 40–60 mins after initiating methane production with formate (–S<300 mM).     

利用固定化米根霉在三相流化床中发酵生成L-乳酸

用聚氨酯泡沫吸附固定米根霉菌丝,在三相流化床中对葡萄糖、木糖以及木糖渣的纤维素酶解液等不同碳源进行Ｌ－乳酸发酵研究,并对游离菌丝和固定化菌丝发酵Ｌ－乳酸进行了比较。结果表明,聚氨酯泡沫是米根霉的良好载体,具有经济、高效等特点。实验条件下,不同碳源的乳酸转化率分别为：葡萄糖,８２．５％;木糖,５３．８％;木糖渣酶水解液,７１．９％。三相流化床中固定化米根霉产酸速率（对葡萄糖）为１９．１ｇ．ｈ＾－１．     

Sorption isotherms and kinetics in the primary biodegradation of anionic surfactants by immobilized bacteria: I. Pseudomonas C12B

The surfactant-degrading biocatalyst Pseudomonas C12B was immobilized by covalent linking on silanized inorganic supports and by physical entrapment of cells within reticulated polyurethane foam. Both immobilized biocatalysts have been shown to be appropriate for the effective primary biodegradation of the anionic surfactants sodium dodecyl sulphate (SDS), dodecylbenzene sulphonic acid (DBS), dioctyl sulphosuccinate (DOSS) and dihexyl sulphosuccinate (DHSS). The overall surfactant removal from water by cells entrapped in reticulated polyurethane foam exhibits a biphasic process, a rapid sorption step of the surfactant onto the cell-loaded support and the intrinsic primary biodegradation slower step, both acting cooperatively. The optimization of variables for the adsorption and the biodegradation processes (flow rate, particle size, substrate concentration) have been studied. Sorption isotherms for the surfactants on reticulated polyurethane foam have been established as type II of the Brunauer, Deming, Deming and Teller (BDDT) classification. The kinetics of the primary biodegradation of SDS by cells covalent linked on sepiolite treated with 3-aminopropyl triethoxysilane (APTS) were found to be first-order. In this case, surfactant adsorption does not exist.