Continuous protein recovery from whey using liquid-solid circulating fluidized bed ion-exchange extraction

A liquid-solid circulating fluidized bed (LSCFB) continuous ion-exchange extraction system has been investigated for total protein recovery from whey solutions under various operating conditions. The effectiveness of a dynamic seal was evaluated between the riser and the downcomer, and the best conditions for the establishment of this seal were established. Start-up studies indicated that the system is robust and stable. Under optimal conditions, a productivity of 8.2 g of total protein removed per hour per kilogram of resin was achieved with a protein removal efficiency of 78.4%. However, higher overall protein recovery of up to 90% was also achieved under other conditions, with lower protein concentration in the effluent and a lower overall productivity.     

A solid phase adsorption method for preparation of bovine serum albumin-bovine hemoglobin conjugate

A solid phase adsorption method was proposed to prepare well-defined bovine serum albumin–bovine hemoglobin (Hb) conjugate. After adsorption by the solid phase, Q Sepharose Fast Flow media, bovine serum albumin (BSA) molecules were allowed to react with glutaraldehyde. The spacing out of BSA molecules on the solid phase was assumed to limit polymerization of BSA molecules, except some molecules bound closely on the solid phase resulting in minor dimer formation. Following the elution procedure, the activated monomeric BSA was separated from the dimers by gel filtration chromatography on Superdex 200 and then reacted with bovine Hb at 4 °C and pH 9.5. The 1:1 (BSA:Hb) conjugate was obtained with the yield of 64%. The P₅₀ values of the conjugates, prepared under anaerobic and aerobic conditions, were 19.1 and 14.2 mmHg, respectively. The dependence of the P₅₀ on chloride ions for the conjugate was slightly diminished, presumably due to covalent attachment of BSA to bovine Hb.     

Interfacial effect of extremely low frequency electromagnetic fields (EM-ELF) on the vaporization step of carbon dioxide from aqueous solutions of body simulated fluid (SBF)

Spontaneous processes in an aqueous solution of body simulated fluid (SBF) were monitored in closed vessel for a period of 1 month at 310 K, at atm pressure, and initial pH of 7.2, both with and without exposure to a square pulsed extremely low frequency electromagnetic fields (EM-ELF) of 250 microT, repeated at 75 Hz. The most important findings are that the SBF surface tension (gamma), evaluated under the EM-ELF field, is lower than the corresponding value measured without EM-ELF at any time. Furthermore, the pH of the exposed SBF is always more basic than that of the unexposed solution. As a consequence, when the EM-ELF is applied, calcium phosphate salts do not precipitate from the SBF solution for a period as long as 30 days. Behind all these experimental evidences there is only one mechanism: the vaporisation from the SBF-air interface of the CO(2)(aq) dissolved into the aqueous electrolyte solution. Thermodynamic analysis of these results establish that, at any given time, the difference, Delta, between the measured surface tensions with and without EM-ELF applied, gives the work of the electromagnetic forces to change the extent at which the CO(2)(aq) adsorbs at the liquid-air interface. It has been demonstrated that the work supply per second and per unit of area by the electromagnetic forces, 3.73 x 10(-10) mJ/s cm(2), is very near to the experimental slope in the plot Delta vs. t 1.7 x 10(-10) mJ/s cm(2). This leads to the conclusion that the EM-ELF fields have an interfacial effect on the concentration value of the CO(2) (aq) at the SBF-air interface. Because of that, the EM-ELF field is enhancing the CO(2) vaporisation rate; thus any other steps, which are a consequence of this mechanism, are changing. These results allow explanation of previous experiments concerning the precipitation of calcium carbonate from flowing hydrogen carbonate aqueous solution in the temperature range 353-373 K at a pressure of 0.1 MPa under the effect of static magnetic fields.     

Soil phosphorus fractions and adsorption as affected by organic and inorganic sources

The effect of organic and inorganic sources of phosphorus (P) on soil P fractions and P adsorption was studied in a field without plant growth on a Kandiudalf in western Kenya. A high-quality organic source, Tithonia diversifolia (Hemsley) A. Gray leaves, and a low-quality source, maize (Zea mays L.) stover, were applied alone or in combination with triple superphosphate (TSP). The P rate was kept constant at 15 kg P ha^-1. Soil extractable P (resin, bicarbonate and sodium hydroxide), microbial biomass P and C and P adsorption isotherms were determined during 16 weeks after application of treatments. Application of tithonia either alone or with TSP increased resin P, bicarbonate P, microbial P, and sodium hydroxide inorganic P. Tithonia alone reduced P adsorption at 2–16 weeks. Maize stover had no effect on any of the P fractions or P adsorption. At 8 weeks, the application of tithonia reduced microbial C-to-P ratio (20) as compared to maize stover, TSP and the control (31–34). The reduction in P adsorption by tithonia was accompanied by increases in all measured P fractions, the sum of P in those fractions (resin, bicarbonate and sodium hydroxide) being larger than the P added. The reduction in P adsorption apparently resulted from competition for adsorption sites, probably by organic anions produced during decomposition of the high quality tithonia. Integration of inorganic P (TSP) with organic materials had little added benefit compared to sole application of TSP, except that combination of tithonia with TSP increased microbial biomass. The results indicate that a high quality organic input can be comparable to or more effective than inorganic P in increasing P availability in the soil.     

Rate-limiting mass transfer in immunosorbents: characterisation of the adsorption of paraquat-protein conjugates to anti-paraquat Sepharose 4B

A series of paraquat-protein conjugates of different molecular size has been prepared by the coupling of paraquat hexanoate to the proteins lysozyme, ovalbumin, bovine serum albumin. The characteristics of the adsorption of these conjugates to an immunosorbent consisting of monoclonal anti-paraquat antibodies covalently immobilised to Sepharose 4B have been determined. Equilibrium adsorption isotherms were found to obey the Langmuir equation and indicated that 80% or more of the antibody binding sites were accessible to the conjugates. The rates of mass transfer of the conjugates to their adsorption sites on the immobilised antibodies was well described by a model in which mass transfer is controlled by transfer across the external film and diffusion within the porous adsorbent bead. The effective diffusivities of the conjugates within the immunosorbent were measured and has allowed the effect of the size of the adsorbing molecule on the rate of adsorption to be considered. The amount of paraquat that could be adsorbed and the rates of adsorption decreased as the size of the protein to which it is conjugated increased. The diffusivity of the conjugates within the pores of the adsorbent is reduced between two and five times compared to their diffusivities in free solution. The reduction is greater for the larger proteins and the variations of the effective diffusivities and the pore diffusivities with the molecular weight of the conjugate can be well described with simple correlations.     

Alcoholic fermentation by agar-immobilized yeast cells

Immobilized yeast cells in agar gel beads were used in a packed bed reactor for the production of ethanol from cane molasses at 30°C, pH 4.5. The maximum productivity, 79.5g ethanol/l.h was obtained with 195g/l reducing sugar as feed. Substrate (64.2%) was utilized at a dilution of 1.33h^-1. The immobilized cell reactor was operated continuously at a constant dilution rate of 0.67h^-1 for 100 days. The maximum specific ethanol productivity and specific sugar uptake rate were 0.610g ethanol/g cell.h and 1.275g sugar/g cell.h, respectively.     

用膨胀床金属亲和层析从淡菜匀浆液中分离纯化纤维素酶

研究了一种新的膨胀床金属亲和层析技术,即将金属亲和层析结合膨胀床层析,直接从淡菜(Blue mussel)匀浆液中纯化纤维素酶。研究了金属亲和配基种类、pH、离子强度及流速对酶吸附和解吸的影响,确定了酶洗脱条件和介质再生条件。一步可纯化纤维素酶194倍,酶收率达82%。本方法不需要预先去除细胞碎片,而且处理速率比传统层析技术高3～4倍。     

吸附树脂AB—8对甘草酸的吸附性能及其在提取纯化中的应用

考察了大孔吸附树脂AB-8对甘草酸的吸附性能和原液浓度pH值、流速、洗脱剂的种类对树脂吸附性能的影响。结果表明,AB-8树脂对甘草酸吸附量高,易于洗脱,分离效果较好,回收率较一般方法高,达70％～80％,纯度达90％以上。     

Pilot scale production of a recombinant human epidermal growth factor, secreted by Bacillus brevis, using expanded bed adsorption

Recombinant Bacillus brevis which carried an expression plasmid encoding the human epidermal growth factor (EGF) gene on a cryptic high-copy number plasmid, pHT926, extracellularly produced EGF in its biologically active form at a concentration of over 1.5 g L⁻¹ in the culture broth in a 30-L jar fermenter. The culture broth also contained some other EGF compounds, which mainly consisted of oligomeric and polymeric forms with disulfide bonds. We developed a simple purification method for EGF, without prior cell removal from the culture broth, comprising cation exchange expanded bed adsorption followed by ultrafiltration with UF 10 000 and 3000 membranes. The EGF compounds were efficiently separated from the EGF in its native form in the expanded bed adsorption step. With this purification method, only EGF in its native form was recovered from the culture broth, with a yield of nearly 80%, and 90% purity. This efficient and economic system has made it possible to use EGF as a pharmaceutical in the livestock industry. Received 10 June 1998/ Accepted in revised form 10 September 1998     

Prepared Bed Land Treatment of Soils Containing Diesel and Crude Oil Hydrocarbons

An ex situ, field-scale, prepared bed land treatment unit (LTU) was used to bio-remediate soils containing petroleum hydrocarbons. Two soils were treated in side-by-side units to compare performance: (1) a clayey silt containing crude oil hydrocarbons from releases 30 to 40 years ago and (2) a silty sand containing diesel fuel hydrocarbons from a leak about three years prior to the bioremediation. The effectiveness of the bioremediation in the LTU was evaluated over a period of 18 months. The results indicated that: (1) prepared bed bioremediation reduced the hydrocarbon concentration, mobility, and relative toxicity in the soil with the diesel fuel, and (2) chemical bioavailability appeared to limit bioremediation of the soil containing the crude oil hydrocarbons. Although the soils containing the crude oil hydrocarbons contained an average of 10,000?mg TPH/kg dry soil, these soils had limited hydrocarbon availability, nontoxic conditions, and low potential for chemical migration. For the soils containing the diesel fuel, active prepared bed bioremediation of about 15 weeks was adequate to reach an environmentally acceptable endpoint. At that time, there was little further TPH loss, no Microtox^TM toxicity, and limited hydrocarbon mobility.