Sterilization-in-place of concentrated nutrient solutions

Sterilization-in-place (batch sterilization) behavior of concentrated nutrient solutions was quantified for various nutrient solutions of fermentation processing interest. Experimental observations of sterilization temperatures and corresponding pressures suggested that sterilization pressures were substantially lower for concentrated nutrient solutions than for water. This effect was believed to be directly related to the lower vapor pressure and lower activity coefficient of these concentrated nutrient solutions. Using thermodynamic data for the specific nutrient, pressures and temperatures, calculated as a function of nutrient concentration, compared favorably with observed values. This method permits estimation of the expected sterilization pressure for concentrated nutrients for which no experimental observations have yet been made at the sterilization temperature of interest. Estimate accuracy, ranging from 0.006 to 0.06 bar, can be enhanced if one set of experimental temperature/pressure values is known at the expected sterilization scale. The potential impact of the nature of concentrated nutrient solutions on steam-in-place vessel headspace temperature distributions, D-values of B. stearothermophilus, and overall sterilization effectiveness is also discussed. The low vapor pressure of concentrated nutrient solutions was hypothesized to directly impact air removal effectiveness and sterilization performance during batch sterilization. Copyright 1999 John Wiley & Sons, Inc.     

DNA gelation in concentrated solutions

Sonicated calf-thymus DNA (200 ± 30 base pairs) spontaneously forms viscoelastic gels over a wide range of concentration, temperature, and buffer conditions. Quasielastic light scattering (QLS) can be used to monitor this process, because the ratio of dynamic-to-static scattering intensity decreases dramatically as gelation occurs. Using QLS, we have explored the effects of DNA concentration and mono- and divalent cations on the thermal stability of DNA gels. We found that the gel–sol transition temperature (T_gel) varies linearly with [DNA] from 7.5 to 17 mg/mL. Both Na⁺ and Mg²⁺ strongly stabilize the gel state. The sharpness of the transition increases with increasing ionic and DNA concentrations. Analysis of the Na⁺-dependent gelation indicates that the process requires the association of one Na⁺ per 118 base pairs. Mg²⁺ effectively stabilizes the gel at concentrations 10-fold below those required for Na⁺. The unexpectedly large effect of Mg²⁺ suggests that ion-specific interactions may play an important role in determining gel stability.     

Hydrodynamics of concentrated proteoglycan solutions

The dynamics of water transport in proteoglycan compartments has been studied in relation to osmotic flow (proteoglycan diffusion) and hydraulic permeability (proteoglycan sedimentation) in concentrated solutions of proteoglycan subunit and native proteoglycan aggregate isolated from Swarm rat chondrosarcoma. A central parameter that describes the kinetics of both types of water movement is the hydrodynamic frictional coefficient of water with proteoglycan. The frictional coefficient is markedly concentration dependent, increasing with increasing concentration, and highlights important structural features and types of organization of the proteoglycans in concentrated solutions. These include the requirements that proteoglycans in the extracellular matrix not to be immobilized but to have translational diffusive mobility and concentration gradients to be osmotically active, that chondroitin sulfate segmental mobility describing translational motion largely determines osmotic flow and hydraulic permeability of the proteoglycans, and that the proteoglycans exhibit an enhanced ability to resist flow as compared to other macromolecules. Additional dynamic studies suggest the formation of transient super-aggregate structures may occur at high concentrations which endows the proteoglycan subunit hydrodynamic properties similar to proteoglycan aggregate.     

Aeration of concentrated activated sludge

A multiphase project has been planned to develop a new biological process capable of economically treating high BOD wastes. Herein is presented the results of the first phase of the program, in which the feasibility of growing concentrated microbial cultures was investigated and the oxygen and power requirements for maintaining such cultures were determined. An example is given of the scale-up of power requirements for oxygen transfer in a prototype system.     

Yeast strains for concentrated substrates

Summary The screening of twenty yeast strains for ethanol productivity at high osmotic pressure at temperatures ranging from 32°C to 45°C is described. Shake flask fermentations of 30°, 40°, and 50° Bx cane molasses were performed. The effect of temperature on productivity at a non-inhibitory ethanol level is weakly pronounced. Most strains fermented poorly at 50° Bx molasses but two Schizosaccharomyces pombe and one commercial baker's yeast, Saccharomyces cerevisiae performed well at all concentrations of molasses. In an extended study with Schizosaccharomyces pombe (CBS 352) and Saccharomyces cerevisiae (SJAB, fresh yeast), simulating a continuous run it was shown that Schizosaccharomyces pombe was less sensitive to high DS than Saccharomyces cerevisiae. At 25% DS the productivity of Schizosaccharomyces pombe is almost twice that of Saccharomyces cerevisiae.     

Microalgae growth on concentrated human urine

In this study, for the first time, a microalga was grown on non-diluted human urine. The essential growth requirements for the species Chlorella sorokiniana were determined for different types of human urine (fresh, hydrolysed, male and female). Batch experimental results using microtiter plates showed that both fresh and synthetic urine supported rapid growth of this species, provided additional trace elements (Cu, Fe, Mn, and Zn) were added. When using hydrolysed urine instead of fresh urine, additional magnesium had to be added as it precipitates during hydrolysis of urea. C. sorokiniana was able to grow on non-diluted urine with a specific growth rate as high as 0.104 h^?1 under light-limited conditions (105 μmol photons m^?2 s^?1), and the growth was not inhibited by ammonium up to a concentration of 1,400 mg NH₄ ⁺-N L^?1. The highest growth rate on human urine was as high as 0.158 h^?1. Because it was demonstrated that concentrated urine is a rich and good nutrient source for the production of microalgae, its application for a large-scale economical and sustainable microalgae production for biochemicals, biofuels and biofertilizers becomes feasible.     

Material properties of concentrated pectin networks

We have examined the mechanical behaviour of different types of pectin at high concentrations (> 30% w/w), relevant to the behaviour of pectin in the plant cell wall, and as a film-forming agent. Mechanical properties were examined as a function of counterion type (K(+), Ca(2+), Mg(2+)), concentration and extent of hydration. Hydration was controlled in an osmotic stress experiment where pectin films were exposed to concentrated polyethylene glycol [PEG] solutions of known osmotic pressure. We investigated the mechanical behaviour under simple extension. The results show that the swelling and stiffness of the films are strongly dependent on pectin source and ionic environment. At a fixed osmotic stress, both Ca(2+) or Mg(2+) counterions reduce swelling and increase the stiffness of the film.     

Protein-protein interactions in concentrated electrolyte solutions

Protein-protein interactions were measured for ovalbumin and for lysozyme in aqueous salt solutions. Protein-protein interactions are correlated with a proposed potential of mean force equal to the free energy to desolvate the protein surface that is made inaccessible to the solvent due to the protein-protein interaction. This energy is calculated from the surface free energy of the protein that is determined from protein-salt preferential-interaction parameter measurements. In classical salting-out behavior, the protein-salt preferential interaction is unfavorable. Because addition of salt raises the surface free energy of the protein according to the surface-tension increment of the salt, protein-protein attraction increases, leading to a reduction in solubility. When the surface chemistry of proteins is altered by binding of a specific ion, salting-in is observed when the interactions between (kosmotrope) ion-protein complexes are more repulsive than those between the uncomplexed proteins. However, salting-out is observed when interactions between (chaotrope) ion-protein complexes are more attractive than those of the uncomplexed proteins.     

Induction of polyploidy by concentrated thymidine

Normal human skin fibroblast cells exposed for more than about 17 h to thymidine at concentrations above 1 mM showed an increase in polyploid index over control levels. This increase, in which cells with even polyploid numbers predominated, varied up to a maximum of 36%. The thymidine-treated cultures also show considerable chromosome damage in both diploid and polyploid cells.     

Preparation of concentrated lactic streptococcus starters

Single-strain cultures of Streptococcus cremoris were grown in a semisynthetic medium with automatic pH control. After centrifugation, the cells were resuspended in sterile nonfat milk (2% of the original volume). There was no significant difference in the maximum population attained when cultures were grown at pH values of 5.5, 6.0, or 6.5 with sodium hydroxide as the neutralizer. With ammonium hydroxide as the neutralizer, maximum populations obtained were increased about twofold. In most cases, the acid-producing ability of the culture concentrates was comparable to that of fresh-milk cultures. There was some variation among strains of S. cremoris with respect to the effects of different neutralizers and levels of pH control on the biological activity of the culture concentrates. The culture concentrates were stored in liquid nitrogen for as long as 231 days without significant loss in biological activity.     

Amylose crystallization from concentrated aqueous solution

Maize amylose, separated from granular starch by means of an aqueous leaching process, was used to investigate spherulite formation from concentrated mixtures of starch in water. Amylose (10-20%, w/w) was found to form a spherulitic semicrystalline morphology over a wide range of cooling rates (1-250 degrees C/min), provided it was first heated to >170 degrees C. This is explained through the effect of temperature on chain conformation. A maximum quench temperature of approximately 70 degrees C was required to produce spherulitic morphology. Quench temperatures between 70 and 110 degrees C produced a gel-like morphology. This is explained on the basis of the relative kinetics of liquid-liquid phase separation vis-à-vis crystallization. The possibility of the presence of a liquid crystalline phase affecting the process of spherulite formation is discussed.     

Inactivation of concentrated biomasses of Rickettsia prowazekii

The methods used for the sparing inactivation of highly concentrated R. prowazekii biomass and for the decrease of its infectious activity are described. These methods are recommended for use in experiments in the field of molecular biology, as well as for disinfection of different materials contaminated with rickettsiae. As conditions for complete inactivation, incubation at 50 degrees C for 1 hour without chemical disinfectants, treatment with 0.5% phenol solution at 30 degrees C for 12 hours and with 0.1% formaldehyde solution at 4 degrees C for 24 hours have been selected. Treatment with 0.5% phenol solution at 36 degrees C for 1 hour or incubation at 45 degrees C without the use of disinfectants ensures an essential decrease in the infectivity of the material if the work with viable infective agents is necessary. Ultraviolet irradiation for 1.5 hours and exposure to the action of 0.1-0.5% sodium azide are less effective.     

Enthalpy relaxation of freeze concentrated sucrose-water glass

The enthalpy relaxation of freeze concentrated sucrose-water glass was investigated using 40% sucrose, differential scanning calorimetry (DSC) with isothermal ageing for 1-6 days at various temperatures (-70, -65, -60, and -55 degrees C). The enthalpy relaxation was observed as an endothermic peak superimposed on the endothermic step-wise change due to the glass transition around -47 degrees C. The enthalpy relaxation was found to increase with ageing time and temperature. An 80% sucrose glass was also investigated at ageing temperatures of -60 and -65 degrees C, and this material exhibited a similar glass transition and enthalpy relaxation to that observed with the frozen 40% sucrose solution. The calculated activation energy of the enthalpy relaxation of the sucrose-water glass was smaller than that reported for pure sucrose. These results suggest that the freeze concentrated sucrose-water glass could have a higher molecular mobility and less stability than pure sucrose glass.     

Immunogenicity of a chromatographically concentrated influenza vaccine

The immunogenicity of the Czechoslovak-made commercial vaccine Adinvira as well as that of the chromatographically concentrated vaccine Leningrad of Soviet origin was evaluated in an immunization project. The both vaccines were well tolerated after administration, the Soviet-made Leningrad brand featuring greater efficiency in eliciting an antibody response against influenza virus haemagglutinin.