Bioreaction and separation in batch chromatographic columns

The objective of this work has been to study the behaviour and performance of a batch chromatographic column under simultaneous bioreaction and separation conditions for several carbohydrate feedstocks. Four bioreactions were chosen, namely the hydrolysis of sucrose to glucose and fructose using the enzyme invertase, the hydrolysis of inulin to fructose and glucose using inulinase, the hydrolysis of lactose to glucose and galactose using lactase and the isomerization of glucose to fructose using glucose isomerase. The chromatographic columns employed were jacketed glass columns ranging from 1 m to 2 m long and the internal diameter ranging from 0.97 cm to 1.97 cm. The stationary phase used was a cation exchange resin (PUROLITE PCR-833) in the Ca2+ form for the hydrolysis and the Mg2+ form for the isomerization reactions. The mobile phase used was a diluted enzyme solution which was continuously pumped through the chromatographic bed. The substrate was injected at the top of the bed as a pulse. The effect of the parameters pulse size, the amount of substrate solution introduced into the system corresponding to a percentage of the total empty column volume (% TECV), pulse concentration, eluent flowrate and the enzyme activity of the eluent were investigated. For the system sucrose-invertase complete conversions of substrate were achieved for pulse sizes and pulse concentrations of up to 20% TECV and 60% w/v, respectively. Products with purity above 90% were obtained. The enzyme consumption was 45% of the amount theoretically required to produce the same amount of product as in a conventional batch reactor. A value of 27 kg sucrose/m3 resin/h for the throughput of the system was achieved. The systematic investigation of the factors affecting the performance of the batch chromatographic bioreactor-separator was carried out by employing a factorial experimental procedure. The main factors affecting the performance of the system were the flowrate and enzyme activity. For the system inulin-inulinase total conversions were also obtained for pulses sizes of up to 20 % TECV and a pulse concentration of 10 % w/v. Fructose rich fractions with 100 % purity and representing up to 99.4 % of the total fructose generated were obtained with an enzyme consumption of 32 % of the amount theoretically required to produce the same amount of product in a conventional batch reactor. The hydrolysis of lactose by lactase was studied in the glass columns and also in an SCCR-S unit adapted for batch operation, in co-operation with Dr. Shieh, a fellow researcher in the Chemical Engineering and Applied Chemistry Department at Aston University. By operating at up to 30 % w/v lactose feed concentrations complete conversions were obtained and the purities of the products generated were above 90%. An enzyme consumption of 48 % of the amount theoretically required to produce the same amount of product in a conventional batch reactor was achieved. On working with the system glucose-glucose isomerase, which is a reversible reaction, the separation obtained with the stationary phase conditioned in the magnesium form was very poor although the conversion obtained was compatible with those for conventional batch reactors. By working with a mixed pulse of enzyme and substrate, up to 82.5 % of the fructose generated with a purity of 100 % was obtained. The mathematical modelling and computer simulation of the batch chromatographic bioreaction-separation has been performed on a personal computer. A finite difference method was used to solve the partial differential equations and the simulation results showed good agreement with the experimental results.

Combined bioreaction and separation in centrifugal fields

The aim of this work has been to investigate the principle of combined centrifugal bioreaction-separation. The production of dextran and fructose by the action of the enzyme dextransucrase on sucrose was employed to elucidate some of the principles of this type of process. Dextran is a valuable pharmaceutical product used mainly as a blood volume expander and blood flow improver whilst fructose is an important dietary product. The development of a single step process capable of the simultaneous biosynthesis of dextran and the separation of the fructose by-product should improve dextran yields whilst reducing capital and processing costs. This thesis shows for the first time that it is possible to conduct successful bioreaction-separations using a rate-zonal centrifugation technique. By layering thin zones of dextrasucrase enzyme onto sucrose gradients and centrifuging, very high molecular weight (MW) dextran-enzyme complexes were formed that rapidly sedimented through the sucrose substrate gradients under the influence of the applied centrifugal field. The low MW fructose by-product sedimented at reduced rates and was thus separated from the enzyme and dextran during the reaction. The MW distribution of dextran recovered from the centrifugal bioreactor was compared with that from a conventional batch bioreactor. The results indicated that the centrifugal bioreactor produced up to 100% more clinical dextran with MWs of between 12 000 and 98 000 at 20% w/w sucrose concentrations than conventional bioreactors. This was due to the removal of acceptor fructose molecules from the sedimenting reaction zone by the action of the centrifugal field. Higher proportions of unwanted lower MW dextran were found in the conventional bioreactor than in the centrifugal bioreactor-separator. The process was studied on a number of alternative centrifugal systems. A zonal rotor fitted with a reorienting gradient core proved most successful for the evaluation of bioreactor performance. Results indicated that viscosity build-up in the reactor must be minimised in order to increase the yields of dextran per unit time and improve product separation. A preliminary attempt at modelling the process has also been made.

Biosynthesis and separation of dextran fructose mixtures in a chromatographic reactor

A literature review of work carried out on batch and continuous chromatographic biochemical reactor-separators has been made. The major part of this work has involved the development of a batch chromatographic reactor-separator for the production of dextran and fructose by the enzymatic action of the enzyme dextransucrase on sucrose. In this reactor, simultaneous reaction and separation occurs thus reducing downstream processing and isolation of products as compared to the existing industrial process. The chromatographic reactor consisted of a glass column packed with a stationary phase consisting of cross linked polysytrene resin in the calcium form. The mobile phase consisted of diluted dextransucrase in deionised water. Initial experiments were carried out on a reactor separtor which had an internal diameter of 0.97cm and length of 1.5m. To study the effect of scale up the reactor diameter was doubled to 1.94cm and length increased to 1.75m. The results have shown that the chromatographic reactor uses more enzyme than a conventional batch reactor for a given conversion of sucrose and that an increase in void volume results in higher conversions of sucrose. A comparison of the molecular weight distribution of dextran produced by the chromatographic reactor was made with that from a conventional batch reactor. The results have shown that the chromatographic reactor produces 30% more dextran of molecular weight greater than 150,000 daltons at 20% w/v sucrose concentration than conventional reactors. This is because some of the fructose molecules are prevented as acting as acceptors in the chromatographic reactor due to their removal from the reaction zone. In the conventional reactor this is not possible and therefore a greater proportion of low molecular weight dextran is produced which does not have much clinical use. A theoretical model was developed to describe the behaviour of the reactor separator and this model was simulated using a computer. The simulation predictions showed good agreement with experimental results at high eluent flowrates and low conversions.

Influence of downstream processing on the breakage of whey protein precipitates

Whey proteins may be fractionated by isoelectric precipitation followed by centrifugal recovery of the precipitate phase. Transport and processing of protein precipitates may alter the precipitate particle properties, which may affect how they behave in subsequent processes. For example, the transport of precipitate solution through pumps, pipes and valves and into a centrifugal separator may cause changes in particle size and density, which may affect the performance of the separator. This work investigates the effect of fluid flow intensity, flow geometry and exposure time on the breakage of whey protein precipitates: Computational fluid dynamics (CFD) was used to quantify the flow intensity in different geometries. Flow geometry can have a critical impact on particle breakage. Sharp geometrical transitions induce large increases in turbulence that can result in substantial particle breakage. As protein precipitate particles break, they tend to form denser more compact structures. The reduction in particle size and increase in compaction is due to breakage. This makes the particles become more resistant to further breakage as particle compactness increases. The effect of flow intensity on particle breakage is coupled to exposure time, with greater exposure time producing more breakage. However, it is expected that the particles will attain an equilibrium particle size and density after prolonged exposure in a constant flow field where no further breakage will occur with exposure time. © 2005 Institution of Chemical Engineers.

How unlucky is 25-sigma?

Purpose - To identify the likelihood of a 25-standard deviation occurring in stock prices over several successive days, in the light of comments by David Viniar, chief financial officer of Goldman Sachs.. Design/methodology/approach - Assumes a bell-curve of market losses and graphs the probability of an event relative to the number of deviations. Calculates using MATLAB for sigmas over 7. Considers whether the losses of US investment banks in 2008 were the result of bad luck or of incompetence. Findings - Finds that the probability of a 25-sigma event is every 100,000 +130 decimal points years. Practical implications - Argues that bad luck is usually associated with incompetence, and investors need not choose between them. Originality/value - Presents the mathematical absurdity of a finance officer's statement, and its implications.

Anterior eye health recording

Aims: To survey eye care practitioners from around the world regarding their current practice for anterior eye health recording to inform guidelines on best practice. Methods: The on-line survey examined the reported use of: word descriptions, sketching, grading scales or photographs; paper or computerised record cards and whether these were guided by proforma headings; grading scale choice, signs graded, level of precision, regional grading; and how much time eye care practitioners spent on average on anterior eye health recording. Results: Eight hundred and nine eye care practitioners from across the world completed the survey. Word description (p <. 0.001), sketches (p = 0.002) and grading scales (p <. 0.001) were used more for recording the anterior eye health of contact lens patients than other patients, but photography was used similarly (p = 0.132). Of the respondents, 84.5% used a grading scale, 13.5% using two, with the original Efron (51.6%) and CCLRU/Brien-Holden-Vision-Institute (48.5%) being the most popular. The median features graded was 11 (range 1-23), frequency from 91.6% (bulbar hyperaemia) to 19.6% (endothelial blebs), with most practitioners grading to the nearest unit (47.4%) and just 14.7% to one decimal place. The average time taken to report anterior eye health was reported to be 6.8. ±. 5.7. min, with the maximum time available 14.0. ±. 11. min. Conclusions: Developed practice and research evidence allows best practice guidelines for anterior eye health recording to be recommended. It is recommended to: record which grading scale is used; always grade to one decimal place, record what you see live rather than based on how you intend to manage a condition; grade bulbar and limbal hyperaemia, limbal neovascularisation, conjunctival papillary redness and roughness (in white light to assess colouration with fluorescein instilled to aid visualisation of papillae/follicles), blepharitis, meibomian gland dysfunction and sketch staining (both corneal and conjunctival) at every visit. Record other anterior eye features only if they are remarkable, but indicate that the key tissue which have been examined.

Data on the biogeochemical cycle of methane in the ocean

Geological, mineralogical and microbiological aspects of the methane cycle in water and sediments of different areas in the oceans are under consideration in the monograph. Original and published estimations of formation- and oxidation rates of methane with use of radioisotope and isotopic methods are given. The role of aerobic and anaerobic microbial oxidation of methane in production of organic matter and in formation of authigenic carbonates is considered. Particular attention is paid to processes of methane transformation in areas of its intensive input to the water column from deep-sea hydrothermal sources, mud volcanoes, and cold methane seeps.

Variabilidad de la inactivación microbiana y de la fase de latencia de los microorganismos supervivientes a un proceso de acidificación

A la industria alimentaria se le exigen productos seguros, nutritivos, apetecibles y de uso cómodo y rápido. Aunar todos esos calificativos en un solo alimento es ardua tarea. Valgan dos ejemplos. Un tratamiento conservante intenso, de buenas perspectivas sanitarias, suele conllevar una pérdida de valor nutritivo y unas características sensoriales poco atractivas. El manejo de los alimentos para transformarlos en productos listos pare el consumo implica la asunción de ciertos riesgos microbiológicos, mayores que los asumidos en productos sin manipulación. ¿Cómo responder ante el incremento de riesgos y peligros que se ciernen sobre los “nuevos alimentos”? Una alternativa que ha ganado correligionarios es la microbiología predictiva. Es una herramienta útil, a disposición de cualquier entidad interesada en los alimentos, que predice, mediante modelos matemáticos, el comportamiento microbiano bajo ciertas condiciones. La mayoría de los modelos disponibles predicen valores únicos (a cada valor de la variable independiente le corresponde un único valor de la dependiente); han demostrado su eficacia durante décadas a base de tratamientos sobredimensionados para salvaguardar la calidad microbiológica de los alimentos y predicen una media, sin considerar la variabilidad. Considérese un valor de reducción decimal, D, de 1 minuto. Si el producto contiene 103 ufc/g, un envase de 1 Kg que haya pasado por un tratamiento 6D, contendrá 1 célula viable. Hasta aquí la predicción de un modelo clásico. Ahora piénsese en una producción industrial, miles de envases de 1 Kg/h. ¿Quién puede creerse que en todos ellos habrá 1 microorganismo superviviente? ¿No es más creíble que en unos no quedará ningún viable, en muchos 1, en otros 2, 3 y quizás en los menos 5 ó 6? Los modelos que no consideran la variabilidad microbiana predicen con precisión la tasa de crecimiento pero han fracasado en la predicción de la fase de latencia...

El orden de los libros en el estante : Un estudio sobre la signatura topográfica en las bibliotecas universitarias de la Universidad Nacional de La Plata

Partiendo de la hipótesis de que el patrón común que vincula a las distintas bibliotecas de la Universidad Nacional de La Plata (UNLP) para la construcción de la signatura de clase es el uso de la Clasificación Decimal Universal (CDU), y que existe una dispersión importante en los elementos que componen la signatura librística, se realizó un relevamiento en las bibliotecas universitarias para indagar sobre las formas en que se construye la signatura topográfica para el ordenamiento del material bibliográfico en el estante. Se tomó una muestra de signaturas topográficas del catálogo colectivo Roble. Se analizaron las diferencias y semejanzas encontradas. Se entrevistó a los responsables de los procesos técnicos de cada biblioteca para indagar sobre las causas de las diferencias encontradas. Se comprobó que, si bien la mayoría de las bibliotecas usan CDU para formar la signatura de clase, emplean ediciones diferentes con criterios de uso disímiles. Además, en la formación de la signatura librística, los criterios empleados varían notablemente de una biblioteca a otra, haciendo aún más difícil la coincidencia de signaturas topográficas asignadas. Se concluye que el factor determinante en el resultado es la forma en que se desarrollan las prácticas laborales dentro de estas instituciones.

El orden de los libros en el estante : Un estudio sobre la signatura topográfica en las bibliotecas universitarias de la Universidad Nacional de La Plata

Partiendo de la hipótesis de que el patrón común que vincula a las distintas bibliotecas de la Universidad Nacional de La Plata (UNLP) para la construcción de la signatura de clase es el uso de la Clasificación Decimal Universal (CDU), y que existe una dispersión importante en los elementos que componen la signatura librística, se realizó un relevamiento en las bibliotecas universitarias para indagar sobre las formas en que se construye la signatura topográfica para el ordenamiento del material bibliográfico en el estante. Se tomó una muestra de signaturas topográficas del catálogo colectivo Roble. Se analizaron las diferencias y semejanzas encontradas. Se entrevistó a los responsables de los procesos técnicos de cada biblioteca para indagar sobre las causas de las diferencias encontradas. Se comprobó que, si bien la mayoría de las bibliotecas usan CDU para formar la signatura de clase, emplean ediciones diferentes con criterios de uso disímiles. Además, en la formación de la signatura librística, los criterios empleados varían notablemente de una biblioteca a otra, haciendo aún más difícil la coincidencia de signaturas topográficas asignadas. Se concluye que el factor determinante en el resultado es la forma en que se desarrollan las prácticas laborales dentro de estas instituciones.

El orden de los libros en el estante : Un estudio sobre la signatura topográfica en las bibliotecas universitarias de la Universidad Nacional de La Plata

Partiendo de la hipótesis de que el patrón común que vincula a las distintas bibliotecas de la Universidad Nacional de La Plata (UNLP) para la construcción de la signatura de clase es el uso de la Clasificación Decimal Universal (CDU), y que existe una dispersión importante en los elementos que componen la signatura librística, se realizó un relevamiento en las bibliotecas universitarias para indagar sobre las formas en que se construye la signatura topográfica para el ordenamiento del material bibliográfico en el estante. Se tomó una muestra de signaturas topográficas del catálogo colectivo Roble. Se analizaron las diferencias y semejanzas encontradas. Se entrevistó a los responsables de los procesos técnicos de cada biblioteca para indagar sobre las causas de las diferencias encontradas. Se comprobó que, si bien la mayoría de las bibliotecas usan CDU para formar la signatura de clase, emplean ediciones diferentes con criterios de uso disímiles. Además, en la formación de la signatura librística, los criterios empleados varían notablemente de una biblioteca a otra, haciendo aún más difícil la coincidencia de signaturas topográficas asignadas. Se concluye que el factor determinante en el resultado es la forma en que se desarrollan las prácticas laborales dentro de estas instituciones.

Hydrocarbons in water, suspended matter, and bottom sediments of the White Sea

Data are presented on concentration and composition of aliphatic and polycyclic aromatic hydrocarbons (HC) in water, suspended matter (collected with a Juday net and by a separator), and in bottom sediments of the White Sea. It was found that during the last years the level of aliphatic HC concentrations in waters of the White Sea (aver. 18 µg/l) practically did not change and was comparable with average concentrations in shelf areas of the World Ocean. In water and bottom sediments distribution of HC is determined by discharge of river marginal filters. Here sedimentation of the bulk of anthropogenic HC occurs. That is why a grain-size controlling factor is not active in the zone of the river depocenter (in particular, of the North Dvina River). The same reasons most probably may explain differences in degree of geochemical relationships between contents of TOC and HC in suspended matter and bottom sediments. After passing through marginal filters natural HC are dominant in all migration forms.