Bioreaction and separation in preparative batch chromatographic columns : the hydrology of lactose to yield glucose,galactose and oligosaccharides

The initial aim of this project was to improve the performance of a chromatographic bioreactor-separator (CBRS). In such a system, a dilute enzyme solution is pumped continuously through a preparative chromatographic column, while pulses of substrate are periodically injected on to the column. Enzymic reaction and separation are therefore performed in a single unit operation. The chromatographic columns used were jacketed glass columns ranging from 1 to 2 metres long with an internal diameter of 1.5 cm. Linking these columns allowed 1, 2, 3 and 4 metre long CBRS systems to be constructed. The hydrolysis of lactose in the presence of β~galactosidase was the reaction of study. From previous work at Aston University, there appeared to be no difficulties in achieving complete lactose hydrolysis in a CBRS. There did, however, appear to be scope for improving the separative performance, so this was adopted as an initial goal. Reducing the particle size of the stationary phase was identified as a way of achieving this improvement. A cation exchange resin was selected which had an average particle size of around half that previously used when studying this reaction. A CBRS system was developed which overcame the operational problems (such as high pressure drop development) associated with use of such a particle size. A significant improvement in separative power was achieved. This was shown by an increase in the number of theoretical plates (N) from about 500 to about 3000 for a 2 metre long CBRS, coupled with higher resolution. A simple experiment with the 1 metre column showed that combined bioreaction and separation was achievable in this system. Having improved the separative performance of the system, the factors affecting enzymic reaction in a CBRS were investigated; including pulse volume and the degree of mixing between enzyme and substrate. The progress of reaction in a CBRS was then studied. This information was related to the interaction of reaction and separation over the reaction zone. The effect of injecting a pulse over a length of time as in CBRS operation was simulated by fed batch experiments. These experiments were performed in parallel with normal batch experiments where the substrate is mixed almost instantly with the enzyme. The batch experiments enabled samples to be taken every minute and revealed that reaction is very rapid. The hydrodynamic characteristics of the two injector configurations used in CBRS construction were studied using Magnetic Resonance Imaging, combined with hydrodynamic calculations. During the optimisation studies, galactooligosaccharides (GOS) were detected as intermediates in the hydrolysis process. GOS are valuable products with potential and existing applications in food manufacture (as nutraceuticals), medicine and drug targeting. The focus of the research was therefore turned to GOS production. A means of controlling reaction to arrest break down of GOS was required. Raising temperature was identified as a possible means of achieving this within a CBRS. Studies were undertaken to optimise the yield of oligosaccharides, culminating in the design, construction and evaluation of a Dithermal Chromatographic Bioreactor-separator.

Remote sensing applications to flood hydrology in Belize

The research compares the usefullness of four remote sensing information sources, these being LANDSAT photographic prints, LANDSAT computer compatible tapes, Metric Camera and SIR-A photographic prints. These sources provide evaluations of the catchment characteristics of the Belize and Sibun river basins in Central America. Map evaluations at 1:250,000 scale are compared to the results of the same scale, remotely sensed information sources. The values of catchment characteristics for both maps and LANDSAT prints are used in multiple regression analysis, providing flood flow formulae, after investigations to provide a suitable dependent variable discharge series are made for short term records. The use of all remotely sensed information sources in providing evaluations of catchment characteristics is discussed. LANDSAT prints and computer compatible tapes of a post flood scene are used to estimate flood distributions and volumes. These are compared to values obtained from unit hydrograph analysis, using the dependent discharge series and evaluate the probable losses from the Belize river to the floodplain, thereby assessing the accuracy of LANDSAT estimates. Information relating to flood behaviour is discussed in terms of basic image presentation as well as image processing. A cost analysis of the purchase and use of all materials is provided. Conclusions of the research indicate that LANDSAT print material may provide information suitable for regression analysis at levels of accuracy as great as those of topographic maps, that the differing information sources are uniquely applicable and that accurate estimates of flood volumes may be determined even by post flood imagery.