Online detection of feed demand in high cell density cultures of Escherichia coli by measurement of changes in dissolved oxygen transients in complex media

A starvation-based dissolved oxygen (DO) transient controller was developed to supply growth-limiting substrate to high cell density fed-batch cultures of recombinant Escherichia coli. The algorithm adjusted a preexisting feed rate in proportion to the culture's oxygen demand, which was estimated from transients in the DO concentration after short periods of feed interruption. In this manner, the addition of glucose feed was precisely controlled at a rate that did not exceed the acetate production threshold, thus preventing acetate accumulation. In comparison to exponential feed algorithms commonly used in industry, the implementation of the new feeding strategy increased the final cell density from 32 to 44 g (dry cell weight).L -1, with less than 16 mM acetate accumulated, producing an ideal culture for subsequent induction. Despite a constant starvation level and relatively low levels of acetate, experimental cultivations still tended to produce acetate towards the end of the process. The use of a simple Monod model provided an explanation as to why this may occur in high cell density cultivations and suggests how it may be overcome

Microbial biocementation

A method of forming a high strength cement in a permeable starting material, the method comprising the step of combining the starting material with effective amounts of (i) a urease producing micro-organism; (ii) urea; and (iii) calcium ions and wherein the effective amount of the urease producing organism provides a urea hydrolysis rate, under standard conditions, of 0.5-50 mM urea hydrolysed.min-1

Microbial phytase-induced calcium-phosphate precipitation: A potential soil stabilization method

Two hypotheses were tested: (1) microbial dephosphorylation of phytate in the presence of Ca2+ ions will result in the precipitation of hydroxyapatite-like crystals and (2) precipitation of calcium-phosphate crystals on and between sand-like particles can cause cementation. A growing culture of the dimorphic phytase-active yeast Arxula adeninivorans was introduced into a column filled with quartz particles and subsequently a liquid growth medium amended with calcium phytate was pumped through the column resulting in increased strength and stiffness of the quartz particle matrix. Environmental scanning electron microscope analysis combined with energy-dispersive X-ray measurement revealed cementation of the quartz particles by calcium-phosphate crystals. This microbial mineralization process could provide a novel approach to improving the mechanical properties like strength and stiffness of sandy soils.BT/BiotechnologyApplied Science

A mathematical model and analytical solution for the fixation of bacteria in biogrout

Biogrout is a new method for soil reinforcement, which is based on microbialinduced carbonate precipitation. Bacteria and reactants are flushed through the soil, resulting in calcium carbonate precipitation and consequent soil reinforcement. Bacteria are crucially important in the Biogrout process since they catalyse the reaction. Hence, to control the process, it is essential to know where the bacteria are located. The bacteria are possibly in suspension but can also be adsorbed or fixated on the matrix of the porous structure. In this article, a model is derived for the placement of bacteria. The model contains three phases of bacteria: bacteria in suspension, adsorbed bacteria and fixed bacteria. An analytical solution is derived for instantaneous reactions between these three phases. The analytical solution is compared to numerical simulations for finite reaction rates. For the numerical simulations the standard Galerkin Finite Element Method is used.Delft Institute of Applied MathematicsElectrical Engineering, Mathematics and Computer Scienc