Antifoam addition to shake flask cultures of recombinant Pichia pastoris increases yield

Pichia pastoris is a widely-used host for recombinant protein production. Initial screening for both suitable clones and optimum culture conditions is typically carried out in multi-well plates. This is followed by up-scaling either to shake-flasks or continuously stirred tank bioreactors. A particular problem in these formats is foaming, which is commonly prevented by the addition of chemical antifoaming agents. Intriguingly, antifoams are often added without prior consideration of their effect on the yeast cells, the protein product or the influence on downstream processes such as protein purification. In this study we characterised, for the first time, the effects of five commonly-used antifoaming agents on the total amount of recombinant green fluorescent protein (GFP) secreted from shake-flask cultures of this industrially-relevant yeast.

From HAZAN to WASAN

The article discusses the waste and source-matter analysis (WASAN) group workshop methodology designed to minimize the generation of avoidable hazardous waste. Several areas analyzed using WASAN are the behavior of a process, waste minimization inside that process, and the consequences for waste production from upstream and downstream processes falling outside of design. A group from the Enriched Uranium Residues Recovery Plant (EURRP) in Springfields, England used WASAN to analyze the generation of plastic bag waste.

Flow processes in a radiant tube burner: isothermal flow

This paper presents the first part of a study of the combustion processes in an industrial radiant tube burner (RTB). The RTB is used typically in heat-treating furnaces. The work was initiated because of the need for improvements in burner lifetime and performance. The present paper is concerned with the flow of combustion air; a future paper will address the combusting flow. A detailed three-dimensional computational fluid dynamics model of the burner was developed, validated with experimental air flow velocity measurements using a split-film probe. Satisfactory agreement was achieved using the k-e turbulence model. Various features along the air inlet passage were subsequently analysed. The effectiveness of the air recuperator swirler was found to be significantly compromised by the need for a generous assembly tolerance. Also, a substantial circumferential flow maldistribution introduced by the swirler is effectively removed by the positioning of a constriction in the downstream passage.

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.