ABSTRACT
The structure of the capsular polysaccharide of Escherichia coli K5 is identical to that of N-acetyl-heparosan, a nonsulfated precursor of heparin, which makes this E. coli antigen an attractive starting point for the chemical synthesis of analogs of low-molecular-weight heparin. This polysaccharide is synthesized as a high-molecular-weight molecule that can be depolymerized by an enzyme displaying endo-beta-eliminase activity. The eliminase-encoding gene, designated elmA, has been cloned from E. coli K5 by expression in E. coli K-12. The K-12 genome is devoid of the elmA sequence. The elmA gene product is 820 amino acids long. Active recombinant eliminase is produced by K-12 cells in both cell-bound and secreted forms. Deletion analyses have shown that the C terminus and the N terminus are required for activity and secretion, respectively.
Subject(s)
Escherichia coli/enzymology , Polysaccharide-Lyases/genetics , Polysaccharides, Bacterial/metabolism , Amino Acid Sequence , Base Sequence , Cloning, Molecular , Culture Media , DNA, Bacterial , Escherichia coli/genetics , Gene Expression , Molecular Sequence Data , Polysaccharide-Lyases/metabolism , Species Specificity , Transformation, GeneticABSTRACT
We present an algorithm for the adaptive control of dissolved oxygen concentration in a bioreactor, based on the agitation rate. The dynamics are represented by an incremental first-order model with variable dead-time and parameters. These are estimated on-line by a recursive least-squares identification method with a forgetting factor and a constant sensitivity. The model is employed to predict the behaviour of the dissolved oxygen concentration over a finite horizon, using an original method which requires little computation. Then, a Generalized Predictive Control optimisation strategy computes the agitation rate from the predictions and the desired set point, while gradually updating the controller smoothness. This algorithm, which requires little preliminary knowledge, has been implemented on a laboratory-scale fed-batch bioreactor for which the use of conventional controllers showed limited performance, due to the unpredictable and evolutive nature of the dynamics. The new controller proved to be robust and effective over a wide range of operating conditions, while requiring no operator adjustments.
