Acute effects of extreme pH and its influences on the survival and biochemical biomarkers of juvenile White Shrimp,Litopenaeus vannamei

The White Shrimp (Litopenaeus vannamei) is reared in several types of systems. Changes in pH in some can produce sub-lethal effects that result in poor growth and survival. The aim of the present study was to determine the acidic and basic pH 50% lethal limits after 96 h exposure (pH_50–96 h) and to evaluate the sub-lethal effects of pH on oxidative stress parameters in juvenile L. vannamei. The experimental design comprised nine treatments with three replications of each. The nine treatments consisted of pH 10.5, 10.0, 9.5, 9.0, 7.0, 5.0, 4.5, 4.0 and 3.5. The treatment at 7.0 was used as the control. The sub-lethal pH tests were: basic pH 9.5, neutral pH 7.0 (control) and acidic pH 4.5. For each experimental pH and sampling point, the hemolymph of six shrimp was collected to determine catalase and glutathione S-transferase’s activities and total antioxidant capacity. The values of lethal pH₅₀ at 24, 48, 72 and 96 h for juvenile L. vannamei had basic pH values equal to 9.82, 9.62, 9.59 and 9.58 and acidic pH values of 3.86, 3.92, 3.94 and 4.04, respectively. The shrimp exposed to basic and acidic pH levels showed antioxidant responses with changes in antioxidant activity.     

Secretion and in vivo folding of the Fab fragment of the antibody McPC603 in Escherichia coli: influence of disulphides and cis-prolines.

Using the well-characterized antibody McPC603 as a model, we had found that the Fv fragment can be isolated from Escherichia coli as a functional protein in good yields, whereas the amount of the correctly folded Fab fragment of the same antibody produced under identical conditions is significantly lower. In this paper, we analyse the reasons for this difference. We found that a variety of signal sequences function in the secretion of the isolated chains of the Fab fragment or in the co-secretion of both chains in E.coli. The low yield of functional Fab fragment is not caused by inefficient expression or secretion in E.coli, but by inefficient folding and/or assembly in the periplasm. We compared the folding yields for the Fv and the Fab fragment in the periplasm under various conditions. Several diagnostic framework variants were constructed and their folding yields measured. The results show that substitutions affecting cis-proline residues and those affecting various disulphide bonds in the protein are by themselves insufficient to dramatically change the partitioning of the folding pathway to the native structure, and the cause must lie in a facile aggregation of folding intermediates common to all structural variants. However, all structural variants could be obtained in native form, demonstrating the general utility of the secretory expression strategy.