Activity Recovery Yield Research Articles

Optimised immobilisation of microbial lipase on hydrous niobium oxide

AbstractThe aim of this work was to establish appropriate conditions for immobilising Candida rugosa lipase on a low‐cost inorganic matrix, hydrous niobium oxide, using a multivariate statistical approach. A 23 full factorial design was employed to determine the effects of support activation with glutaraldehyde (concentration 2.5–4.5%, pH 7–10) and lipase loading (200‐700 U g−1 matrix) on the hydrolytic and synthetic activities of the immobilised derivatives. From the results the following conditions were established: lipase loading of 450 U g−1 matrix and niobium oxide activation with glutaraldehyde at a concentration of 2.5% and pH 8. Under these conditions, high activity recovery (47.21%) and esterification yield (86.90%) were attained. The results also show that hydrous niobium oxide can be a valid alternative to replace high‐cost, commercially available inorganic matrices such as controlled pore silica. Copyright © 2006 Society of Chemical Industry

The role of detergent in refolding of GdnHCl-denatured arginine kinase from shrimpFenneropenaeus Chinensis: the solubilization of aggregate and refolding in detergent solutions

Strong aggregation occurred in the refolding route of arginine kinase (AK) denatured with 3 mol GdnHCl/L (GdnHCl, guanidine hydrochloride). The activity recovery of GdnHCl-denatured AK was very low and dependent on the protein concentration in the process of refolding. For denatured AK at 1.2 micromol/L concentration, the recovered activity yield was about 45.2% of the native enzyme, whereas at 5.2 micromol/L the activity recovery yield was only 20% of native activity. The nonionic detergent Triton X-100 and Tween 20 (< or = 100 mmol/L concentration) not only effectively blocked the aggregation but also enabled the denatured AK to recover most of its native activity. The kinetics of aggregate solubilization showed that there was an induction phase dependent on the detergent, but there was no dependency when detergent was absent. The apparent activity recovery had a cooperative relation with detergents in the process of refolding, which suggested the existence of some interaction between the detergent and the refolding intermediate. On the basis of the study results, a scheme of refolding was proposed.

Study of the factors affecting the forward and back extraction of yeast-lipase and its activity by reverse micelles

The experimental results of yeast-lipase extracted by AOT-reverse-micelles in isooctane are reported. The effects of pH values and ionic strength in the aqueous phase, surfactant concentration, phase volume ratio, temperature, stirring time, and cosolvent concentration on the phase transfer of the lipase by the reverse micelles are studied. One hundred percent of forward extraction yield, 68% of back extraction yield, and 45% of activity recovery yield for the lipase are obtained. The phase transfer of the lipase is controlled by electrostatic and hydrophobic interactions.