Schlieben_2004_Protein.Expr.Purif_34_103

Reference

Title : Expression, purification, and aggregation studies of His-tagged thermoalkalophilic lipase from Bacillus thermocatenulatus - Schlieben_2004_Protein.Expr.Purif_34_103
Author(s) : Schlieben NH , Niefind K , Schomburg D
Ref : Protein Expr Purif , 34 :103 , 2004
Abstract : The His-tagged lipase BTL2 from Bacillus thermocatenulatus was expressed in Escherichia coli and purified to homogeneity by a simple, one-step purification protocol using immobilized metal affinity chromatography. The success of protein separation and purification was pH-dependent and increased with decreasing pH. The purified BTL2 lipase showed a strong tendency to aggregate upon concentration, which prevented a reproducible crystallization. Aggregation studies using dynamic light-scattering (DLS) analysis were performed to improve the purification and concentration of BTL2 lipase. Different chemical classes of additives were tested to manipulate the aggregation behaviour of BTL2 lipase with the aim of obtaining a monodisperse sample to use for crystallization. For the process of concentration of BTL2 lipase in monomeric form, the alcohol 2-propanol and the ionic detergent dodecyl dimethylamine-N-oxide (LDAO) were found to be necessary. For the concentrated lipase, the availability of 5% 2-propanol was sufficient to hold the lipase in monomeric form and no additional detergent was needed.
ESTHER : Schlieben_2004_Protein.Expr.Purif_34_103
PubMedSearch : Schlieben_2004_Protein.Expr.Purif_34_103
PubMedID: 14766305
Gene_locus related to this paper: bactc-lipas

Related information

Gene_locus related to this paper: bactc-lipas

Citations formats

Schlieben NH, Niefind K, Schomburg D (2004)
Expression, purification, and aggregation studies of His-tagged thermoalkalophilic lipase from Bacillus thermocatenulatus
Protein Expr Purif 34 :103

Schlieben NH, Niefind K, Schomburg D (2004)
Protein Expr Purif 34 :103