What is it about?
Production of hyper-thermostable enzymes in mesophilic hosts frequently causes undesired aggregation of these proteins. During production of Pyrococcus furiosus endo-β-1,3 glucanase (LamA) in Escherichia coli, soluble and insoluble species form. Here, we show that more than 90% of the LamA protein that is present in solution can be reactivated by treatment with 3 M guanidinium hydrochloride at 80 °C. Also, a large amount of insoluble LamA aggregates can be converted into soluble native monomer by application of this procedure. Thus, chaotropic heat treatment can improve the yield and quality of hyperthermostable proteins that form aberrant species during production in E. coli.
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Why is it important?
Production of hyperthermostable enzymes in mesophilic hosts frequently causes undesired aggregation of these proteins and result in low yield and low specific activity. Finding ways to improve the quality of these enzymes is of main relevance in biotechnology. Furthermore, such studies provide important information about the folding landscape of hyper-thermostable proteins.
Perspectives
The chaotropic heat treatment procedure described in this article is an easy adaptable method that can be used to screen the folding landscape of hyper-thermostable proteins and may add to the success of producing these proteins in mesophilic hosts.
Professor Willem J.H. van Berkel
Wageningen University
Read the Original
This page is a summary of: Chaotropic heat treatment resolves native-like aggregation of a heterologously produced hyperthermostable laminarinase, Biotechnology Journal, May 2017, Wiley,
DOI: 10.1002/biot.201700007.
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