Abstract:
:The application of artificial chaperones in biotechnology has been inspired by the mechanism of molecular chaperones like GroEL/GroES. It involves addition of a capturing detergent during dilution of the chaotropic reagent, that prevents protein aggregation, and finally, addition of a oligosaccharide that removes the detergent allowing the protein to refold. Here, guanidinium hydrochloride-denatured hexameric leucine aminopeptidase is shown to be efficiently refolded by using the cationic detergent cetyltrimethylammonium bromide and the linear polysaccharide dextrin-10 as artificial chaperones. The effect of these additives and the time dependence on the recovery of total enzymatic activity, kinetic parameters (K(M), k(cat)), intrinsic steady-state tryptophan fluorescence and oligomeric structure is presented. The method described is very promising since 92% of fully active and correct folded LAP could be produced. Moreover, we showed that the stripping process is relatively slow, it allows the protein to refold almost entirely to its native state.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Laslo AC,Ganea E,Obinger Cdoi
10.1016/j.jbiotec.2009.01.023subject
Has Abstractpub_date
2009-03-25 00:00:00pages
162-8issue
3-4eissn
0168-1656issn
1873-4863pii
S0168-1656(09)00040-6journal_volume
140pub_type
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journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2005.04.005
更新日期:2005-09-29 00:00:00
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更新日期:2016-02-20 00:00:00
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journal_title:Journal of biotechnology
pub_type: 杂志文章
doi:10.1016/j.jbiotec.2018.07.007
更新日期:2018-09-10 00:00:00
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journal_title:Journal of biotechnology
pub_type: 杂志文章,评审
doi:10.1016/j.jbiotec.2005.12.006
更新日期:2006-06-25 00:00:00
abstract::Xylanase T-6 is a thermostable alkaline-tolerant enzyme that is produced by Bacillus stearothermophilus T-6. Xylanase T-6 was found to bleach pulp effectively at pH 9 and 65 degrees C and was used successfully on an industrial-scale mill trial. To facilitate the future characterization of the protein via X-ray analysi...
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pub_type: 杂志文章
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pub_type: 杂志文章
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