Enzymatic characterization of a soluble aggregate induced by N-terminal extension to a lipolytic enzyme.
J Biotechnol
; 281: 130-136, 2018 Sep 10.
Article
em En
| MEDLINE
| ID: mdl-29981449
ABSTRACT
A self-assembling peptide (27PEP) was isolated from an open reading frame (ORF). The ORF consisted of an unknown functional domain and a catalytic (lipolytic and phospholipolytic) domain (MPlaG) on metagenomic fosmid clone. This extension of 27 amino acids prior to the N-terminus of the catalytic domain (27PEP-MPlaG), starting at Met261, produced an aggregate of high molecular weight (> 700â¯kDa). Compared with MPlaG, 27PEP-MPlaG showed the same temperature and pH effect for maximum activity but was stable in the presence of inhibitors such as EDTA and PMSF. The 27PEP-MPlaG exhibited lower specific activity than that of MPlaG, but when pre-incubated for 30â¯min at temperatures between 4 and 100⯰C, its activity increased at temperatures greater than 40⯰C under alkaline conditions and eventually reached the specific activity level of MPlaG at 60⯰C. We experimentally determined that the aggregate caused by 27PEP was dissociated at elevated temperatures resulting in an active catalytic monomer. The 27PEP-indued aggregation may be attractive as application tool for improving or engineering of biocatalysts and biomaterials.
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Texto completo:
1
Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Peptídeos
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Fosfolipases A1
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Agregados Proteicos
Idioma:
En
Revista:
J Biotechnol
Ano de publicação:
2018
Tipo de documento:
Article