Production of extracellular PETase from Ideonella sakaiensis using sec-dependent signal peptides in E. coli.
Biochem Biophys Res Commun
; 508(1): 250-255, 2019 01 01.
Article
em En
| MEDLINE
| ID: mdl-30477746
ABSTRACT
Poly(ethylene terephthalate) (PET) is the most commonly used polyester polymer resin in fabrics and storage materials, and its accumulation in the environment is a global problem. The ability of PET hydrolase from Ideonella sakaiensis 201-F6 (IsPETase) to degrade PET at moderate temperatures has been studied extensively. However, due to its low structural stability and solubility, it is difficult to apply standard laboratory-level IsPETase expression and purification procedures in industry. To overcome this difficulty, the expression of IsPETase can be improved by using a secretion system. This is the first report on the production of an extracellular IsPETase, active against PET film, using Sec-dependent translocation signal peptides from E. coli. In this work, we tested the effects of fusions of the Sec-dependent and SRP-dependent signal peptides from E. coli secretory proteins into IsPETase, and successfully produced the extracellular enzyme using pET22b-SPMalEIsPETase and pET22b-SPLamBIsPETase expression systems. We also confirmed that the secreted IsPETase has PET-degradation activity. The work will be used for development of a new E. coli strain capable of degrading and assimilating PET in its culture medium.
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MEDLINE
Assunto principal:
Sinais Direcionadores de Proteínas
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Polietilenotereftalatos
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Proteínas de Escherichia coli
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Escherichia coli
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Burkholderiales
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Hidrolases
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article