High level expression in Escherichia coli of a fungal gene under the control of strong promoters.
FEMS Microbiol Lett
; 56(1-2): 45-51, 1990 Mar 01.
Article
en En
| MEDLINE
| ID: mdl-2185127
ABSTRACT
A recent report (Patino et al., (1989) FEMS Microbiol. Lett. 58, 139-144) described the low level expression, in Escherichia coli, of the Isopenicillin N Synthase (IPNS) gene from Cephalosporium acremonium under the control of strong promoters. We report here our work on the expression of the IPNS gene. Plasmids containing the IPNS gene under the control of the trp or trc promoters directed synthesis of high levels of active IPNS in E. coli. Constitutive and inductive high level IPNS expression systems have been developed. Importantly, the expression vectors do not encode beta-lactamase so IPNS activity can be determined directly by biological assays. Analysis by nmr verified that the IPNS produced from these expression systems catalysed the conversion of delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-valine (LLD-ACV) to isopenicillin N in high yield.
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Banco de datos:
MEDLINE
Asunto principal:
Oxidorreductasas
/
Acremonium
/
Regulación Fúngica de la Expresión Génica
/
Regiones Promotoras Genéticas
/
Escherichia coli
/
Genes Fúngicos
Idioma:
En
Revista:
FEMS Microbiol Lett
Año:
1990
Tipo del documento:
Article
País de afiliación:
Reino Unido