Cloning and expression in Escherichia coli of the Clostridium thermoaceticum gene encoding thermostable formyltetrahydrofolate synthetase View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1988-02

AUTHORS

Charles R. Lovell, Alan Przybyla, Lars G. Ljungdahl

ABSTRACT

Formyltetrahydrofolate synthetase (FTHFS) (EC 6.3.4.3), a thermostable protein of four identical subunits from Clostridium thermoaceticum was cloned into Escherichia coli SK1592. The clone (CRL47) contained a 9.5 kb EcoRI fragment of C. thermoaceticum DNA ligated into pBR322. It produced catalytically active, thermostable FTHFS, that was not found in E. coli SK1592 containing native pBR322. The identity of the expressed enzyme was confirmed by specific binding of rabbit polyclonal anti-FTHFS serum produced against C. thermoaceticum FTHFS. The specific activities (mumol.min-1.mg-1) of FTHFS in cell free extracts of CRL47 were 28-89 when assayed at 50 degrees C and pH 8. This was from 3-10-fold higher than in C. thermoaceticum extracts. FTHFS was purified to homogeneity from CRL47. The purified enzyme behaved during electrophoresis and gel chromatography and it had similar specific activity and thermostability as the enzyme purified from C. thermoaceticum. More... »

PAGES

280-285

Journal

TITLE

Archives of Microbiology

ISSUE

4

VOLUME

149

Author Affiliations

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/bf00411642

    DOI

    http://dx.doi.org/10.1007/bf00411642

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1013659480

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/2833195


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