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Expression in Escherichia coli of a sub‐gene encoding the lipoyl domain of the pyruvate dehydrogenase complex of Bacillus stearothermophilus
Author(s) -
Dardel Frédéric,
Packman Leonard C.,
Perham Richard N.
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)80249-i
Subject(s) - pyruvate dehydrogenase complex , escherichia coli , chemistry , gene , biochemistry , dihydrolipoyl transacetylase , microbiology and biotechnology , biology , enzyme , pyruvate dehydrogenase phosphatase
A sub‐gene encoding the lipoyl domain (residues 1–85) of the lipoate acetyltransferase chain of the pyruvate dehydrogenase complex of Bacillus stearothermophilus was over‐expressed in Escherichia coli . Approx. 80% of the domain was unlipoylated but most of the remainder was correctly lipoylated on Lys‐42 and could be reductively acetylated by the B stearothermophilus enzyme complex. A small proportion (approx. 4%) of the domain carried an aberrant substituent, possibly an octanoyl group, on Lys‐42. The 400 MHz 1 H NMR spectra of the lipoylated and unlipoylated domains were essentially identical and closely resembled that of the native lipoyl domain.