Molecular cloning, nucleotide sequence and expression of a Sulfolobus solfataricus gene encoding a class II fumarase.
Fumarase catalyzes the interconversion of L-malate and fumarate. A Sulfolobus solfataricus fumarase gene (fumC) was cloned and sequenced. Typical archaebacterial regulatory sites were identified in the region flanking the fumC open reading frame. The fumC gene encodes a protein of 438 amino acids (47,899 Da) which shows several significant similarities with class II fumarases from both eubacterial and eukariotic sources as well as with aspartases. S. solfataricus fumarase expressed in Escherichia coli retains enzymatic activity and its thermostability is comparable to that of S. solfataricus purified enzyme despite a 11 amino acid C-terminal deletion.
Colombo
S
S
Dipartimento di Fisiologia e Biochimica Generali, Università degli Studi di Milano, Italy.
Grisa
M
M
Tortora
P
P
Vanoni
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D004269:DNA, Bacterial / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D005649:Fumarate Hydratase / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D012045:Regulatory Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D016944:Sulfolobus / Q000235:genetics*; D014169:Transformation, Bacterial
D003062:Codon; D004269:DNA, Bacterial; D046911:Macromolecular Substances; D005649:Fumarate Hydratase