Cloning, expression, and sequencing of squalene-hopene cyclase, a key enzyme in triterpenoid metabolism.
The pentacyclic hopanoids, a class of eubacterial lipids, are synthesized by squalene-hopene cyclase and side chain-elongating enzymes. With the aid of DNA probes based on the amino-terminal sequence of purified squalene-hopene cyclase from Bacillus acidocaldarius, clones of Escherichia coli that express this enzyme in the cytoplasmic membrane were isolated. According to the DNA sequence, the cyclase contained 627 amino acids with a molecular mass of 69,473 Da. A high percentage of the amino acids were basic. No significant similarity to existing sequenced proteins was found.
Ochs
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Botanisches Institut, Mikrobiologie, Universität Tübingen, Germany.
Kaletta
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C
Entian
K D
KD
Beck-Sickinger
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A
Poralla
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K
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D019751:Intramolecular Transferases; D007535:Isomerases / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D015345:Oligonucleotide Probes / Q000138:chemical synthesis; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012689:Sequence Homology, Nucleic Acid
D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D007535:Isomerases; D019751:Intramolecular Transferases; C066630:squalene-hopene cyclase