PMID: 9555026May 23, 1998Paper

Squalene-hopene cyclase from Methylococcus capsulatus (Bath): a bacterium producing hopanoids and steroids

Biochimica Et Biophysica Acta
A TippeltK Poralla

Abstract

We report the cloning and characterisation of the Methylococcus capsulatus shc gene, which encodes the squalene-hopene cyclase (SHC). This enzyme catalyses the complex cyclization of squalene to the pentacyclic triterpene skeleton of hopanoids and represents the key reaction in this biosynthesis. Using a combination of PCR amplification and DNA hybridization, two overlapping 2.6 kb PstI and 3.3 kb SalI DNA fragments were cloned bearing a 1962 bp open reading frame encoding a 74 kDa protein with 654 amino acids and a predicted isoelectric point at about pH 6.3. The deduced amino acid sequence of the M. capsulatus shc gene showed significant similarity to known prokaryotic SHCs and to a lesser degree to the related eukaryotic oxidosqualene cyclases (OSCs). Like other triterpene cyclases, the M. capsulatus SHC contains seven so-called QW-motifs as well as an aspartate-rich domain. The recombinant M. capsulatus SHC was expressed in Escherichia coli and in vitro activity of the recombinant cyclase was demonstrated using crude cell-free lysate or solubilized membrane preparation. The cyclization products hop-22-ene and hopan-22-ol (diplopterol) were identified by GC and GC-MS.

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Citations

Nov 28, 2008·The ISME Journal·Ann Pearson, Douglas B Rusch
Mar 15, 2014·PloS One·Elias EnglundPia Lindberg
Dec 25, 2007·FEBS Letters·Junichi ShinozakiYutaka Ebizuka
Sep 16, 2008·Phytochemistry·Junichi ShinozakiYutaka Ebizuka
Apr 11, 2012·Proteins·Silvia RacoltaJürgen Pleiss
Aug 10, 2007·Environmental Microbiology·Ann PearsonMeytal B Higgins
Dec 6, 2014·Frontiers in Microbiology·Charlène Leneveu-JenvrinSylvie Chevalier
Apr 12, 2014·Frontiers in Plant Science·Meriyem AktasFranz Narberhaus
Sep 1, 2007·HFSP Journal·Patrick Forterre, Simonetta Gribaldo
May 3, 2011·Applied and Environmental Microbiology·Gabriele Siedenburg, Dieter Jendrossek
Jun 4, 2005·Applied and Environmental Microbiology·Luan TaoQiong Cheng
Jul 26, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Junichi ShinozakiAkihito Takano
Jun 2, 2020·Frontiers in Bioengineering and Biotechnology·Zhen LiuXiangzhao Mao
Feb 5, 1999·Journal of Molecular Biology·K U WendtG E Schulz

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