ArticleJournal of bacteriology2008
Cloning, solubilization, and characterization of squalene synthase from Thermosynechococcus elongatus BP-1.
Article in Journal of bacteriology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 62 citations in OpenAlex.
- Total and engineered biosynthesis of cinatrins: inhibitors of phospholipase AChemical science · 2026Article
- Bioassay-Guided Fractionation Leads to the Detection of Cholic Acid Generated by the RareMarine drugs · 2022Article
- Structure-guided product determination of the bacterial type II diterpene synthase Tpn2.Communications chemistry · 2022Article
- Cloning, Expression Analysis and Functional Characterization of Squalene Synthase (SQS) from Tripterygium wilfordii.Molecules (Basel, Switzerland) · 2018Article
- Exploring functional membrane microdomains in bacteria: an overview.Current opinion in microbiology · 2017Review
- Pheno- and Genotyping of Hopanoid Production inFrontiers in microbiology · 2017Article
- Production of squalene by microbes: an update.World journal of microbiology & biotechnology · 2016Review
- Molecular Cloning and Functional Analysis of Squalene Synthase 2(SQS2) in Salvia miltiorrhiza Bunge.Frontiers in plant science · 2016Article
- Directed optimization of a newly identified squalene synthase from Mortierella alpine based on sequence truncation and site-directed mutagenesis.Journal of industrial microbiology & biotechnology · 2015Article
- Triterpenoid profiling and functional characterization of the initial genes involved in isoprenoid biosynthesis in neem (Azadirachta indica).BMC plant biology · 2015Article
- Exploring the existence of lipid rafts in bacteria.Microbiology and molecular biology reviews : MMBR · 2015Article
- Terpenoids and their biosynthesis in cyanobacteria.Life (Basel, Switzerland) · 2015Review
- Cloning and characterization of squalene synthase gene from Poria cocos and its up-regulation by methyl jasmonate.World journal of microbiology & biotechnology · 2014Article
- Production of squalene in Synechocystis sp. PCC 6803.PloS one · 2014Article
- Characterization of Squalene synthase gene from Chlorophytum borivilianum (Sant. and Fernand.).Molecular biotechnology · 2013Article
- Statistical experimental design guided optimization of a one-pot biphasic multienzyme total synthesis of amorpha-4,11-diene.PloS one · 2013Article
- Functional characterization and differential expression studies of squalene synthase from Withania somnifera.Molecular biology reports · 2012Article
- Article
- Cloning and characterization of squalene synthase gene from Fusarium fujikuroi (Saw.) Wr.Journal of industrial microbiology & biotechnology · 2010Article
- Functional microdomains in bacterial membranes.Genes & development · 2010Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Squalene synthase (SQS) is a bifunctional enzyme that catalyzes the condensation of two molecules of farnesyl diphosphate (FPP) to give presqualene diphosphate (PSPP) and the subsequent rearrangement of PSPP to squalene. These reactions constitute the first pathway-specific steps in hopane biosynthesis in Bacteria and sterol biosynthesis in Eukarya. The genes encoding SQS were isolated from the hopane-producing bacteria Thermosynechococcus elongatus BP-1, Bradyrhizobium japonicum, and Zymomonas mobilis and cloned into an Escherichia coli expression system. The expressed proteins with a His(6) tag were found exclusively in inclusion bodies when no additives were used in the buffer. After extensive optimization, soluble recombinant T. elongatus BP-1 SQS was obtained when cells were disrupted and purified in buffers containing glycerol. The recombinant B. japonicum and Z. mobilis SQSs could not be solubilized under any of the expression and purification conditions used. Purified T. elongatus His(6)-SQS gave a single band at 42 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and molecular ion at m/z 41886 by electrospray mass spectrometry. Incubation with FPP and NADPH gave squalene as the sole product. Incubation of the enzyme with [(14)C]FPP in the absence of NADPH gave PSPP. The enzyme requires Mg(2+) for activity, has an optimum pH of 7.6, and is strongly stimulated by detergent. Under optimal conditions, the K(m) of FPP is 0.97 +/- 0.10 microM and the k(cat) is 1.74 +/- 0.04 s(-1). Zaragozic acid A, a potent inhibitor of mammalian, fungal, and Saccharomyces cerevisiae SQSs, also inhibited recombinant T. elongatus BP-1 SQS, with a 50% inhibitory concentration of 95.5 +/- 13.6 nM.
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