Isolation of an Ustilago maydis gene encoding 3-hydroxy-3-methylglutaryl-coenzyme A reductase and expression of a C-terminal-truncated form in Escherichia coli

Cloning and Characterization of a Gene Encoding HMG-CoA Reductase fromGanoderma lucidumand Its Functional Identification in Yeast

“…These data indicate that Gl-HMGR mediated mevalonate biosynthesis in S. cerevisiae, strain JRY1130, and that Gl-HMGR was a functional fungal gene encoding HMGR in G. lucidum. Furthermore, our results indicate that the hydrophobic region is not essential to Gl-HMGR functioning, which is consistent with the data of Croxen et al, 15) Ruiz-Albert et al, 29) and Ferrer et al, 30) and also confirms the accuracy of the structural prediction of the Gl-HMGR protein.…”

“…15,29) In order to determine whether the hydrophobic N-terminal domain of Gl-HMGR is necessary for enzyme catalytic activity, we used HMGR cDNA excluding the N-terminal hydrophobic region to complement the HMGR-deficient S. cerevisiae, strain JRY1130. A 1,782-bp cDNA gene fragment (excluding the N-terminal hydrophobic region) was inserted, within the correct reading frame based on the structural prediction of Gl-HMGR, into expression vector pYF1845.…”

“…Genes encoding HMGR have been cloned and characterized from several sources, including U. maydis, 15) S. cerevisiae, 12) Drosophila melanogaster, 22) and Arabidopsis thaliana. 19) In view of a possible quantitative correlation between HMGR expression levels and triterpene production, it is of interest to study the HMGR-catalyzed committed step at the molecular level.…”

“…In fungi (viz., Saccharomyces cerevisiae and the smut fungus Ustilago maydis) and in animals, the membrane domain is large and complex, containing seven or eight transmembrane segments. [14][15][16][17] In contrast, the membrane domains of plant HMGR proteins have only one or two transmembrane segments. [18][19][20] The N-terminal membranebound domain, although not necessary for catalytic activity, is required for sterol-regulated degradation of the protein 17) and for membrane proliferation.…”

Enzymatic Total Synthesis of Gibberellin A4from Acetate

“…It has been reported that the membranespanning domain near the N-terminal region of HMGR was not necessary for the enzyme activity. 23) Hence, N. crassa HMGR in soluble protein form (the residual 510 amino acids near the C-terminal, ÁHMGR) was produced in E. coli by truncating the N-terminal region (Á2-664). The resulting eight ORFs and the bacterial GGPS gene amplified by PCR were introduced into the pQE-30 vector.…”