PMID: 2115771Jul 1, 1990Paper

Homocitrate synthase from Penicillium chrysogenum. Localization, purification of the cytosolic isoenzyme, and sensitivity to lysine

The Biochemical Journal
W M Jaklitsch, C P Kubicek

Abstract

Subcellular fractionation of cell-free extracts obtained by nitrogen cavitation showed that Penicillium chrysogenum Q176 contains a cytosolic as well as a mitochondrial homocitrate synthase activity. The cytosolic isoenzyme was purified about 500-fold, and its kinetic and molecular properties were investigated. Native homocitrate synthase shows a molecular mass of 155 +/- 10 kDa as determined by gel filtration and a pH of 4.9 +/- 0.1 as determined by chromatofocusing. The kinetic behaviour towards 2-oxoglutarate is hyperbolic, with Km = 2.2 mM; with respect to acetyl-CoA the enzyme shows sigmoidal saturation kinetics, with [S]0.5 = 41 microM and h = 2.6. The enzyme was inhibited strongly by L-lysine (Ki = 8 +/- 2 microM; 50% inhibition by 53 microM at 6 mM-2-oxoglutarate), competitively with 2-oxoglutarate, in protamine sulphate-treated and desalted cell-free extracts and in partially purified preparations. The extent of this inhibition was strongly pH-dependent. Both isoenzymes are equally susceptible to inhibition by lysine. The same inhibition pattern is shown by the enzyme from strain D6/1014A, which is a better producer of penicillin than strain Q176.

Citations

Jan 1, 1996·Critical Reviews in Biotechnology·C P Kubicek
Jan 25, 2011·Microbial Biotechnology·Juan F MartínCarlos García-Estrada
Jun 5, 2013·Journal of Molecular Graphics & Modelling·Xiang ShengChengbu Liu
Apr 21, 2012·International Journal of Evolutionary Biology·Hiromi Nishida
Sep 13, 2015·Protein Expression and Purification·Iwona Gabriel, Sławomir Milewski
Jul 4, 2002·FEBS Letters·Asri Peni WulandariHisakazu Yamane
May 7, 2011·Natural Product Reports·Tohru DairiIsao Fujii
Apr 18, 1997·The Journal of Biological Chemistry·S ChenJ P Aris
Jan 22, 2010·The Journal of Biological Chemistry·Stacie L BulferRaymond C Trievel
Sep 25, 2009·The Journal of Biological Chemistry·Stacie L BulferRaymond C Trievel
May 20, 2009·Biochemistry·Ashwani Kumar VashishthaPaul F Cook
Sep 8, 1998·Microbiology and Molecular Biology Reviews : MMBR·A A Brakhage

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