PMID: 16615855Apr 18, 2006Paper

L-methionine gamma-lyase from Citrobacter freundii: cloning of the gene and kinetic parameters of the enzyme

Biochemistry. Biokhimii︠a︡
Ilya V ManukhovGennadii B Zavilgelsky

Abstract

It is shown for the first time for the Enterobacteriaceae family that a gene encoding L-methionine gamma-lyase (MGL) is present in the genome of Citrobacter freundii. Homogeneous enzyme has been purified from C. freundii cells and its N-terminal sequence has been determined. The hybrid plasmid pUCmgl obtained from the C. freundii genomic library contains an EcoRI insert of about 3000 bp, which ensures the appearance of MGL activity when expressed in Escherichia coli TG1 cells. The nucleotide sequence of the EcoRI fragment contains two open reading frames. The first frame (the megL gene) encodes a protein of 398 amino acid residues that has sequence homology with MGLs from different sources. The second frame encodes a protein with sequence homology with proteins belonging to the family of permeases. To overexpress the megL gene it was cloned into pET-15b vector. Recombinant enzyme has been purified and its kinetic parameters have been determined. It is demonstrated that a presence of a hybrid plasmid pUCmgl, containing the megL gene in the E. coli K12 cells, leads to a decrease in efficiency of EcoKI-restriction. It seems likely that decomposition of L-methionine under the action of MGL leads to a decrease in the intracellular c...Continue Reading

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Citations

Dec 17, 2009·Applied Microbiology and Biotechnology·Pei-xia JiangXin-Hui Xing
Dec 21, 2010·Applied and Environmental Microbiology·Marie-Pierre ForquinIsabelle Martin-Verstraete
Sep 4, 2012·Doklady. Biochemistry and Biophysics·S V RevtovichT V Demidkina
Jan 12, 2016·Acta Crystallographica. Section F, Structural Biology Communications·Svetlana RevtovichTatyana Demidkina
Dec 23, 2015·Biotechnology Research International·Mohsen Helmy SelimAmany Ahmed Hassabo Anise
Jan 15, 2015·Biochimica Et Biophysica Acta·Natalya V AnufrievaTatyana V Demidkina
Sep 25, 2014·BioMed Research International·Bhupender SharmaShamsher S Kanwar
Apr 8, 2015·Molekuliarnaia biologiia·E A BatuevV I Pol'shakov
May 29, 2016·Analytical Biochemistry·Timothy C FooKallidaikurichi V Venkatachalam
Oct 11, 2017·Current Medicinal Chemistry·Vadim S PokrovskyElena V Lukasheva
Nov 6, 2014·Acta Crystallographica. Section D, Biological Crystallography·Elena A MorozovaTatyana V Demidkina
Jun 5, 2019·Analytical Biochemistry·Shiro KatoTadao Oikawa
Apr 12, 2021·International Journal of Biological Macromolecules·E MorozovaT Demidkina

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