Cloning, sequence analysis, expression, and inactivation of the Corynebacterium glutamicum icd gene encoding isocitrate dehydrogenase and biochemical characterization of the enzyme.

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  • Additional Information
    • Source:
      Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 2985120R Publication Model: Print Cited Medium: Print ISSN: 0021-9193 (Print) Linking ISSN: 00219193 NLM ISO Abbreviation: J Bacteriol Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, DC : American Society for Microbiology
    • Subject Terms:
      Corynebacterium/*genetics ; Isocitrate Dehydrogenase/*genetics; Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; Cloning, Molecular ; Corynebacterium/enzymology ; Gene Expression ; Glutamic Acid/biosynthesis ; Isocitrate Dehydrogenase/chemistry ; Isocitrate Dehydrogenase/isolation & purification ; Molecular Sequence Data
    • Abstract:
      NADP(+)-dependent isocitrate dehydrogenase (ICD) is an important enzyme of the intermediary metabolism, as it controls the carbon flux within the citric acid cycle and supplies the cell with 2-oxoglutarate and NADPH for biosynthetic purposes. In the amino acid-producing organism Corynebacterium glutamicum, the specific activity of ICD was independent of the growth substrate and of the growth phase at approximately 1 U/mg, indicating that this enzyme is constitutively formed. The ICD gene, icd, was isolated, subcloned on a plasmid, and introduced into C. glutamicum. Compared with the wild type, the recombinant strains showed up to 10-fold-higher specific ICD activities. The nucleotide sequence of a 3,595-bp DNA fragment containing the icd gene was determined. The predicted gene product of icd consists of 739 amino acids (M(r) = 80.091) and showed 58.5% identity with the monomeric ICD isozyme II from Vibrio sp. strain ABE-1 but no similarity to any known ICD of the dimeric type. Inactivation of the chromosomal icd gene led to glutamate auxotrophy and to the absence of any detectable ICD activity, suggesting that only a single ICD is present in C. glutamicum. From an icd-overexpressing C. glutamicum strain, ICD was purified and biochemically characterized. The native ICD was found to be a monomer; to be specific for NADP+; to be weakly inhibited by oxaloacetate, 2-oxoglutarate, and citrate; and to be severely inhibited by oxaloacetate plus glyoxylate. The data indicate that ICD from C. glutamicum is structurally similar to ICDs from bacteria of the genera Vibrio, Rhodomicrobium, and Azotobacter but different from all other known procaryotic and eucaryotic ICDs.
    • References:
      J Biol Chem. 1970 Jun;245(12):3186-94. (PMID: 4393512)
      Biochemistry. 1969 Aug;8(8):3175-84. (PMID: 4390163)
      Nat New Biol. 1973 Nov 14;246(150):40-1. (PMID: 4519026)
      Biochemistry. 1974 Feb 12;13(4):677-83. (PMID: 4149369)
      J Gen Microbiol. 1975 Mar;87(1):37-51. (PMID: 1094097)
      Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7. (PMID: 271968)
      Science. 1979 Mar 16;203(4385):1111-2. (PMID: 34215)
      Gene. 1982 Oct;19(3):259-68. (PMID: 6295879)
      Methods Enzymol. 1983;100:243-55. (PMID: 6353143)
      J Biol Chem. 1985 Jul 15;260(14):8430-7. (PMID: 2861202)
      J Biol Chem. 1985 Sep 5;260(19):10563-8. (PMID: 3897222)
      Biochem J. 1986 Mar 1;234(2):317-23. (PMID: 3521584)
      J Biol Chem. 1987 Aug 5;262(22):10422-5. (PMID: 3112144)
      Biochim Biophys Acta. 1988 Feb 10;952(3):282-9. (PMID: 3337829)
      J Biochem. 1987 Dec;102(6):1489-98. (PMID: 3129416)
      Nature. 1990 Jan 4;343(6253):38-43. (PMID: 2296288)
      FEMS Microbiol Lett. 1989 Dec;53(3):299-303. (PMID: 2612892)
      J Bacteriol. 1990 Mar;172(3):1663-6. (PMID: 2106514)
      Comp Biochem Physiol B. 1990;95(3):431-58. (PMID: 2184988)
      Science. 1990 Aug 31;249(4972):1012-6. (PMID: 2204109)
      J Biol Chem. 1991 Feb 5;266(4):2339-45. (PMID: 1989987)
      Biotechnology (N Y). 1991 Jan;9(1):84-7. (PMID: 1367217)
      Appl Microbiol Biotechnol. 1991 Feb;34(5):617-22. (PMID: 1369320)
      Gene. 1991 Jun 15;102(1):93-8. (PMID: 1864513)
      Biochemistry. 1991 Sep 3;30(35):8671-8. (PMID: 1888729)
      Eur J Biochem. 1991 Nov 15;202(1):85-93. (PMID: 1935983)
      Appl Environ Microbiol. 1992 Jan;58(1):93-8. (PMID: 1539996)
      Mol Microbiol. 1992 Feb;6(3):317-26. (PMID: 1552846)
      Biochemistry. 1992 Jul 14;31(27):6219-23. (PMID: 1627563)
      J Biol Chem. 1992 Aug 15;267(23):16417-23. (PMID: 1644826)
      J Bacteriol. 1994 Jun;176(12):3474-83. (PMID: 8206824)
      J Bacteriol. 1994 Aug;176(15):4669-79. (PMID: 8045898)
      Microbiology. 1994 Aug;140 ( Pt 8):1817-28. (PMID: 7522844)
      J Lab Clin Med. 1963 Jul;62:148-58. (PMID: 13937022)
      J Bacteriol. 1992 Oct;174(19):6076-86. (PMID: 1400158)
      J Cell Biochem. 1993 Jan;51(1):14-8. (PMID: 8381789)
      J Biochem. 1992 Dec;112(6):849-55. (PMID: 1295895)
      Plant Mol Biol. 1993 Mar;21(5):739-52. (PMID: 8467073)
      Biochem J. 1993 Jun 15;292 ( Pt 3):705-10. (PMID: 8318002)
      Biochemistry. 1993 Sep 14;32(36):9317-22. (PMID: 8369301)
      J Bacteriol. 1993 Nov;175(21):6873-80. (PMID: 8226630)
      J Bacteriol. 1994 May;176(9):2718-26. (PMID: 8169222)
      J Biochem. 1968 Jul;64(1):45-53. (PMID: 4974271)
      J Biochem. 1968 Sep;64(3):355-63. (PMID: 5707822)
      Biochem J. 1972 Nov;130(1):211-9. (PMID: 4144072)
    • Gene Symbol:
      icd
    • Molecular Sequence:
      GENBANK X71489
    • Accession Number:
      3KX376GY7L (Glutamic Acid)
      EC 1.1.1.41 (Isocitrate Dehydrogenase)
    • Publication Date:
      Date Created: 19950201 Date Completed: 19950227 Latest Revision: 20220408
    • Publication Date:
      20221213
    • Accession Number:
      PMC176656
    • Accession Number:
      10.1128/jb.177.3.774-782.1995
    • Accession Number:
      7836312