Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Rodrigues, V.d.a..S.; Basso, L.A.; Santos, D.S.
    Homogeneous recombinant Mycobacterium tuberculosis shikimate dehydrogenase production: an essential step towards target-based drug design (2009), Int. J. Biol. Macromol., 45, 200-205.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.25 plasmid pET23a(+)::aroE (either wild-type or mutants) transformed into Escherichia coli C41(DE3) Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.25 D105A the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid Mycobacterium tuberculosis
1.1.1.25 D105N the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid Mycobacterium tuberculosis
1.1.1.25 K69A the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid. Best recombinant protein expression protocol of the K69A mutant in insoluble form is using Escherichia coli C41(DE3) strain, grown at 37°C for 24 h after induction with 1 mM IPTG, and cell disruption by sonication Mycobacterium tuberculosis
1.1.1.25 K69H the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid Mycobacterium tuberculosis
1.1.1.25 K69I the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid Mycobacterium tuberculosis
1.1.1.25 K69Q the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.25 0.011
-
 NADPH
-
 Mycobacterium tuberculosis
1.1.1.25 0.029
-
 3-dehydroshikimate
-
 Mycobacterium tuberculosis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.1.1.25 additional information the freeze-thaw method is able to yield the mutant protein in soluble form, after growth at 37°C for 24 h with IPTG induction of Escherichia coli C41 (DE3) cells harboring the recombinant plasmid. Best recombinant protein expression protocol of the K69A mutant in insoluble form is using Escherichia coli C41(DE3) strain, grown at 37°C for 24 h after induction with 1 mM IPTG, and cell disruption by sonication. Solubilization of recombinant wild-type and K69A enzymes in inclusion bodies is achieved by either treating the pellet with the denaturant agent urea (3 or 6 M) or by treating with the non ionic detergent zwittergent 3-14 (5%) Mycobacterium tuberculosis
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.25 27000
-
 x * 27000, SDS-PAGE Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.25 Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.25 homogeneous recombinant mutant K69A and wild-type proteins are obtained by a three-step purification protocol, by gel filtration, 11.8fold Mycobacterium tuberculosis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.25 1.2
-
 crude extract Mycobacterium tuberculosis
1.1.1.25 14.1
-
 11.8fold purified enzyme Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.25 3-dehydroshikimate + NADPH + H+
-
 Mycobacterium tuberculosis shikimate + NADP+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.25 ? x * 27000, SDS-PAGE Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.25 50
-
 NADPH
-
 Mycobacterium tuberculosis