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Literature summary for 1.1.1.25 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
Search for more literature for 1.1.1.25

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
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
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
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
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
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
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]

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

Localization

Localization Comment Organism GeneOntology No. Textmining
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]

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

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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]

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

Substrates and Products (Substrate)

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

Subunits

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

Turnover Number [1/s]

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