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Literature summary extracted from

  • Eustaquio, A.S.; Pojer, F.; Noel, J.P.; Moore, B.S.
    Discovery and characterization of a marine bacterial SAM-dependent chlorinase (2008), Nat. Chem. Biol., 4, 69-74.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.94 expressed in Escherichia coli BL21(DE3) cells Salinispora tropica CNB-440

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.94 hanging drop vapor diffusion method, using 28% (w/v) PEG 3350, 0.3 M KCl, 2 mM dithiothreitol, 0.1 M MOPSO, pH 7.0 Salinispora tropica CNB-440

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.94 G131S the mutation leads to the complete loss of all SalL halogenase activities and the inability to crystallize SalL Salinispora tropica CNB-440
2.5.1.94 W129F the mutant shows 52% chlorinase activity, 76% brominase activity, and 3% iodidase activity compared to the wild type enzyme Salinispora tropica CNB-440
2.5.1.94 Y70T the mutation results in a two-orders-of-magnitude reduction of SalL activity, the mutant shows 0.07% chlorinase activity, 0.1% brominase activity, and 0.4% iodidase activity compared to the wild type enzyme Salinispora tropica CNB-440
2.5.1.94 Y70T/G131S the mutant shows 0.3% chlorinase activity, 0.5% brominase activity, and no iodidase activity compared to the wild type enzyme Salinispora tropica CNB-440

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.94 0.0002
-
 5'-chloro-5'-deoxyadenosine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.001
-
 S-adenosyl-L-methionine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.0046
-
 5'-deoxy-5'-iodoadenosine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 45
-
 chloride in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 150
-
 bromide in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 260
-
 Iodide in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.94 30000
-
 3 * 30000, in solution as determined by sedimentation velocity studies Salinispora tropica CNB-440

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.94 Salinispora tropica CNB-440 A4X3Q0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.94
-
 Salinispora tropica CNB-440

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.94 5-deoxy-5'-chloroadenosine + L-methionine
-
 Salinispora tropica CNB-440 S-adenosyl-L-methionine + chloride
-
 r
2.5.1.94 additional information the enzyme does not accept fluoride and 5-deoxy-5'-fluoroadenosine as substrates Salinispora tropica CNB-440 ?
-
 ?
2.5.1.94 S-adenosyl-L-methionine + bromide the enzyme also accepts bromide as substrate (with 90% relative activity compared to chloride) but the brominase activity is probably not biologically relevant in this marine bacterium Salinispora tropica CNB-440 5'-deoxy-5'-bromoadenosine + L-methionine
-
 r
2.5.1.94 S-adenosyl-L-methionine + chloride
-
 Salinispora tropica CNB-440 5'-deoxy-5'-chloroadenosine + L-methionine
-
 r
2.5.1.94 S-adenosyl-L-methionine + iodide the enzyme also accepts iodide as substrate (with 33% relative activity compared to chloride) but the iodinase activity is probably not biologically relevant in this marine bacterium Salinispora tropica CNB-440 5'-deoxy-5'-iodoadenosine + L-methionine
-
 r

Subunits

EC Number Subunits Comment Organism
2.5.1.94 homotrimer 3 * 30000, in solution as determined by sedimentation velocity studies Salinispora tropica CNB-440

Synonyms

EC Number Synonyms Comment Organism
2.5.1.94 chlorinase
-
 Salinispora tropica CNB-440
2.5.1.94 SalL
-
 Salinispora tropica CNB-440

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.94 0.005
-
 Iodide in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.013
-
 bromide in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.015
-
 S-adenosyl-L-methionine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.015
-
 chloride in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.2
-
 L-methionine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.2
-
 5'-chloro-5'-deoxyadenosine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.83
-
 5'-deoxy-5'-iodoadenosine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440

General Information

EC Number General Information Comment Organism
2.5.1.94 physiological function SalL functions in an orthogonal manner to biological chlorination reactions to initiate the biosynthesis of a new halogenated polyketide synthase building block Salinispora tropica CNB-440

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.5.1.94 0.0000192
-
 Iodide in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.0000883
-
 bromide in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.0003
-
 chloride in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 0.077
-
 L-methionine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 15
-
 S-adenosyl-L-methionine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 181.1
-
 5'-deoxy-5'-iodoadenosine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440
2.5.1.94 1000
-
 5'-chloro-5'-deoxyadenosine in 50 mM phosphate buffer (pH 7.9) at 37°C Salinispora tropica CNB-440