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Literature summary extracted from

  • Damnjanovic, J.; Nakano, H.; Iwasaki, Y.
    Deletion of a dynamic surface loop improves stability and changes kinetic behavior of phosphatidylinositol-synthesizing Streptomyces phospholipase D (2014), Biotechnol. Bioeng., 111, 674-682.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.4.4 gene pld, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Streptomyces antibioticus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.4.4 D40H/T291Y site-directed mutagenesis of residues D40 and T291 located within dynamic surface loops, the mutant is able to synthesize phosphatidylinositol by transphosphatidylation Streptomyces antibioticus
3.1.4.4 W187D/Y191Y/Y385R site-directed mutagenesis, the mutant is able to synthesize phosphatidylinositol by transphosphatidylation, thermostabilization of enzyme mutant W187D/Y191Y/Y385R, termed DYR, is attempted by rational design based on deletion of the D40 loop, generating two variants, DELTA37-45 DYR and DELTA38-46 DYR PLD. DELTA38-46 DYR shows highest thermostability as its activity half-life at 70°C proves 11.7 and 8.0 times longer than that of the DYR mutant and mutant DELTA37-45 DYR, respectively, molecular dynamics, overview Streptomyces antibioticus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.4.4 phosphatidylcholine + H2O Streptomyces antibioticus
-
 choline + phosphatidate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.4.4 Streptomyces antibioticus Q53728 gene pld
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.4.4 phosphatidylcholine + H2O
-
 Streptomyces antibioticus choline + phosphatidate
-
 ?
3.1.4.4 phosphatidylcholine + myo-inositol the wild-type enzyme is capable of synthesizing phosphatidylinositol by transphosphatidylation. Increase in phosphatidylinositol yield is possible by providing excess of solvated myo-inositol, which is achievable at high temperatures due to its highly temperature-dependent solubility, especially by enzyme variants with increased thermostability, e.g. mutant W187D/Y191Y/Y385R Streptomyces antibioticus phosphatidylinositol + choline
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.1.4.4 PLD
-
 Streptomyces antibioticus
3.1.4.4 SaPLD
-
 Streptomyces antibioticus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.4.4 30
-
 assay at Streptomyces antibioticus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.4.4 5.6
-
 assay at Streptomyces antibioticus