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Literature summary extracted from

  • Yang, Y.; Ko, T.P.; Liu, L.; Li, J.; Huang, C.H.; Chen, J.; Guo, R.T.; Du, G.
    Roles of tryptophan residue and disulfide bond in the variable lid region of oxidized polyvinyl alcohol hydrolase (2014), Biochem. Biophys. Res. Commun., 452, 509-514.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.7.1.7 gene oph, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Sphingopyxis sp.
3.7.1.7 recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 trxB (DE3) Pseudomonas sp.

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.7.1.7 purified recombinant detagged wild-type enzyme and mutants S172A and S172C, free and in complexes with PEG, butyrate, or DMSO, respectively, by sitting drop method using 30% PEG 5000 MME, 0.1 M MES pH 6.5. and 0.2 M (NH4)2SO4 for the wild-type, and 0.1 M tri-sodium citrate, pH 5.6, 30% w/v PEG 4000, and 0.3% w/v n-octyl-beta-D-glucoside for the mutants, X-ray diffraction structure determination and analysis at 1.49-2.10 A resolution, molecular replacement Pseudomonas sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.7.1.7 C241A/C248A site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme Sphingopyxis sp.
3.7.1.7 additional information the mutants show reduced kcat/Km for 4-nitrophenyl acetate, indicating the importance of Trp255 in sequestering the active site from solvent. The significantly lower activity for 4-nitrophenyl butyrate can be a result of product inhibition. The mutant activity is retained with 4-nitrophenyl caprylate and 4-nitrophenyl laurate as the substrates, reflecting the amphipathic nature of the cleft Pseudomonas sp.
3.7.1.7 S172A site-directed mutagenesis, the mutant shows 20% activity compared to the wild-type enzyme Pseudomonas sp.
3.7.1.7 S172C site-directed mutagenesis Pseudomonas sp.
3.7.1.7 W255A site-directed mutagenesis Pseudomonas sp.
3.7.1.7 W255F site-directed mutagenesis Pseudomonas sp.
3.7.1.7 W255Y site-directed mutagenesis Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.7.1.7 additional information
-
 additional information Michaelis-Menten kinetics of the wild-type enzyme with 4-nitrophenyl fatty acid (C2-C16) substrates, overview Sphingopyxis sp.
3.7.1.7 additional information
-
 additional information Michaelis-Menten kinetics of wild-type and mutant enzymes with 4-nitrophenyl fatty acid (C2-C16) substrates, overview Pseudomonas sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.7.1.7 additional information Pseudomonas sp. the enzyme catalyzes the cleavage of C-C bond in beta-diketone ?
-
 ?
3.7.1.7 additional information Sphingopyxis sp. the enzyme catalyzes the cleavage of C-C bond in beta-diketone ?
-
 ?
3.7.1.7 additional information Pseudomonas sp. VM15C the enzyme catalyzes the cleavage of C-C bond in beta-diketone ?
-
 ?
3.7.1.7 oxidized polyvinyl alcohol + H2O Pseudomonas sp.
-
 ?
-
 ?
3.7.1.7 oxidized polyvinyl alcohol + H2O Sphingopyxis sp.
-
 ?
-
 ?
3.7.1.7 oxidized polyvinyl alcohol + H2O Pseudomonas sp. VM15C
-
 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.7.1.7 Pseudomonas sp. Q9LCQ7
-
-
3.7.1.7 Pseudomonas sp. VM15C Q9LCQ7
-
-
3.7.1.7 Sphingopyxis sp. Q588Z2 gene oph
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.7.1.7 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 trxB (DE3) by nickel affinity chromatography and digestion by TEV protease to remove the His-tagged thioredoxin, followed by a second nickel affinity chromatography, and anion exchange chromatography and ultrafiltration Pseudomonas sp.
3.7.1.7 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Sphingopyxis sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.7.1.7 additional information the enzyme catalyzes the cleavage of C-C bond in beta-diketone Pseudomonas sp. ?
-
 ?
3.7.1.7 additional information the enzyme catalyzes the cleavage of C-C bond in beta-diketone Sphingopyxis sp. ?
-
 ?
3.7.1.7 additional information the enzyme shows lipase activity with 4-nitrophenyl esters as the substrates. The wild-type enzyme shows increased Km and decreased kcat/Km with the acyl chain length Sphingopyxis sp. ?
-
 ?
3.7.1.7 additional information the enzyme shows lipase activity with 4-nitrophenyl esters as the substrates. The wild-type enzyme shows increased Km and decreased kcat/Km with the acyl chain length, and the mutants show reduced kcat/Km for 4-nitrophenyl acetate, indicating the importance of Trp255 in sequestering the active site from solvent. The significantly lower activity for 4-nitrophenyl butyrate can be a result of product inhibition Pseudomonas sp. ?
-
 ?
3.7.1.7 additional information the enzyme catalyzes the cleavage of C-C bond in beta-diketone Pseudomonas sp. VM15C ?
-
 ?
3.7.1.7 additional information the enzyme shows lipase activity with 4-nitrophenyl esters as the substrates. The wild-type enzyme shows increased Km and decreased kcat/Km with the acyl chain length, and the mutants show reduced kcat/Km for 4-nitrophenyl acetate, indicating the importance of Trp255 in sequestering the active site from solvent. The significantly lower activity for 4-nitrophenyl butyrate can be a result of product inhibition Pseudomonas sp. VM15C ?
-
 ?
3.7.1.7 oxidized polyvinyl alcohol + H2O
-
 Pseudomonas sp. ?
-
 ?
3.7.1.7 oxidized polyvinyl alcohol + H2O
-
 Sphingopyxis sp. ?
-
 ?
3.7.1.7 oxidized polyvinyl alcohol + H2O
-
 Pseudomonas sp. VM15C ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.7.1.7 OPH
-
 Pseudomonas sp.
3.7.1.7 OPH
-
 Sphingopyxis sp.
3.7.1.7 oxidized polyvinyl alcohol hydrolase
-
 Pseudomonas sp.
3.7.1.7 oxidized polyvinyl alcohol hydrolase
-
 Sphingopyxis sp.
3.7.1.7 oxidized PVA hydrolase
-
 Pseudomonas sp.
3.7.1.7 oxidized PVA hydrolase
-
 Sphingopyxis sp.
3.7.1.7 pOPH
-
 Pseudomonas sp.
3.7.1.7 sOPH
-
 Sphingopyxis sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.7.1.7 37
-
 assay at Pseudomonas sp.
3.7.1.7 37
-
 assay at Sphingopyxis sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.7.1.7 8
-
 assay at Pseudomonas sp.
3.7.1.7 8
-
 assay at Sphingopyxis sp.

General Information

EC Number General Information Comment Organism
3.7.1.7 evolution the enzyme belongs to the alpha/beta-hydrolase family and contains a unique lid region that covers the active site Pseudomonas sp.
3.7.1.7 evolution the enzyme belongs to the alpha/beta-hydrolase family and contains a unique lid region that covers the active site Sphingopyxis sp.
3.7.1.7 additional information roles of tryptophan residue and disulfide bond in the variable lid region of oxidized polyvinyl alcohol hydrolase, the lid is the most variable region of the enzyme. The disulfide bond formation of Cys257/267 is important for the activity of pOPH Pseudomonas sp.
3.7.1.7 additional information roles of tryptophan residue and disulfide bond in the variable lid region of oxidized polyvinyl alcohol hydrolase, the lid is the most variable region of the enzyme. The disulfide bond formation of Cys257/267 is not essential for sOPH, which has a shorter lid structure Sphingopyxis sp.
3.7.1.7 physiological function the enzyme is involved in degradation of polyvinyl alcohol Pseudomonas sp.
3.7.1.7 physiological function the enzyme is involved in degradation of polyvinyl alcohol Sphingopyxis sp.