Structural determination of phosphatidylinositol-synthesizing engineered phospholipase D from streptomyces antibioticus

Ariela Samantha

Research output: ThesisMaster's Thesis

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Abstract

The substrate specificity of phospholipase D (PLO) from Streptomyces antibioticus (SaPLD) has been altered through protein engineering techniques. The mutant enzyme, namely TNYR SaPLD, is capable of producing phosphatidylinositol (Pl) using phosphatidylcholine (PC) and myo-inositol as substrates. TNYR SaPLD has been expressed, purified and crystallised for structural determination by X-ray crystallography to gain a better understanding of the substrate binding features of the mutant enzyme. The work gave insight into the role of the enzyme's catalytic residues and the mutated residues. The structure of the enzyme as well as its conformational changes during substrate binding also have been described.
Original languageEnglish
QualificationDoctor of Philosophy
Awarding Institution
  • The University of Western Australia
Award date23 Jan 2019
DOIs
Publication statusUnpublished - 2019

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Phospholipase D
Phosphatidylinositols
Substrates
Enzymes
X ray crystallography
Inositol
Phosphatidylcholines
Proteins

Cite this

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title = "Structural determination of phosphatidylinositol-synthesizing engineered phospholipase D from streptomyces antibioticus",
abstract = "The substrate specificity of phospholipase D (PLO) from Streptomyces antibioticus (SaPLD) has been altered through protein engineering techniques. The mutant enzyme, namely TNYR SaPLD, is capable of producing phosphatidylinositol (Pl) using phosphatidylcholine (PC) and myo-inositol as substrates. TNYR SaPLD has been expressed, purified and crystallised for structural determination by X-ray crystallography to gain a better understanding of the substrate binding features of the mutant enzyme. The work gave insight into the role of the enzyme's catalytic residues and the mutated residues. The structure of the enzyme as well as its conformational changes during substrate binding also have been described.",
keywords = "phospholipase D, X-ray crystallography, Streptomyces antibioticus, Protein engineering, phosphatidylinositol",
author = "Ariela Samantha",
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doi = "10.26182/5c5bc4559211c",
language = "English",
school = "The University of Western Australia",

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PY - 2019

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AB - The substrate specificity of phospholipase D (PLO) from Streptomyces antibioticus (SaPLD) has been altered through protein engineering techniques. The mutant enzyme, namely TNYR SaPLD, is capable of producing phosphatidylinositol (Pl) using phosphatidylcholine (PC) and myo-inositol as substrates. TNYR SaPLD has been expressed, purified and crystallised for structural determination by X-ray crystallography to gain a better understanding of the substrate binding features of the mutant enzyme. The work gave insight into the role of the enzyme's catalytic residues and the mutated residues. The structure of the enzyme as well as its conformational changes during substrate binding also have been described.

KW - phospholipase D

KW - X-ray crystallography

KW - Streptomyces antibioticus

KW - Protein engineering

KW - phosphatidylinositol

U2 - 10.26182/5c5bc4559211c

DO - 10.26182/5c5bc4559211c

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