Abstract
We have compared the purification procedures as well as the biochemical and kinetic properties of wild type (wt-SAL3), untagged recombinant (rec(-His)SAL3), and tagged recombinant (rec(+His)SAL3) purified forms of Staphylococcus aureus lipase (SAL3). We used the pH-stat method (with emulsified tributyrin and olive oil as substrates) and the monomolecular film technique (with the three dicaprin isomers spread in the form of monomolecular films at the air-water interface). The data obtained showed that the recombinant expression process as well as the presence of a his-tag at the N-terminus of recombinant SAL3 affects significantly many biochemical and catalytic properties. The effects of the heterologous expression process on the catalytic properties of the staphylococcal lipases are three times more deleterious than the presence of an N-terminal tag extension.
MeSH terms
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Bacterial Proteins / chemistry
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism*
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Cloning, Molecular
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Diglycerides / metabolism
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Emulsions
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Escherichia coli
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Histidine / chemistry
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Hydrolysis
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Industrial Microbiology*
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Kinetics
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Lipase / chemistry
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Lipase / isolation & purification
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Lipase / metabolism*
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Models, Molecular
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Oligopeptides / chemistry
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Olive Oil
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Plant Oils / chemistry
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
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Staphylococcus aureus / chemistry
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Staphylococcus aureus / enzymology*
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Structure-Activity Relationship
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Substrate Specificity
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Surface Properties
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Triglycerides / metabolism
Substances
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Bacterial Proteins
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Diglycerides
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Emulsions
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His-His-His-His-His-His
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Oligopeptides
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Olive Oil
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Plant Oils
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Recombinant Fusion Proteins
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Triglycerides
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Histidine
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Lipase
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tributyrin