Abstract
Though synthesized with a cleavable signal peptide and devoid of membrane anchors, the 262-amino-acid-residue Streptomyces K15 DD-transpeptidase/penicillin-binding protein is membrane-bound. Overexpression in Streptomyces lividans resulted in the export of an appreciable amount of the synthesized protein (4 mg/litre of culture supernatant). The water-soluble enzyme was purified close to protein homogeneity with a yield of 75%. It requires the presence of 0.5 M-NaCl to remain soluble. It is indistinguishable from the detergent-extract wild-type enzyme with respect to molecular mass, thermostability, transpeptidase activity and penicillin-binding capacity.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins*
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Carrier Proteins / genetics*
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Carrier Proteins / isolation & purification
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Carrier Proteins / metabolism
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Cell Membrane / enzymology
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Enzyme Stability
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Gene Expression*
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Hexosyltransferases*
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Hot Temperature
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Molecular Sequence Data
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Molecular Weight
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Muramoylpentapeptide Carboxypeptidase / genetics*
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Muramoylpentapeptide Carboxypeptidase / isolation & purification
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Muramoylpentapeptide Carboxypeptidase / metabolism
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Penicillin-Binding Proteins
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Penicillins / metabolism
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Peptidyl Transferases*
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Plasmids
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Sodium Chloride
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Solubility
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Streptomyces / enzymology*
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Streptomyces / genetics
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Transformation, Bacterial
Substances
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Bacterial Proteins
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Carrier Proteins
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Penicillin-Binding Proteins
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Penicillins
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Sodium Chloride
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Peptidyl Transferases
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Hexosyltransferases
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Muramoylpentapeptide Carboxypeptidase