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Expression and Purification ofRecombinantlysostaphin inEscherichia coli

✍ Scribed by Err-Cheng Chan

Publisher
Springer Netherlands
Year
1996
Tongue
English
Weight
284 KB
Volume
18
Category
Article
ISSN
0141-5492

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✦ Synopsis

A 1.5 kb plasmid-encoded lysostaphin gene fragment of Staphylococcus staphylolyticus was amplified by polymerase chain reaction OPCR) and cloned in Escherichia coli by using plasmid pET29b(+) as an expression vector. By optimizing culture conditions, the activities of lysostaphin were expressed as 66 %, 30 %, and 4 % in extracellular, intraeeilular, and periplasmic fractions of recombinant E. coli, respectively. The enzyme was purified to homogeneity by using a simple one-step fractionation on bacterial cells of lysostaphin-resistant

Staphylococcus aureus mutant. The recombinant enzyme had an Mr of approximate 27 kDa, and its bacteriolytic activity was indistinguishable to the authentic lysostaphin purified from Staphylococcus staphylolytlcus.

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