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Related Experiment Videos

A pure enzyme catalyzing penicillin biosynthesis.

I J Hollander, Y Q Shen, J Heim

    Science (New York, N.Y.)
    |May 11, 1984
    PubMed
    Summary

    Isopenicillin N synthetase (cyclase) was purified from Cephalosporium acremonium. The homogeneous enzyme, with a molecular weight of 40-42 kDa, is stable for weeks when stored at -20°C with sucrose.

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    Editorial.

    Microbial ecology·2013

    Area of Science:

    • Biochemistry
    • Enzymology
    • Microbial Biotechnology

    Background:

    • Isopenicillin N synthetase (IPNS) is a key enzyme in penicillin biosynthesis.
    • Understanding IPNS structure and function is crucial for metabolic engineering of antibiotic production.

    Purpose of the Study:

    • To purify Isopenicillin N synthetase (cyclase) to homogeneity from Cephalosporium acremonium.
    • To characterize the basic properties of the purified enzyme.

    Main Methods:

    • Enzyme purification using protamine sulfate and ammonium sulfate precipitations.
    • Gel filtration and ion-exchange high-performance liquid chromatography (HPLC).
    • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for molecular weight determination.

    Main Results:

    • Purified Isopenicillin N synthetase (cyclase) to homogeneity from Cephalosporium acremonium strain C-10.
    • Determined enzyme molecular weight to be 40,000-42,000 Da, confirmed by a single band on SDS-PAGE.
    • Achieved a 10% yield in the purification process.

    Conclusions:

    • The purified Isopenicillin N synthetase is stable when stored at -20°C with sucrose.
    • This stable, purified enzyme provides a valuable resource for further biochemical and structural studies.

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