Isolation of cyclase, epimerase and a ring expansion enzyme for

Details Isolation of cyclase, epimerase and a ring expansion enzyme for Isolation of cyclase, epimerase and a ring expansion enzyme for

1982-08-23

1985-04-09

Naff, David M.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

435 43, 435 47, 435 49, 435190, 435191, 435814, 435815, 435816, C12N 900, C12N 906, C12P 3500, C12P 3506

Patent

active

045102465

ABSTRACT:
The enzymes, cyclase, epimerase and a ring expansion enzyme, are isolated separately from a cell-free extract of a prokaryotic beta-lactam producing organism. The isolated enzymes are used to produce unnatural cephalosporins from polypeptide precursors. Isolation is carried out by adding ammonium sulfate to 40% saturation to the cell-free extract to precipitate contaminating proteins, adding ammonium sulfate to 70% saturation to the resultant supernatant to precipitate the enzymes, suspending the precipitated enzymes in a buffer, separating epimerase from the suspension by gel filtration, and separating cyclase and the ring expansion enzyme from each other by ion exchange chromatography.

REFERENCES:
patent: 4075061 (1978-02-01), Fleming et al.
patent: 4178210 (1979-12-01), Demain et al.
patent: 4248966 (1981-02-01), Demain et al.
patent: 4307192 (1981-12-01), Demain et al.

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