The Ohio State University

Battelle Memorial Institute

Charles L Desanti

Michael L Dickens

William R Strohl

John Bartholomew

Ryan W Daly

Patrick Norris

Sherry K Blight

David Charles Seibert, Jr

A biological culture unit comprises a chamber body and a valve plate. The chamber body has defined therein a growth chamber and an aliquot chamber. The valve plate is disposed on the top surface of the chamber body and is movable to define selectable configurations of the biological culture unit for: (1) loading the growth chamber; (3) transferring an aliquot from the growth chamber to the aliquot chamber via a path defined within the biological culture unit in the transfer position; and (4) extracting the aliquot from the aliquot chamber. The selectable positions may further include (2) a growth position, and (5) a termination position for putting a termination agent into the growth and aliquot chambers. The valve plate may further include a neutralizer port that is aligned with the aliquot chamber in the loading position, for loading a neutralizing agent into the aliquot chamber.

Biological culture unit

The present invention provides novel methods for producing doxorubicin using daunomycin as a substrate. One method employs a genetically engineered host microorganism which is transformed with a vector, preferably a plasmid, which contains the doxA gene. Preferably, the doxA gene, also referred to herein as 'doxA', is cloned into a plasmid which is then introduced into the host microorganism, preferably a bacterial host, more preferably Streptomyces, to provide a transformed host microorganism. The doxA gene, when present on a plasmid, confers on the transformed host the ability to convert daunomycin and 13-dihydrodaunomycin, to doxorubicin. The doxA gene encodes a P450-like enzyme which catalyzes the hydroxylation of daunomycin and 13-dihydrodaunomycin at C-14 to form doxorubicin; such enzyme is designated 'daunomycin C-14 hydroxylase'. Thus, the expression of doxA in the transformed host using a plasmid which contains doxA enables the transformed host to convert daunomycin to doxorubicin. The doxorubicin is then extracted from host microorganism cultures.

Methods of producing doxorubicin

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Columbus, OH, United States of America

Charles L Desanti