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Fermentation and isolation of C10‐deacetylase for the production of 10‐deacetylbaccatin III from baccatin III
Author(s) -
Nanduri Venkata B.,
Hanson Ronald L.,
LaPorte Thomas L.,
Ko Raphael Y.,
Patel Ramesh N.,
Szarka Laszlo J.
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260480518
Subject(s) - fermentation , isolation (microbiology) , chemistry , biochemistry , stereochemistry , biology , microbiology and biotechnology
10‐Deacetylabaccatin III (10 DAB), an important precursor for paclitaxel semisynthesis, is enhanced in yew extracts using C10‐deacetylase and C13‐deacylase enzymes. 4 C10‐deacetylase is an intracellular enzyme produced by the fermentation of a soil microorganism, Nocardioides luteus (SC 13912). During the fermentation of Nocardioides luteus , the growth of cells reaches a maximum growth at 28 h. C10‐deacetylase enzyme activity starts at 26 h and peaks at 38 h of the fermentation. The cells are recovered by centrifugation. The C10‐deacetylase enzyme was purified from the Nocardioides luteus cells. The enzyme was purified 190‐fold to near homogeneity. The purified enzyme appeared as a single band on 12.5% SDS‐PAGE analysis with a molecular weight of 40,000 daltons. © 1995 John Wiley & Sons, Inc.