Selection of a subtilisin-hyperproducing Bacillus in a highly structured environment |
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Authors: | D Naki C Paech G Ganshaw V Schellenberger |
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Institution: | (1) Genencor Intl. Inc., 925 Page Mill Rd., Palo Alto, CA 94304, USA Tel.: +1 650 846 7646 Fax: +1 650 845 6509 e-mail: vschellenberger@genencor.com, US |
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Abstract: | Mutants that secrete increased amounts of enzyme into a selection medium can be efficiently enriched from large populations
of mutagenized microorganisms during growth in hollow fibers. Under these conditions, each colony grows in its own microenvironment
and cross-feeding between neighboring colonies is limited. We applied the technique to B. subtilis carrying a plasmid-encoded protease gene. The plasmid was subjected to random mutagenesis and clones secreting up to fivefold-increased
amounts of enzyme were selected using a medium containing bovine serum albumin as the sole nitrogen source.
Received: 22 May 1997 / Received revision: 21 October 1997 / Accepted: 7 November 1997 |
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