Accelerated process development for protease production in continuous multi-stage cultures.
A fermentation process was developed and optimized for the production of a specific protease from Bacillus licheniformis PWD-1. Media formulations were constructed and crucial environmental parameters were optimized to enhance growth and product formation. Process dynamics of substrate consumption, biomass-, product-, as well as by-product formation were determined under controlled conditions in a bioreactor. Using kinetic data from batch- and continuous-culture experiments, a fed-batch process was developed producing proteolytic activities 10 times those found during regular batch culture. In one stage continuous stirred tank culture protease formation was completely decoupled from sporulation. Shift experiments in one-stage continuous cultures led to the development of a two-stage continuous stirred tank fermentation process using optimized conditions for growth in the first stage and protease formation in the second stage. Accordingly, the basis for a continuous production of the enzyme on a pilot scale was accomplished.
Raninger
A
A
Department of Biochemical Reaction Engineering, Institute of Biotechnology-SFB Biocatalysis, University of Technology Graz, Petersgasse 12, 8010 Graz, Austria. alexander.raninger@gmx.net
Steiner
W
W
D001407:Bacillus / Q000145:classification / Q000201:enzymology* / Q000254:growth & development*; D019149:Bioreactors / Q000382:microbiology*; D002455:Cell Division; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods*; D010450:Endopeptidases / Q000096:biosynthesis*; D004867:Equipment Design; D005947:Glucose / Q000378:metabolism*; D013045:Species Specificity; D013696:Temperature
D010450:Endopeptidases; D005947:Glucose