Parallel Bubble Columns with Fed-Batch Technique for Microbial Process Development on a Small Scale

Autor: Matthias Arnold, Jutta Altenbach-Rehm, Ch. Nell, Dirk Weuster-Botz
Rok vydání: 1999
Předmět:
Zdroj: Chemical Engineering & Technology. 22:1051-1058
ISSN: 1521-4125
0930-7516
DOI: 10.1002/(sici)1521-4125(199912)22:12<1051::aid-ceat1051>3.0.co;2-c
Popis: 12 small-scale bubble columns, each and every column with a total volume of 500 ml, individually controlled distribution of sterile and humidified air and a pH-probe, were operated in an incubation chamber with temperature control. Intermittent feeding of substrate or inducer, as well as of base for parallel pH-control was achieved by connecting a precision syringe pump to a valve distributor with one 2/2-way miniature valve for each bubble column. This new parallel bio-reactor technique was applied for optimization of the feed profile of the chemical inducer isopropyl-β-D-thiogalactoside (IPTG) to improve the expression of a foreign protein (guanosin-5'-diphosphate-α-D-mannose-pyrophosphorylase, GDP-manPP) under the control of the lac promoter in Escherichia coli. A mean cell density of 8 g l -1 of recombinant E. coli was achieved in parallel fed-batch fermentations within 9.5 h due to a sufficient oxygen transfer (k L a of up to 0.2 s -1 ) and parallel pH-control. The GDP-manPP activity was improved by more than 100 % compared to the standard IPTG pulse within 3 sets of parallel fed-batch fermentations. A Genetic Algorithm was used for optimization of the IPTG feed profile. Scale-up of the optimized fed-batch process into the stirred tank reactor (scale-up factor of 20) was possible, if all reaction conditions were copied exactly with respect to the reactor volume. A final GDP-manPP activity of 715 U l -1 was measured 3 h after IPTG induction was finished.
Databáze: OpenAIRE
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