Escherichia coli BL21 in a BIOSTAT® B Reactor - 4 Pages

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Escherichia coli BL21 in a BIOSTAT®  B Reactor

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sartoriusstedim biotech Robust Batch Cultivation Process for Recombinant Protein Production with Escherichia coli BL21 in a BIOSTAT® B Reactor turning science

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Pohle D, Beckmann B, and Sanders EA* Hamburg University of Applied Sciences Faculty of Life Sciences, Dep. Biotechnology Lohbruegger Kirchstr. 65 21033 Hamburg | Germany * Corresponding author: ernst.sanders@haw-hamburg.de 1. Introduction Worldwide and since decades recombinant protein production is often done with wellknown apathogenic strains of E coli, e.g. K12 derivatives like DH5, TG1, W3110 or members of the B family like B834, BL21, TKB1 (Casali 2003). Up to now E coli kept its importance as host strain due to the ease of transformation and genetically modification, its fast growing,...

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36 °C while no pH control was applied. The dissolved oxygen concentration DO was maintained at 30% saturation by increasing agitation speed (500 – 2,000 rpm) and O2-enrichment if required. A constant air flow of 2 l min-1 was applied through a ring sparger. The cultivation conditions were maintained by the BIOSTAT® B controller. Within 2 hours the induction cell density of OD600 = 19 ± 2 was reached and expression of the recombinant protein was induced by the addition of L-arabinose. Subsequently the cells grew with a specific growth rate of 0.5 h–1 for 1.5 h, then the specific growth rate...

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4. Summary An extremely easy-to-use process for cultivation of E. coli cells and production of recombinant proteins in BIOSTAT® B type bioreactors was demonstrated. With just a two-component-medium and without any effort to control the pH a final cell density of OD600 ≈ 56 (BDM ≈ 20 g l–1) was achieved. Direct inoculation with thawed cryocultures allows abandonment of individual and unequal seed cultures from shaking flasks and a process length of not more than a working day. The production of GFP was induced by addition of various concentrations of L-arabinose at OD600 ≈ 20. Maximum...

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