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Smart sustainable bottle (SSB) system for E. coli based recombinant protein production.

Li Z, Carstensen B, Rinas U - Microb. Cell Fact. (2014)

Bottom Line: Oxygen transfer capacities are in the range as in conventional bioreactors operated at intermediate aeration rates and by far exceed those found in conventional shaking flasks and disposable bioreactors.The production performance regarding amount and solubility of proteins with robust and delicate properties was as good as in state-of-the-art stirred tank commercial bioreactors.The SSB system represents a low cost protein production device applicable for easy, effective, and reproducible recombinant protein production.

View Article: PubMed Central - PubMed

Affiliation: Leibniz University of Hannover, Technical Chemistry - Life Science, Hannover, Germany. Zhaopeng.Li@iftc.uni-hannover.de.

ABSTRACT

Background: Recombinant proteins are usually required in laboratories interested in the protein but not in the production process itself. Thus, technical equipment which is easy to handle and straight forward protein production procedures are of great benefit to those laboratories. Companies selling single use cultivation bags and bioreactors are trying to satisfy at least part of these needs. However, single-use systems can contribute to major costs which might be acceptable when "good manufacturing practices" are required but not acceptable for most laboratories facing tight funding.

Results: The assembly and application of a simple self-made "smart sustainable bottle" (SSB) system for E. coli based protein production is presented. The core of the SSB system is a 2-L glass bottle which is operated at constant temperature, air flow, and stirrer speed without measurement and control of pH and dissolved oxygen. Oxygen transfer capacities are in the range as in conventional bioreactors operated at intermediate aeration rates and by far exceed those found in conventional shaking flasks and disposable bioreactors. The SSB system was applied for the production of various recombinant proteins using T7-based expression systems and a defined autoinduction medium. The production performance regarding amount and solubility of proteins with robust and delicate properties was as good as in state-of-the-art stirred tank commercial bioreactors.

Conclusions: The SSB system represents a low cost protein production device applicable for easy, effective, and reproducible recombinant protein production.

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Related in: MedlinePlus

Schematic diagram and photos of the SSB system. Schematic diagram of SSB system (A): flowmeter (1), 0.2 μm venting filter (2), 1-L pre-wetting bottle (3), 2-L cultivation bottle (4), magnetic stirrer (5), 1-L safety bottle for off-gas (6), optional second 0.5-L safety bottle for off-gas (7*), optional off-gas analyzer (8*), circulating thermostat (9), optional sample port (10*), optional liquid inlet port (11*). Devices marked with an asterisk are optional and not required for successful operation of the SSB system. Overview of the SSB system (B), sparger and cooling finger (C), and pre-wetting and cultivation bottles (D). For details of the assembly of the SSB system please refer to Additional file 1.
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Fig1: Schematic diagram and photos of the SSB system. Schematic diagram of SSB system (A): flowmeter (1), 0.2 μm venting filter (2), 1-L pre-wetting bottle (3), 2-L cultivation bottle (4), magnetic stirrer (5), 1-L safety bottle for off-gas (6), optional second 0.5-L safety bottle for off-gas (7*), optional off-gas analyzer (8*), circulating thermostat (9), optional sample port (10*), optional liquid inlet port (11*). Devices marked with an asterisk are optional and not required for successful operation of the SSB system. Overview of the SSB system (B), sparger and cooling finger (C), and pre-wetting and cultivation bottles (D). For details of the assembly of the SSB system please refer to Additional file 1.

Mentions: The core of the SSB system is a 2-L glass bottle which is operated at constant temperature, constant air flow, and constant stirrer speed without measurement and control of pH and dissolved oxygen (Figure 1). The temperature is controlled via a self-made cooling finger connected to a conventional circulating thermostat. The airflow is controlled via a conventional flow meter and the inlet air passed through a 0.2 μm venting sterile filter and subsequently moistened by passing through a bottle containing sterile water to prevent water loss from the main bottle during long-term cultivation. The outlet air from the main bottle is passed through a 1-L empty safety bottle and if there is interest to connect an off-gas analyzer to the SSB system it is recommended to add a second safety bottle to protect the analyzer in case excessive foaming occurs. Aeration of the culture broth occurs by passing the air through a self-made sparger and by mixing using a conventional magnetic stirrer. For details of the assembly of the SSB system please refer to Additional file 1.Figure 1


Smart sustainable bottle (SSB) system for E. coli based recombinant protein production.

Li Z, Carstensen B, Rinas U - Microb. Cell Fact. (2014)

Schematic diagram and photos of the SSB system. Schematic diagram of SSB system (A): flowmeter (1), 0.2 μm venting filter (2), 1-L pre-wetting bottle (3), 2-L cultivation bottle (4), magnetic stirrer (5), 1-L safety bottle for off-gas (6), optional second 0.5-L safety bottle for off-gas (7*), optional off-gas analyzer (8*), circulating thermostat (9), optional sample port (10*), optional liquid inlet port (11*). Devices marked with an asterisk are optional and not required for successful operation of the SSB system. Overview of the SSB system (B), sparger and cooling finger (C), and pre-wetting and cultivation bottles (D). For details of the assembly of the SSB system please refer to Additional file 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4226889&req=5

Fig1: Schematic diagram and photos of the SSB system. Schematic diagram of SSB system (A): flowmeter (1), 0.2 μm venting filter (2), 1-L pre-wetting bottle (3), 2-L cultivation bottle (4), magnetic stirrer (5), 1-L safety bottle for off-gas (6), optional second 0.5-L safety bottle for off-gas (7*), optional off-gas analyzer (8*), circulating thermostat (9), optional sample port (10*), optional liquid inlet port (11*). Devices marked with an asterisk are optional and not required for successful operation of the SSB system. Overview of the SSB system (B), sparger and cooling finger (C), and pre-wetting and cultivation bottles (D). For details of the assembly of the SSB system please refer to Additional file 1.
Mentions: The core of the SSB system is a 2-L glass bottle which is operated at constant temperature, constant air flow, and constant stirrer speed without measurement and control of pH and dissolved oxygen (Figure 1). The temperature is controlled via a self-made cooling finger connected to a conventional circulating thermostat. The airflow is controlled via a conventional flow meter and the inlet air passed through a 0.2 μm venting sterile filter and subsequently moistened by passing through a bottle containing sterile water to prevent water loss from the main bottle during long-term cultivation. The outlet air from the main bottle is passed through a 1-L empty safety bottle and if there is interest to connect an off-gas analyzer to the SSB system it is recommended to add a second safety bottle to protect the analyzer in case excessive foaming occurs. Aeration of the culture broth occurs by passing the air through a self-made sparger and by mixing using a conventional magnetic stirrer. For details of the assembly of the SSB system please refer to Additional file 1.Figure 1

Bottom Line: Oxygen transfer capacities are in the range as in conventional bioreactors operated at intermediate aeration rates and by far exceed those found in conventional shaking flasks and disposable bioreactors.The production performance regarding amount and solubility of proteins with robust and delicate properties was as good as in state-of-the-art stirred tank commercial bioreactors.The SSB system represents a low cost protein production device applicable for easy, effective, and reproducible recombinant protein production.

View Article: PubMed Central - PubMed

Affiliation: Leibniz University of Hannover, Technical Chemistry - Life Science, Hannover, Germany. Zhaopeng.Li@iftc.uni-hannover.de.

ABSTRACT

Background: Recombinant proteins are usually required in laboratories interested in the protein but not in the production process itself. Thus, technical equipment which is easy to handle and straight forward protein production procedures are of great benefit to those laboratories. Companies selling single use cultivation bags and bioreactors are trying to satisfy at least part of these needs. However, single-use systems can contribute to major costs which might be acceptable when "good manufacturing practices" are required but not acceptable for most laboratories facing tight funding.

Results: The assembly and application of a simple self-made "smart sustainable bottle" (SSB) system for E. coli based protein production is presented. The core of the SSB system is a 2-L glass bottle which is operated at constant temperature, air flow, and stirrer speed without measurement and control of pH and dissolved oxygen. Oxygen transfer capacities are in the range as in conventional bioreactors operated at intermediate aeration rates and by far exceed those found in conventional shaking flasks and disposable bioreactors. The SSB system was applied for the production of various recombinant proteins using T7-based expression systems and a defined autoinduction medium. The production performance regarding amount and solubility of proteins with robust and delicate properties was as good as in state-of-the-art stirred tank commercial bioreactors.

Conclusions: The SSB system represents a low cost protein production device applicable for easy, effective, and reproducible recombinant protein production.

Show MeSH
Related in: MedlinePlus