Prediction of recombinant protein production by Escherichia coli derived online from indicators of metabolic burden
Background: The oxygen transfer rate (OTR) and the biomass concentration are two important parameters describing a microbial fermentation. It has been shown before that from the course of these parameters over time information on metabolic burden during heterologous protein production can be obtaine...
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| Veröffentlicht in: | Biotechnology progress 2018-11, Vol.34 (6), p.1543-1552 |
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| creator | Mühlmann, Martina Julia Forsten, Eva Noack, Saskia Büchs, Jochen |
| description | Background: The oxygen transfer rate (OTR) and the biomass concentration are two important parameters describing a microbial fermentation. It has been shown before that from the course of these parameters over time information on metabolic burden during heterologous protein production can be obtained. While online monitoring in large fermenters is ubiquitously established, it is still not a common practice in small‐scale cultures. Nevertheless, several techniques like the Respiration Activity MOnitoring System (RAMOS) device for online monitoring of the OTR in shake flasks and the BioLector device for measuring scattered light (ScL) representing biomass in microtiter plates have been developed. Results: A new microtiter plate‐based method is presented that reveals how online derived ScL signals can be transformed into signals that are proportional to the courses of OTR over time for Escherichia coli. The transformed signal is obtained by simply taking the first derivative of ScL (dScL/dt). The proportionality of both parameters is successfully validated for the strains E. coli BL21(DE3) and Tuner(DE3) expressing cellulases and the fluorescent protein FbFP, respectively. Relative amounts of produced heterologous proteins are predicted exclusively based on the course of the transformed ScL signal. A variety of induction conditions with varying inducer concentration and induction time were investigated with this method. Conclusion: The presented method based on ScL measurement allows for high‐throughput online determination of signals proportional to OTR courses. They enable the interpretation of physiological states and offer the possibility to predict the recombinant protein production in E. coli. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1543–1552, 2018 |
| doi_str_mv | 10.1002/btpr.2704 |
| format | Article |
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It has been shown before that from the course of these parameters over time information on metabolic burden during heterologous protein production can be obtained. While online monitoring in large fermenters is ubiquitously established, it is still not a common practice in small‐scale cultures. Nevertheless, several techniques like the Respiration Activity MOnitoring System (RAMOS) device for online monitoring of the OTR in shake flasks and the BioLector device for measuring scattered light (ScL) representing biomass in microtiter plates have been developed. Results: A new microtiter plate‐based method is presented that reveals how online derived ScL signals can be transformed into signals that are proportional to the courses of OTR over time for Escherichia coli. The transformed signal is obtained by simply taking the first derivative of ScL (dScL/dt). The proportionality of both parameters is successfully validated for the strains E. coli BL21(DE3) and Tuner(DE3) expressing cellulases and the fluorescent protein FbFP, respectively. Relative amounts of produced heterologous proteins are predicted exclusively based on the course of the transformed ScL signal. A variety of induction conditions with varying inducer concentration and induction time were investigated with this method. Conclusion: The presented method based on ScL measurement allows for high‐throughput online determination of signals proportional to OTR courses. They enable the interpretation of physiological states and offer the possibility to predict the recombinant protein production in E. coli. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1543–1552, 2018</description><identifier>ISSN: 8756-7938</identifier><identifier>EISSN: 1520-6033</identifier><identifier>DOI: 10.1002/btpr.2704</identifier><identifier>PMID: 30248250</identifier><language>eng</language><publisher>Hoboken, USA: John Wiley & Sons, Inc</publisher><subject>Bacteria ; Biomass ; Cell culture ; E coli ; Escherichia coli ; Fermentation ; Fermenters ; Flasks ; Fluorescence ; Internet ; Laboratory equipment ; Measuring instruments ; Metabolism ; Microorganisms ; microtiter plate ; Monitoring ; On-line systems ; Organic chemistry ; Oxygen transfer ; oxygen transfer rate ; Parameters ; Predictions ; Proteins ; recombinant protein ; Scattered light</subject><ispartof>Biotechnology progress, 2018-11, Vol.34 (6), p.1543-1552</ispartof><rights>2018 American Institute of Chemical Engineers</rights><rights>2018 American Institute of Chemical Engineers.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3904-d4296f957b37a3226a405dd180ad5b3ad960f7a074ea5d4a8260ba614fbf06e03</citedby><cites>FETCH-LOGICAL-c3904-d4296f957b37a3226a405dd180ad5b3ad960f7a074ea5d4a8260ba614fbf06e03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fbtpr.2704$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fbtpr.2704$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,778,782,1414,27907,27908,45557,45558</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30248250$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mühlmann, Martina Julia</creatorcontrib><creatorcontrib>Forsten, Eva</creatorcontrib><creatorcontrib>Noack, Saskia</creatorcontrib><creatorcontrib>Büchs, Jochen</creatorcontrib><title>Prediction of recombinant protein production by Escherichia coli derived online from indicators of metabolic burden</title><title>Biotechnology progress</title><addtitle>Biotechnol Prog</addtitle><description>Background: The oxygen transfer rate (OTR) and the biomass concentration are two important parameters describing a microbial fermentation. It has been shown before that from the course of these parameters over time information on metabolic burden during heterologous protein production can be obtained. While online monitoring in large fermenters is ubiquitously established, it is still not a common practice in small‐scale cultures. Nevertheless, several techniques like the Respiration Activity MOnitoring System (RAMOS) device for online monitoring of the OTR in shake flasks and the BioLector device for measuring scattered light (ScL) representing biomass in microtiter plates have been developed. Results: A new microtiter plate‐based method is presented that reveals how online derived ScL signals can be transformed into signals that are proportional to the courses of OTR over time for Escherichia coli. The transformed signal is obtained by simply taking the first derivative of ScL (dScL/dt). The proportionality of both parameters is successfully validated for the strains E. coli BL21(DE3) and Tuner(DE3) expressing cellulases and the fluorescent protein FbFP, respectively. Relative amounts of produced heterologous proteins are predicted exclusively based on the course of the transformed ScL signal. A variety of induction conditions with varying inducer concentration and induction time were investigated with this method. Conclusion: The presented method based on ScL measurement allows for high‐throughput online determination of signals proportional to OTR courses. They enable the interpretation of physiological states and offer the possibility to predict the recombinant protein production in E. coli. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1543–1552, 2018</description><subject>Bacteria</subject><subject>Biomass</subject><subject>Cell culture</subject><subject>E coli</subject><subject>Escherichia coli</subject><subject>Fermentation</subject><subject>Fermenters</subject><subject>Flasks</subject><subject>Fluorescence</subject><subject>Internet</subject><subject>Laboratory equipment</subject><subject>Measuring instruments</subject><subject>Metabolism</subject><subject>Microorganisms</subject><subject>microtiter plate</subject><subject>Monitoring</subject><subject>On-line systems</subject><subject>Organic chemistry</subject><subject>Oxygen transfer</subject><subject>oxygen transfer rate</subject><subject>Parameters</subject><subject>Predictions</subject><subject>Proteins</subject><subject>recombinant protein</subject><subject>Scattered light</subject><issn>8756-7938</issn><issn>1520-6033</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><recordid>eNp1kUtrFUEQRhtRzPXqwj8gDW50MUn1c2aWGuIDAgaJ66EfNaTDTPe1e0a5_94eb3QhuPooOByq6iPkJYNzBsAv7HLI57wF-YjsmOLQaBDiMdl1rdJN24vujDwr5R4AOtD8KTkTwGXHFexIucnog1tCijSNNKNLsw3RxIUeclowxC39eiLskV4Vd4c5uLtgqEtToL5OP9DTFKcQkY45zTTE6jRLymWTzrgYW1FH7Zo9xufkyWimgi8eck--fbi6vfzUXH_5-Pny3XXjRA-y8ZL3euxVa0VrBOfaSFDesw6MV1YY32sYWwOtRKO8NB3XYI1mcrQjaASxJ29O3nrB9xXLMsyhOJwmEzGtZeCMcdZ1vH5oT17_g96nNce6XaWUVKrnLa_U2xPlciol4zgccphNPg4Mhq2JYWti2Jqo7KsH42pn9H_JP6-vwMUJ-BkmPP7fNLy_vfn6W_kLwE2UKQ</recordid><startdate>201811</startdate><enddate>201811</enddate><creator>Mühlmann, Martina Julia</creator><creator>Forsten, Eva</creator><creator>Noack, Saskia</creator><creator>Büchs, Jochen</creator><general>John Wiley & Sons, Inc</general><general>Wiley Subscription Services, Inc</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>201811</creationdate><title>Prediction of recombinant protein production by Escherichia coli derived online from indicators of metabolic burden</title><author>Mühlmann, Martina Julia ; 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The proportionality of both parameters is successfully validated for the strains E. coli BL21(DE3) and Tuner(DE3) expressing cellulases and the fluorescent protein FbFP, respectively. Relative amounts of produced heterologous proteins are predicted exclusively based on the course of the transformed ScL signal. A variety of induction conditions with varying inducer concentration and induction time were investigated with this method. Conclusion: The presented method based on ScL measurement allows for high‐throughput online determination of signals proportional to OTR courses. They enable the interpretation of physiological states and offer the possibility to predict the recombinant protein production in E. coli. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1543–1552, 2018</abstract><cop>Hoboken, USA</cop><pub>John Wiley & Sons, Inc</pub><pmid>30248250</pmid><doi>10.1002/btpr.2704</doi><tpages>10</tpages></addata></record> |
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| subjects | Bacteria Biomass Cell culture E coli Escherichia coli Fermentation Fermenters Flasks Fluorescence Internet Laboratory equipment Measuring instruments Metabolism Microorganisms microtiter plate Monitoring On-line systems Organic chemistry Oxygen transfer oxygen transfer rate Parameters Predictions Proteins recombinant protein Scattered light |
| title | Prediction of recombinant protein production by Escherichia coli derived online from indicators of metabolic burden |
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