Metabolic characterisation of E. coli citrate synthase and phosphoenolpyruvate carboxylase mutants in aerobic cultures
E. coli is still one of the most commonly used hosts for protein production. However, when it is grown with excess glucose, acetate accumulation occurs. Elevated acetate concentrations have an inhibitory effect on growth rate and recombinant protein yield, and thus elimination of acetate formation i...
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| Veröffentlicht in: | Biotechnology letters 2006-12, Vol.28 (23), p.1945-1953 |
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| container_issue | 23 |
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| container_title | Biotechnology letters |
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| creator | Maeseneire, S. L. De Mey, M. De Vandedrinck, S Vandamme, E. J |
| description | E. coli is still one of the most commonly used hosts for protein production. However, when it is grown with excess glucose, acetate accumulation occurs. Elevated acetate concentrations have an inhibitory effect on growth rate and recombinant protein yield, and thus elimination of acetate formation is an important aim towards industrial production of recombinant proteins. Here we examine if over-expression of citrate synthase (gltA) or phosphoenolpyruvate carboxylase (ppc) can eliminate acetate production. Knock-out as well as over-expression mutants were constructed and characterized. Knocking out ppc or gltA decreased the maximum cell density by 14% and increased the acetate excretion by 7%, respectively decreased it by 10%. Over-expression of ppc or gltA increased the maximum cell dry weight by 91% and 23%, respectively. No acetate excretion was detected at these increased cell densities (35 and 23 g/l, respectively). |
| doi_str_mv | 10.1007/s10529-006-9182-8 |
| format | Article |
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L. De ; Mey, M. De ; Vandedrinck, S ; Vandamme, E. J</creator><creatorcontrib>Maeseneire, S. L. De ; Mey, M. De ; Vandedrinck, S ; Vandamme, E. J</creatorcontrib><description>E. coli is still one of the most commonly used hosts for protein production. However, when it is grown with excess glucose, acetate accumulation occurs. Elevated acetate concentrations have an inhibitory effect on growth rate and recombinant protein yield, and thus elimination of acetate formation is an important aim towards industrial production of recombinant proteins. Here we examine if over-expression of citrate synthase (gltA) or phosphoenolpyruvate carboxylase (ppc) can eliminate acetate production. Knock-out as well as over-expression mutants were constructed and characterized. Knocking out ppc or gltA decreased the maximum cell density by 14% and increased the acetate excretion by 7%, respectively decreased it by 10%. Over-expression of ppc or gltA increased the maximum cell dry weight by 91% and 23%, respectively. No acetate excretion was detected at these increased cell densities (35 and 23 g/l, respectively).</description><identifier>ISSN: 0141-5492</identifier><identifier>EISSN: 1573-6776</identifier><identifier>DOI: 10.1007/s10529-006-9182-8</identifier><identifier>PMID: 17028777</identifier><identifier>CODEN: BILED3</identifier><language>eng</language><publisher>Dordrecht: Dordrecht : Kluwer Academic Publishers</publisher><subject>Acetate reduction ; Acetates - chemistry ; Biological and medical sciences ; Biotechnology ; Biotechnology - methods ; Citrate (si)-Synthase - chemistry ; Citrate (si)-Synthase - genetics ; Cloning, Molecular ; E coli ; Escherichia coli ; Escherichia coli - enzymology ; Escherichia coli - genetics ; Excretion ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation, Bacterial ; Genetic Techniques ; gltA ; Glucose - metabolism ; Hydrogen-Ion Concentration ; Industrial production ; Metabolic engineering ; Metabolism ; Microbiological Techniques ; Models, Biological ; Mutation ; Phosphoenolpyruvate Carboxylase - chemistry ; Phosphoenolpyruvate Carboxylase - genetics ; ppc ; Proteins</subject><ispartof>Biotechnology letters, 2006-12, Vol.28 (23), p.1945-1953</ispartof><rights>2007 INIST-CNRS</rights><rights>Springer Science+Business Media, Inc. 2006</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c443t-af4ea19dd1325bf41322cd38c9717317f84ff7fd749eff5224db16c75b1d7bf33</citedby><cites>FETCH-LOGICAL-c443t-af4ea19dd1325bf41322cd38c9717317f84ff7fd749eff5224db16c75b1d7bf33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18314371$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17028777$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Maeseneire, S. L. De</creatorcontrib><creatorcontrib>Mey, M. De</creatorcontrib><creatorcontrib>Vandedrinck, S</creatorcontrib><creatorcontrib>Vandamme, E. J</creatorcontrib><title>Metabolic characterisation of E. coli citrate synthase and phosphoenolpyruvate carboxylase mutants in aerobic cultures</title><title>Biotechnology letters</title><addtitle>Biotechnol Lett</addtitle><description>E. coli is still one of the most commonly used hosts for protein production. However, when it is grown with excess glucose, acetate accumulation occurs. Elevated acetate concentrations have an inhibitory effect on growth rate and recombinant protein yield, and thus elimination of acetate formation is an important aim towards industrial production of recombinant proteins. Here we examine if over-expression of citrate synthase (gltA) or phosphoenolpyruvate carboxylase (ppc) can eliminate acetate production. Knock-out as well as over-expression mutants were constructed and characterized. Knocking out ppc or gltA decreased the maximum cell density by 14% and increased the acetate excretion by 7%, respectively decreased it by 10%. Over-expression of ppc or gltA increased the maximum cell dry weight by 91% and 23%, respectively. No acetate excretion was detected at these increased cell densities (35 and 23 g/l, respectively).</description><subject>Acetate reduction</subject><subject>Acetates - chemistry</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Biotechnology - methods</subject><subject>Citrate (si)-Synthase - chemistry</subject><subject>Citrate (si)-Synthase - genetics</subject><subject>Cloning, Molecular</subject><subject>E coli</subject><subject>Escherichia coli</subject><subject>Escherichia coli - enzymology</subject><subject>Escherichia coli - genetics</subject><subject>Excretion</subject><subject>Fundamental and applied biological sciences. 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Here we examine if over-expression of citrate synthase (gltA) or phosphoenolpyruvate carboxylase (ppc) can eliminate acetate production. Knock-out as well as over-expression mutants were constructed and characterized. Knocking out ppc or gltA decreased the maximum cell density by 14% and increased the acetate excretion by 7%, respectively decreased it by 10%. Over-expression of ppc or gltA increased the maximum cell dry weight by 91% and 23%, respectively. No acetate excretion was detected at these increased cell densities (35 and 23 g/l, respectively).</abstract><cop>Dordrecht</cop><pub>Dordrecht : Kluwer Academic Publishers</pub><pmid>17028777</pmid><doi>10.1007/s10529-006-9182-8</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Acetate reduction Acetates - chemistry Biological and medical sciences Biotechnology Biotechnology - methods Citrate (si)-Synthase - chemistry Citrate (si)-Synthase - genetics Cloning, Molecular E coli Escherichia coli Escherichia coli - enzymology Escherichia coli - genetics Excretion Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Bacterial Genetic Techniques gltA Glucose - metabolism Hydrogen-Ion Concentration Industrial production Metabolic engineering Metabolism Microbiological Techniques Models, Biological Mutation Phosphoenolpyruvate Carboxylase - chemistry Phosphoenolpyruvate Carboxylase - genetics ppc Proteins |
| title | Metabolic characterisation of E. coli citrate synthase and phosphoenolpyruvate carboxylase mutants in aerobic cultures |
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