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The conclusions from these simulations were confirmed by experiments with recombinant strains of the wild types and polyhydroxyalkanoate (PHA)\u2010negative mutants of <jats:italic>Ralstonia eutropha<\/jats:italic> and <jats:italic>Pseudomonas putida<\/jats:italic> expressing CGP synthetase genes (<jats:italic>cphA<\/jats:italic>) of <jats:italic>Synechocystis<\/jats:italic> sp. strain PCC6308 or <jats:italic>Anabaena<\/jats:italic> sp. strain PCC7120. In particular, the effects of suitable precursor substrates and of oxygen supply as well as of the capability to accumulate PHA in addition to CGP biosynthesis were investigated. Since CGP consists of the amino acids aspartate and arginine, the tricarboxylic acid cycle (TCC), which provides intermediates for biosynthesis of these amino acids, seems to be important. Excretion of intermediates of the TCC upon cultivation at restricted oxygen supply and conversion of fumarate mainly to malate and to only little succinate in the absence of oxygen indicated that TCC intermediates for arginine and aspartate biosynthesis were provided by the oxidative or reductive parts of the TCC, respectively. The following important conclusions were made from the experiments and the simulations: (i) external arginine additionally supplied to the medium, (ii) oxygen limitation, and (iii) absence of PHA accumulation exerted positive effects on CGP accumulation. These conclusions were utilized to obtain CGP contents in the cells of as high as 17.9% (w\u2009\u00b7\u2009w<jats:sup>\u22121<\/jats:sup>) during cultivation of the investigated bacteria at the 30\u2010L scale using mineral salts medium. Such high CGP contents were previously not obtained with these bacteria at a 30\u2010L scale, even if complex media were used. \u00a9 2006 Wiley Periodicals, Inc.<\/jats:p>","DOI":"10.1002\/bit.20760","type":"journal-article","created":{"date-parts":[[2006,1,24]],"date-time":"2006-01-24T20:18:39Z","timestamp":1138133919000},"page":"698-717","source":"Crossref","is-referenced-by-count":46,"title":["Optimization of cyanophycin production in recombinant strains of <i>Pseudomonas putida<\/i> and <i>Ralstonia eutropha<\/i> employing elementary mode analysis and statistical experimental design"],"prefix":"10.1002","volume":"93","author":[{"given":"Simone Cardoso","family":"Diniz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ingo","family":"Voss","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Steinb\u00fcchel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2006,1,24]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s002030000206"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.67.5.2176-2182.2001"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/bm010075a"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1327.2000.01622.x"},{"key":"e_1_2_1_6_1","first-page":"1624","volume-title":"Methoden der enzymatischen Analyse","author":"Bergmeyer HU","year":"1974"},{"key":"e_1_2_1_7_1","unstructured":"Boehringer GmbH.1980.Methoden der enzymatischen Lebensmittelanalytik. 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