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Current kinetic parameters for\n            <jats:named-content content-type=\"genus-species\">Lactococcus lactis<\/jats:named-content>\n            are scattered through different studies and performed under different assay conditions. Furthermore, assay conditions often diverge from conditions prevailing in the intracellular environment. To establish uniform assay conditions that resemble intracellular conditions, we analyzed the intracellular composition of anaerobic glucose-limited chemostat cultures of\n            <jats:named-content content-type=\"genus-species\">L. lactis<\/jats:named-content>\n            subsp.\n            <jats:named-content content-type=\"genus-species\">cremoris<\/jats:named-content>\n            MG 1363. Based on this, we designed a new assay medium for enzyme activity measurements of growing cells of\n            <jats:named-content content-type=\"genus-species\">L. lactis<\/jats:named-content>\n            , mimicking as closely as practically possible its intracellular environment. Procedures were optimized to be carried out in 96-well plates, and the reproducibility and dynamic range were checked for all enzyme activity measurements. The effects of freezing and the carryover of ammonium sulfate from the addition of coupling enzymes were also established. Activities of all 10 glycolytic and 4 fermentative enzymes were measured. Remarkably, most\n            <jats:italic>in vivo<\/jats:italic>\n            -like activities were lower than previously published data. Yet, the ratios of\n            <jats:italic>V<\/jats:italic>\n            <jats:sub>max<\/jats:sub>\n            over measured\n            <jats:italic>in vivo<\/jats:italic>\n            fluxes were above 1. With this work, we have developed and extensively validated standard protocols for enzyme activity measurements for\n            <jats:named-content content-type=\"genus-species\">L. lactis<\/jats:named-content>\n            .\n          <\/jats:p>","DOI":"10.1128\/aem.05276-11","type":"journal-article","created":{"date-parts":[[2011,10,22]],"date-time":"2011-10-22T04:07:56Z","timestamp":1319256476000},"page":"134-143","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":55,"title":["Standardized Assay Medium To Measure Lactococcus lactis Enzyme Activities while Mimicking Intracellular Conditions"],"prefix":"10.1128","volume":"78","author":[{"given":"Anisha","family":"Goel","sequence":"first","affiliation":[{"name":"Centre for Integrative Bioinformatics, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands"},{"name":"Laboratory of Microbiology, Wageningen University and Research Centre, Wageningen, The Netherlands"},{"name":"Kluyver Centre for Genomics of Industrial Fermentation\/NCSB, Delft, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Filipe","family":"Santos","sequence":"additional","affiliation":[{"name":"Centre for Integrative Bioinformatics, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands"},{"name":"Kluyver Centre for Genomics of Industrial Fermentation\/NCSB, Delft, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Willem M.","family":"de Vos","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, Wageningen University and Research Centre, Wageningen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bas","family":"Teusink","sequence":"additional","affiliation":[{"name":"Centre for Integrative Bioinformatics, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands"},{"name":"Kluyver Centre for Genomics of Industrial Fermentation\/NCSB, Delft, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Douwe","family":"Molenaar","sequence":"additional","affiliation":[{"name":"Centre for Integrative Bioinformatics, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands"},{"name":"Kluyver Centre for Genomics of Industrial Fermentation\/NCSB, Delft, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.compbiolchem.2008.08.001"},{"key":"e_1_3_3_3_2","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1016\/S0005-2736(96)00199-X","article-title":"Regulation of cellular Mg2+ by Saccharomyces cerevisiae.","volume":"1323","author":"Beeler T","year":"1997","unstructured":"BeelerTBruceKDunnT. 1997. 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