{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T19:44:44Z","timestamp":1787168684348,"version":"build-2736575974"},"reference-count":35,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2015,9,3]],"date-time":"2015-09-03T00:00:00Z","timestamp":1441238400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["GM077596"],"award-info":[{"award-number":["GM077596"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2016,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:label\/>\n                    <jats:p>\n                      As resistance to current anti\u2010malarial therapeutics spreads, new compounds to treat malaria are increasingly needed. One promising compound is FR900098, a naturally occurring phosphonate. Due to limitations in both chemical synthesis and biosynthetic methods for FR900098 production, this potential therapeutic has yet to see widespread implementation. Here we applied a combinatorial pathway engineering strategy to improve the production of FR900098 in\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                      by modulating each of the pathway's nine genes with four promoters of different strengths. Due to the large size of the library and the low screening throughput, it was necessary to develop a novel screening strategy that significantly reduced the sample size needed to find an optimal strain. This was done by using biased libraries that localize searching around top hits and home in on high\u2010producing strains. By incorporating this strategy, a significantly improved strain was found after screening less than 3% of the entire library. When coupled with culturing optimization, a strain was found to produce 96\u2009mg\/L, a 16\u2010fold improvement over the original strain. We believe the enriched library method developed here can be used on other large pathways that may be difficult to engineer by combinatorial methods due to low screening throughput. Biotechnol. Bioeng. 2016;113: 384\u2013392. \u00a9 2015 Wiley Periodicals, Inc.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/bit.25719","type":"journal-article","created":{"date-parts":[[2015,8,4]],"date-time":"2015-08-04T08:19:15Z","timestamp":1438676355000},"page":"384-392","source":"Crossref","is-referenced-by-count":21,"title":["Combinatorial pathway engineering for optimized production of the anti\u2010malarial FR900098"],"prefix":"10.1002","volume":"113","author":[{"given":"Todd S.","family":"Freestone","sequence":"first","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering and Institute for Genomic Biology University of Illinois at Urbana\u2010Champaign Urbana Illinois"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huimin","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering and Institute for Genomic Biology University of Illinois at Urbana\u2010Champaign Urbana Illinois"},{"name":"Departments of Chemistry, Biochemistry, and Bioengineering, Center for Biophysics and Computational Biology, School of Molecular and Cellular Biology University of Illinois at Urbana\u2010Champaign Urbana Illinois 61801"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,9,3]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1191652"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.263533"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja00224a054"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1039\/C4CC09181G"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMp1108322"},{"key":"e_1_2_6_7_1","first-page":"40","article-title":"Customized optimization of metabolic pathways by combinatorial transcriptional engineering","author":"Du J","year":"2012","journal-title":"Nucleic Acids Res"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-004-3381-z"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2008.07.010"},{"key":"e_1_2_6_10_1","first-page":"4","article-title":"Golden Gate Shuffling: A One\u2010Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes","author":"Engler C","year":"2009","journal-title":"Plos One"},{"key":"e_1_2_6_11_1","first-page":"10","article-title":"Efficient search, mapping, and optimization of multi\u2010protein genetic systems in diverse bacteria","author":"Farasat I","year":"2014","journal-title":"Mol Syst Biol"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/ol702082k"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1318"},{"key":"e_1_2_6_14_1","volume-title":"Directed evolution of phosphite dehydrogenase and engineered biosynthesis of FR\u2010900098 [Dissertation]","author":"Johannes TW","year":"2008"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2009.12.009"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.285.5433.1573"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02736-12"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(97)01138-4"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymben.2014.06.002"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt809"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.78.091707.100215"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-013-5361-4"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.33.13"},{"key":"e_1_2_6_24_1","first-page":"3553","article-title":"A novel short and efficient synthetic route to the antimalarial agent FR900098 and derivatives","author":"Perruchon J","year":"2007","journal-title":"Synthesis\u2010Stuttgart"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/335457a0"},{"key":"e_1_2_6_26_1","first-page":"37","article-title":"DNA assembler, an in vivo genetic method for rapid construction of biochemical pathways","author":"Shao ZY","year":"2009","journal-title":"Nucleic Acids Res"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3063"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tet.2012.11.049"},{"key":"e_1_2_6_29_1","doi-asserted-by":"crossref","first-page":"214","DOI":"10.4269\/ajtmh.2007.77.214","article-title":"Molecular epidemiology of malaria in Cameroon. 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