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In this study, the full potential of <jats:italic>Chlorella<\/jats:italic> sp. HS2 is explored through optimization of the parameters for its heterotrophic cultivation. First, carbon and nitrogen sources were screened in PhotobioBox. Initial screening using the Plackett-Burman design (PBD) was then adopted and the concentrations of the major nutrients (glucose, sodium nitrate, and dipotassium phosphate) were optimized via response surface methodology (RSM) with a central composite design (CCD). Upon validation of the model via flask-scale cultivation, the optimized BG11 medium was found to result in a three-fold improvement in biomass amounts, from 5.85 to 18.13\u2009g\/L, in comparison to a non-optimized BG11 medium containing 72\u2009g\/L glucose. Scaling up the cultivation to a 5-L fermenter resulted in a greatly improved biomass concentration of 35.3 g\/L owing to more efficient oxygenation of the culture. In addition, phosphorus feeding fermentation was employed in an effort to address early depletion of phosphate, and a maximum biomass concentration of 42.95 g\/L was achieved, with biomass productivity of 5.37\u2009g\/L\/D.<\/jats:p>","DOI":"10.1038\/s41598-019-55854-9","type":"journal-article","created":{"date-parts":[[2019,12,18]],"date-time":"2019-12-18T11:02:55Z","timestamp":1576666975000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":48,"title":["Optimization of heterotrophic cultivation of Chlorella sp. HS2 using screening, statistical assessment, and validation"],"prefix":"10.1038","volume":"9","author":[{"given":"Hee Su","family":"Kim","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Won-Kun","family":"Park","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bongsoo","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gyeongho","family":"Seon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"William I.","family":"Suh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Myounghoon","family":"Moon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yong Keun","family":"Chang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2019,12,18]]},"reference":[{"key":"55854_CR1","doi-asserted-by":"publisher","first-page":"1485","DOI":"10.1007\/s10811-014-0459-3","volume":"27","author":"J Lowrey","year":"2014","unstructured":"Lowrey, J., Brooks, M. 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