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This study intended to optimize the carotenoid synthesis pathway in Aurantiochytrium sp. by metabolic engineering to increase the carotenoid content. Multi-sourced key enzyme genes involved in lipid synthesis (LPAAT and DGAT) and astaxanthin synthesis (crtZ and crtW) were selected to construct single-gene expression vectors and transformed into Aurantiochytrium sp. The results showed that the overexpression of LPAAT of Phaeodactylum tricornutum in Aurantiochytrium sp. caused an increase of 39.3% in astaxanthin, 424.7% in \u03b2-carotene, 901.8% in canthaxanthin, and 575.9% in lutein, as well as a down-regulation of 15.3% in the fatty acid content. Transcriptomics analysis revealed enhanced expression of genes involved in purine and amino acid metabolism in the transformed strains, and the down-regulation of the citric acid cycle led to an increase in the source of acetyl coenzyme A for the production of fatty acids. This study provides strong experimental evidence to support the application of increasing carotenoid levels in Aurantiochytrium sp.<\/jats:p>","DOI":"10.3390\/md23040164","type":"journal-article","created":{"date-parts":[[2025,4,11]],"date-time":"2025-04-11T05:38:26Z","timestamp":1744349906000},"page":"164","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Effect of Heterologous Expression of Key Enzymes Involved in Astaxanthin and Lipid Synthesis on Lipid and Carotenoid Production in Aurantiochytrium sp."],"prefix":"10.3390","volume":"23","author":[{"given":"Yaping","family":"Shao","sequence":"first","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5953-8441","authenticated-orcid":false,"given":"Zhengquan","family":"Gao","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3672-8610","authenticated-orcid":false,"given":"Fengjie","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, School of Science and Technology, Georgia Gwinnett College, Lawrenceville, GA 30043, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulin","family":"Cui","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Zou","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinju","family":"Ma","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiaolei","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuyong","family":"Wu","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunxiao","family":"Meng","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Binzhou Medical University, Yantai 264003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e01243-19","DOI":"10.1128\/AEM.01243-19","article-title":"Effects of methanol on carotenoids as well as biomass and fatty acid biosynthesis in Schizochytrium limacinum B4D1","volume":"85","author":"Du","year":"2019","journal-title":"Appl. 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