Matthäus Falk, Ketelhot Markus, Gatter Michael, Barth Gerold
Institute of Microbiology, Dresden University of Technology, Dresden, Germany.
Appl Environ Microbiol. 2014 Mar;80(5):1660-9. doi: 10.1128/AEM.03167-13. Epub 2013 Dec 27.
The codon-optimized genes crtB and crtI of Pantoea ananatis were expressed in Yarrowia lipolytica under the control of the TEF1 promoter of Y. lipolytica. Additionally, the rate-limiting genes for isoprenoid biosynthesis in Y. lipolytica, GGS1 and HMG1, were overexpressed to increase the production of lycopene. All of the genes were also expressed in a Y. lipolytica strain with POX1 to POX6 and GUT2 deleted, which led to an increase in the size of lipid bodies and a further increase in lycopene production. Lycopene is located mainly within lipid bodies, and increased lipid body formation leads to an increase in the lycopene storage capacity of Y. lipolytica. Growth-limiting conditions increase the specific lycopene content. Finally, a yield of 16 mg g(-1) (dry cell weight) was reached in fed-batch cultures, which is the highest value reported so far for a eukaryotic host.
将解脂耶氏酵母的密码子优化基因crtB和crtI在解脂耶氏酵母TEF1启动子的控制下在解脂耶氏酵母中表达。此外,解脂耶氏酵母中类异戊二烯生物合成的限速基因GGS1和HMG1被过表达以增加番茄红素的产量。所有基因也在缺失了POX1至POX6和GUT2的解脂耶氏酵母菌株中表达,这导致脂质体大小增加以及番茄红素产量进一步提高。番茄红素主要位于脂质体内,脂质体形成增加导致解脂耶氏酵母的番茄红素储存能力增强。生长限制条件会增加番茄红素的比含量。最后,在补料分批培养中达到了16 mg g(-1)(干细胞重量)的产量,这是迄今为止报道的真核宿主的最高值。