Center for Infectious Diseases and Vaccinology, and Center for Environmental Biotechnology, the Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
Proc Natl Acad Sci U S A. 2011 Apr 26;108(17):6899-904. doi: 10.1073/pnas.1103014108. Epub 2011 Apr 11.
To avoid costly biomass recovery in photosynthetic microbial biofuel production, we genetically modified cyanobacteria to produce and secrete fatty acids. Starting with introducing an acyl-acyl carrier protein thioesterase gene, we made six successive generations of genetic modifications of cyanobacterium Synechocystis sp. PCC6803 wild type (SD100). The fatty acid secretion yield was increased to 197 ± 14 mg/L of culture in one improved strain at a cell density of 1.0 × 10(9) cells/mL by adding codon-optimized thioesterase genes and weakening polar cell wall layers. Although these strains exhibited damaged cell membranes at low cell densities, they grew more rapidly at high cell densities in late exponential and stationary phase and exhibited less cell damage than cells in wild-type cultures. Our results suggest that fatty acid secreting cyanobacteria are a promising technology for renewable biofuel production.
为避免在光合微生物生物燃料生产中耗费大量生物质能回收,我们对蓝藻进行基因改造以生产和分泌脂肪酸。我们从引入酰基辅酶 A-酰基载体蛋白硫酯酶基因开始,对野生型集胞藻 PCC6803(SD100)进行了六代连续遗传修饰。通过添加密码子优化的硫酯酶基因和削弱极性细胞壁层,在一个改良的菌株中,脂肪酸分泌产量在 1.0×10(9)个细胞/mL 的细胞密度下提高到 197±14mg/L 的培养物。尽管这些菌株在低细胞密度下表现出细胞膜受损,但它们在后期指数和静止期在高细胞密度下生长更快,并且细胞损伤比野生型培养物中的细胞少。我们的研究结果表明,分泌脂肪酸的蓝藻是可再生生物燃料生产的一种很有前途的技术。