期刊论文详细信息
Biotechnology for Biofuels
Engineering cyanobacteria to improve photosynthetic production of alka(e)nes
Weihua Wang2  Xufeng Liu1  Xuefeng Lu2 
[1] University of Chinese Academy of Sciences, Beijing, 100049, China
[2] Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao, 266101, China
关键词: Metabolic engineering;    Fatty acid;    Alka(e)ne;    Synechocystis sp. PCC6803;    Cyanobacteria;   
Others  :  798057
DOI  :  10.1186/1754-6834-6-69
 received in 2012-12-01, accepted in 2013-04-29,  发布年份 2013
PDF
【 摘 要 】

Background

Cyanobacteria can utilize solar energy and convert carbon dioxide into biofuel molecules in one single biological system. Synechocystis sp. PCC 6803 is a model cyanobacterium for basic and applied research. Alkanes are the major constituents of gasoline, diesel and jet fuels. A two-step alkane biosynthetic pathway was identified in cyanobacteria recently. It opens a door to achieve photosynthetic production of alka(e)nes with high efficiency by genetically engineering cyanobacteria.

Results

A series of Synechocystis sp. PCC6803 mutant strains have been constructed and confirmed. Overexpression of both acyl-acyl carrier protein reductase and aldehyde-deformylating oxygenase from several cyanobacteria strains led to a doubled alka(e)ne production. Redirecting the carbon flux to acyl- ACP can provide larger precursor pool for further conversion to alka(e)nes. In combination with the overexpression of alkane biosynthetic genes, alka(e)ne production was significantly improved in these engineered strains. Alka(e)ne content in a Synechocystis mutant harboring alkane biosynthetic genes over-expressed in both slr0168 and slr1556 gene loci (LX56) was 1.3% of cell dry weight, which was enhanced by 8.3 times compared with wildtype strain (0.14% of cell dry weight) cultivated in shake flasks. Both LX56 mutant and the wildtype strain were cultivated in column photo-bioreactors, and the alka(e)ne production in LX56 mutant was 26 mg/L (1.1% of cell dry weight), which was enhanced by 8 times compared with wildtype strain (0.13% of cell dry weight).

Conclusions

The extent of alka(e)ne production could correlate positively with the expression level of alkane biosynthetic genes. Redirecting the carbon flux to acyl-ACP and overexpressing alkane biosynthetic genes simultaneously can enhance alka(e)ne production in cyanobacteria effectively.

【 授权许可】

   
2013 Wang et al.; licensee BioMed Central Ltd.

【 预 览 】
附件列表
Files Size Format View
20140706095232407.pdf 660KB PDF download
Figure 5. 17KB Image download
Figure 4. 83KB Image download
Figure 3. 80KB Image download
Figure 2. 86KB Image download
Figure 1. 70KB Image download
【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】
  • [1]Atsumi S, Higashide W, Liao JC: Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde. Nat Biotechnol 2009, 27:1177-1180.
  • [2]Dexter J, Fu PC: Metabolic engineering of cyanobacteria for ethanol production. Energ Environ Sci 2009, 2:857-864.
  • [3]Lindberg P, Park S, Melis A: Engineering a platform for photosynthetic isoprene production in cyanobacteria, using Synechocystis as the model organism. Metab Eng 2010, 12:70-79.
  • [4]Liu XY, Sheng J, Curtiss R: Fatty acid production in genetically modified cyanobacteria. P Natl Acad Sci USA 2011, 108:6899-6904.
  • [5]Tan XM, Yao L, Gao QQ, Wang WH, Qi FX, Lu XF: Photosynthesis driven conversion of carbon dioxide to fatty alcohols and hydrocarbons in cyanobacteria. Metab Eng 2011, 13:169-176.
  • [6]Ungerer J, Tao L, Davis M, Ghirardi M, Maness PC, Yu JP: Sustained photosynthetic conversion of CO2 to ethylene in recombinant cyanobacterium Synechocystis 6803. Energ Environ Sci 2012, 5:8998-9006.
  • [7]Lu XF: A perspective: Photosynthetic production of fatty acid-based biofuels in genetically engineered cyanobacteria. Biotechnol Adv 2010, 28:742-746.
  • [8]Ducat DC, Way JC, Silver PA: Engineering cyanobacteria to generate high-value products. Trends Biotechnol 2011, 29:95-103.
  • [9]Kaneko T, Sato S, Kotani H, Tanaka A, Asamizu E, Nakamura Y, Miyajima N, Hirosawa M, Sugiura M, Sasamoto S: Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res 1996, 3:109-136.
  • [10]Shih PM, Wu DY, Latifi A, Axen SD, Fewer DP, Talla E, Calteau A, Cai F, de Marsac NT, Rippka R: Improving the coverage of the cyanobacterial phylum using diversity-driven genome sequencing. Proc Natl Acad Sci U S A 2013, 110:1053-1058.
  • [11]Huang HH, Camsund D, Lindblad P, Heidorn T: Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology. Nucleic Acids Res 2010, 38:2577-2593.
  • [12]Peralta-Yahya PP, Zhang FZ, del Cardayre SB, Keasling JD: Microbial engineering for the production of advanced biofuels. Nature 2012, 488:320-328.
  • [13]Schirmer A, Rude MA, Li XZ, Popova E, del Cardayre SB: Microbial Biosynthesis of Alkanes. Science 2010, 329:559-562.
  • [14]Han J, McCarthy ED, Hoeven WV, Calvin M, Bradley WH: Organic geochemical studies, ii. A preliminary report on the distribution of aliphatic hydrocarbons in algae, in bacteria, and in a recent lake sediment. Proc Natl Acad Sci U S A 1968, 59:29-33.
  • [15]Li N, Chang WC, Warui DM, Booker SJ, Krebs C, Bollinger JM: Evidence for Only Oxygenative Cleavage of Aldehydes to Alk(a/e)nes and Formate by Cyanobacterial Aldehyde Decarbonylases. Biochemistry-US 2012, 51:7908-7916.
  • [16]Davis MS, Solbiati J, Cronan JE: Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli. J Biol Chem 2000, 275:28593-28598.
  • [17]Kaczmarzyk D, Fulda M: Fatty acid activation in cyanobacteria mediated by acyl-acyl carrier protein synthetase enables fatty acid recycling. Plant Physiol 2010, 152:1598-1610.
  • [18]Gao QQ, Wang WH, Zhao H, Lu XF: Effects of fatty acid activation on photosynthetic production of fatty acid-based biofuels in Synechocystis sp PCC6803. Biotechnology for Biofuels 2012., 5
  • [19]Ladygina N, Dedyukhina EG, Vainshtein MB: A review on microbial synthesis of hydrocarbons. Process Biochem 2006, 41:1001-1014.
  • [20]Hu P, Borglin S, Kamennaya NA, Chen L, Park H, Mahoney L, Kijac A, Shan G, Chavarría KL, Zhang C: Metabolic phenotyping of the cyanobacterium Synechocystis 6803 engineered for production of alkanes and free fatty acids. Applied Energy 2013, 102:850-859.
  • [21]Reppas NB, Ridley CP, Reppas N, Ridley C, Rodley CP: Producing hydrocarbons comprises culturing engineered cyanobacterium in culture medium and exposing engineered cyanobacterium to light and carbon dioxide. US: JOULE UNLIMITED INC; 2010. 7794969-B1
  • [22]Eser BE, Das D, Han J, Jones PR, Marsh ENG: Oxygen-Independent Alkane Formation by Non-Heme Iron-Dependent Cyanobacterial Aldehyde Decarbonylase: Investigation of Kinetics and Requirement for an External Electron Donor. Biochemistry-US 2011, 50:10743-10750.
  • [23]Hein S, Tran H, Steinbuchel A: Synechocystis sp. PCC6803 possesses a two-component polyhydroxyalkanoic acid synthase similar to that of anoxygenic purple sulfur bacteria. Arch Microbiol 1998, 170:162-170.
  • [24]Liu X, Curtiss R 3rd: Thermorecovery of cyanobacterial fatty acids at elevated temperatures. J Biotechnol 2012, 161:445-449.
  • [25]Gao ZX, Zhao H, Li ZM, Tan XM, Lu XF: Photosynthetic production of ethanol from carbon dioxide in genetically engineered cyanobacteria. Energ Environ Sci 2012, 5:9857-9865.
  • [26]Kamarainen J, Knoop H, Stanford NJ, Guerrero F, Akhtar MK, Aro EM, Steuer R, Jones PR: Physiological tolerance and stoichiometric potential of cyanobacteria for hydrocarbon fuel production. J Biotechnol 2012, 162:67-74.
  • [27]Yin C, Li W, Du Y, Kong R, Xu X: Identification of a gene, ccr-1 (sll1242), required for chill-light tolerance and growth at 15°C in Synechocystis sp. PCC 6803. Microbiology 2007, 153:1261-1267.
  • [28]Williams JGK: Construction of specific mutations in photosystem II photosynthetic reaction center by genetic engineering methods in Synechocystis-6803. Methods Enzymol 1988, 167:766-778.
  • [29]Bligh EG, Dyer WJ: A rapid method of total lipid extraction and purification. Can J Biochem Physiol 1959, 37:911-917.
  文献评价指标  
  下载次数:50次 浏览次数:21次