1. 1,3-Propanediol Group 2 生科 陳麗如 生科 林嘉慧 資工 陳奎昊 資工 劉盈詮

2. Outlines Set up genetic-modified experiment to produce 1,3-propanediol. Two ways to maximize yield of PDO. –Linear programming –Copasi

3. PDO 生合成：結構式 C 6 H 12 O 6 -> 1.5 C 3 H 8 O 2 + 1.5 CO 2 Metabolic Engineering of Propanediol Pathways. Biotechnol. 1998, 14, 116-125

4. Design specific primer pairs to amplify target genes glycerol-3-phosphate dehydrogenase (NAD + ) glycerol dehydratase 1,3-propanediol dehydrogenase (dhaT) (dhaB) Construct Ins1 promoter:G3P dehydrogenase –pTC vector Construct trc promoter:dhaB-dhaT-pTC vector Ins1 promoter Transform to E. coli K-12 Evaluate efficiency of PDO production From Escherichia coli K-12 MG1655

5. Pathway Metabolic Engineering of Propanediol Pathways. Biotechnol. 1998, 14, 116-125

6. Case 1 Anaerobic 1ATP Glucose 1 Propanediol + 1 Acetate + 1CO 2 Metabolic Engineering of Propanediol Pathways. Biotechnol. 1998, 14, 116-125

7. Experimental result from linear programming No.Column name ActivityLower bound 1X610 2X110 3X210 4X310 5X410

8. Simulation result from Copasi http://www.copasi.org/tiki-index.php Conc. Time PDO:1

9. Simulation result from Copasi (Cont.) PDO DHAP

10. Case 2 Aerobic Glucose + 0.18 O 2 1.45 Propanediol + 0.2 H 2 O +1.63 CO 2

11. Experimental Result from Linear Programming No.Column nameActivityLower bound 1X61.379310 2X1 10 3X22.222220 4X1400 5X1600 6X1500 7X1700 8X1900 9X1800 10X31.219510 11X41.219510 12X51.56250

12. Simulation Result from Copasi 1.45 Conc. Time

13. Case 3 Aerobic 1ATP Glucose + 0.31 O 2 1.42 Propanediol + 0.32 H 2 O +1.73 CO 2 Metabolic Engineering of Propanediol Pathways. Biotechnol. 1998, 14, 116-125

14. Experimental Result from Linear Programming No.Column nameActivityLower bound 1X61.408450 2X110 3X210 4X310 5X410 6X52.380950 7X1400 8X1500 9X1600 10X1700 11X1800 12X1900

15. Simulation Result from Copasi 1.42 Conc. Time

16. Conclusions Our proposed experiment resembles case1 (Anaerobic). Case1 is not the best procedure to maximize the yeild of PDO. The results of linear programming technique and Copasi tool are similar to biological data.