1,3-Propanediol1,3-Propanediol

Group 2生科 陳麗如生科 林嘉慧資工 陳奎昊資工 劉盈詮

Outlines

• Set up genetic-modified experiment to produce 1,3-propanediol.

• Two ways to maximize yield of PDO.– Linear programming– Copasi

PDO 生合成：結構式

C6H12O6 -> 1.5 C3H8O2 + 1.5 CO2

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

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

Pathway

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

Case 1

Anaerobic

1ATP

Glucose 1 Propanediol + 1 Acetate + 1CO2

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

Experimental result from linear programming

No. Column name

Activity Lower bound

1 X6 1 0

2 X1 1 0

3 X2 1 0

4 X3 1 0

5 X4 1 0

Simulation result from Copasi

http://www.copasi.org/tiki-index.php

Conc.

Time

PDO:1

Simulation result from Copasi (Cont.)

PDO

DHAP

Case 2

Aerobic

Glucose + 0.18 O2

1.45 Propanediol + 0.2 H2O

+1.63 CO2

Experimental Result fromLinear Programming

No. Column name Activity Lower bound

1 X6 1.37931 0

2 X1 1 0

3 X2 2.22222 0

4 X14 0 0

5 X16 0 0

6 X15 0 0

7 X17 0 0

8 X19 0 0

9 X18 0 0

10 X3 1.21951 0

11 X4 1.21951 0

12 X5 1.5625 0

Simulation Result from Copasi

1.45

Conc.

Time

Case 3

Aerobic

1ATP

Glucose + 0.31 O2

1.42 Propanediol + 0.32 H2O

+1.73 CO2

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

Experimental Result from Linear Programming

No. Column name Activity Lower bound

1 X6 1.40845 0

2 X1 1 0

3 X2 1 0

4 X3 1 0

5 X4 1 0

6 X5 2.38095 0

7 X14 0 0

8 X15 0 0

9 X16 0 0

10 X17 0 0

11 X18 0 0

12 X19 0 0

Simulation Result from Copasi

1.42Conc.

Time

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.