D Make purified P(3HB) sheets

We made P(3HB) sheets. To make the sheets, we cultured E.coli JM109 in erlenmeyer flasks at 37°C for 72h. Contact angle is an indicator to represent the strength of the water-repellent. The angle shows the physical properties, especially surface tension. When contact angle of sheets is larger than 90°, from Young equation, the sheets would have more strong water-repellent by increasing real surface area. Contact angle of P(3HB) sheets is about 100° from literature data. [Protocol]

Fig2-2-4-4-1, P(3HB) sheet

Protocol

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1. Move dried cells into an airtight container.

2. Add chloroform, at rate of 2mg dried cells for 1 ml chloroform.

3. Incubate and stir the chloroform solution for more than 72 hrs at 20~25℃.

4. Filter the chloroform solution.

5. Concentrate by evaporation.

6. Dropwise the solution in methanol.

7. Filter the polymer in methanol, and dry the polymer.

8. Add a little chloroform to dissolve the polymer.

9. Poured the chloroform solution into a Petri dish.