Team:Tokyo Tech/Experiment/PHB2

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=P(3HB) production by <I>E.coli</I> & Confirmation of P(3HB)=
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=Making purified P(3HB) sheets=
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<div id="tokyotech" style=" font:Arial ;left ; font-size: 15px; color: #000000; padding: 30px;">
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We made P(3HB) sheets. To make the sheets, we cultured <I>E.coli</I> 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.
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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.
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[[http://2012.igem.org/Team:Tokyo_Tech/Experiment/PHB#D_Polymer_extraction_and_purification Protocol]]
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[[File:tokyotech PHB6.png|600px|thumb|center|Fig2-2-4-4-1, P(3HB) sheet]]
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==Protocol==
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[[http://2012.igem.org/Team:Tokyo_Tech/Projects/PHAs/index.htm#Application Go to the P(3HB) production page "Application"]]
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1. Move dried cells into an airtight container.
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2. Add chloroform, at rate of 2mg dried cells for 1 ml chloroform.
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3. Incubate and stir the chloroform solution for more than 72 hrs at 20~25℃.
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4. Filter the chloroform solution.
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5. Concentrate by evaporation.
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6. Dropwise the solution in methanol.
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7. Filter the polymer in methanol, and dry the polymer.
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8. Add a little chloroform to dissolve the polymer.
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9. Poured the chloroform solution into a Petri dish.
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10. Dry at room temperature.

Latest revision as of 19:56, 26 October 2012

bar



Making 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

[Go to the P(3HB) production page "Application"]


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.

10. Dry at room temperature.