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Team:NCTU Formosa
From 2012.igem.org
Abstract
Greenhouse Effect and the limitation of the fossil fuels have always been a concern for people on Earth. Our team, NCTU_Formosa, constructed the synthetic pathways in E. coli to achieved high-yield, high-specificity production of isobutanol as biofuels. The Ecofuel E.coline project provides several optimization methods to improve the production rate of butanol as follows: (1) A suitable medium was selected. (2) Five E. coli strains were tested for isobutanol production rate. (3)Temperature-controlled RBS was used to control the expression levels of a series of metabolic proteins at the precise times by culture temperature shifts. (4) The fusion protein of a target enzyme and a zinc finger protein was constructed to increase the specificity of chemical reaction. The experimental data reveals that the E.coli strain DH5α grows in M9TY medium for 24 hours at 42℃ have highest production of isobutanol (~0.75%). E.coline can provide more affordable and higher quantity of biofuel, moreover, lower the burden of our Earth. Futhermore, the biosynthesis design methods can be implemented to control the expression of a series of metabolic proteins required for the maximal synthesis of a drug or energetic compound.
Chinese Version:
溫室效應和化石燃料是世界各國近年以來所熱切關注的議題，因此 NCTU_Formosa團隊希望建立一條能達到高產率、高專一性生產，並且可作為生質能源-異丁醇的生物合成路徑，遂有“ Ecofuel E.coline ”的誕生。
“ Ecofuel E.coline ”提供下列數個方法來增進異丁醇的生產速率：(1)篩選出更佳的培養液配方(2)比較五種E.coli菌株產生異丁醇的速率，並擇其最優者(3)不同的培養溫度變換條件下，以溫度調控的核醣體結合位來精準操控一系列代謝所需之蛋白質(4)建立由生化合成酵素和鋅指蛋白之複合體以增進化學反應的專一性。我們的實驗結果顯示在M9TY 培養液下， E.coli 菌種 DH5α 於42℃培養24 小時後，有最佳的異丁醇產率 (~0.75%)。
概括來說“ Ecofuel E.coline ”能夠提供經濟實惠且高品質的生質燃料，以求降低我們對地球的傷害。此外，溫度調控生物合成的設計還可以應用在量產一系列合成藥物或提供能源之化合物代謝所需的蛋白質。
News
25 Sep
In order to make sure our tolerance data, we used totally three days to do this experiment. Also we collect the concentrate of isobutal in our test experiment every 4 hours in a day.And we are continuing update our wiki and part wiki.
19 Sep
This week, we try to assemble our main parts together and incubate bacteria to test the productivity of isobutanol. Team work tops everything during our cooperation. So, to finish our wiki, each of our teammates are taking charge of different parts of wiki edition, and we took turn to collect tubes for 24 hours in the isobutanol tolerance test. Except for working in the wet lab, we also had fun times taking group photos and new films! By the way, a reporter wrote a special column for us about iGem & Synthesis Biology on Sep. 17. We're now processing the information. Please stay tune and check it out then!
14 Sep
For the experiment, there are new works in for everyone. We have to measure the O.D. on 42 degree and 30 degree, test all the different system we have. The most important is that we should find the G3 part which was made last year, insert into our recent part, and measure the yield. For the wiki, all the deadline of very page sould complete by 9/24. Last, we set up a new group for devising the poster.
