Journal

Milestones

The iGEM-Team Bielefeld 2013 started working and decided to deal with the project 'Microbial Fuel Cell'.

Initiating labwork on the sub-project endogenous mediator Glycerol dehydrogenase.
Starting lab work with the first successful PCR's. The main work however is still in the planning of our project. Designing experiments and a lot of research.
Finding sponsors goes ahead. Many companies like our project and want to support us.

Starting labwork on the sub-project Porins.
Successful PCR on OprF gene from Pseudomonas fluorescens. OprF with Pre- and Suffix overlaps could be amplified from genome.
Planning of our Human Practice projects started and the first participations are fixed.

Poster presentation at the congress ‘[http://www.biotechnologie2020plus.de Next generation of biotechnological processes 2020+]’ in Berlin.
Sponsoring acquisition is nearly finished.
Problem with cloning delaying the progress in the laboratory.
We have published our [http://ekvv.uni-bielefeld.de/blog/uniaktuell/entry/mit_bakterien_batterie_strom_erzeugen first press release] with a great response. The press release was picked up worldwide.

Our great expert Dr. Falk Harnisch has answered numerous questions about our project and helped us very well.
GldA BioBrick (<bbpart>BBa_K1172201</bbpart>) was examined.
- GldA Biobrick Devices with different promotors and RBS could be added: (<bbpart>BBa_K1172203</bbpart>, <bbpart>BBa_K1172204</bbpart>, <bbpart>BBa_K1172205</bbpart>)
OprF BioBrick (<bbpart>BBa_K1172501</bbpart>) is available.
- OprF Biobrick was functionalized with different promotors and RBS: (<bbpart>BBa_K1172502</bbpart>, <bbpart>BBa_K1172503</bbpart>, <bbpart>BBa_K1172504</bbpart>, <bbpart>BBa_K1172505</bbpart>, <bbpart>BBa_K1172507</bbpart>)
Successful protein expression and overproduction of glycerol dehydrogenase. SDS-PAGE shows GldA at expected size.

Successful characterization of Glycerol dehydrogenase
- NADH-Assays show an increasing intra- and extracellular NADH concentration for Escherichia coli KRX with overexpression of glycerol dehydrogenase.
- Glycerol dehydrogenase was examined by MALDI-TOF MS/MS with a Mascot Score of 266 against Escherichia coli database.

Great characterization of outer membrane porin OprF
- Successful Hexadecan-Assay for characterization of hydrophobicity of the outer membrane. The hydrophobicity increases continuously with increasing promoter strength up to 221% hydrophobizity in contrast to the wild type.
- ONPG and NPN uptake assays were performed. Membrane permeability is continuously increasing for Escherichia coli with heterologeous expression of OprF.
- According to AFM images, E. coli KRX with OprF shows a slightly rougher cell surface morphology in contrast to Escherichia coli KRX wild type, which confirms the results.
- We can see a strong overexpression band on SDS-PAGE at the expected OprF size of about 36 kDa which is equated with a strong expression and overproduction of OprF. Furthermore outer membrane porin was examined by MALDI-TOF MS/MS with a Mascot Score of 222 against bacteria database.

We created the MtrCAB BioBrick <bbpart>K1172401</bbpart>
- - MtrCAB Biobrick Devices with different promotors and RBS could be added: (<bbpart>BBa_K1172403</bbpart>, <bbpart>BBa_K1172404</bbpart>, <bbpart>BBa_K1172405</bbpart>)

We supported the CeBiTec student academy with supervising the ‘Synthetic Biology’ experiment and the day of synthetic biology in the city of Bielefeld.
Our team assigned the Track Food & Energy in this year, with the project title ‘Ecolectricity – currently available’.

Successful cultivations of Escherichia coli KRX with OprF and GldA in our Microbial Fuel Cell.
- NADH overproduction for the GldA strain improves extracellular electron transfer mediated by NADH and resulting in an 20 % increased bioelectricity output.
- Escherichia coli KRX with OprF shows 100 % higher voltage and electric charge than E. coli wild type. Electron shuttle-mediated electron transfer across the membrane is greatly improved by heterologous expression of outer membrane porin OprF.