Journal

Milestones

Overview Prologue

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

Overview May

• 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.

Overview June

• 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.

Overview July

• Poster presentation at the congress ‘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 first press release with a great response. The press release was picked up worldwide.

Overview August

• 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.

Overview September

• 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 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’.

Overview October

• 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.