Engineering a minimal cloning vector from a pUC18 plasmid backbone with an extended multiple cloning site

Minimal plasmids play an essential role in many intermediate steps in molecular biology. They can for example be used to assemble building blocks in synthetic biology or be used as intermediate cloning plasmids that are ideal for PCR-based mutagenesis methods. A small backbone also opens up for additional unique restriction enzyme cloning sites. Here we describe the generation of pICOz, a 1185 bp fully functional high-copy cloning plasmid with an extended multiple cloning site (MCS). To our knowledge, this is the smallest high-copy cloning vector ever described

Geobacter, Anaeromyxobacter and Anaerolineae populations are enriched on anodes of root exudate-driven microbial fuel cells in rice field soil

Plant-based sediment microbial fuel cells (PMFCs) couple the oxidation of root exudates in living rice plants to current production. We analysed the composition of the microbial community on anodes from PMFC with natural rice field soil as substratum for rice by analysing 16S rRNA as an indicator of microbial activity and diversity. Terminal restriction fragment length polymorphism (TRFLP) analysis indicated that the active bacterial community on anodes from PMFCs differed strongly compared with controls. Moreover, clones related to Deltaproteobacteria and Chloroflexi were highly abundant (49% and 21%, respectively) on PMFCs anodes. Geobacter (19%), Anaeromyxobacter (15%) and Anaerolineae (17%) populations were predominant on anodes with natural rice field soil and differed strongly from those previously detected with potting soil. In open circuit (OC) control PMFCs, not allowing electron transfer, Deltaproteobacteria (33%), Betaproteobacteria (20%), Chloroflexi (12%), Alphaproteobacteria (10%) and Firmicutes (10%) were detected. The presence of an electron accepting anode also had a strong influence on methanogenic archaea. Hydrogenotrophic methanogens were more active on PMFC (21%) than on OC controls (10%), whereas acetoclastic Methanosaetaceae were more active on OC controls (31%) compared with PMFCs (9%). In conclusion, electron accepting anodes and rice root exudates selected for distinct potential anode-reducing microbial populations in rice soil inoculated PMFC

Integrating microalgae production with anaerobic digestion: a biorefinery approach

This is the peer reviewed version of the following article: [Uggetti, E. , Sialve, B. , Trably, E. and Steyer, J. (2014), Integrating microalgae production with anaerobic digestion: a biorefinery approach. Biofuels, Bioprod. Bioref, 8: 516-529. doi:10.1002/bbb.1469], which has been published in final form at https://doi.org/10.1002/bbb.1469. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-ArchivingIn the energy and chemical sectors, alternative production chains should be considered in order to simultaneously reduce the dependence on oil and mitigate climate change. Biomass is probably the only viable alternative to fossil resources for production of liquid transportation fuels and chemicals since, besides fossils, it is one of the only available sources of carbon-rich material on Earth. Over recent years, interest in microalgae biomass has grown in both fundamental and applied research fields. The biorefinery concept includes different technologies able to convert biomass into added-value chemicals, products (food and feed) and biofuels (biodiesel, bioethanol, biohydrogen). As in oil refinery, a biorefinery aims at producing multiple products, maximizing the value derived from differences in biomass components, including microalgae. This paper provides an overview of the various microalgae-derived products, focusing on anaerobic digestion for conversion of microalgal biomass into methane. Special attention is paid to the range of possible inputs for anaerobic digestion (microalgal biomass and microalgal residue after lipid extraction) and the outputs resulting from the process (e.g. biogas and digestate). The strong interest in microalgae anaerobic digestion lies in its ability to mineralize microalgae containing organic nitrogen and phosphorus, resulting in a flux of ammonium and phosphate that can then be used as substrate for growing microalgae or that can be further processed to produce fertilizers. At present, anaerobic digestion outputs can provide nutrients, CO2 and water to cultivate microalgae, which in turn, are used as substrate for methane and fertilizer generation.Peer ReviewedPostprint (author's final draft

Engineering a minimal cloning vector from a pUC18 plasmid backbone with an extended multiple cloning site

Minimal plasmids play an essential role in many intermediate steps in molecular biology. For example, they can be used to assemble building blocks in synthetic biology or be used as intermediate cloning plasmids that are ideal for PCR-based mutagenesis methods. A small backbone also opens up for additional unique restriction enzyme cloning sites. Here we describe the generation of pICOz, a 1185-bp fully functional high-copy cloning plasmid with an extended multiple cloning site. We believe that this is the smallest high-copy cloning vector ever described. METHOD SUMMARY: We eliminated all superfluous sequences in a commonly used cloning vector in order to generate as small a cloning plasmid as possible by simple iterative PCR mutagenesis

A Bioelectrochemical Approach to Characterize Extracellular Electron Transfer by Synechocystis sp. PCC6803

Biophotovoltaic devices employ photosynthetic organisms at the anode of a microbial fuel cell to generate electrical power. Although a range of cyanobacteria and algae have been shown to generate photocurrent in devices of a multitude of architectures, mechanistic understanding of extracellular electron transfer by phototrophs remains minimal. Here we describe a mediatorless bioelectrochemical device to measure the electrogenic output of a planktonically grown cyanobacterium, Synechocystis sp. PCC6803. Light dependent production of current is measured, and its magnitude is shown to scale with microbial cell concentration and light intensity. Bioelectrochemical characterization of a Synechocystis mutant lacking Photosystem II demonstrates conclusively that production of the majority of photocurrent requires a functional water splitting aparatus and electrons are likely ultimately derived from water. This shows the potential of the device to rapidly and quantitatively characterize photocurrent production by genetically modified strains, an approach that can be used in future studies to delineate the mechanisms of cyanobacterial extracellular electron transport

Phosphorus–iron interaction in sediments : can an electrode minimize phosphorus release from sediments?

All restoration strategies to mitigate eutrophication depend on the success of phosphorus (P) removal from the water body. Therefore, the inputs from the watershed and from the enriched sediments, that were the sink of most P that has been discharged in the water body, should be controlled. In sediments, iron (hydr)oxides minerals are potent repositories of P and the release of P into the water column may occur upon dissolution of the iron (hydr)oxides mediated by iron reducing bacteria. Several species of these bacteria are also known as electroactive microorganisms and have been recently identified in lake sediments. This capacity of bacteria to transfer electrons to electrodes, producing electricity from the oxidation of organic matter, might play a role on P release in sediments. In the present work it is discussed the relationship between phosphorus and iron cycling as well as the application of an electrode to work as external electron acceptor in sediments, in order to prevent metal bound P dissolution under anoxic conditions.The authors are grateful to two anonymous reviewers of a previous version of the manuscript for the constructive comments and suggestions. The authors also acknowledge the Grant SFRH/BPD/80528/2011 from the Foundation for Science and Technology, Portugal, awarded to Gilberto Martins

Potential of Electric Power Production from Microbial Fuel Cell (MFC) in Evapotranspiration Reactor for Leachate Treatment Using Alocasia macrorrhiza Plant and Eleusine indica Grass

Microbial fuel cell is one of attractive electric power generator from nature bacterial activity. While, Evapotranspiration is one of the waste water treatment system which developed to eliminate biological weakness that utilize the natural evaporation process and bacterial activity on plant roots and plant media. This study aims to determine the potential of electrical energy from leachate treatment using evapotranspiration reactor. The study was conducted using local plant, namely Alocasia macrorrhiza and local grass, namely Eleusine Indica. The system was using horizontal MFC by placing the cathodes and anodes at different chamber (i.e. in the leachate reactor and reactor with plant media). Carbon plates was used for chatode-anodes material with size of 40 cm x 10 cm x1 cm. Electrical power production was measure by a digital multimeter for 30 days reactor operation. The result shows electric power production was fluctuated during reactor operation from all reactors. The electric power generated from each reactor was fluctuated, but from the reactor using Alocasia macrorrhiza plant reach to 70 μwatt average. From the reactor using Eleusine Indica grass was reached 60 μwatt average. Electric power production fluctuation is related to the bacterial growth pattern in the soil media and on the plant roots which undergo the adaptation process until the middle of the operational period and then in stable growth condition until the end of the reactor operation. The results indicate that the evapotranspiration reactor using Alocasia macrorrhiza plant was 60-95% higher electric power potential than using Eleusine Indica grass in short-term (30-day) operation. Although, MFC system in evapotranspiration reactor system was one of potential system for renewable electric power generation