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Nostoc Nostoc Nostoc punctiforme punctiforme punctiforme punctiforme ATCC ATCC ATCC ATCC 29133 29133 29133 29133

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Functional

Functional Functional Functional G G G Genetic enetic enetic enetic IIIInvestigation nvestigation nvestigation nvestigation of of of of Dps Dps Dps Dps proteins proteins proteins proteins in in in in the the the the C C C Cyanobacterium yanobacterium yanobacterium yanobacterium Nostoc

Nostoc Nostoc Nostoc punctiforme punctiforme punctiforme punctiforme ATCC ATCC ATCC ATCC 29133 29133 29133 29133

Xue Li

In recent years, the energy crisis and climate change have become of globally great concern, along with an increased demand of energy. To reduce the greenhouse gas emissions caused by fossil fuels, alternative renewable fuels are recruited for the progressive replacement of fossil fuels. One of the most promising energy carrier is hydrogen, which is CO

2

neutral.

Cyanobacteria are photosynthetic bacteria which have the ability to produce hydrogen by using light with water as the electron donor.

Nostoc punctiforme ATCC 29133 is a filamentous nitrogen-fixing cyanobacterium, during nitrogen deprivation, the vegetative cells differentiate to heterocysts, where nitrogenase reduces N

2

to NH

3

coupled by the production of hydrogen. The uptake hydrogenase then recycles the hydrogen produced in the nitrogen fixation. Both of the enzymes are oxygen intolerant and the formation of heterocyst is considered as a solution to the incompatibility of photosynthesis and nitrogen fixation (or hydrogen production). However, the cellular environment in heterocyst is still suboptimal due to the enzyme activity inhibited by reactive oxygen species (ROS) (e.g. hydroxyl radical (OH•), hydrogen peroxide (H

2

O

2

), etc.) generated during cellular metabolism such as respiration. The study of O

2

scavenging and protection against oxidative stress processes in heterocyst is of specific value.

A group of ferritin-like proteins named Dps (DNA-binding Protein from Starved cells) were discovered and have the ability of protecting cells against oxidative stress. By large scale quantitative proteomics, five putative Dps-like proteins - Npun_F3730, Npun_F6212, Npun_R3258, Npun_R5701 and Npun_5799 were found in Nostoc punctiforme ATCC 29133.

Some of them are specifically expressed in heterocysts but the specific function of them are still unclear.

In this project, to study their genetic function, we made knock-outs as well as over expressed specific dps gene in E. coli, phenotypical changes of the E. coli cells could be observed and investigated while treated with H

2

O

2

. In a long-term perspective, we are aimed to produce a transgenic cyanobacteria which has optimal hydrogen productivity in the future.

Degree project in biology, Master of Science (2 years), 2012 Examensarbete i biologi 30 hp till masterexamen, 2012

Department of Photochemistry and Molecular Science, Uppsala University

Supervisor: Karin Stensjö

References

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