349765 High-Throughput Chemical Lysis of Synechocystis Sp. PCC 6803
Due to anthropogenic global warming, Antarctic ice melted five times more quickly from 2001-2011 than in the previous decade. To avert a global climate crisis, lawmakers must quickly substitute renewable energy resources for the CO2-emitting fossil fuels that account for 81.1% of our global primary energy supply. Microalgae and cyanobacteria engineered to produce carbon-neutral biofuels provide a sustainable solution to fossil fuels that will not interfere with food markets. While oleaginous algae are superior oil producers, cyanobacteria are much easier to genetically alter to produce a variety of fuels. However, unusually complex multi-layered cell walls render cyanobacteria difficult to lyse chemically and lower transformation efficiency dramatically compared to that of E. coli. In order to make these organisms practical to work with in high-throughput industrial settings, chemical lysis and transformation protocols must be improved.
Synechocystis cyanobacteria were lysed to 90% of mechanical lysis levels using several chemical additives. It is hypothesized these additives work in concert to break down different layers of the complex cyanobacterial cell wall, each of which must be breached to yield total lysis. A high-throughput variation of the above method achieved greater lysis of E. coli than mechanical methods. Cellulases and a reducing agent were helpful against cyanobacteria but not E. coli. This suggests that these two additives may act against the cyanobacterial exopolysaccharide or S-layer.
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