Evaluation of cell-disruption effects of pulsed-electric-field treatment of Synechocystis PCC 6803.

In order to use Synechocystis PCC 6803 as feedstock of nonpetroleum-based diesel fuel, pulsed electric field (PEF) technology was used for cell disruption prior to extraction of intracellular lipids. Severe cell disruption was evident after PEF treatment, especially with treatment intensity (TI) > 35 kWh/m(3). Temperature increase during the treatment brought about most of the destruction of autofluorescence compounds, as well as a fraction of inactivation and the destruction of the plasma and thylakoid membranes. However, the forces associated with the pulsing electric field caused significant damage to the plasma membrane, cell wall, and thylakoid membrane, and it even led to complete disruption of some cells into fragments, which resulted in biomass loss. Treatment by PEF enhanced the potential for the low-toxicity solvent isopropanol to access lipid molecules during subsequent solvent extraction, leading to lower usage of isopropanol for the same extraction efficiency. Thus, PEF shows promise for lowering the costs and environmental effects of the lipid-extraction step.

[1]  B. Rittmann.  Opportunities for renewable bioenergy using microorganisms. , 2008, Biotechnology and bioengineering.

[2]  Ahmed E. Yousef,et al.  Spectrofluorimetric assessment of bacterial cell membrane damage by pulsed electric field , 2002 .

[3]  G. McPherson,et al.  The Impending Peak and Decline of Petroleum Production: an Underestimated Challenge for Conservation of Ecological Integrity , 2010, Conservation biology : the journal of the Society for Conservation Biology.

[4]  J. Heuser,et al.  Ultrastructure of the membrane systems in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 , 2006, Protoplasma.

[5]  P. Parameswaran,et al.  Feasibility of Focused‐Pulsed Treated Waste Activated Sludge as a Supplemental Electron Donor for Denitrification , 2010, Water environment research : a research publication of the Water Environment Federation.

[6]  G. Evrendilek,et al.  Pulsed Electric Fields: Processing System, Microbial and Enzyme Inhibition, and Shelf Life Extension of Foods , 2007, IEEE Transactions on Plasma Science.

[7]  Y. Chisti.  Biodiesel from microalgae. , 2007, Biotechnology advances.

[8]  Michael J. Cooney,et al.  Extraction of Bio‐oils from Microalgae , 2009 .

[9]  P. Parameswaran,et al.  Using a Pulsed Electric Field as a Pretreatment for Improved Biosolids Digestion and Methanogenesis , 2009, Water environment research : a research publication of the Water Environment Federation.

[10]  E. Torres,et al.  Biodiesel: an overview , 2005 .

[11]  M. Poot,et al.  Bacterial viability and antibiotic susceptibility testing with SYTOX green nucleic acid stain , 1997, Applied and environmental microbiology.

[12]  Q. Hu,et al.  Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. , 2008, The Plant journal : for cell and molecular biology.

[13]  S. Condón,et al.  Relationship between Sublethal Injury and Inactivation of Yeast Cells by the Combination of Sorbic Acid and Pulsed Electric Fields , 2007, Applied and Environmental Microbiology.

[14]  D. Knorr,et al.  Impact of pulsed electric field treatment on the recovery and quality of plant oils , 2005 .

[15]  N. Murata,et al.  Synechocystis PCC6803 Mutants Defective in Desaturation of Fatty Acids , 1989 .

[16]  E. Hoiczyk,et al.  Cyanobacterial Cell Walls: News from an Unusual Prokaryotic Envelope , 2000, Journal of bacteriology.

[17]  W. Vermaas,et al.  The three-dimensional structure of the cyanobacterium Synechocystis sp. PCC 6803 , 2005, Archives of Microbiology.

[18]  Patrick C. Wouters,et al.  Effects of Pulsed Electric Fields on Inactivation Kinetics of Listeria innocua , 1999, Applied and Environmental Microbiology.

[19]  Ravindra P. Joshi,et al.  Bacterial decontamination of liquids with pulsed electric fields , 2000 .

[20]  Charlotte K. Williams,et al.  The Path Forward for Biofuels and Biomaterials , 2006, Science.

[21]  W. Vermaas.  Molecular genetics of the cyanobacteriumSynechocystis sp. PCC 6803: Principles and possible biotechnology applications , 2004, Journal of Applied Phycology.

[22]  M. Frentzen,et al.  Characterisation of acyltransferases from Synechocystis sp. PCC6803. , 2005, Biochemical and biophysical research communications.

[23]  W. Hamilton,et al.  Effects of high electric fields on microorganisms: II. Mechanism of action of the lethal effect , 1967 .

[24]  Bruce E Rittmann,et al.  Evaluation of methods to extract and quantify lipids from Synechocystis PCC 6803. , 2011, Bioresource technology.

[25]  M Barjenbruch,et al.  Sludge pre-treatment with pulsed electric fields. , 2004, Water science and technology : a journal of the International Association on Water Pollution Research.

[26]  Feng Chen,et al.  Fatty acid profile of the edible filamentous cyanobacterium Nostoc flagelliforme at different temperatures and developmental stages in liquid suspension culture , 2005 .

[27]  Prathap Parameswaran,et al.  Focused-Pulsed sludge pre-treatment increases the bacterial diversity and relative abundance of acetoclastic methanogens in a full-scale anaerobic digester. , 2009, Water research.

[28]  B. Rittmann,et al.  Photoautotrophic nutrient utilization and limitation during semi‐continuous growth of Synechocystis sp. PCC6803 , 2010, Biotechnology and bioengineering.

[29]  J. Waterbury,et al.  Generic assignments, strain histories, and properties of pure cultures of cyanobacteria , 1979 .

[30]  O. Koksharova.  Application of molecular genetic and microbiological techniques in ecology and biotechnology of cyanobacteria , 2010, Microbiology.

[31]  J. Weaver,et al.  Theory of electroporation: A review , 1996 .

[32]  Takakazu Kaneko,et al.  CyanoBase, a www database containing the complete nucleotide sequence of the genome of Synechocystis sp. strain PCC6803 , 1998, Nucleic Acids Res..

[33]  J. Folch,et al.  A simple method for the isolation and purification of total lipides from animal tissues. , 1957, The Journal of biological chemistry.

[34]  P. C. Wouters,et al.  Membrane Permeabilization in Relation to Inactivation Kinetics of Lactobacillus Species due to Pulsed Electric Fields , 2001, Applied and Environmental Microbiology.

[35]  B. Rittmann,et al.  Full-scale application of focused-pulsed pre-treatment for improving biosolids digestion and conversion to methane. , 2008, Water science and technology : a journal of the International Association on Water Pollution Research.

[36]  Filiz Karaosmanoglu,et al.  Vegetable oil fuels: A review , 1999 .

[37]  M. M. Allen,et al.  The acid stress response of the cyanobacterium Synechocystis sp. strain PCC 6308 , 2002, Archives of Microbiology.

[38]  V. Zverlov,et al.  Biofuels from microbes , 2007, Applied Microbiology and Biotechnology.