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Increased Lipid Accumulation in the Chlamydomonas reinhardtiista7-10 Starchless Isoamylase Mutant and Increased Carbohydrate Synthesis in Complemented Strains

Victoria H. Work, Randor Radakovits, Robert E. Jinkerson, Jonathan E. Meuser, Lee G. Elliott, David J. Vinyard, Lieve M. L. Laurens, G. Charles Dismukes, Matthew C. Posewitz
Victoria H. Work
1Division of Environmental Science and Engineering, Colorado School of Mines, 1500 Illinois St., Golden, Colorado 80401
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Randor Radakovits
2Department of Chemistry and Geochemistry, Colorado School of Mines, 1500 Illinois St., Golden, Colorado 80401
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Robert E. Jinkerson
2Department of Chemistry and Geochemistry, Colorado School of Mines, 1500 Illinois St., Golden, Colorado 80401
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Jonathan E. Meuser
1Division of Environmental Science and Engineering, Colorado School of Mines, 1500 Illinois St., Golden, Colorado 80401
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Lee G. Elliott
1Division of Environmental Science and Engineering, Colorado School of Mines, 1500 Illinois St., Golden, Colorado 80401
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David J. Vinyard
3Department of Chemistry & Chemical Biological and Waksman Institute, Rutgers University, Piscataway, New Jersey 08854
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Lieve M. L. Laurens
4National Renewable Energy Laboratory, Golden, Colorado 80401
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G. Charles Dismukes
3Department of Chemistry & Chemical Biological and Waksman Institute, Rutgers University, Piscataway, New Jersey 08854
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Matthew C. Posewitz
2Department of Chemistry and Geochemistry, Colorado School of Mines, 1500 Illinois St., Golden, Colorado 80401
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  • For correspondence: mposewit@mines.edu
DOI: 10.1128/EC.00075-10
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ABSTRACT

The accumulation of bioenergy carriers was assessed in two starchless mutants of Chlamydomonas reinhardtii (the sta6 [ADP-glucose pyrophosphorylase] and sta7-10 [isoamylase] mutants), a control strain (CC124), and two complemented strains of the sta7-10 mutant. The results indicate that the genetic blockage of starch synthesis in the sta6 and sta7-10 mutants increases the accumulation of lipids on a cellular basis during nitrogen deprivation relative to that in the CC124 control as determined by conversion to fatty acid methyl esters. However, this increased level of lipid accumulation is energetically insufficient to completely offset the loss of cellular starch that is synthesized by CC124 during nitrogen deprivation. We therefore investigated acetate utilization and O2 evolution to obtain further insights into the physiological adjustments utilized by the two starchless mutants in the absence of starch synthesis. The results demonstrate that both starchless mutants metabolize less acetate and have more severely attenuated levels of photosynthetic O2 evolution than CC124, indicating that a decrease in overall anabolic processes is a significant physiological response in the starchless mutants during nitrogen deprivation. Interestingly, two independent sta7-10:STA7 complemented strains exhibited significantly greater quantities of cellular starch and lipid than CC124 during acclimation to nitrogen deprivation. Moreover, the complemented strains synthesized significant quantities of starch even when cultured in nutrient-replete medium.

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Increased Lipid Accumulation in the Chlamydomonas reinhardtiista7-10 Starchless Isoamylase Mutant and Increased Carbohydrate Synthesis in Complemented Strains
Victoria H. Work, Randor Radakovits, Robert E. Jinkerson, Jonathan E. Meuser, Lee G. Elliott, David J. Vinyard, Lieve M. L. Laurens, G. Charles Dismukes, Matthew C. Posewitz
Eukaryotic Cell Aug 2010, 9 (8) 1251-1261; DOI: 10.1128/EC.00075-10

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Increased Lipid Accumulation in the Chlamydomonas reinhardtiista7-10 Starchless Isoamylase Mutant and Increased Carbohydrate Synthesis in Complemented Strains
Victoria H. Work, Randor Radakovits, Robert E. Jinkerson, Jonathan E. Meuser, Lee G. Elliott, David J. Vinyard, Lieve M. L. Laurens, G. Charles Dismukes, Matthew C. Posewitz
Eukaryotic Cell Aug 2010, 9 (8) 1251-1261; DOI: 10.1128/EC.00075-10
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KEYWORDS

Carbohydrate Metabolism
Chlamydomonas reinhardtii
Genetic Complementation Test
Isoamylase
Lipid Metabolism
Mutation
Starch

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