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GTP-binding protein Era: a novel gene target for biofuel production.

Voshol GP, Meyer V, van den Hondel CA - BMC Biotechnol. (2015)

Bottom Line: Moreover, the cellular size was significantly smaller than that of the inactivated transposon mutant.We also found that this protein is important for hydrocarbon and fatty acid metabolism as well as determination of the cell size in PCC 7942.Our results suggest that the GTP-binding protein Era can be used as a novel target for further improvement of biofuel precursors production.

View Article: PubMed Central - PubMed

Affiliation: Molecular Microbiology and Biotechnology, Institute of Biology Leiden, Leiden University, Leiden, The Netherlands. g.p.voshol@biology.leidenuniv.nl.

ABSTRACT

Background: Biodiesel production using cyanobacteria is a promising alternative to fossil fuels. In this study we created a transposon library of Synechococcus elongatus PCC 7942 in order to identify novel gene targets for enhanced fatty acid and hydrocarbon production. The transposon library was subsequently screened for desirable traits using macro- and microscopic observations as well as staining with the lipophilic dye Nile Red.

Results: Based on the screening results, we selected a single mutant, which has an insertion in the gene encoding for the GTP-binding protein Era. We subsequently verified the phenotype-genotype relation by overexpression, reintroducing and complementing the mutation. Overexpression of era caused a reduction in the cell size in the late exponential phase of growth and an increase in the total amount of intracellular fatty acids. Reintroduction of the inactivated transposon caused a significant increase in the cellular length as well as changes in the amounts of individual hydrocarbons and fatty acids. Ectopic complementation of this mutation fully restored the hydrocarbon production profile to that of wild-type and partially restored the fatty acid production. Moreover, the cellular size was significantly smaller than that of the inactivated transposon mutant.

Conclusions: The GTP-binding protein Era has never been studied in cyanobacteria and proved to be an essential gene for S. elongatus PCC 7942. We also found that this protein is important for hydrocarbon and fatty acid metabolism as well as determination of the cell size in PCC 7942. Our results suggest that the GTP-binding protein Era can be used as a novel target for further improvement of biofuel precursors production.

No MeSH data available.


Related in: MedlinePlus

Quantitative measurement of intracellular fatty acids in PCC 7942, strain Se:era, Se:∆era and Se:∆era + era. Metabolites indicated with an asterisk * are significantly different (p < 0.05) from those of strain PCC 7942. Error bars represent the standard deviation of four replicates.
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Fig7: Quantitative measurement of intracellular fatty acids in PCC 7942, strain Se:era, Se:∆era and Se:∆era + era. Metabolites indicated with an asterisk * are significantly different (p < 0.05) from those of strain PCC 7942. Error bars represent the standard deviation of four replicates.

Mentions: PCC 7942 contains a mixture of saturated and unsaturated fatty acids ranging from C14 to C18. Palmitic (C16:0) and palmitoleic acid (C16:1) represent the main intracellular fatty acids of PCC 7942 (Figure 7 and Additional file 2: Table S1). Together these fatty acids represent approximately 89% (35.7 ± 1.2 μg C16:0/mg DW and 30.1 ± 1.2 μg C16:1/mg DW) of the total fatty acids (73.6 ± 2.7 μg/mg DW). The remainder mainly consists of saturated and unsaturated C14 and C18 fatty acids followed by trace amounts of odd chain C15:0, C17:0 and C17:1.Figure 7


GTP-binding protein Era: a novel gene target for biofuel production.

Voshol GP, Meyer V, van den Hondel CA - BMC Biotechnol. (2015)

Quantitative measurement of intracellular fatty acids in PCC 7942, strain Se:era, Se:∆era and Se:∆era + era. Metabolites indicated with an asterisk * are significantly different (p < 0.05) from those of strain PCC 7942. Error bars represent the standard deviation of four replicates.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4380250&req=5

Fig7: Quantitative measurement of intracellular fatty acids in PCC 7942, strain Se:era, Se:∆era and Se:∆era + era. Metabolites indicated with an asterisk * are significantly different (p < 0.05) from those of strain PCC 7942. Error bars represent the standard deviation of four replicates.
Mentions: PCC 7942 contains a mixture of saturated and unsaturated fatty acids ranging from C14 to C18. Palmitic (C16:0) and palmitoleic acid (C16:1) represent the main intracellular fatty acids of PCC 7942 (Figure 7 and Additional file 2: Table S1). Together these fatty acids represent approximately 89% (35.7 ± 1.2 μg C16:0/mg DW and 30.1 ± 1.2 μg C16:1/mg DW) of the total fatty acids (73.6 ± 2.7 μg/mg DW). The remainder mainly consists of saturated and unsaturated C14 and C18 fatty acids followed by trace amounts of odd chain C15:0, C17:0 and C17:1.Figure 7

Bottom Line: Moreover, the cellular size was significantly smaller than that of the inactivated transposon mutant.We also found that this protein is important for hydrocarbon and fatty acid metabolism as well as determination of the cell size in PCC 7942.Our results suggest that the GTP-binding protein Era can be used as a novel target for further improvement of biofuel precursors production.

View Article: PubMed Central - PubMed

Affiliation: Molecular Microbiology and Biotechnology, Institute of Biology Leiden, Leiden University, Leiden, The Netherlands. g.p.voshol@biology.leidenuniv.nl.

ABSTRACT

Background: Biodiesel production using cyanobacteria is a promising alternative to fossil fuels. In this study we created a transposon library of Synechococcus elongatus PCC 7942 in order to identify novel gene targets for enhanced fatty acid and hydrocarbon production. The transposon library was subsequently screened for desirable traits using macro- and microscopic observations as well as staining with the lipophilic dye Nile Red.

Results: Based on the screening results, we selected a single mutant, which has an insertion in the gene encoding for the GTP-binding protein Era. We subsequently verified the phenotype-genotype relation by overexpression, reintroducing and complementing the mutation. Overexpression of era caused a reduction in the cell size in the late exponential phase of growth and an increase in the total amount of intracellular fatty acids. Reintroduction of the inactivated transposon caused a significant increase in the cellular length as well as changes in the amounts of individual hydrocarbons and fatty acids. Ectopic complementation of this mutation fully restored the hydrocarbon production profile to that of wild-type and partially restored the fatty acid production. Moreover, the cellular size was significantly smaller than that of the inactivated transposon mutant.

Conclusions: The GTP-binding protein Era has never been studied in cyanobacteria and proved to be an essential gene for S. elongatus PCC 7942. We also found that this protein is important for hydrocarbon and fatty acid metabolism as well as determination of the cell size in PCC 7942. Our results suggest that the GTP-binding protein Era can be used as a novel target for further improvement of biofuel precursors production.

No MeSH data available.


Related in: MedlinePlus