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Enhancement of glycerol metabolism in the oleaginous marine diatom Fistulifera solaris JPCC DA0580 to improve triacylglycerol productivity.

Muto M, Tanaka M, Liang Y, Yoshino T, Matsumoto M, Tanaka T - Biotechnol Biofuels (2015)

Bottom Line: Overexpression of the endogenous glycerol kinase (GK) gene in an oleaginous marine diatom, Fistulifera solaris JPCC DA0580, accelerates glycerol metabolism and improves lipid and biomass productivities.We have demonstrated the potential of metabolic engineering in oleaginous microalgae to improve lipid productivity.Metabolic engineering techniques can be used to optimize BDF production.

View Article: PubMed Central - PubMed

Affiliation: Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 Japan ; JST, CREST, Sanbancho 5, Chiyoda-ku, Tokyo 102-0075 Japan.

ABSTRACT

Background: Microalgal oil is a promising alternative feedstock for biodiesel fuel (BDF). Mixotrophic cultivation with glycerol, the primary byproduct of BDF production, may be used to optimize BDF production. This strategy would reduce costs through glycerol recycling and improve lipid productivity and biomass productivity by overcoming the growth retardation caused by decreased light penetration in high-density culture.

Results: Overexpression of the endogenous glycerol kinase (GK) gene in an oleaginous marine diatom, Fistulifera solaris JPCC DA0580, accelerates glycerol metabolism and improves lipid and biomass productivities. Two candidates were selected from a collection of 90 G418-resistant clones, based on growth and confirmation of genome integration. GK gene expression was higher in the selected clones (GK1_7 and GK2_16) than in the wild-type culture. The GK2_16 clone achieved a 12% increase in lipid productivity.

Conclusion: We have demonstrated the potential of metabolic engineering in oleaginous microalgae to improve lipid productivity. Metabolic engineering techniques can be used to optimize BDF production.

No MeSH data available.


Related in: MedlinePlus

Final cell densities of the transformants (optical density at 750 nm). The glycerol kinase (GK) transformants were generated with pSP-GK1/fcpB or pSP-GK2/fcpB. The white bar indicates the cell density of the negative control transformed with pSP-NPT/H4 (n = 5). The black bar indicates 10 clones with cell densities similar to that of the negative control.
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Fig1: Final cell densities of the transformants (optical density at 750 nm). The glycerol kinase (GK) transformants were generated with pSP-GK1/fcpB or pSP-GK2/fcpB. The white bar indicates the cell density of the negative control transformed with pSP-NPT/H4 (n = 5). The black bar indicates 10 clones with cell densities similar to that of the negative control.

Mentions: After transformation with pSP-GK1 and pSP-GK2, 48 GK1 clones (GK1_1 to GK1_48) and 42 GK2 clones (GK2_1 to GK2_42) were obtained from an agar plate containing the antibiotic reagent G418. These clones were cultured in 96-well microtiter plates, and the final cell density was evaluated by determining the optical density to estimate the effect of genetic transformation on cell growth. The final cell densities of the transformants varied from 1% to 102% of the mean density of the control strains (n = 5), which were transformed with the empty pSP-NPT/H4 vector (Figure 1). The variation of transformant cell density may be derived from different loci and copy numbers of the integrated transgenic fragments [5,26,27]. The final cell densities of 10 pSP-GK1 and pSP-GK2 transformants, including GK2_16, GK2_40, GK2_41, GK1_7, GK2_24, GK2_38, GK2_39, GK1_16, GK1_24, and GK1_14 (Figure 1, black columns), were not significantly varied as compared with the mean ± SD (OD = 0.157 ± 0.015) of the control strains (n = 5), and were selected for subsequent experiments.Figure 1


Enhancement of glycerol metabolism in the oleaginous marine diatom Fistulifera solaris JPCC DA0580 to improve triacylglycerol productivity.

Muto M, Tanaka M, Liang Y, Yoshino T, Matsumoto M, Tanaka T - Biotechnol Biofuels (2015)

Final cell densities of the transformants (optical density at 750 nm). The glycerol kinase (GK) transformants were generated with pSP-GK1/fcpB or pSP-GK2/fcpB. The white bar indicates the cell density of the negative control transformed with pSP-NPT/H4 (n = 5). The black bar indicates 10 clones with cell densities similar to that of the negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4308894&req=5

Fig1: Final cell densities of the transformants (optical density at 750 nm). The glycerol kinase (GK) transformants were generated with pSP-GK1/fcpB or pSP-GK2/fcpB. The white bar indicates the cell density of the negative control transformed with pSP-NPT/H4 (n = 5). The black bar indicates 10 clones with cell densities similar to that of the negative control.
Mentions: After transformation with pSP-GK1 and pSP-GK2, 48 GK1 clones (GK1_1 to GK1_48) and 42 GK2 clones (GK2_1 to GK2_42) were obtained from an agar plate containing the antibiotic reagent G418. These clones were cultured in 96-well microtiter plates, and the final cell density was evaluated by determining the optical density to estimate the effect of genetic transformation on cell growth. The final cell densities of the transformants varied from 1% to 102% of the mean density of the control strains (n = 5), which were transformed with the empty pSP-NPT/H4 vector (Figure 1). The variation of transformant cell density may be derived from different loci and copy numbers of the integrated transgenic fragments [5,26,27]. The final cell densities of 10 pSP-GK1 and pSP-GK2 transformants, including GK2_16, GK2_40, GK2_41, GK1_7, GK2_24, GK2_38, GK2_39, GK1_16, GK1_24, and GK1_14 (Figure 1, black columns), were not significantly varied as compared with the mean ± SD (OD = 0.157 ± 0.015) of the control strains (n = 5), and were selected for subsequent experiments.Figure 1

Bottom Line: Overexpression of the endogenous glycerol kinase (GK) gene in an oleaginous marine diatom, Fistulifera solaris JPCC DA0580, accelerates glycerol metabolism and improves lipid and biomass productivities.We have demonstrated the potential of metabolic engineering in oleaginous microalgae to improve lipid productivity.Metabolic engineering techniques can be used to optimize BDF production.

View Article: PubMed Central - PubMed

Affiliation: Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 Japan ; JST, CREST, Sanbancho 5, Chiyoda-ku, Tokyo 102-0075 Japan.

ABSTRACT

Background: Microalgal oil is a promising alternative feedstock for biodiesel fuel (BDF). Mixotrophic cultivation with glycerol, the primary byproduct of BDF production, may be used to optimize BDF production. This strategy would reduce costs through glycerol recycling and improve lipid productivity and biomass productivity by overcoming the growth retardation caused by decreased light penetration in high-density culture.

Results: Overexpression of the endogenous glycerol kinase (GK) gene in an oleaginous marine diatom, Fistulifera solaris JPCC DA0580, accelerates glycerol metabolism and improves lipid and biomass productivities. Two candidates were selected from a collection of 90 G418-resistant clones, based on growth and confirmation of genome integration. GK gene expression was higher in the selected clones (GK1_7 and GK2_16) than in the wild-type culture. The GK2_16 clone achieved a 12% increase in lipid productivity.

Conclusion: We have demonstrated the potential of metabolic engineering in oleaginous microalgae to improve lipid productivity. Metabolic engineering techniques can be used to optimize BDF production.

No MeSH data available.


Related in: MedlinePlus