Abstract

BackgroundBiodiesel 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.ResultsBased 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.ConclusionsThe 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-015-0132-1) contains supplementary material, which is available to authorized users.

Highlights

  • Biodiesel production using cyanobacteria is a promising alternative to fossil fuels

  • Isolation of a putative fatty acid overproducing mutant by random mutagenesis To obtain a mutant library, we haven chosen to construct a transposon library using plasmid pRL1063a, which contains a Tn5 transposon with a kanamycin resistance marker, promoterless luxAB reporter genes and an origin of replication which functions in E. coli, but not in PCC 7942

  • This activity is most likely linked with the K homology (KH) domain which is well conserved in all GTP-binding protein Era homologs including that of S. elongatus PCC 7942 [18]

Read more

Summary

Introduction

Biodiesel production using cyanobacteria is a promising alternative to fossil fuels. The transposon library was subsequently screened for desirable traits using macro- and microscopic observations as well as staining with the lipophilic dye Nile Red. Biodiesel produced by photosynthetic microorganisms, such as eukaryotic algae and cyanobacteria, provides a promising alternative to reduce our reliance on fossil fuels. The relatively low productivity and the high cost of harvesting the biomass present mayor limitations for commercialization of cyanobacteria-derived biodiesels [1,2]. These limitations have been mainly addressed by using a rational approach [3,4,5]. The rational approach involves designing strains with enhanced biodiesel precursor

Methods
Results
Discussion
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call