The use of fossil fuels is now widely accepted as unsustainable due to depleting resources and the accumulation of greenhouse gases in the environment that have already exceeded the “dangerously high” threshold of 450 ppm CO2-e. To achieve environmental and economic sustainability, fuel production processes are required that are not only renewable, but also capable of sequestering atmospheric CO2. Currently, nearly all renewable energy sources (e.g. hydroelectric, solar, wind, tidal, geothermal) target the electricity market, while fuels make up a much larger share of the global energy demand (∼66%). Biofuels are therefore rapidly being developed. Second generation microalgal systems have the advantage that they can produce a wide range of feedstocks for the production of biodiesel, bioethanol, biomethane and biohydrogen. Biodiesel is currently produced from oil synthesized by conventional fuel crops that harvest the sun’s energy and store it as chemical energy. This presents a route for renewable and carbon-neutral fuel production. However, current supplies from oil crops and animal fats account for only approximately 0.3% of the current demand for transport fuels. Increasing biofuel production on arable land could have severe consequences for global food supply. In contrast, producing biodiesel from algae is widely regarded as one of the most efficient ways of generating biofuels and also appears to represent the only current renewable source of oil that could meet the global demand for transport fuels. The main advantages of second generation microalgal systems are that they: (1) Have a higher photon conversion efficiency (as evidenced by increased biomass yields per hectare): (2) Can be harvested batch-wise nearly all-year-round, providing a reliable and continuous supply of oil: (3) Can utilize salt and waste water streams, thereby greatly reducing freshwater use: (4) Can couple CO2-neutral fuel production with CO2 sequestration: (5) Produce non-toxic and highly biodegradable biofuels. Current limitations exist mainly in the harvesting process and in the supply of CO2 for high efficiency production. This review provides a brief overview of second generation biodiesel production systems using microalgae.

https://www.springer.com/cda/content/document/cda_downloaddocument/BioEnergyResearch_Second+Generation+Biofuels+High-Efficiency+Microalgae+for+Biodiesel+Production.pdf?SGWID=0-0-45-1347846-0

Second generation biofuels: high-efficiency microalgae for biodiesel production

Biohydrogen production: prospects and limitations to practical application

Following scrutiny of present biofuels, algae are seriously considered as feedstocks for next-generation biofuels production. Their high productivity and the associated high lipid yields make them attractive options. In this review, we analyse a number aspects of large-scale lipid and overall algal biomass production from a biochemical and energetic standpoint. We illustrate that the maximum conversion efficiency of total solar energy into primary photosynthetic organic products falls in the region of 10%. Biomass biochemical composition further conditions this yield: 30 and 50% of the primary product mass is lost on producing cell protein and lipid. Obtained yields are one third to one tenth of the theoretical ones. Wasted energy from captured photons is a major loss term and a major challenge in maximising mass algal production. Using irradiance data and kinetic parameters derived from reported field studies, we produce a simple model of algal biomass production and its variation with latitude and lipid content. An economic analysis of algal biomass production considers a number of scenarios and the effect of changing individual parameters. Our main conclusions are that: (i) the biochemical composition of the biomass influences the economics, in particular, increased lipid content reduces other valuable compounds in the biomass; (ii) the “biofuel only” option is unlikely to be economically viable; and (iii) among the hardest problems in assessing the economics are the cost of the CO2 supply and uncertain nature of downstream processing. We conclude by considering the pressing research and development needs.

Microalgae as biodiesel & biomass feedstocks: Review & analysis of the biochemistry, energetics & economics

There are currently intensive global research efforts aimed at increasing and modifying the accumulation of lipids, alcohols, hydrocarbons, polysaccharides, and other energy storage compounds in photosynthetic organisms, yeast, and bacteria through genetic engineering. Many improvements have been realized, including increased lipid and carbohydrate production, improved H2 yields, and the diversion of central metabolic intermediates into fungible biofuels. Photosynthetic microorganisms are attracting considerable interest within these efforts due to their relatively high photosynthetic conversion efficiencies, diverse metabolic capabilities, superior growth rates, and ability to store or secrete energy-rich hydrocarbons. Relative to cyanobacteria, eukaryotic microalgae possess several unique metabolic attributes of relevance to biofuel production, including the accumulation of significant quantities of triacylglycerol; the synthesis of storage starch (amylopectin and amylose), which is similar to that found in higher plants; and the ability to efficiently couple photosynthetic electron transport to H2 production. Although the application of genetic engineering to improve energy production phenotypes in eukaryotic microalgae is in its infancy, significant advances in the development of genetic manipulation tools have recently been achieved with microalgal model systems and are being used to manipulate central carbon metabolism in these organisms. It is likely that many of these advances can be extended to industrially relevant organisms. This review is focused on potential avenues of genetic engineering that may be undertaken in order to improve microalgae as a biofuel platform for the production of biohydrogen, starch-derived alcohols, diesel fuel surrogates, and/or alkanes.

Genetic Engineering of Algae for Enhanced Biofuel Production

Biological hydrogen production; fundamentals and limiting processes

DNA insertional mutagenesis and screening of the green alga Chlamydomonas reinhardtii was employed to isolate tla1, a stable transformant having a truncated light-harvesting chlorophyll antenna size. Molecular analysis showed a single plasmid insertion into an open reading frame of the nuclear genome corresponding to a novel gene (Tla1) that encodes a protein of 213 amino acids. Genetic analysis showed co-segregation of plasmid and tla1 phenotype. Biochemical analyses showed the tla1 mutant to be chlorophyll deficient, with a functional chlorophyll antenna size of photosystem I and photosystem II being about 50% and 65% of that of the wild type, respectively. It contained a correspondingly lower amount of light-harvesting proteins than the wild type and had lower steady-state levels of Lhcb mRNA. The tla1 strain required a higher light intensity for the saturation of photosynthesis and showed greater solar conversion efficiencies and a higher photosynthetic productivity than the wild type under mass culture conditions. Results are discussed in terms of the tla1 mutation, its phenotype, and the role played by the Tla1 gene in the regulation of the photosynthetic chlorophyll antenna size in C. reinhardtii.

https://www.jstor.org/stable/pdfplus/23387801.pdf

tla1, a DNA insertional transformant of the green alga Chlamydomonas reinhardtii with a truncated light-harvesting chlorophyll antenna size.

Truncated chlorophyll antenna size of the photosystems—a practical method to improve microalgal productivity and hydrogen production in mass culture

Publisher Summary
This chapter illustrates that photosynthetic efficiencies and hydrogen production by microalgal cultures may be increased upon minimizing the number of the light-harvesting chlorophyll (Chl) antenna pigments of photosynthesis. This research employs a mutagenesis approach, based on the random insertion of tagged DNA into C. reinhardtii cells, by which to impair the Chl antenna size regulation mechanism. This procedure, along with the stringent screening employed may help to unlock the “black box” of the developmental regulation of the Chl antenna size in microalgae. Thus, it is expected that mutants with a permanently truncated Chl antenna size, as well as mutants with a permanently large Chl antenna, are isolated. The advantage of this molecular genetic approach is that it leads to the identification of genes responsible for the operation of this highly conserved regulatory mechanism.

Maximizing photosynthetic efficiencies and hydrogen production in microalga cultures

DNA insertional transformants of Chlamydomonas reinhardtii were screened chemochromically for attenuated H2 production. One mutant, displaying low H2 gas photoproduction, has a nonfunctional copy of a gene that shows high homology to the family of isoamylase genes found in several photosynthetic organisms. DNA gel blotting and gene complementation were used to link this isoamylase gene to previously characterized nontagged sta7 mutants. This mutant is therefore denoted sta7-10. In C. reinhardtii, the STA7 isoamylase gene is important for the accumulation of crystalline starch, and the sta7-10 mutant reported here contains 7 h under anaerobic incubation. Complementation of mutant cells with genomic DNA corresponding to the STA7 gene restores both the starch accumulation and H2 production phenotypes. The results indicate that STA7 and starch metabolism play an important role in C. reinhardtii H2 photoproduction. Moreover, the results indicate that mere anaerobiosis is not sufficient to maintain hydrogenase gene expression without the underlying physiology, an important aspect of which is starch metabolism.

/pdf/hydrogen-photoproduction-is-attenuated-by-disruption-of-an-390mj51fhm.pdf

Hydrogen Photoproduction Is Attenuated by Disruption of an Isoamylase Gene in Chlamydomonas reinhardtii

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tla1, a DNA insertional transformant of the green alga Chlamydomonas reinhardtii with a truncated light-harvesting chlorophyll antenna size.

Truncated chlorophyll antenna size of the photosystems—a practical method to improve microalgal productivity and hydrogen production in mass culture

Maximizing photosynthetic efficiencies and hydrogen production in microalga cultures

Hydrogen Photoproduction Is Attenuated by Disruption of an Isoamylase Gene in Chlamydomonas reinhardtii