Search

The heterologous expression of terpene synthases in microbial hosts has opened numerous possibilities for bioproduction of desirable metabolites. Photosynthetic microbial hosts present a sustainable alternative to traditional fermentative systems, using freely available (sun)light and carbon dioxide as inputs for bio-production. Here, we report the expression of a patchoulol synthase from Pogostemon cablin Benth in the model green microalga Chlamydomonas reinhardtii. The sesquiterpenoid patchoulol was produced from the alga and was used as a marker of sesquiterpenoid production capacity. A novel strategy for gene loading was employed and patchoulol was produced up to 922±242µgg -1 CDW in six days. We additionally investigated the effect of carbon source on sesquiterpenoid productivity from C. reinhardtii in scale-up batch cultivations. It was determined that up to 1.03mgL -1 sesquiterpenoid products could be produced in completely photoautotrophic conditions and that the alga exhibited altered sesquiterpenoid production metabolism related to carbon source., (Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)

Photobiological hydrogen production using microalgae is being developed into a promising clean fuel stream for the future. In this study, microarray analyses were used to obtain global expression profiles of mRNA abundance in the green alga Chlamydomonas reinhardtii at different time points before the onset and during the course of sulfur-depleted hydrogen production. These studies were followed by real-time quantitative reverse transcription-PCR and protein analyses. The present work provides new insights into photosynthesis, sulfur acquisition strategies, and carbon metabolism-related gene expression during sulfur-induced hydrogen production. A general trend toward repression of transcripts encoding photosynthetic genes was observed. In contrast to all other LHCBM genes, the abundance of the LHCBM9 transcript (encoding a major light-harvesting polypeptide) and its protein was strongly elevated throughout the experiment. This suggests a major remodeling of the photosystem II light-harvesting complex as well as an important function of LHCBM9 under sulfur starvation and photobiological hydrogen production. This paper presents the first global transcriptional analysis of C. reinhardtii before, during, and after photobiological hydrogen production under sulfur deprivation.

The development of clean borderless fuels is of vital importance to human and environmental health and global prosperity. Currently, fuels make up approximately 67% of the global energy market (total market = 15 TW year(-1)) (Hoffert et al. 1998). In contrast, global electricity demand accounts for only 33% (Hoffert et al. 1998). Yet, despite the importance of fuels, almost all CO(2) free energy production systems under development are designed to drive electricity generation (e.g. clean-coal technology, nuclear, photovoltaic, wind, geothermal, wave and hydroelectric). In contrast, and indeed almost uniquely, biofuels also target the much larger fuel market and so in the future will play an increasingly important role in maintaining energy security (Lal 2005). Currently, the main biofuels that are at varying stages of development include bio-ethanol, liquid carbohydrates [e.g. biodiesel or biomass to liquid (BTL) products], biomethane and bio-H(2). This review is focused on placing bio-H(2) production processes into the context of the current biofuels market and summarizing advances made both at the level of bioengineering and bioreactor design.

Solar energy capture, conversion into chemical energy and biopolymers by photoautotrophic organisms, is the basis for almost all life on Earth. A broad range of organisms have developed complex molecular machinery for the efficient conversion of sunlight to chemical energy over the past 3 billion years, which to the present day has not been matched by any man-made technologies. Chlorophyll photochemistry within photosystem II (PSII) drives the water-splitting reaction efficiently at room temperature, in contrast with the thermal dissociation reaction that requires a temperature of ca. 1550 K. The successful elucidation of the high-resolution structure of PSII, and in particular the structure of its Mn(4)Ca cluster provides an invaluable blueprint for designing solar powered biotechnologies for the future. This knowledge, combined with new molecular genetic tools, fully sequenced genomes, and an ever increasing knowledge base of physiological processes of oxygenic phototrophs has inspired scientists from many countries to develop new biotechnological strategies to produce renewable CO(2)-neutral energy from sunlight. This review focuses particularly on the potential of use of cyanobacteria and microalgae for biohydrogen production. Specifically this article reviews the predicted size of the global energy market and the constraints of global warming upon it, before detailing the complex set of biochemical pathways that underlie the photosynthetic process and how they could be modified for improved biohydrogen production.

Background: Oleaginous microalgae are promising production hosts for the sustainable generation of lipid-based bioproducts and as bioenergy carriers such as biodiesel. Transcriptomics of the lipid accumulation phase, triggered efficiently by nitrogen starvation, is a valuable approach for the identification of gene targets for metabolic engineering. Results: An explorative analysis of the detailed transcriptional response to different stages of nitrogen availability was performed in the oleaginous green alga Monoraphidium neglectum. Transcript data were correlated with metabolic data for cellular contents of starch and of different lipid fractions. A pronounced transcriptional down-regulation of photosynthesis became apparent in response to nitrogen starvation, whereas glucose catabolism was found to be up-regulated. An in-depth reconstruction and analysis of the pathways for glycerolipid, central carbon, and starch metabolism revealed that distinct transcriptional changes were generally found only for specific steps within a metabolic pathway. In addition to pathway analyses, the transcript data were also used to refine the current genome annotation. The transcriptome data were integrated into a database and complemented with data for other microalgae which were also subjected to nitrogen starvation. It is available at https://tdbmn.cebitec.uni-bielefeld.de. Conclusions: Based on the transcriptional responses to different stages of nitrogen availability, a model for triacylglycerol and lipid hyperaccumulation is proposed, which involves transcriptional induction of thioesterases, differential regulation of lipases, and a re-routing of the central carbon metabolism. Over-expression of distinct thioesterases was identified to be a potential strategy to increase the oleaginous phenotype of M. neglectum, and furthermore specific lipases were identified as potential targets for future metabolic engineering approaches. [ABSTRACT FROM AUTHOR]

The main function of the photosynthetic process is to capture solar energy and to store it in the form of chemical ‘fuels’. Increasingly, the photosynthetic machinery is being used for the production of biofuels such as bio-ethanol, biodiesel and bio-H2. Fuel production efficiency is directly dependent on the solar photon capture and conversion efficiency of the system. Green algae (e.g. Chlamydomonas reinhardtii) have evolved genetic strategies to assemble large light-harvesting antenna complexes (LHC) to maximize light capture under low-light conditions, with the downside that under high solar irradiance, most of the absorbed photons are wasted as fluorescence and heat to protect against photodamage. This limits the production process efficiency of mass culture. We applied RNAi technology to down-regulate the entire LHC gene family simultaneously to reduce energy losses by fluorescence and heat. The mutant Stm3LR3 had significantly reduced levels of LHCI and LHCII mRNAs and proteins while chlorophyll and pigment synthesis was functional. The grana were markedly less tightly stacked, consistent with the role of LHCII. Stm3LR3 also exhibited reduced levels of fluorescence, a higher photosynthetic quantum yield and a reduced sensitivity to photoinhibition, resulting in an increased efficiency of cell cultivation under elevated light conditions. Collectively, these properties offer three advantages in terms of algal bioreactor efficiency under natural high-light levels: (i) reduced fluorescence and LHC-dependent heat losses and thus increased photosynthetic efficiencies under high-light conditions; (ii) improved light penetration properties; and (iii) potentially reduced risk of oxidative photodamage of PSII. [ABSTRACT FROM AUTHOR]

The development of clean borderless fuels is of vital importance to human and environmental health and global prosperity. Currently, fuels make up approximately 67% of the global energy market (total market = 15 TW year−1) ( Hoffert et al. 1998 ). In contrast, global electricity demand accounts for only 33% ( Hoffert et al. 1998 ). Yet, despite the importance of fuels, almost all CO2 free energy production systems under development are designed to drive electricity generation (e.g. clean-coal technology, nuclear, photovoltaic, wind, geothermal, wave and hydroelectric). In contrast, and indeed almost uniquely, biofuels also target the much larger fuel market and so in the future will play an increasingly important role in maintaining energy security ( Lal 2005 ). Currently, the main biofuels that are at varying stages of development include bio-ethanol, liquid carbohydrates [e.g. biodiesel or biomass to liquid (BTL) products], biomethane and bio-H2. This review is focused on placing bio-H2 production processes into the context of the current biofuels market and summarizing advances made both at the level of bioengineering and bioreactor design. [ABSTRACT FROM AUTHOR]

The rapid development of clean fuels for the future is a critically important global challenge for two main reasons. First, new fuels are needed to supplement and ultimately replace depleting oil reserves. Second, fuels capable of zero CO2 emissions are needed to slow the impact of global warming. This review summarizes the development of solar powered bio-H2 production processes based on the conversion of photosynthetic products by fermentative bacteria, as well as using photoheterotrophic and photoautrophic organisms. The use of advanced bioreactor systems and their potential and limitations in terms of process design, efficiency, and cost are also briefly reviewed. [ABSTRACT FROM AUTHOR]

Photosynthetic microalgae hold great promise as non-food feedstocks for the sustainable production of a range of bio-products and genetic engineering is an increasingly important strategy to improve the natural cellular features. For the vast majority of microalgal strains, however, genetic engineering is not yet possible and the establishment of efficient genetic tools for a broad range of microalgal species is an important task to reach biotechnological success. Stable DNA transformation is one of the crucial steps for most genetic engineering approaches. In this context, we report the first successful and stable nuclear genetic transformation of the biotechnologically promising oleaginous microalga M. neglectum . Transformation was achieved by electroporation and the efficiency could progressively be improved by implementation of an additional cell pretreatment step, aiming at weakening the rigid natural cell wall. Furthermore, a first reporter transgene was established for M. neglectum . As a result of this work, genetic engineering strategies now become accessible which are required to successfully establish M. neglectum as a novel host for biotechnological applications. [ABSTRACT FROM AUTHOR]

Abstract: Phototrophic organisms use photosynthesis to convert solar energy into chemical energy. In nature, the chemical energy is stored in a diverse range of biopolymers. These sunlight-derived, energy-rich biopolymers can be converted into environmentally clean and CO2 neutral fuels. A select group of photosynthetic microorganisms have developed the ability to extract and divert protons and electrons derived from water to chloroplast hydrogenase(s) to produce molecular H2 fuel. Here, we describe the development and characterization of C. reinhardtii strains, derived from the high H2 production mutant Stm6, into which the HUP1 (hexose uptake protein) hexose symporter from Chlorella kessleri was introduced. The isolated cell lines can use externally supplied glucose for heterotrophic growth in the dark. More importantly, external glucose supply (1mM) was shown to increase the H2 production capacity in strain Stm6Glc4 to ∼150% of that of the high-H2 producing strain, Stm6. This establishes the foundations for a new fuel production process in which H2O and glucose can simultaneously be used for H2 production. It also opens new perspectives on future strategies for improving bio-H2 production efficiency under natural day/night regimes and for using sugar waste material for energy production in green algae as photosynthetic catalysts. [Copyright &y& Elsevier]