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At least 145 records · Page 8

Optimization of Energy Flow through Synthetic Metabolic Modules and Regulatory Networks in a Model Photosynthetic Eukaryotic Microbe

Photosynthetic organisms have recently gained considerable attention for a role in development of renewable energy sources. Genome-enabled systems biology methods, coupled with functional and synthetic genomics, present opportunities to develop sustainable and economical applications such as fuel production within the next 10 to 15 years. However, optimization of light-driven metabolism for biomass or biofuel production will require a detailed systems biology understanding of photosynthetic processes and cellular metabolism. Genome-scale metabolic models (GEMs) are at the core of systems analysis of cellular processes and form a common organizational framework for analyses of data resulting from functional genomics experimental work and computational studies. Therefore, there is a clear demand for high quality photosynthetic model organisms and the appropriate computational tools that enable systems analysis of light-driven metabolism. Through research conducted we expanded the currently available repertoire of photosynthetic GEMs to include the commercially valuable model diatom Phaeoctylum tricornutum. Diatoms have a peculiar and distinct evolutionary footprint and represent a major eukaryotic lineage that is taxonomically and functionally distinct from green and red algae and vascular plants. Therefore, the true potential for light-driven metabolism aimed at biofuel production remains poorly understood at a systems level for a large subset of the global diversity of photosynthetic organisms. The metabolic capabilities of P. tricornutum were comparatively modeled with those from other photosynthetic groups in order to elucidate the occurrence of metabolic traits within and between phototrophs. Additionally, this research resulted in significant extension of the COnstraints Based Reconstruction and Analysis (COBRA) Toolbox to accommodate the crucial need for infrastructure required for ‘omics data integration and analysis in the context of genome-scale models. Therefore, the proposed research achieved two important goals. First, within the broad scope of photosynthetic organisms, we functionally compared and, as a result, identified cellular processes that require optimization in order to enable deployment as biofuel feedstock. Second, the proposed research resulted in development of key computational infrastructure, which can be further extended to other biological systems, that is currently lacking but necessary for multiple ‘omics data integration.

59 BASIC BIOLOGICAL SCIENCES↗

KBase Narrative - Complete Genome Sequence of Acidovorax temperans strain LMJ

We have isolated a new strain of Acidovorax temperans strain LMJ (hereafter called strain LMJ) from a contaminated Tris-Acetate-Phosphate (TAP) medium plate of a green micro-alga Chlamydomonas reinhardtii strain LMJ.SG0182 (a Chlamydomonas Library project (CLiP) strain). We sequenced the whole genome of the strain LMJ using the PacBio Sequel II technology and have submitted it to NCBI along with the SRA and PacBio methylation motif data. We present the whole genome sequence of this strain that offer insights into its coding and non-coding genes and its nearest taxonomic neighbors.

Mitra, Mautusi↗

KBase Narrative - Complete genome sequence of a novel Microbacterium sp. strain Clip185.

We have isolated a new species of Microbacterium, an Actinobacterium. We have temporarily named this bacterium as Microbacterium sp. strain Clip185 (hereafter called strain Clip185) from a contaminated Tris-Acetate-Phosphate (TAP) medium culture plate of a green micro-alga Chlamydomonas reinhardtii strain LMJ.RY0402.185141 (a Chlamydomonas Library project CLiP strain). We sequenced the whole genome of strain Clip185 using the PacBio Sequel II Continuous Long Read technology and have submitted it to NCBI along with the SRA and PacBio methylation motif data. Additionally, we have submitted the PacBio methylome to REBASE, Ref#35996. We present the whole genome sequence of this new Microbacterium species that offers insights into its coding and non-coding genes and its nearest taxonomic neighbors.

Mitra, Mautusi↗

Complete genome sequence of Sphingobium yanoikuyae strain CC4533

We have isolated a new strain of Sphingobium yanoikuyae , which belongs to the class Alphaproteobacteria, order Sphingomonadales, and family Sphingomonadaceae. This carotenoid-producing strain is capable of degrading xenobiotics and is tolerant to toxic levels of six heavy metals. We have designated the newly isolated strain of S. yanoikuyae as S. yanoikuyae strain CC4533 (hereafter called strain CC4533) because it was isolated from a contaminated Tris-Acetate-Phosphate (TAP) medium culture plate of a green micro-alga Chlamydomonas reinhardtii wild type strain CC4533. We sequenced the whole genome of strain CC4533 using the PacBio Sequel II Continuous Long Read technology and have submitted it to NCBI along with the SRA and PacBio methylation motif data. Additionally, we have submitted the PacBio methylome to REBASE, Ref#35996. We present the whole genome sequence of S. yanoikuyae strain CC4533 that offers insights into its coding and non-coding genes and its nearest taxonomic neighbors.

59 BASIC BIOLOGICAL SCIENCES↗

Regionalized Life-Cycle Water Impacts of Microalgal-Based Biofuels in the United States

While algal biofuels have the potential to reduce the national reliance on fossil fuels, high water consumption associated with algal biomass cultivation represents a major concern potentially compromising the sustainable commercialization of this technology. This study focuses on quantifying the water footprint (WF) and water scarcity footprint (WSF) of renewable diesel derived from algal biomass and provides insights into where algal cultivation is less water-intensive than traditional ethanol and biodiesel feedstocks. Results are generated with an engineering process model developed to predict the life-cycle water consumption, considering green, blue, and gray water, of algae facilities across the United States at a high spatiotemporal resolution. The total WFs for Florida and Arizona are determined to be 13.1 and 17.6 m 3 GJ –1 , respectively. The blue WF in Arizona is shown to be 8.5 times larger than in Florida, while the green WF is 4.5 times smaller, but when combined into a total WF, there is just a 26% difference between the two locations. The analysis reveals that the total life-cycle WFs of algal renewable diesel are smaller than the optimal WFs of corn ethanol and soybean biodiesel. Algal systems benefit from higher growth rates and offer the opportunity to manage wastewater streams, therefore generating smaller green and gray WFs than those of conventional biofuels. Here, the WSF analysis identifies the Gulf Coast as the most suitable region for algal cultivation, with cultivation in the western US shown to exacerbate local water stress levels.

54 ENVIRONMENTAL SCIENCES↗

Synthesis of Hydrophilic Derivative Surfactants From Algae-Derived Unsaponifiable Lipids

In the context of decarbonizing the economy, the utilization of biologically sourced feedstocks to produce replacements for petroleum-derived materials is becoming more urgent. Improving renewable biomass production and utilization is imperative for commercializing future biorefineries. Algae-derived biomass is a particularly promising feedstock thanks to its attractive oil content and composition; specifically, the high-value products in the unsaponifiable lipids have not been included in a conversion process. Here we demonstrate surfactant synthesis from a complex oil fraction as the hydrophobic donor moieties, yielding products that are similar to commercially available surfactants such as the linear alkyl benzene sulfonates. Unsaponifiable lipids extracted from algae were derivatized to non-ionic surfactants using a green chemical synthesis route based on a double esterification with succinic acid and polyethylene glycol. The in-depth molecular and structural surfactant characterization is included and indicates that the resulting properties fall between those of pure cholesterol and phytol used as surrogates for the reaction synthesis demonstration. This is the first demonstration of an effective and potentially high-value synthesis of functional surfactants with properties that can be tailored based on the relative composition of the resulting hydrocarbon alcohol components in the mixture. This novel green chemistry synthesis approach provides a route to high-value product synthesis from algae.

09 BIOMASS FUELS↗

The Elements of Life, Photosynthesis and Genomics

I am a Professor of Biochemistry, Biophysics and Structural Biology and Plant and Microbial Biology at the University of California in Berkeley. I was born and raised in India, emigrated to the United States to attend university, earning a B.S. in Molecular Biology and a Ph.D. in Biochemistry at the University of Wisconsin in Madison. Following post-doctoral studies with Lawrence Bogorad at Harvard University where I became interested in genetic control of trace element quotas, I joined the department of Chemistry and Biochemistry at UCLA. One of the first to appreciate essential trace metals as potential regulators of gene expression, I articulated the details of the nutritional Cu regulon in Chlamydomonas. In parallel, I used genetic approaches to discover the genes governing missing steps in tetrapyrrole metabolism, including the attachment of heme to apocytochromes in the thylakoid lumen and the factors catalyzing the formation of ring V in chlorophyll. After biochemistry and classical genetics, I embraced genomics, taking a leadership role on the Joint Genome Institute’s efforts on the Chlamydomonas genome and more recently, contributing to high quality assemblies of several genomes in the green algal radiation, and large transcriptomic and proteomic datasets — focusing on the diel metabolic cycle in synchronized cultures and acclimation to key environmental and nutritional stressors — that are well-used and appreciated by the community. Finally, a new venture in Berkeley is the promotion of Auxenochlorella protothecoides as the true “green yeast” and as a platform for engineering algae to produce useful bioproducts.

59 BASIC BIOLOGICAL SCIENCES↗

Identification and preliminary characterization of conserved uncharacterized proteins from Chlamydomonas reinhardtii , Arabidopsis thaliana , and Setaria viridis

Abstract The rapid accumulation of sequenced plant genomes in the past decade has outpaced the still difficult problem of genome‐wide protein‐coding gene annotation. A substantial fraction of protein‐coding genes in all plant genomes are poorly annotated or unannotated and remain functionally uncharacterized. We identified unannotated proteins in three model organisms representing distinct branches of the green lineage (Viridiplantae): Arabidopsis thaliana (eudicot), Setaria viridis (monocot), and Chlamydomonas reinhardtii (Chlorophyte alga). Using similarity searching, we identified a subset of unannotated proteins that were conserved between these species and defined them as Deep Green proteins. Bioinformatic, genomic, and structural predictions were performed to begin classifying Deep Green genes and proteins. Compared to whole proteomes for each species, the Deep Green set was enriched for proteins with predicted chloroplast targeting signals predictive of photosynthetic or plastid functions, a result that was consistent with enrichment for daylight phase diurnal expression patterning. Structural predictions using AlphaFold and comparisons to known structures showed that a significant proportion of Deep Green proteins may possess novel folds. Though only available for three organisms, the Deep Green genes and proteins provide a starting resource of high‐value targets for further investigation of potentially new protein structures and functions conserved across the green lineage.

59 BASIC BIOLOGICAL SCIENCES↗

Green diatom mutants reveal an intricate biosynthetic pathway of fucoxanthin

Fucoxanthin is a major light-harvesting pigment in ecologically important algae such as diatoms, haptophytes, and brown algae (Phaeophyceae). Therefore, it is a major driver of global primary productivity. Species of these algal groups are brown colored because the high amounts of fucoxanthin bound to the proteins of their photosynthetic machineries enable efficient absorption of green light. While the structure of these fucoxanthin-chlorophyll proteins has recently been resolved, the biosynthetic pathway of fucoxanthin is still unknown. Here, we identified two enzymes central to this pathway by generating corresponding knockout mutants of the diatom Phaeodactylum tricornutum that are green due to the lack of fucoxanthin. Complementation of the mutants with the native genes or orthologs from haptophytes restored fucoxanthin biosynthesis. We propose a complete biosynthetic path to fucoxanthin in diatoms and haptophytes based on the carotenoid intermediates identified in the mutants and in vitro biochemical assays. It is substantially more complex than anticipated and reveals diadinoxanthin metabolism as the central regulatory hub connecting the photoprotective xanthophyll cycle and the formation of fucoxanthin. Moreover, our data show that the pathway evolved by repeated duplication and neofunctionalization of genes for the xanthophyll cycle enzymes violaxanthin de-epoxidase and zeaxanthin epoxidase. Brown algae lack diadinoxanthin and the genes described here and instead use an alternative pathway predicted to involve fewer enzymes. Our work represents a major step forward in elucidating the biosynthesis of fucoxanthin and understanding the evolution, biogenesis, and regulation of the photosynthetic machinery in algae.

59 BASIC BIOLOGICAL SCIENCES↗

Controllable Phycobilin Modification: An Alternative Photoacclimation Response in Cryptophyte Algae

Cryptophyte algae are well-known for their ability to survive under low light conditions using their auxiliary light harvesting antennas, phycobiliproteins. Mainly acting to absorb light where chlorophyll cannot (500-650 nm), phycobiliproteins also play an instrumental role in helping cryptophyte algae respond to changes in light intensity through the process of photoacclimation. Until recently, photoacclimation in cryptophyte algae was only observed as a change in the cellular concentration of phycobiliproteins; however, an additional photoacclimation response was recently discovered that causes shifts in the phycobiliprotein absorbance peaks following growth under red, blue, or green light. Here, we reproduce this newly identified photoacclimation response in two species of cryptophyte algae and elucidate the origin of the response on the protein level. We compare isolated native and photoacclimated phycobiliproteins for these two species using spectroscopy and mass spectrometry, and we report the X-ray structures of each phycobiliprotein and the corresponding photoacclimated complex. We find that neither the protein sequences nor the protein structures are modified by photoacclimation. We conclude that cryptophyte algae change one chromophore in the phycobiliprotein β subunits in response to changes in the spectral quality of light. Ultrafast pump-probe spectroscopy shows that the energy transfer is weakly affected by photoacclimation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Marine Circular Bioeconomy Workshop

Funds were used to support a Marine Circular Bioeconomy (MCB) Workshop. The purpose of the workshop was to explore the potential of marine macroalgae and microalgae production to meet the global climate, energy, and food challenges confronting society during the 21st century.

54 ENVIRONMENTAL SCIENCES↗

BETO 2021 Peer Review - 1.3.5.270 - Rewiring Algal Carbon Energetics for Renewables (RACER)

Critically needed improvements in biomass and biofuel intermediate productivity can be made by addressing fundamental inefficiencies in algal carbon conversion efficiency (CCE) to biofuel intermediates. Algae photosynthesis is, at best, able to convert 5-7% of incident light energy to biomass, while conversion to fuel intermediates falls 15-25% short of its maximum potential due to inefficiencies along the pathways. Recent progress in the Rewiring Algal Carbon Energetics (RACER) project consortium focused on a means to address the above inefficiencies in a pathway from algal biomass to a trifecta of fuel intermediates, ethanol, 2,3-butane diol, lipids and green biocrude. This project engineered a production-relevant algal species Desmodesmus armatus (SE 00107), to demonstrate biomass productivity improvements, with a doubling of the fuel intermediate yields. The new algae biorefinery paradigm embodied in RACER opens opportunities for algae engineering beyond efforts typically targeted solely at lipid content or improved light harvesting efficiency. Parallel approaches showed improved CCE through elimination of wasted energy during photosynthesis and increased carbon flux to transitory carbohydrate storage in the cells. Outdoor operation and nutrient management strategies with improvements in pretreatment, fermentation and extraction in a Combined Algal Processing approach showed a 40% reduction in MFSP, with a combined biofuel productivity of > 3700 gal/acre.

algal↗

Decarbonization of Wastewater Treatment with Microalgae Processes.

Microalgae can contribute to the decarbonization of wastewater treatment by municipalities, industries and agriculture, by substituting sunlight for energy intensive conventional treatment processes, while capturing nutrients and carbon in the algal biomass. The carbon fixed into the algal biomass can be converted to renewable natural gas (RNG) using anaerobic digestion or into renewable diesel and sustainable aviation fuel (SAF) using hydrothermal liquefaction (HTL), with greatly reduce the carbon footprints compared to conventional fuels. Bioplastics, biofertilizers and other bioproducts from microalgae biomass also provide potential benefits in greenhouse gas (GHG) reduction and domestic supply chains. Microalgae technologies can counter eutrophication caused by harmful algal blooms by recovering nutrients, N and P, from wastewaters, and even from very low nutrient concentrations found in surface waters. CO2 is required in microalgae cultivation and wastewater treatment to support maximal rates of the photosynthesis, with the CO2 obtained from the wastes themselves, other local sources or even atmospheric CO2. Greenhouse gas mitigation with microalgae processes is based on life cycle assessments, comparing such green processes with current energy intensive wastewater treatment, and nutrient reduction technologies. Microalgae technologies are specifically relevant to smaller, often disadvantaged, communities, where currently about 5,000 algae wastewater treatment ponds are operated by public utilities with many more are operated by industries. However, many, if not most, of these pond facilities require urgent technology upgrades to achieve the potential and goals of low cost GHG mitigation and efficient nutrient recycling. MicroBio Engineering Inc. is developing and has demonstrated several technologies that combine innovative carbon mitigation and decarbonization technologies for a circular economy, with longer-term potential for large-scale biofuels and biofertilizer production.

09 BIOMASS FUELS↗

Decarbonization of Wastewater Treatment with Microalgae Processes.

Microalgae can contribute to the decarbonization of wastewater treatment by municipalities, industries and agriculture, by substituting sunlight for energy intensive conventional treatment processes, while capturing nutrients and carbon in the algal biomass. The carbon fixed into the algal biomass can be converted to renewable natural gas (RNG) using anaerobic digestion or into renewable diesel and sustainable aviation fuel (SAF) using hydrothermal liquefaction (HTL), with greatly reduce the carbon footprints compared to conventional fuels. Bioplastics, biofertilizers and other bioproducts from microalgae biomass also provide potential benefits in greenhouse gas (GHG) reduction and domestic supply chains. Microalgae technologies can counter eutrophication caused by harmful algal blooms by recovering nutrients, N and P, from wastewaters, and even from very low nutrient concentrations found in surface waters. CO2 is required in microalgae cultivation and wastewater treatment to support maximal rates of the photosynthesis, with the CO2 obtained from the wastes themselves, other local sources or even atmospheric CO2. Greenhouse gas mitigation with microalgae processes is based on life cycle assessments, comparing such green processes with current energy intensive wastewater treatment, and nutrient reduction technologies. Microalgae technologies are specifically relevant to smaller, often disadvantaged, communities, where currently about 5,000 algae wastewater treatment ponds are operated by public utilities with many more are operated by industries. However, many, if not most, of these pond facilities require urgent technology upgrades to achieve the potential and goals of low cost GHG mitigation and efficient nutrient recycling. MicroBio Engineering Inc. is developing and has demonstrated several technologies that combine innovative carbon mitigation and decarbonization technologies for a circular economy, with longer-term potential for large-scale biofuels and biofertilizer production.

09 BIOMASS FUELS↗

Decarbonization of Wastewater Treatment with Microalgae Processes.

Microalgae can contribute to the decarbonization of wastewater treatment by municipalities, industries and agriculture, by substituting sunlight for energy intensive conventional treatment processes, while capturing nutrients and carbon in the algal biomass. The carbon fixed into the algal biomass can be converted to renewable natural gas (RNG) using anaerobic digestion or into renewable diesel and sustainable aviation fuel (SAF) using hydrothermal liquefaction (HTL), with greatly reduce the carbon footprints compared to conventional fuels. Bioplastics, biofertilizers and other bioproducts from microalgae biomass also provide potential benefits in greenhouse gas (GHG) reduction and domestic supply chains. Microalgae technologies can counter eutrophication caused by harmful algal blooms by recovering nutrients, N and P, from wastewaters, and even from very low nutrient concentrations found in surface waters. CO2 is required in microalgae cultivation and wastewater treatment to support maximal rates of the photosynthesis, with the CO2 obtained from the wastes themselves, other local sources or even atmospheric CO2. Greenhouse gas mitigation with microalgae processes is based on life cycle assessments, comparing such green processes with current energy intensive wastewater treatment, and nutrient reduction technologies. Microalgae technologies are specifically relevant to smaller, often disadvantaged, communities, where currently about 5,000 algae wastewater treatment ponds are operated by public utilities with many more are operated by industries. However, many, if not most, of these pond facilities require urgent technology upgrades to achieve the potential and goals of low cost GHG mitigation and efficient nutrient recycling. MicroBio Engineering Inc. is developing and has demonstrated several technologies that combine innovative carbon mitigation and decarbonization technologies for a circular economy, with longer-term potential for large-scale biofuels and biofertilizer production.

09 BIOMASS FUELS↗

Metabolic rewiring and biomass redistribution enable optimized mixotrophic growth in Chlamydomonas

Aquatic photosynthetic systems account for approximately one-half of all global carbon assimilation and could be a significant source of renewable fuels and feedstocks. However, rapid growth and biomass production in algae have not always translated into high product yields, partly because central metabolism is context specific, with metabolic fluxes being influenced by nutrient conditions and other environmental factors. In the green microalga Chlamydomonas reinhardtii (Chlamydomonas), mixotrophic cultures (acetate + light) grow far faster than phototrophic (light only) or heterotrophic (acetate + dark) cultures, even though acetate partially suppresses photosynthesis. Here, an isotopic dilution strategy with unlabeled acetate was combined with 13 CO 2 transient labeling to perform isotopically nonstationary metabolic flux analysis (INST-MFA) and to directly compare autotrophic and mixotrophic metabolism in Chlamydomonas supported by data from transcriptomics, proteomics, and metabolomics. INST-MFA indicated that acetate induces a synergistic rewiring of metabolism, conserving carbon by using the glyoxylate cycle and suppressing gluconeogenesis, the latter of which was discordant with omics results and prior models. Additionally, our data provide a plausible rationale for the well-known suppression of photosynthesis by acetate. We propose that reduced total protein content in mixotrophic versus phototrophic cells, much of which is attributed to reduced levels of photosynthetic proteins, decreases the costly metabolic burden of protein synthesis and represents a growth rate optimization strategy.

59 BASIC BIOLOGICAL SCIENCES↗