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Improved Method for Isolation of Microbial RNA from Biofuel Feedstock for Metatranscriptomics

Advances in Microbiology 2013年第1期3卷 101-107页

作者： Hailan Piao Lye Meng Markillie David E. Culley Roderick I. Mackie Matthias Hess Department of Animal Sciences Institute for Genomic Biology and Energy Biosciences University of Illinois Urbana USA DOE Joint Genome Institute Walnut Creek USA Pacific Northwest National Laboratory Richland USA School of Molecular Biosciences Washington State University Richland USA

Metatranscriptomics—gene express profiling via DNA sequencing—is a powerful tool to identify genes that are actively expressed and might contribute to the phenotype of individual organisms or the phenome (the sum of several phenotypes) of a microbial community. Furthermore, metatranscriptome studies can result in extensive catalogues of genes that encode for enzymes of industrial relevance. In both cases, a major challenge for generating a high quality metatranscriptome is the extreme lability of RNA and its susceptibility to ubiquitous RNAses. The microbial community (the microbiome) of the cow rumen efficiently degrades lignocelullosic biomass, generates significant amounts of methane, a greenhouse gas twenty times more potent than carbon dioxide, and is of general importance for the physiological wellbeing of the host animal. Metatranscriptomes of the rumen microbiome from animals kept under different conditions and from various types of rumen-incubated biomass can be expected to provide new insights into these highly interesting phenotypes and subsequently provide the framework for an enhanced understanding of this socioeconomically important ecosystem. The ability to isolate large amounts of intact RNA will significantly facilitate accurate transcript annotation and expression profiling. Here we report a method that combines mechanical disruption with chemical homogenization of the sample material and consistently yields 1 mg of intact RNA from 1 g of rumen-incubated biofuel feedstock. The yield of total RNA obtained with our method exceeds the RNA yield achieved with previously reported isolation techniques, which renders RNA isolated with the method presented here as an ideal starting material for metatranscriptomic analyses and other molecular biology applications that require significant amounts of starting material.

关键词： RNA Extraction biofuels CAZymes Metatranscriptomics Systems Microbiology Microbial Ecology

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Growth and Production of Crambe Submitted to Doses of Nitrogen and Potassium

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American Journal of Plant Sciences 2016年第1期7卷 8-16页

作者： Ramara Sena de Souza Lúcia Helena Garófalo Chaves Josely Dantas Fernandes Ana Carolina Feitosa de Vasconcelos Federal University of Campina Grande Campina Grande Brazil State University of Paraiba Campus II Lagoa Seca Brazil

The dependence on non-renewable fuels of countries around the world increased the interest for the gradual replacement of fossil fuels with renewable ones. Thus, several development programs for the use of renewable sources for power generation have been created and, in this perspective, biofuels have been highlighted. Crambe is an alternative crop for the production of biodiesel due to its rusticity, precocity, high oil content and adaptability to different climates. Optimum management practices of fertilizer use in crambe cultivation are little known and there are no specific recommendations for its fertilization. Therefore, the objective of this research was to study the effects of mineral fertilizers concerning the nitrogen and potassium elements in the growth and production of crambe. The experiment was installed in a greenhouse, with a completely randomized design. Nitrogen doses used were 30, 60, 90 and 120 kg·ha-1and 50, 75, 100 to 125 kg·ha-1 for potassium. The increase in production was directly proportional to the increase in nitrogen levels. The potassium doses did not cause significant effect on the growth of crambe.

关键词： biofuels Fertilization Crambe abyssinica N K

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Confirmation of Pearl Millet-Napiergrass Hybrids Using EST-Derived Simple Sequence Repeat (SSR) Markers

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American Journal of Plant Sciences 2013年第5期4卷 1004-1012页

作者： Charlie D. Dowling Byron L. Burson Jamie L. Foster Lee Tarpley Russell W. Jessup Department of Soil and Crop Sciences Texas A&M University College Station USA Department of Soil and Crop Sciences Texas A&M University College Station USA Texas A&M Agricultural Research and Extension Center Texas A&M AgriLife Beaumont USA. Texas A&M Agricultural Research Center Texas A&M AgriLife Beeville USA USDA-ARS Crop Germplasm Research Unit Texas A&M University College Station USA

Prospects for deploying perennial grasses that are currently considered leading candidates for dedicated energy crops over large acreages are debatable because of several limitations, including vegetative propagation or small seed size, low biomass production during the first growing season, and incomplete assessments of crop invasiveness risk. Pearl Millet-Napiergrass hybrids (“PMN”;Pennisetum glaucum [L.] R. Br. × P. purpureum Schumach.), in contrast, are large-seeded, sterile feedstocks capable of high biomass production during establishment year. Novel methods are warranted for confirmation of PMN hybrids, as traditional morphological observations can be inconclusive and chromosome number determination using cytological methods is laborious and time consuming. Six putative PMN lines were produced in this study, and 10 progeny from each line were evaluated using morphological traits, seed fertility, flow cytometry, and expressed sequence tag-simple sequence repeat (EST-SSR) markers. All putative hybrid lines were sterile and failed to produce seed. The PMN hybrids could not be distinguished from either parent using flow cytometry due to highly similar nuclear genome DNA contents. A number of paternal napiergrass-specific EST-SSRs were identified for each PMN line, and four paternal-specific EST-SSRs conserved across all napiergrass accessions were selected to screen the putative PMN hybrids. These EST-SSRs confirmed that all F1 individuals analyzed were PMN hybrids. The use of paternal-specific markers therefore provides a valuable tool in the development of both “Seeded-yet-Sterile” biofuel PMN feedstocks and additional PMN cultivar-and parental species-specific markers.

关键词： Pennisetum glaucum Pennisetum purpureum Bulked Segregant Analysis Marker-Assisted Selection Marker-Assisted Breeding EST-SSR Expressed Sequence Tag Simple Sequence Repeat Microsatellites Biofuel biofuels Pearl Millet × Napiergrass Pearl Millet Napiergrass Interspecific Hybrid PCR Polymerase Chain Reaction Comparative Genomics

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Bioenergy: Examining the Efficient Utilization of Agricultural Biomass as a Source of Sustainable Renewable Energy in Louisiana

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Journal of Sustainable Bioenergy Systems 2023年第3期13卷 99-115页

作者： Priscilla M. Loh Yaw A. Twumasi Zhu H. Ning Matilda Anokye Recheal N. D. Armah Caroline Y. Apraku Judith Oppong John B. Namwamba Lucinda Kangwana Janeth Mjema Department of Urban Forestry Environment and Natural Resources Southern University and A&M College Baton Rouge USA

The use of renewable energy is steadily being adopted as a mitigative measure for reducing greenhouse gas emissions. By assessing biomass production and consumption estimates from Louisiana parishes, this study examines the utilization of agricultural biomass as a convenient renewable energy source, and the potential of marginal lands for growing bioenergy crops in Louisiana. This was achieved by retrieving parish-level acreage production of some biofuel crops recorded in 2021 using the Quick Stats Database, to map out the spatial locations and distribution of the biofuel crops. To examine the potential of Louisiana’s marginal lands in bioenergy crop production, data was obtained from the Soil Survey Geographic (SSURGO) database and mapped-out according to the eight Land Capability Classes numbered I-VIII. The results of the mapped-out acreage data revealed that 25% of the 64 parishes including Morehouse recorded high corn production estimates, while 43%, such as East Carroll, recorded high soybean production. Meanwhile, cotton production estimates were relatively low, as recorded in only 9 parishes, with one parish, Tensas, having the highest acreage production of around 23,000. Although the identified marginal lands in parishes such as Allen and Vernon had no records of corn, soybean, or cotton production, the soil survey database revealed that these marginal lands have high nutrient soils like Alfisols, Entisols and Inceptisols with optimal nutrient balance essential for high yield bioenergy crop production. Hence, this paper highlights Louisiana’s agricultural biomass to be leveraged as sustainable renewable sources while adhering to clear production guidelines, biofuel sustainability certification, and internationally agreed sustainability criteria.

关键词： biofuels Agricultural Crops Ethanol Biodiesel Marginal Lands Louisiana

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Lipid Production by Rhodotorula glutinisfrom Pulp and Paper Wastewater for Biodiesel Production

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Journal of Sustainable Bioenergy Systems 2015年第3期5卷 114-125页

作者： Marta Amirsadeghi Sara Shields-Menard W. Todd French Rafael Hernandez Dave C. Swalm School of Chemical Engineering Mississippi State University Starkville Mississippi USA Department of Biological Science Mississippi State University Starkville Mississippi USA College of Engineering University of Louisiana at Lafayette Lafayette Louisiana USA

This study investigated the potential of oleaginous yeast Rhodotorula glutinis utilizing pulp and paper wastewater effluents as cultivation media for the sustainable production of microbial lipids as biodiesel feedstock. R. glutinis is oleaginous yeast, which has the ability to produce significant quantities of intercellular lipids in the form of triacylglycerols. Yeast lipids are a promising potential feedstock for biodiesel production due to similar fatty acid composition to plant oils. The effect of various carbon sources on biomass production, lipid accumulation, substrate utilization, and fatty acid composition using R. glutinis in the pulp and paper wastewater media was studied. The pulp and paper wastewater was supplemented with glucose, xylose, and glycerol as carbon sources under nitrogen-limited conditions. The maximum lipid productions of 1.3 - 2.9 g•L–1, which corresponded to the intracellular lipid contents of 8% - 15% cell dry weight (CDW), were obtained under various carbon substrates. A kinetic study of the batch fermentation was performed in a 3 L aerobic batch fermenter to describe the cell growth, lipid accumulation, and substrate utilization process, and the kinetic parameter was estimated. The fatty acid profile of oleaginous yeast was rich in palmitic, oleic, and linoleic acids and comparable to vegetable oils. Thus, the results of this study indicated that pulp and paper wastewater could be used to produce lipids as biodiesel feedstock.

关键词： biofuels Lipid Production Biodiesel Feedstock Rhodotorula glutinis Pulp and Paper Wastewater

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Characterization of a Thermostable, Recombinant Carboxylesterase from the Hyperthermophilic Archaeon Metallosphaera sedulaDSM5348

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Advances in Enzyme Research 2014年第1期2卷 1-13页

作者： Rushyannah Killens-Cade Rachel Turner Christine MacInnes Amy Grunden Department of Biology North Carolina State University Raleigh USA Department of Plant and Microbial Biology North Carolina State University Raleigh USA

Lipid-producing microalgae are emerging as the leading platform for producing alternative biofuels in response to diminishing petroleum reserves. Optimization of fatty acid production is required for efficient conversion of microalgal fatty acids into usable transportation fuels. Microbial lipases/esterases can be used to enhance fatty acid production because of their efficacy in catalyzing hydrolysis of esters into alcohols and fatty acids while minimizing the potential poisoning of catalysts needed in the biofuel production process. Although studies have extensively focused on lipases/esterases produced by mesophilic organisms, an understanding of lipases/esterases produced by thermophilic, acidic tolerant microbes, such as Metallosphaera sedula, is limited. In this work, the carboxylesterase from Metallosphaera sedula DSM5348 encoded by Msed_1072 was recombinantly expressed in Escherichia coli strain BL21 (λDE3). The purified enzyme either with a hexahistidine (His6)-tag (Msed_1072Nt and Msed_1072Ct) or without the hexahistidine (His6)-tag (Msed_1072) was biochemically characterized using a variety of substrates over a range of temperatures and pH and in the presence of metal ions, organic solvents, and detergents. In this study, the fusion of the protein with a hexahistidine (His6)-tag did not result in a change in substrate specificity, but the findings provide information on which enzyme variant can hydrolyze fatty acid esters in the presence of various chemicals, and this has important implication for their use in industrial processes. It also demonstrates that Metallosphaera sedula Msed_1072 can have application in microalgae-based biofuel production systems.

关键词： Carboxylesterase Metallosphaera sedula Hyperthermoacidophile biofuels Algae Lipids

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Metabolic Engineering of Thermoanaerobacterium thermosaccharolyticumfor Increased n-Butanol Production

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Advances in Microbiology 2013年第1期3卷 46-51页

作者： Ashwini Bhandiwad Anna Guseva Lee Lynd Dartmouth College Hanover USA Mascoma Corporation Lebanon USA

Thermoanaerobacterium thermosaccharolyticum shows promise as a host for n-butanol production since it natively has the required genes involved in the n-butanol biosynthetic pathway. Overexpression of the natively occurring bcs operon containing the genes thl, hbd, crt, bcd, etfA, and etfB responsible for the formation of butyryl CoA increased the n-butanol production by 180% compared to the wild type from a n-butanol titer of 1.8 mM to 5.1 mM. The deletion of one of the six alcohol dehydrogenase genes confirmed that it was the primary gene responsible for ethanol and n-butanol production from acetyl CoA and butyryl CoA respectively.

关键词： biofuels n-Butanol Thermophile Metabolic Engineering

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Trehalose and Sucrose Osmolytes Accumulated by Algae as Potential Raw Material for Bioethanol

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Natural Resources 2011年第3期2卷 173-179页

作者： Ma.del Pilar Bremauntz Luis G.Torres-Bustillos Rosa-Olivia Canizares-Villanueva Enrique Duran-Paramo Luis Fernández-Linares Laboratory of Bioprocesses Department of BioprocessesUnidad Profesional Interdisciplinaria de Biotecnologia UPIBI-Instituto Politecnico NacionalMexico CityMexico Departamento de Biotecnologia y Bioingenieria Centro de Investigacion y de Estudios Avanzados-IPNMexico CityMexico

Currently, obtaining sustainable fuels, such as biodiesel and bioethanol, from cheap and renewable materials is a challenge. In recent years, a new approach being developed consists of producing, sugars from algae by photosynthesis. Sugar accumulation can be increased under osmotic stress (osmoregulation). The aim of this study is to show the pro-duction of sugars from algae, isolated from natural sources, and the effect of osmotic stress on fermentable sugars ac-cumulation. Strain isolation, production of sugars from each alga and the effect of osmotic stress on growth and sugar production are described. Twelve algal strains were isolated, showing growths between 0.6 and 1.8 g of biomass dry weight /L, all with production of intracellular and extracellular sugars. The strain identified as Chlorella sp. showed an increase in sugar production from 23.64 to 421 mg of sugars/g of biomass dry weight after 24 h of osmotic stress with 0.4 M NaCl. Sucrose and trehalose, both fermentable sugars, were the compatible osmolytes accumulated in response to the osmotic stress. The isolated strains are potential producers of fermentable sugars, using the photosynthetic pathway and osmotic stress.

关键词： Algae biofuels Osmoregulators Carbohydrates Bioethanol

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A Life Cycle Assessment Based Evaluation of a Coupled Wastewater Treatment and Biofuel Production Paradigm

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Journal of Environmental Protection 2013年第9期4卷 1018-1033页

作者： Monica C. Rothermel Amy E. Landis William J. Barr Kullapa Soratana Kayla M. Reddington Matthew K. Weschler Grace Witter Willie F. Harper De- partment of Systems Engineering and Management Air Force Institute of Technology WPAFB USA Department of Civil and Environmental Engineering University of Pittsburgh Pittsburgh USA School of Sustainable Engineering and the Built Environment Global Institute of Sustainability Arizona State University Engineering G-Wing Tempe USA

A laboratory experiment was performed to determine the feasibility of coupling a conventional wastewater treatment system with an algal photobioreactor (PBR) for the removal of nutrients from wastewater and production of renewable resources. An activated sludge batch reactor was set up in series with an algal PBR to feed synthetic wastewater to Chlorella vulgaris. The nutrient concentration in the water as well as lipid content, carbohydrate content, and growth rate of the algal biomass were tested over 10 cycles to determine the capabilities of the coupled system. The study revealed complete nutrient removal in some cycles, with the average final nutrient content of 2 mg-P/L and 3 mg-N/L in effluent of the PBR. The algae biomass contained 24% ± 3% lipids and 26% ± 7% carbohydrates by dry weight. A life cycle assessment revealed the highest energy demand occurred during harvesting of the algal mixture through centrifugation or filtration, but the highest global warming and eutrophication impacts were due to CO2 use and PBR construction material production. It is feasible for the system to treat wastewater while generating renewable resources, but the system must be optimized to reduce life cycle environmental impacts and result in a net energy gain before large-scale implementation is possible.

关键词： Wastewater biofuels Life Cycle Assessment Nitrogen Carbohydrate

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China’s bioenergy industry development roadmap

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Engineering Sciences 2009年第2期7卷 57-63页

作者： Shi Yuanchun Li Shizhong Liu Xuejun China Agricultural University Beijing 100094 China Tsinghua University Beijing 100084 China

Positive development of renewable energy, saving and substitution of fossil energy, promotion of the energy structure adjustment are the inevitable strategy choices of China’s sustainable development. This paper discussed the China’s bioenergy resources status, development targets and technology development roadmaps. China has 136.140 million hm2 of marginal land, which distribute mainly in western and northern regions. There are 1 billion t of crop residues and forestry waste annually, and 300 million t can be used to produce different kinds of bioenergies. And organic waste and manure can generate 50 billion m3 of biogas. The discussed development target indicated that it can construct a biomass oilfield with the capacity of 100 million t/year and reduce 200 million t of CO_2 emission by 2020. The bioenergy technology development roadmap indicated that the bioethanol mainly uses non grain starch and hemicellulose products as raw materials in the near-term (2006-2010). The biodiesel technology will focus on the advanced production technology, FT diesel, liquefaction of biomass and raw material production technology.

关键词： bioenergy development roadmap biofuels bio-pellets biopower

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