Journal articles: 'Genetically engineered feedstocks'

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Endres, A. Bryan. "New hope for dedicated genetically engineered bioenergy feedstocks?" GCB Bioenergy 4, no. 2 (October 17, 2011): 127–29. http://dx.doi.org/10.1111/j.1757-1707.2011.01134.x.

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Heinrich, Daniel, Matthias Raberg, Philipp Fricke, Shane T. Kenny, Laura Morales-Gamez, Ramesh P. Babu, Kevin E. O'Connor, and Alexander Steinbüchel. "Synthesis Gas (Syngas)-Derived Medium-Chain-Length Polyhydroxyalkanoate Synthesis in Engineered Rhodospirillum rubrum." Applied and Environmental Microbiology 82, no. 20 (August 12, 2016): 6132–40. http://dx.doi.org/10.1128/aem.01744-16.

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ABSTRACTThe purple nonsulfur alphaproteobacteriumRhodospirillum rubrumS1 was genetically engineered to synthesize a heteropolymer of mainly 3-hydroxydecanoic acid and 3-hydroxyoctanoic acid [P(3HD-co-3HO)] from CO- and CO2-containing artificial synthesis gas (syngas). For this, genes fromPseudomonas putidaKT2440 coding for a 3-hydroxyacyl acyl carrier protein (ACP) thioesterase (phaG), a medium-chain-length (MCL) fatty acid coenzyme A (CoA) ligase (PP_0763), and an MCL polyhydroxyalkanoate (PHA) synthase (phaC1) were cloned and expressed under the control of the CO-inducible promoter PcooFfromR. rubrumS1 in a PHA-negative mutant ofR. rubrum. P(3HD-co-3HO) was accumulated to up to 7.1% (wt/wt) of the cell dry weight by a recombinant mutant strain utilizing exclusively the provided gaseous feedstock syngas. In addition to an increased synthesis of these medium-chain-length PHAs (PHAMCL), enhanced gene expression through the PcooFpromoter also led to an increased molar fraction of 3HO in the synthesized copolymer compared with the Placpromoter, which regulated expression on the original vector. The recombinant strains were able to partially degrade the polymer, and the deletion ofphaZ2, which codes for a PHA depolymerase most likely involved in intracellular PHA degradation, did not reduce mobilization of the accumulated polymer significantly. However, an amino acid exchange in the active site of PhaZ2 led to a slight increase in PHAMCLaccumulation. The accumulated polymer was isolated; it exhibited a molecular mass of 124.3 kDa and a melting point of 49.6°C. With the metabolically engineered strains presented in this proof-of-principle study, we demonstrated the synthesis of elastomeric second-generation biopolymers from renewable feedstocks not competing with human nutrition.IMPORTANCEPolyhydroxyalkanoates (PHAs) are natural biodegradable polymers (biopolymers) showing properties similar to those of commonly produced petroleum-based nondegradable polymers. The utilization of cheap substrates for the microbial production of PHAs is crucial to lower production costs. Feedstock not competing with human nutrition is highly favorable. Syngas, a mixture of carbon monoxide, carbon dioxide, and hydrogen, can be obtained by pyrolysis of organic waste and can be utilized for PHA synthesis by several kinds of bacteria. Up to now, the biosynthesis of PHAs from syngas has been limited to short-chain-length PHAs, which results in a stiff and brittle material. In this study, the syngas-utilizing bacteriumRhodospirillum rubrumwas genetically modified to synthesize a polymer which consisted of medium-chain-length constituents, resulting in a rubber-like material. This study reports the establishment of a microbial synthesis of these so-called medium-chain-length PHAs from syngas and therefore potentially extends the applications of syngas-derived PHAs.

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Manandhar, Ashish, and Ajay Shah. "Techno-Economic Analysis of Bio-Based Lactic Acid Production Utilizing Corn Grain as Feedstock." Processes 8, no. 2 (February 6, 2020): 199. http://dx.doi.org/10.3390/pr8020199.

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Lactic acid is an important chemical with numerous commercial applications that can be fermentatively produced from biological feedstocks. Producing lactic acid from corn grain could complement the use of already existing infrastructure for corn grain-based ethanol production with a higher value product. The objective of this study was to evaluate the techno-economic feasibility of producing 100,000 metric tons (t) of lactic acid annually from corn grain in a biorefinery. The study estimated the resources (equipment, raw materials, energy, and labor) requirements and costs to produce lactic acid from bacteria, fungi and yeast-based fermentation pathways. Lactic acid production costs were $1181, $1251 and $844, for bacteria, fungi and yeast, respectively. Genetically engineered yeast strains capable of producing lactic acid at low pH support significantly cheaper processes because they do not require simultaneous neutralization and recovery of lactic acid, resulting in lower requirements for chemical, equipment, and utilities. Lactic acid production costs were highly sensitive to sugar-to-lactic-acid conversion rates, grain price, plant size, annual operation hours, and potential use of gypsum. Improvements in process efficiencies and lower equipment and chemical costs would further reduce the cost of lactic acid production from corn grain.

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Sato, Trey K., Tongjun Liu, Lucas S. Parreiras, Daniel L. Williams, Dana J. Wohlbach, Benjamin D. Bice, Irene M. Ong, et al. "Harnessing Genetic Diversity in Saccharomyces cerevisiae for Fermentation of Xylose in Hydrolysates of Alkaline Hydrogen Peroxide-Pretreated Biomass." Applied and Environmental Microbiology 80, no. 2 (November 8, 2013): 540–54. http://dx.doi.org/10.1128/aem.01885-13.

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ABSTRACTThe fermentation of lignocellulose-derived sugars, particularly xylose, into ethanol by the yeastSaccharomyces cerevisiaeis known to be inhibited by compounds produced during feedstock pretreatment. We devised a strategy that combined chemical profiling of pretreated feedstocks, high-throughput phenotyping of genetically diverseS. cerevisiaestrains isolated from a range of ecological niches, and directed engineering and evolution against identified inhibitors to produce strains with improved fermentation properties. We identified and quantified for the first time the major inhibitory compounds in alkaline hydrogen peroxide (AHP)-pretreated lignocellulosic hydrolysates, including Na+, acetate, andp-coumaric (pCA) and ferulic (FA) acids. By phenotyping these yeast strains for their abilities to grow in the presence of these AHP inhibitors, one heterozygous diploid strain tolerant to all four inhibitors was selected, engineered for xylose metabolism, and then allowed to evolve on xylose with increasing amounts ofpCA and FA. After only 149 generations, one evolved isolate, GLBRCY87, exhibited faster xylose uptake rates in both laboratory media and AHP switchgrass hydrolysate than its ancestral GLBRCY73 strain and completely converted 115 g/liter of total sugars in undetoxified AHP hydrolysate into more than 40 g/liter ethanol. Strikingly, genome sequencing revealed that during the evolution from GLBRCY73, the GLBRCY87 strain acquired the conversion of heterozygous to homozygous alleles in chromosome VII and amplification of chromosome XIV. Our approach highlights that simultaneous selection on xylose andpCA or FA with a wildS. cerevisiaestrain containing inherent tolerance to AHP pretreatment inhibitors has potential for rapid evolution of robust properties in lignocellulosic biofuel production.

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Li, Yan-Feng, Hong Jiang, Zhong Hu, Guang-Lei Liu, Zhen-Ming Chi, and Zhe Chi. "Overexpression of an Inulinase Gene in an Oleaginous Yeast, Aureobasidium melanogenum P10, for Efficient Lipid Production from Inulin." Journal of Molecular Microbiology and Biotechnology 28, no. 4 (2018): 190–200. http://dx.doi.org/10.1159/000493139.

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In this study, in order to directly and efficiently convert inulin into a single-cell oil (SCO), an INU1 gene encoding inulinase from Kluyveromyces marxianus was integrated into the genomic DNA and actively expressed in an SCO producer Aureobasidium melanogenum P10. The transformant API41 obtained produced 28.5 U/mL of inulinase and its wild-type strain P10 yielded only 8.62 U/mL. Most (97.5%) of the inulinase produced by the transformant API41 was secreted into the culture. During a 10-L fermentation, 66.2% (w/w) lipid in the yeast cells of the transformant API41 and 14.38 g/L of cell dry weight were attained from inulin of 80.0 g/L within 120 h, high inulinase activity (23.7 U/mL) was also produced within 72 h, and the added inulin was actively hydrolyzed. This confirmed that the genetically engineered yeast of A. melanogenum P10 is suitable for direct production of lipids from inulin. The lipids produced could be used as feedstocks for biodiesel production.

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Dashtban, Mehdi, Greg Kepka, Bernhard Seiboth, and Wensheng Qin. "Xylitol Production by Genetically Engineered Trichoderma reesei Strains Using Barley Straw as Feedstock." Applied Biochemistry and Biotechnology 169, no. 2 (December 18, 2012): 554–69. http://dx.doi.org/10.1007/s12010-012-0008-y.

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Abalde-Cela, Sara, Anna Gould, Xin Liu, Elena Kazamia, Alison G. Smith, and Chris Abell. "High-throughput detection of ethanol-producing cyanobacteria in a microdroplet platform." Journal of The Royal Society Interface 12, no. 106 (May 2015): 20150216. http://dx.doi.org/10.1098/rsif.2015.0216.

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Ethanol production by microorganisms is an important renewable energy source. Most processes involve fermentation of sugars from plant feedstock, but there is increasing interest in direct ethanol production by photosynthetic organisms. To facilitate this, a high-throughput screening technique for the detection of ethanol is required. Here, a method for the quantitative detection of ethanol in a microdroplet-based platform is described that can be used for screening cyanobacterial strains to identify those with the highest ethanol productivity levels. The detection of ethanol by enzymatic assay was optimized both in bulk and in microdroplets. In parallel, the encapsulation of engineered ethanol-producing cyanobacteria in microdroplets and their growth dynamics in microdroplet reservoirs were demonstrated. The combination of modular microdroplet operations including droplet generation for cyanobacteria encapsulation, droplet re-injection and pico-injection, and laser-induced fluorescence, were used to create this new platform to screen genetically engineered strains of cyanobacteria with different levels of ethanol production.

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Kang, Qian, Lise Appels, Tianwei Tan, and Raf Dewil. "Bioethanol from Lignocellulosic Biomass: Current Findings Determine Research Priorities." Scientific World Journal 2014 (2014): 1–13. http://dx.doi.org/10.1155/2014/298153.

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“Second generation” bioethanol, with lignocellulose material as feedstock, is a promising alternative for first generation bioethanol. This paper provides an overview of the current status and reveals the bottlenecks that hamper its implementation. The current literature specifies a conversion of biomass to bioethanol of 30 to ~50% only. Novel processes increase the conversion yield to about 92% of the theoretical yield. New combined processes reduce both the number of operational steps and the production of inhibitors. Recent advances in genetically engineered microorganisms are promising for higher alcohol tolerance and conversion efficiency. By combining advanced systems and by intensive additional research to eliminate current bottlenecks, second generation bioethanol could surpass the traditional first generation processes.

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Salamanca-Cardona, Lucia, Ryan A. Scheel, Norman Scott Bergey, Arthur J. Stipanovic, Ken'ichiro Matsumoto, Seiichi Taguchi, and Christopher T. Nomura. "Consolidated bioprocessing of poly(lactate-co-3-hydroxybutyrate) from xylan as a sole feedstock by genetically-engineered Escherichia coli." Journal of Bioscience and Bioengineering 122, no. 4 (October 2016): 406–14. http://dx.doi.org/10.1016/j.jbiosc.2016.03.009.

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Culaba, Alvin B., Aristotle T. Ubando, Phoebe Mae L. Ching, Wei-Hsin Chen, and Jo-Shu Chang. "Biofuel from Microalgae: Sustainable Pathways." Sustainability 12, no. 19 (September 28, 2020): 8009. http://dx.doi.org/10.3390/su12198009.

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As the demand for biofuels increases globally, microalgae offer a viable biomass feedstock to produce biofuel. With abundant sources of biomass in rural communities, these materials could be converted to biodiesel. Efforts are being done in order to pursue commercialization. However, its main usage is for other applications such as pharmaceutical, nutraceutical, and aquaculture, which has a high return of investment. In the last 5 decades of algal research, cultivation to genetically engineered algae have been pursued in order to push algal biofuel commercialization. This will be beneficial to society, especially if coupled with a good government policy of algal biofuels and other by-products. Algal technology is a disruptive but complementary technology that will provide sustainability with regard to the world’s current issues. Commercialization of algal fuel is still a bottleneck and a challenge. Having a large production is technical feasible, but it is not economical as of now. Efforts for the cultivation and production of bio-oil are still ongoing and will continue to develop over time. The life cycle assessment methodology allows for a sustainable evaluation of the production of microalgae biomass to biodiesel.

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Verma, Samakshi, and Arindam Kuila. "Involvement of green technology in microalgal biodiesel production." Reviews on Environmental Health 35, no. 2 (June 25, 2020): 173–88. http://dx.doi.org/10.1515/reveh-2019-0061.

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AbstractAccording to the report of the renewable energy policy network for the 21st century published in 2014, biodiesel and bioethanol are the most used biofuels and are responsible for transportation worldwide. Biodiesel specially has shown an increase in production globally by 15 times by volume from 2002 to 2012. Promising feedstock of biodiesel are cyanobacteria and microalgae as they possess a shorter cultivation time (4 fold lesser) and high oil content (10 fold higher) than corn, jatropha and soybean (conventional oil-producing territorial plants). Various valuable natural chemicals are also produced from these organisms including food supplements such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), pigments, and vitamins. Additionally, cellular components of microalgae and cyanobacteria are connected with therapeutic characteristics such as anti-inflammatory, antioxidant, antiviral and immune stimulating. Commercialization of algal biodiesel (or other products) can be achieved by isolating and identifying the high-yielding strains that possess a faster growth rate. Indigenous strains can be genetically engineered into high-yielding transgenic strains. The present article discusses about the use of nanotechnology and genetic engineering approach for improved lipid accumulation in microalgae for biodiesel production.

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Djukic-Vukovic, Aleksandra, Ljiljana Mojovic, Dusanka Pejin, Maja Vukasinovic-Sekulic, Marica Rakin, Svetlana Nikolic, and Jelena Pejin. "New trends and challenges in lactic acid production on renewable biomass." Chemical Industry 65, no. 4 (2011): 411–22. http://dx.doi.org/10.2298/hemind110114022d.

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Lactic acid is a relatively cheap chemical with a wide range of applications: as a preservative and acidifying agent in food and dairy industry, a monomer for biodegradable poly-lactide polymers (PLA) in pharmaceutical industry, precursor and chemical feedstock for chemical, textile and leather industries. Traditional raw materials for fermentative production of lactic acid, refined sugars, are now being replaced with starch from corn, rice and other crops for industrial production, with a tendency for utilization of agro industrial wastes. Processes based on renewable waste sources have ecological (zero CO2 emission, eco-friendly by-products) and economical (cheap raw materials, reduction of storage costs) advantages. An intensive research interest has been recently devoted to develop and improve the lactic acid production on more complex industrial by-products, like thin stillage from bioethanol production, corncobs, paper waste, straw etc. Complex and variable chemical composition and purity of these raw materials and high nutritional requirements of Lare the main obstacles in these production processes. Media supplementation to improve the fermentation is an important factor, especially from an economic point of view. Today, a particular challenge is to increase the productivity of lactic acid production on complex renewable biomass. Several strategies are currently being explored for this purpose such as process integration, use of Lwith amylolytic activity, employment of mixed cultures of Land/or utilization of genetically engineered microorganisms. Modern techniques of genetic engineering enable construction of microorganisms with desired characteristics and implementation of single step processes without or with minimal pre-treatment. In addition, new bioreactor constructions (such as membrane bioreactors), utilization of immobilized systems are also being explored. Electrodialysis, bipolar membrane separation process, enhanced filtration techniques etc. can provide some progress in purification technologies, although it is still remaining the most expensive phase in the lactic acid production. A new approach of parallel production of lactic bacteria biomass with probiotic activity and lactic acid could provide additional benefit and profit rise in the production process.

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Wei, Liu-Jing, Yu-Tao Zhong, Ming-Yue Nie, Shun-Cheng Liu, and Qiang Hua. "Biosynthesis of α-Pinene by Genetically Engineered Yarrowia lipolytica from Low-Cost Renewable Feedstocks." Journal of Agricultural and Food Chemistry, December 24, 2020. http://dx.doi.org/10.1021/acs.jafc.0c06504.

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Cernak, Paul, Raissa Estrela, Snigdha Poddar, Jeffrey M. Skerker, Ya-Fang Cheng, Annika K. Carlson, Berling Chen, et al. "EngineeringKluyveromyces marxianusas a Robust Synthetic Biology Platform Host." mBio 9, no. 5 (September 25, 2018). http://dx.doi.org/10.1128/mbio.01410-18.

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ABSTRACTThroughout history, the yeastSaccharomyces cerevisiaehas played a central role in human society due to its use in food production and more recently as a major industrial and model microorganism, because of the many genetic and genomic tools available to probe its biology. However,S. cerevisiaehas proven difficult to engineer to expand the carbon sources it can utilize, the products it can make, and the harsh conditions it can tolerate in industrial applications. Other yeasts that could solve many of these problems remain difficult to manipulate genetically. Here, we engineered the thermotolerant yeastKluyveromyces marxianusto create a new synthetic biology platform. Using CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats with Cas9)-mediated genome editing, we show that wild isolates ofK. marxianuscan be made heterothallic for sexual crossing. By breeding two of these mating-type engineeredK. marxianusstrains, we combined three complex traits—thermotolerance, lipid production, and facile transformation with exogenous DNA—into a single host. The ability to crossK. marxianusstrains with relative ease, together with CRISPR-Cas9 genome editing, should enable engineering ofK. marxianusisolates with promising lipid production at temperatures far exceeding those of other fungi under development for industrial applications. These results establishK. marxianusas a synthetic biology platform comparable toS. cerevisiae, with naturally more robust traits that hold potential for the industrial production of renewable chemicals.IMPORTANCEThe yeastKluyveromyces marxianusgrows at high temperatures and on a wide range of carbon sources, making it a promising host for industrial biotechnology to produce renewable chemicals from plant biomass feedstocks. However, major genetic engineering limitations have kept this yeast from replacing the commonly used yeastSaccharomyces cerevisiaein industrial applications. Here, we describe genetic tools for genome editing and breedingK. marxianusstrains, which we use to create a new thermotolerant strain with promising fatty acid production. These results open the door to usingK. marxianusas a versatile synthetic biology platform organism for industrial applications.

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Liu, Fang, Pandi Wang, Xiaojuan Xiong, Xinhua Zeng, Xiaobo Zhang, and Gang Wu. "A Review of Nervonic Acid Production in Plants: Prospects for the Genetic Engineering of High Nervonic Acid Cultivars Plants." Frontiers in Plant Science 12 (March 5, 2021). http://dx.doi.org/10.3389/fpls.2021.626625.

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Nervonic acid (NA) is a very-long-chain monounsaturated fatty acid that plays crucial roles in brain development and has attracted widespread research interest. The markets encouraged the development of a refined, NA-enriched plant oil as feedstocks for the needed further studies of NA biological functions to the end commercial application. Plant seed oils offer a renewable and environmentally friendly source of NA, but their industrial production is presently hindered by various factors. This review focuses on the NA biosynthesis and assembly, NA resources from plants, and the genetic engineering of NA biosynthesis in oil crops, discusses the factors that affect NA production in genetically engineered oil crops, and provides prospects for the application of NA and prospective trends in the engineering of NA. This review emphasizes the progress made toward various NA-related topics and explores the limitations and trends, thereby providing integrated and comprehensive insight into the nature of NA production mechanisms during genetic engineering. Furthermore, this report supports further work involving the manipulation of NA production through transgenic technologies and molecular breeding for the enhancement of crop nutritional quality or creation of plant biochemical factories to produce NA for use in nutraceutical, pharmaceutical, and chemical industries.

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Wu, Xiao-Xi, Jian-Wei Li, Su-Fang Xing, Hui-Ting Chen, Chao Song, Shu-Guang Wang, and Zhen Yan. "Establishment of a resource recycling strategy by optimizing isobutanol production in engineered cyanobacteria using high salinity stress." Biotechnology for Biofuels 14, no. 1 (August 30, 2021). http://dx.doi.org/10.1186/s13068-021-02023-8.

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Abstract Background Isobutanol is an attractive biofuel with many advantages. Third-generation biorefineries that convert CO2 into bio-based fuels have drawn considerable attention due to their lower feedstock cost and more ecofriendly refining process. Although autotrophic cyanobacteria have been genetically modified for isobutanol biosynthesis, there is a lack of stable and convenient strategies to improve their production. Results In this study, we first engineered Synechococcus elongatus for isobutanol biosynthesis by introducing five exogenous enzymes, reaching a production titer of 0.126 g/L at day 20. It was then discovered that high salinity stress could result in a whopping fivefold increase in isobutanol production, with a maximal in-flask titer of 0.637 g/L at day 20. Metabolomics analysis revealed that high salinity stress substantially altered the metabolic profiles of the engineered S. elongatus. A major reason for the enhanced isobutanol production is the acceleration of lipid degradation under high salinity stress, which increases NADH. The NADH then participates in the engineered isobutanol-producing pathway. In addition, increased membrane permeability also contributed to the isobutanol production titer. A cultivation system was subsequently developed by mixing synthetic wastewater with seawater to grow the engineered cyanobacteria, reaching a similar isobutanol production titer as cultivation in the medium. Conclusions High salinity stress on engineered cyanobacteria is a practical and feasible biotechnology to optimize isobutanol production. This biotechnology provides a cost-effective approach to biofuel production, and simultaneously recycles chemical nutrients from wastewater and seawater.

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Journal articles on the topic 'Genetically engineered feedstocks'

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