DOCSLIB.ORG

Gas Fermentation of C1 Feedstocks: Commercialization Status and Future Prospects

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Gas Fermentation of C1 Feedstocks: Commercialization Status and Future Prospects Review Gas fermentation of C1 feedstocks: commercialization status and future prospects Leonardo V. Teixeira, Liza F. Moutinho, and Aline S. Romão-Dumaresq, SENAI Innovation Institute for Biosynthetics, Technology Center for Chemical and Textile Industry, Rio de Janeiro, Brazil Received December 04, 2017; revised June 04, 2018; accepted June 05, 2018 View online at Wiley Online Library (wileyonlinelibrary.com); DOI: 10.1002/bbb.1912; Biofuels, Bioprod. Bioref. (2018) Abstract: The increasing emissions of carbon dioxide, methane and carbon oxide (collectively referred as C1 compounds) are likely to configure a major contribution to global warming and other envi- ronmental issues. The implementation of carbon capture and storage (CCS) is considered a crucial strategy to prevent global warming, but the overall costs of currently available CCS technologies are still prohibitive for its large-scale deployment. Using microorganisms capable of assimilating C1 com- pounds for producing value-added products could be an important driver for mitigating emissions and minimizing their deleterious consequences, while simultaneously deriving additional economic benefits from these compounds. This review summarizes the main microorganisms and metabolic routes being investigated, with special focus on both the products targeted and the current industrial initiatives. There are a number of companies investing in these routes and in some instances commercial deploy- ment was identified. Despite the variety of commercially-appealing products, genetic manipulation of microorganisms to maximize yields and the design of technologies capable of efficiently using the gas- eous feedstocks are major challenges yet to be overcome to fully unlock the potential of C1 microbio- logical routes. © 2018 Society of Chemical Industry and John Wiley & Sons, Ltd Keywords: technology status; carbon dioxide; methane; biotechnology; greenhouse gas; C1 feedstock Introduction effect of countries’ mitigation pledges in terms of annual emissions of greenhouse gases (GHG) does not seem to Many of the world’s ecosystems are already overexploited suffice.2 and unsustainable. With the expected increase in both Based on 2016 figures, carbon dioxide (CO2) accounts global population and its average per capita income, for the largest proportion of GHG emissions on a CO2 demand for natural resources will rise accordingly. In equivalent basis, surpassing 90% of emissions, followed by addition, climate change derived from human activities methane (CH4) (Fig. 1). In absolute values, CO2 emissions, could aggravate environmental issues by affecting agricul- which were at 28.8 Gt in 2007, could reach 40.2 Gt in 2030.5 tural productivity and water supplies.1 Despite the urgent The implementation of carbon capture and storage (CCS) is need to restrain the increase in global average temperature considered a crucial strategy to combat global warming, but emphasized in the 2015 Paris Agreement, the aggregate the overall costs of currently available CCS technologies are Correspondence to: Leonardo V. Teixeira, ISI for Biosynthetics, Technology Center for Chemical and Textile Industry, Rio de Janeiro, Brazil. E-mail: [email protected] © 2018 Society of Chemical Industry and John Wiley & Sons, Ltd 1 LV Teixeira, LF Moutinho, AS Romão-Dumaresq Review: Commercialization status of gas fermentation of C1 feedstocks ecules, the knowledge of attainable products is relevant for informed decision making. In a related matter, it would be desirable to identify meta- bolic pathways in which versatile metabolic intermediates are reached, as a way of unlocking different molecules of interest. Another alternative to achieve versatility is by producing the so-called platform chemicals, that is, chemical intermedi- ates capable of yielding a large set of derivatives through physical and/or chemical transformations, targeting several distinct end uses.14 Note that this definition is similar to that of a metabolic building block, but the former is generally transformed via chemical reactions, whereas the latter is bio- chemically transformed (within the microbial host). The present paper is organized as follows. First, we describe C1 feedstocks, their sources and generation data (focusing on Brazil and the USA). Then, we provide an overview of available biochemical routes and microbial hosts using each feedstock, as well as the product applica- Figure 1. Global greenhouse gas (GHG) emissions tions and players involved. We finalize by outlining the breakdown on a GtCO2 equivalent basis (based on 2016 most advanced initiatives, opportunities and challenges data).3Notes: (1) CO and H are excluded from the chart 2 related to C1 feedstock bioconversion. because they are not direct GHGs. However, they have indirect effects on global warming and climate change.4 C1 Feedstocks still prohibitive for its large-scale deployment.6 Using CO 2 Many of the efforts to attain sustainable production of and other one-carbon feedstocks such as CH and carbon 4 fuels, chemicals, and materials envision the use of biomass monoxide (CO) (collectively referred to as C1 feedstocks) feedstocks.15 First-generation sugars, obtained mainly for obtaining value-added products could be an important from sugarcane and corn, are already well established and driver for mitigating GHG emissions and minimizing their used in fermentative processes such as the production of deleterious consequences. The latter approach is known as ethanol. However, there is a concern that an expansion carbon capture and utilization (CCU). in fermentation processes could impose pressure on food Other than the use of CH as an energy source (in natural 4 supplies.16 Second-generation sugars derived from ligno- gas) and as feedstock for synthesis gas (or syngas, a mixture cellulosic residues are an alternative to overcome this issue of CO, CO and H ), it is also employed as a precursor to 2 2 but their widespread use is still hindered by the general chemicals, especially ammonia and methanol.7 The uses absence of cost-effective technologies for overcoming the of CO include the production of urea, inorganic carbon- 2 residues’ recalcitrance17 and also by difficulties related ates and pigments, methanol, salicylic acid and propylene to logistics and storage.18 Besides the sugars in lignocel- carbonate, besides direct use in carbonated beverages and lulosic residues, they also contain lignin ranging from in food processing and preservation.8,9 Finally, CO is used 10% to 35% of the biomass19 and, despite initiatives to add as a fuel, as a metallurgical reducing agent, and to pro- value to lignin,20 it is mostly burned for energy generation. duce methanol, acetic acid, phosgene, and oxo alcohols.10 Alternative C1-based feedstocks for biochemical processes Biological production systems relying on either photosyn- do not compete with food supplies or underutilize the thetic or nonphotosynthetic microorganisms that assimilate feedstock and also have a significantly lower cost. The C1 feedstocks could significantly broaden this spectrum.11 main ones being investigated are described below. Some excellent reviews focus on the related microorgan- 11–13 isms, metabolic routes and process advances, but there Syngas and CO-rich industrial Off-gases is no current product-oriented review highlighting tech- nologies at commercialization status. Considering the very A possible solution to use nearly all of the biomass content cost-intensive and time-consuming nature of developing a is its gasification to syngas, a mixture ranging from 30% microbial strain capable of feasibly producing target mol- to 60% CO, 25% to 30% H2, 5% to 15% CO2 and 0% to 5% 2 © 2018 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. (2018); DOI: 10.1002/bbb Review: Commercialization status of gas fermentation of C1 feedstocks LV Teixeira, LF Moutinho, AS Romão-Dumaresq 21 3 CH4, followed by its biochemical processing. Syngas is around 52 billion m per year according to recent esti- a traditional raw material for Fischer–Tropsch synthesis, mates.33 The current production of biogas is only between which consists of the catalytic polymerization and hydro- 0.2 and 0.6 billion m3 annually.34 For the sake of com- genation of CO to a multiphase mixture of hydrocarbons, parison, the current US production of CH4 from these oxygenates, and water (syncrude). Syncrude refining yields secondary sources is nearly 13.9 billion m3 per year.12 transportation fuels and other drop-in chemicals.22 The Fischer–Tropsch process has some drawbacks when com- Carbon dioxide (CO2) pared with biological processes, such as high reaction tem- The use of CO as a feedstock for fermentative processes peratures (150–350 °C) and pressures (up to 30 bar), low 2 is also being envisioned. Although being a major, concen- product selectivity, greater susceptibility of the inorganic trated source of CO emissions, fossil-fuel power-plant catalysts to poisoning by sulfur, chlorine and tars, and ele- 2 emissions present low concentrations of CO and many vated costs.23 In contrast, when performing fermentation 2 impurities including NO and SO compounds (especially with syngas, a relevant issue is the presence of hydrogen x x in coal-fired units),35 which makes CCS a more suitable cyanide,24,25 which is toxic to microorganisms. option in this case. Oil refineries emit around 900 mil- Besides agricultural waste, municipal solid waste and lion tons of CO per year, approximately the same amount organic industrial
Load more

Recommended publications
  • Discovery, Taxonomic Distribution, and Phenotypic Characterization of a Gene Required for 3-Methylhopanoid Production
    Discovery, taxonomic distribution, and phenotypic characterization of a gene required for 3-methylhopanoid production The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation Welander, P. V., and R. E. Summons. “Discovery, Taxonomic Distribution, and Phenotypic Characterization of a Gene Required for 3-methylhopanoid Production.” Proceedings of the National Academy of Sciences 109.32 (2012): 12905–12910. CrossRef. Web. As Published http://dx.doi.org/10.1073/pnas.1208255109 Publisher National Academy of Sciences (U.S.) Version Final published version Citable link http://hdl.handle.net/1721.1/77589 Terms of Use Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. Discovery, taxonomic distribution, and phenotypic characterization of a gene required for 3-methylhopanoid production Paula V. Welander1 and Roger E. Summons Department of Earth, Atmospheric, and Planetary Sciences, Massachusetts Institute of Technology, 77 Massachusetts Avenue, E25-629, Cambridge, MA 02139 Edited by John M. Hayes, Woods Hole Oceanographic Institution, Berkeley, CA, and approved July 2, 2012 (received for review May 15, 2012) Hopanoids methylated at the C-3 position are a subset of bacterial majority of C-3 methylated hopanoid producers are aerobic triterpenoids that are readily preserved in modern and ancient se- methanotrophs. However, the production of 3-methylhopanoids diments and in petroleum. The production of 3-methylhopanoids has also been demonstrated in the acetic acid bacteria (12) indi- by extant aerobic methanotrophs and their common occurrence cating that the taxonomic distribution of 3-methylhopanoids is in modern and fossil methane seep communities, in conjunction not restricted to methanotrophs.
    [Show full text]
  • Syngas Derived from Lignocellulosic Biomass Gasification As An
    processes Review Syngas Derived from Lignocellulosic Biomass Gasification as an Alternative Resource for Innovative Bioprocesses Cosetta Ciliberti 1, Antonino Biundo 1,2, Roberto Albergo 3 , Gennaro Agrimi 1,2 , Giacobbe Braccio 3, Isabella de Bari 3 and Isabella Pisano 1,2,* 1 Department of Bioscience, Biotechnology and Biopharmaceutics, University of Bari, Via Edoardo Orabona, 4, 70125 Bari, Italy; [email protected] (C.C.); [email protected] (A.B.); [email protected] (G.A.) 2 Interuniversity Consortium for Biotechnology (CIB), 34100 Trieste, Italy 3 Italian National Agency for New Technologies, Energy and Sustainable Economic Development (ENEA), Division of Bioenergy, Biorefinery and Green Chemistry, C.R. Trisaia S.S. 106 Jonica, 75026 Rotondella (MT), Italy; [email protected] (R.A.); [email protected] (G.B.); [email protected] (I.d.B.) * Correspondence: [email protected] Received: 3 November 2020; Accepted: 24 November 2020; Published: 28 November 2020 Abstract: A hybrid system based on lignocellulosic biomass gasification and syngas fermentation represents a second-generation biorefinery approach that is currently in the development phase. Lignocellulosic biomass can be gasified to produce syngas, which is a gas mixture consisting mainly of H2, CO, and CO2. The major challenge of biomass gasification is the syngas’s final quality. Consequently, the development of effective syngas clean-up technologies has gained increased interest in recent years. Furthermore, the bioconversion of syngas components has been intensively studied using acetogenic bacteria and their Wood–Ljungdahl pathway to produce, among others, acetate, ethanol, butyrate, butanol, caproate, hexanol, 2,3-butanediol, and lactate. Nowadays, syngas fermentation appears to be a promising alternative for producing commodity chemicals in comparison to fossil-based processes.
    [Show full text]
  • Gas Fermentation at Calysta
    Gas Fermentation at Calysta Intro for REMEDY workshop October 20, 2020 Gas Fermentation is the Next Step in Industrial Biotech • Lower cost feedstocks are needed for biological products to compete with petroleum derivatives • C1 feedstocks (CH4, CO, CO2) are accepted to be among the cheapest sources of carbon • C1 feedstocks are generally pollutants, with significant safety and solubility issues compared to traditional biofeedstocks • Calysta owns the world’s only commercially-validated gas fermentation technology allowing use of C1 feedstocks 22/10/2020 2 Calysta’s Methanotroph Platform Methylococcus capsulatus Bath • Gammaproteobacteria, type I methanotroph • Relatively fast growth rate (methane:oxygen mix) • Genome sequence available • Amenable to genetic manipulation • Only methanotroph proven at commercial scale • Variety of formats for strain testing: well plates, pressure bottles, 2L fermenters • Amended media and optimized feeding strategies produce high cell densities in small scale. 22/10/2020 3 3 Calysta’s Platform: Strain Engineering Calysta has developed a set of novel engineering tools for methanotrophs: • Reporter genes Different promoter gene-fusions with synthetic fluorescent and chromogenic proteins (non-Aequorea) expressed in M. capsulatus • Plasmids that replicate both in methanotrophs and in E. coli • Constitutive and inducible (low/med/high) promoters • Techniques for chromosomal knockin and knockouts 4 Calysta Performs Methanotrophic Fermentation at All Scales Fermentors High Throughput TPP NorFerm Commercial Plant
    [Show full text]
  • Comunicación Para Una Sociedad Sostenible Proyecto Profesoral Aprobado Por El Cci, Soberanía Alimentaria Y Comunicación Para La Sostenibilidad
    COMUNICACIÓN PARA UNA SOCIEDAD SOSTENIBLE PROYECTO PROFESORAL APROBADO POR EL CCI, SOBERANÍA ALIMENTARIA Y COMUNICACIÓN PARA LA SOSTENIBILIDAD JOSÉ LUIS GARCÍA RUÍZ CODIGO 211722 UNIVERSIDAD AUTÓNOMA DE OCCIDENTE FACULTAD DE HUMANIDADES Y ARTES DEPARTAMENTO DE DISEÑO PROGRAMA COMUNICACIÓN PUBLICITARIA SANTIAGO DE CALI 2019 COMUNICACIÓN PARA UNA SOCIEDAD SOSTENIBLE PROYECTO PROFESORAL APROBADO POR EL CCI, SOBERANÍA ALIMENTARIA Y COMUNICACIÓN PARA LA SOSTENIBILIDAD JOSÉ LUIS GARCÍA RUÍZ CODIGO 2117228 Pasantía de investigación para optar al título de Publicista Director Ana Lucia Jiménez Maestría en Educación UNIVERSIDAD AUTÓNOMA DE OCCIDENTE FACULTAD DE HUMANIDADES Y ARTES DEPARTAMENTO DE DISEÑO PROGRAMA COMUNICACIÓN PUBLICITARIA SANTIAGO DE CALI 2019 Nota de aceptación: Aprobado por el Comité de Grado en cumplimiento de los requisitos exigidos por la Universidad Autónoma de Occidente para optar al título de Publicista JULIAN HERNANDEZ Jurado LIBARDO MAYA Jurado Santiago de Cali, 20 de Noviembre de 2019 CONTENIDO pág. RESUMEN 6 INTRODUCCIÓN 11 1. PRESENTACIÓN DEL GRUPO DE INVESTIGACIÓN 12 2. JUSTIFICACIÓN 16 3. OBJETIVOS DE LA PASANTIA 17 3.1 OBJETIVO GENERAL 17 3.1.1 Objetivos específicos 17 4. PLAN DE TRABAJO 18 5. ELEMENTOS INNOVADORES DE LA PROPUESTA 20 5.1 BUSCAR EL ESTADO DEL ARTE PARA EVIDENCIAR TODO LO ESCRITO SOBRE LAS PALABRAS CLAVES. 20 5.2 REFORZAR EL MARCO TEORICO 55 5.3 DESARROLLAR TRABAJO DE CAMPO EJECUTANDO LA ENTREVISTA CON BASE AL MARCO TEÓRICO ANTES MENCIONADO. 59 5.3.1 Sociedad de consumo 78 5.3.2 Consumo simbólico 79 5.3.3 La nueva identidad del consumidor 80 6. RECURSOS 82 4 7. CRONOGRAMA DE ACTIVIDADES 83 8.
    [Show full text]
  • Advanced Alternative Fuel Pathways: Technology Overview and Status
    WORKING PAPER 2019-13 Advanced alternative fuel pathways: Technology overview and status Authors: Chelsea Baldino, Rosalie Berg, Nikita Pavlenko, Stephanie Searle Date: July 19, 2019 Keywords: Thermochemical conversion; biochemical conversion; non-food feedstock; lignocellulosic biomass; biofuels; renewable fuel; electrofuels Introduction and summarized in this paper. For Fischer-Tropsch (FT) synthesis and this figure, as well for all the follow- methanation, before it can be used as Policymakers recognize the signifi- ing figures in this paper that illustrate a transportation fuel. Hydrothermal cant greenhouse gas (GHG) savings the individual conversion technology liquefaction and fast pyrolysis are that are possible by transitioning from pathways in more detail, yellow boxes other primary conversion technolo- first-generation, food-based biofuels indicate processes and green boxes gies that process biomass into crude to alternative, non-food based fuels, represent an input to the process. oils, which are then fed into hydropro- i.e., advanced alternative fuels. First- Blue boxes represent the desired cessing or other upgrading technolo- generation biofuels are produced by fuel output, whereas gray boxes are gies to produce drop-in fuel. converting readily available biomass intermediary products. Bold arrows chemicals—sugars, starch or oils—into indicate primary steps in the conver- This paper synthesizes the best avail- transportation fuels such as ethanol sion process; narrow arrows indicate able information on the feedstocks and fatty acid methyl esters (FAME). less common, or less important, steps. that are or could be suitable for each Advanced biofuels, on the other hand, The black, dashed box groups pro- pathway. Overall, these feedstocks are those produced by converting cesses that can be applied to the include materials such as lignocel- additional chemicals contained in oxygen and hydrogen that is produced lulosic biomass, municipal or indus- the biomass feedstocks, specifically in the PtX pathway.
    [Show full text]
  • Syngas Fermentation: Quantification of Assay Techniques, Reaction Kinetics, and Pressure Dependencies of the Clostridial P11 Hydrogenase
    Brigham Young University BYU ScholarsArchive Theses and Dissertations 2010-03-18 Syngas Fermentation: Quantification of Assay Techniques, Reaction Kinetics, and Pressure Dependencies of the Clostridial P11 Hydrogenase Bradley E. Skidmore Brigham Young University - Provo Follow this and additional works at: https://scholarsarchive.byu.edu/etd Part of the Chemical Engineering Commons BYU ScholarsArchive Citation Skidmore, Bradley E., "Syngas Fermentation: Quantification of Assay Techniques, Reaction Kinetics, and Pressure Dependencies of the Clostridial P11 Hydrogenase" (2010). Theses and Dissertations. 2098. https://scholarsarchive.byu.edu/etd/2098 This Thesis is brought to you for free and open access by BYU ScholarsArchive. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of BYU ScholarsArchive. For more information, please contact [email protected], [email protected]. Syngas Fermentation: Quantification of Assay Techniques, Reaction Kinetics, and Pressure Dependencies of the Clostridial P11 Hydrogenase Bradley Skidmore A thesis submitted to the faculty of Brigham Young University in partial fulfillment of the requirements for the degree of Master of Science Randy S. Lewis, Chair Steven R. Herron Thomas A. Knotts Department of Chemical Engineering Brigham Young University April 2010 Copyright © 2010 Bradley Skidmore All Rights Reserved ABSTRACT Syngas Fermentation: Quantification of Assay Techniques, Reaction Kinetics, and Pressure Dependencies of the Clostridial P11 Hydrogenase Bradley Skidmore Department of Chemical Engineering Master of Science Ethanol usage as a transportation fuel is rapidly increasing in the United States. Production of ethanol from cellulose feedstocks via gasification followed by syngas fermentation offers an environmentally friendly approach that mitigates many of the adverse effects associated with production from corn.
    [Show full text]
  • Microbial Fermentation of Syngas/CO
    Microbial fermentation of syngas/CO My PhD project started in April 2009, under the supervision of professor Madalena Alves and professor Diana Sousa, from University of Minho, Portugal. During my third year of PhD program, I’m developing my research activities at the Department of Microbiology, University of Wageningen, Holland, under the supervision of Professor Alfons Stams and Doctor Caroline Plugge. Syngas or synthesis gas is produced during the gasification of different materials, e.g. coal, oil and natural gas, tar sands, recalcitrant wastes, lignocellulosic biomass, and sewage sludge. The principal components of syngas are carbon monoxide (CO), hydrogen (H2), and carbon dioxide (CO2). Both catalytic and biological processes can be used for the production of biofuels and bulk chemicals from syngas. The development of novel bioprocesses for syngas conversion to added-value products is a promising field comprising some advantages over the chemical processes. However, biological conversion of syngas is still rather unexplored within the bioprocess engineering community. The main objectives of my research are to explore the potential of biofuels or other valuable compounds production from syngas, and to get more insights about the microbiology of the syngas fermentation process. The screening of different anaerobic inocula, at different temperatures (mesophilic and thermophilic conditions), the achievement of enriched mixed cultures degrading syngas, and the isolation of new bacterial strains and further characterization is being carried out in order to get more insights into the physiological and biochemical aspects of syngas fermentation. This work results from the cooperation between the Wageningen University, Laboratory of Microbiology and University of Minho, Centre of Biological Engineering, being supported by the Fundação para a Ciência e Tecnologia (FCT), Portugal, through the PhD grant SFRH/BD/48965/2008 given to Joana I.
    [Show full text]
  • A Genome-Scale Metabolic Model for Methylococcus Capsulatus Predicts Reduced Efficiency Uphill Electron Transfer to Pmmo
    bioRxiv preprint doi: https://doi.org/10.1101/329714; this version posted May 24, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 A genome-scale metabolic model for Methylococcus 2 capsulatus predicts reduced efficiency uphill electron 3 transfer to pMMO. 4 Christian Lieven*1, [email protected] 5 Leander A. H. Petersen2, 3, [email protected] 6 Sten Bay Jørgensen3, [email protected] 7 Krist V. Gernaey3, [email protected] 8 Markus J. Herrgard1, [email protected] 9 Nikolaus Sonnenschein1, [email protected] 10 11 1The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark 12 2Unibio A/S, 13 3Department of Chemical Engineering, Technical University of Denmark 14 * Corresponding Author 15 16 17 18 19 20 21 22 1 bioRxiv preprint doi: https://doi.org/10.1101/329714; this version posted May 24, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Abstract 2 Background: 3 Genome-scale metabolic models allow researchers to calculate yields, 4 to predict consumption and production rates, and to study the effect of 5 genetic modifications in silico, without running resource-intensive 6 experiments. While these models have become an invaluable tool for 7 optimizing industrial production hosts like E.
    [Show full text]
  • Development of a CRISPR/Cas9 System for Methylococcus Capsulatus in Vivo Gene Editing
    GENETICS AND MOLECULAR BIOLOGY crossm Development of a CRISPR/Cas9 System for Methylococcus capsulatus In Vivo Gene Editing Timothy Tapscott,a Michael T. Guarnieri,a Calvin A. Henarda aNational Bioenergy Center, National Renewable Energy Laboratory (NREL), Golden, Colorado, USA Downloaded from ABSTRACT Methanotrophic bacteria play a crucial role in the Earth’s biogeochemi- cal cycle and have the potential to be employed in industrial biomanufacturing pro- cesses due to their capacity to use natural gas- and biogas-derived methane as a sole carbon and energy source. Advanced gene-editing systems have the potential to enable rapid, high-throughput methanotrophic genetics and biocatalyst develop- ment. To this end, we employed a series of broad-host-range expression plasmids to construct a conjugatable clustered regularly interspaced short palindromic repeats http://aem.asm.org/ (CRISPR)/Cas9 gene-editing system in Methylococcus capsulatus (Bath). Heterologous coexpression of the Streptococcus pyogenes Cas9 endonuclease and a synthetic sin- gle guide RNA (gRNA) showed efficient Cas9 DNA targeting and double-stranded DNA (dsDNA) cleavage that resulted in cell death. We demonstrated effective in vivo editing of plasmid DNA using both Cas9 and Cas9D10A nickase to convert green fluo- rescent protein (GFP)- to blue fluorescent protein (BFP)-expressing cells with 71% ef- ficiency. Further, we successfully introduced a premature stop codon into the soluble methane monooxygenase (sMMO) hydroxylase component-encoding mmoX gene with D10A the Cas9 nickase, disrupting sMMO function. These data provide proof of concept on July 17, 2019 by guest for CRISPR/Cas9-mediated gene editing in M. capsulatus. Given the broad-host-range replicons and conjugation capability of these CRISPR/Cas9 tools, they have potential util- ity in other methanotrophs and a wide array of Gram-negative microorganisms.
    [Show full text]
  • Evolution of Methanotrophy in the Beijerinckiaceae&Mdash
    The ISME Journal (2014) 8, 369–382 & 2014 International Society for Microbial Ecology All rights reserved 1751-7362/14 www.nature.com/ismej ORIGINAL ARTICLE The (d)evolution of methanotrophy in the Beijerinckiaceae—a comparative genomics analysis Ivica Tamas1, Angela V Smirnova1, Zhiguo He1,2 and Peter F Dunfield1 1Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada and 2Department of Bioengineering, School of Minerals Processing and Bioengineering, Central South University, Changsha, Hunan, China The alphaproteobacterial family Beijerinckiaceae contains generalists that grow on a wide range of substrates, and specialists that grow only on methane and methanol. We investigated the evolution of this family by comparing the genomes of the generalist organotroph Beijerinckia indica, the facultative methanotroph Methylocella silvestris and the obligate methanotroph Methylocapsa acidiphila. Highly resolved phylogenetic construction based on universally conserved genes demonstrated that the Beijerinckiaceae forms a monophyletic cluster with the Methylocystaceae, the only other family of alphaproteobacterial methanotrophs. Phylogenetic analyses also demonstrated a vertical inheritance pattern of methanotrophy and methylotrophy genes within these families. Conversely, many lateral gene transfer (LGT) events were detected for genes encoding carbohydrate transport and metabolism, energy production and conversion, and transcriptional regulation in the genome of B. indica, suggesting that it has recently acquired these genes. A key difference between the generalist B. indica and its specialist methanotrophic relatives was an abundance of transporter elements, particularly periplasmic-binding proteins and major facilitator transporters. The most parsimonious scenario for the evolution of methanotrophy in the Alphaproteobacteria is that it occurred only once, when a methylotroph acquired methane monooxygenases (MMOs) via LGT.
    [Show full text]
  • Production of Alcohols Via Syngas Fermentation Using
    PRODUCTION OF ALCOHOLS VIA SYNGAS FERMENTATION USING ALKALIBACULUM BACCHI MONOCULTURE AND A MIXED CULTURE By KAN LIU Bachelor of Science in Pharmaceutical Engineering Tianjin University of Science & Technology Tianjin, China 2006 Master of Science in Fermentation Engineering Tianjin University of Science & Technology Tianjin, China 2009 Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY December, 2013 PRODUCTION OF ALCOHOLS VIA SYNGAS FERMENTATION USING ALKALIBACULUM BACCHI MONOCULTURE AND A MIXED CULTURE Dissertation Approved: Dr. Hasan K. Atiyeh Dissertation Adviser Dr. Babu Z. Fathepure Dr. Raymond L. Huhnke Dr. Mark R. Wilkins ii ACKNOWLEDGEMENTS I appreciate my advisor Dr. Hasan Atiyeh giving me the opportunity to study in the United States and obtain my Ph.D. at Oklahoma State University. During the four years study, he gave me a lot of encouragements and guidance on my Ph.D. project, making me grow up quickly and become more professional in the way to think, to perform and to present. The experience and knowledge I learned from him will be a precious treasure for my future career. I also acknowledge the financial support for this project from USDA-NIFA 2010-34447-20772, Oklahoma NSF EPSCoR, Oklahoma Bioenergy Center and the Oklahoma Agricultural Experiment Station. I also appreciate Dr. Raymond Huhnke for guidance on my Ph.D. project, presentations and papers. Also, I am very grateful to Dr. Mark Wilkins’s help and support for designing my experiments, reviewing my papers, presentations and providing valuable suggestions, especially giving me a precious opportunity to visit Washington D.C.
    [Show full text]
  • Methanotrophic Bacterial Biomass As Potential Mineral Feed Ingredients for Animals
    International Journal of Environmental Research and Public Health Article Methanotrophic Bacterial Biomass as Potential Mineral Feed Ingredients for Animals Agnieszka Ku´zniar 1,*, Karolina Furtak 2 , Kinga Włodarczyk 1, Zofia St˛epniewska 1 and Agnieszka Woli ´nska 1 1 Department of Biochemistry and Environmental Chemistry, The John Paul II Catholic University of Lublin, Konstantynów St. 1 I, 20-708 Lublin, Poland 2 Department of Agriculture Microbiology, Institute of Soil Sciences and Plant Cultivation State Research Institute, Czartoryskich St. 8, 24-100 Puławy, Poland * Correspondence: [email protected]; Tel.: +48-81-454-5461 Received: 14 June 2019; Accepted: 23 July 2019; Published: 26 July 2019 Abstract: Microorganisms play an important role in animal nutrition, as they can be used as a source of food or feed. The aim of the study was to determine the nutritional elements and fatty acids contained in the biomass of methanotrophic bacteria. Four bacterial consortia composed of Methylocystis and Methylosinus originating from Sphagnum flexuosum (Sp1), S. magellanicum (Sp2), S. fallax II (Sp3), S. magellanicum IV (Sp4), and one composed of Methylocaldum, Methylosinus, and Methylocystis that originated from coalbed rock (Sk108) were studied. Nutritional elements were determined using the flame atomic absorption spectroscopy technique after a biomass mineralization stage, whereas the fatty acid content was analyzed with the GC technique. Additionally, the growth of biomass and dynamics of methane consumption were monitored. It was found that the methanotrophic biomass 1 contained high concentrations of K, Mg, and Fe, i.e., approx. 9.6–19.1, 2.2–7.6, and 2.4–6.6 g kg− , respectively.
    [Show full text]