Impact of Emerging Technologies on the Cell Disruption and Fractionation of Microalgal Biomass Rui Zhang
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Cell Disruption
Application Note Use of Microfluidizer™ technology for cell disruption. They are difficult and time consuming to clean, which This Application Note gives an overview of has to be done for every sample. Most manufacturers the techniques used for cell disruption. In of French Presses have discontinued production but addition this is a summary of why a they are still in use, available from small companies Microfluidizer is best suited for this and second hand. application and the specific advantages Microfluidics technology has over High pressure homogenizers (HPH) These devices alternative cell disruption methods. Also are the next best alternative to Microfluidizers for cell disruption. Prices are typically equal to, or lower, than included are tips for optimal cell processing Microfluidizers. Cooling, cleaning, wear (valves!) and with a Microfluidizer scalability can be issues. In particular if we look All cell disruption methods are not created beyond simply the % of cells ruptured to the quality and usability of the ruptured suspension the equal. Results published in the scientific Microfluidizer is the clear winner. Table 1 highlights literature show that the disruption method the increased yield from a Microfluidizer compared to strongly influences the physical-chemical an HPH. properties of the disintegrate, such as Ultrasonication: utilizes cavitational forces. Often particle size, disruption efficiency, viscosity used for very small sample volumes, the cell and protein release.1,2 For all of these suspension is sonicated with an ultrasonic probe. Local important parameters the Microfluidizer high temperatures, resulting in low yields2,4, scalability comes out tops. and noise are the main issues with this technology. -
Effects of Temperature, Light Intensity and Quality, Carbon Dioxide, and Culture Medium Nutrients on Growth and Lipid Production of Ettlia Oleoabundans
Effects of Temperature, Light Intensity and Quality, Carbon Dioxide, and Culture Medium Nutrients on Growth and Lipid Production of Ettlia oleoabundans by Ying Yang A Dissertation Submitted to the Faculty of WORCESTER POLYTECHNIC INSTITUTE in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biology and Biotechnology by December 2013 Approved by: Dr. Pamela Weathers, Advisor Dr. Robert Thompson, Committee Member Dr. Luis Vidali, Committee Member Dr. Reeta Rao, Committee Member “A journey of a thousand miles begins with a single step.” — Lao Tzu (604 BC – 531 BC) ii Abstract Ettlia oleoabundans, a freshwater green microalga, was grown under different environmental conditions to study its growth, lipid yield and quality for a better understanding of the fundamental physiology of this oleaginous species. E. oleoabundans showed steady increase in biomass under low temperature and low light intensity, and at high temperature lipid cell content significantly increased independent of nitrate depletion. Studies on light quality showed that red light treatment did not change the biomass concentration, but stimulated lipid yield especially oleic acid, the most desirable biodiesel precursor. Moreover, no photoreversibility in lipid production was observed when applying alternating short-term red and far-red lights, which left the phytochrome effect still an open question. In addition, carbon dioxide enrichment via an air sparging system significantly boosted exponential growth and increased carbon conversion efficiency. Finally, a practical study demonstrated the feasibility of growing E. oleoabundans for high lipid production using a diluted agricultural anaerobic waste effluent as the medium. Together, these studies showed the potential of E. oleoabundans as a promising high yield feedstock for the production of high quality biodiesel. -
Transcriptional Landscapes of Lipid Producing Microalgae Benoît M
Transcriptional landscapes of lipid producing microalgae Benoît M. Carrères 2019 Transcriptional landscapes of lipid producing microalgae Benoî[email protected]:~$ ▮ Transcriptional landscapes of lipid producing microalgae Benoît Manuel Carrères Thesis committee Promotors Prof. Dr Vitor A. P. Martins dos Santos Professor of Systems and Synthetic Biology Wageningen University & Research Prof. Dr René H. Wij$els Professor of Bioprocess Engineering Wageningen University & Research Co-promotors Dr Peter J. Schaa% Associate professor* Systems and Synthetic Biology Wageningen University & Research Dr Dirk E. Martens Associate professor* Bioprocess Engineering Wageningen University & Research ,ther mem-ers Prof. Dr Alison Smith* University of Cam-ridge Prof. Dr. Dic+ de Ridder* Wageningen University & Research Dr Aalt D.). van Di#+* Wageningen University & Research Dr Ga-ino Sanche/(Pere/* Genetwister* Wageningen This research 0as cond1cted under the auspices of the .rad1ate School V2A. 3Advanced studies in Food Technology* Agro-iotechnology* Nutrition and Health Sciences). Transcriptional landscapes of lipid producing microalgae Benoît Manuel Carrères Thesis su-mitted in ful8lment of the re9uirements for the degree of doctor at Wageningen University -y the authority of the Rector Magnificus, Prof. Dr A.P.). Mol* in the presence of the Thesis' ommittee a%%ointed by the Academic Board to be defended in pu-lic on Wednesday 2; Novem-er 2;<= at 1.>; p.m in the Aula. Benoît Manuel Carrères 5ranscriptional landsca%es of lipid producing -
Genetic Engineering of Green Microalgae for the Production of Biofuel and High Value Products
GENETIC ENGINEERING OF GREEN MICROALGAE FOR THE PRODUCTION OF BIOFUEL AND HIGH VALUE PRODUCTS Joanna Beata Szaub Department of Structural and Molecular Biology University College London A thesis submitted for the degree of Doctor of Philosophy August 2012 DECLARATION I, Joanna Beata Szaub confirm that the work presented in this thesis is my own. Where information has been derived from other sources, I confirm that this has been indicated in the thesis. Signed: 1 ABSTRACT A major consideration in the exploitation of microalgae as biotechnology platforms is choosing robust, fast-growing strains that are amenable to genetic manipulation. The freshwater green alga Chlorella sorokiniana has been reported as one of the fastest growing and thermotolerant species, and studies in this thesis have confirmed strain UTEX1230 as the most productive strain of C. sorokiniana with doubling time under optimal growth conditions of less than three hours. Furthermore, the strain showed robust growth at elevated temperatures and salinities. In order to enhance the productivity of this strain, mutants with reduced biochemical and functional PSII antenna size were isolated. TAM4 was confirmed to have a truncated antenna and able to achieve higher cell density than WT, particularly in cultures under decreased irradiation. The possibility of genetic engineering this strain has been explored by developing molecular tools for both chloroplast and nuclear transformation. For chloroplast transformation, various regions of the organelle’s genome have been cloned and sequenced, and used in the construction of transformation vectors. However, no stable chloroplast transformant lines were obtained following microparticle bombardment. For nuclear transformation, cycloheximide-resistant mutants have been isolated and shown to possess specific missense mutations within the RPL41 gene. -
Lateral Gene Transfer of Anion-Conducting Channelrhodopsins Between Green Algae and Giant Viruses
bioRxiv preprint doi: https://doi.org/10.1101/2020.04.15.042127; this version posted April 23, 2020. 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-NC-ND 4.0 International license. 1 5 Lateral gene transfer of anion-conducting channelrhodopsins between green algae and giant viruses Andrey Rozenberg 1,5, Johannes Oppermann 2,5, Jonas Wietek 2,3, Rodrigo Gaston Fernandez Lahore 2, Ruth-Anne Sandaa 4, Gunnar Bratbak 4, Peter Hegemann 2,6, and Oded 10 Béjà 1,6 1Faculty of Biology, Technion - Israel Institute of Technology, Haifa 32000, Israel. 2Institute for Biology, Experimental Biophysics, Humboldt-Universität zu Berlin, Invalidenstraße 42, Berlin 10115, Germany. 3Present address: Department of Neurobiology, Weizmann 15 Institute of Science, Rehovot 7610001, Israel. 4Department of Biological Sciences, University of Bergen, N-5020 Bergen, Norway. 5These authors contributed equally: Andrey Rozenberg, Johannes Oppermann. 6These authors jointly supervised this work: Peter Hegemann, Oded Béjà. e-mail: [email protected] ; [email protected] 20 ABSTRACT Channelrhodopsins (ChRs) are algal light-gated ion channels widely used as optogenetic tools for manipulating neuronal activity 1,2. Four ChR families are currently known. Green algal 3–5 and cryptophyte 6 cation-conducting ChRs (CCRs), cryptophyte anion-conducting ChRs (ACRs) 7, and the MerMAID ChRs 8. Here we 25 report the discovery of a new family of phylogenetically distinct ChRs encoded by marine giant viruses and acquired from their unicellular green algal prasinophyte hosts. -
Disruption of Microbial Cells for Intracellular Products
Disruption of microbial cells for intracellular products YUSUF CHISTI and MURRAY MOO-YOUNG* cell either be genetically engineered so that what would normally be an intracellular product is excreted into the Department of Chemical Engineering, University of environment, or it must be disintegrated by physical, Waterloo, Waterloo, Ontario, Canada N2L 3G I chemical or enzymatic means to release its contents into the surrounding medium. The genetic manipulation of Summary. Disintegration of microbial cells is a necessary microbial cells to make them leaky is limited in scope. first step for the production of intracellular enzymes and Making the ceil fully permeable to any significant fraction organelIes. With increasing use of intracellular microbial of the intracellular products and enzymes would not only material in industry and medicine, the cell disruption unit be difficult, but also will imply discontinued existence of operation is gaining in importance. the cell. It is in this context that the unit operation of This review examines the state of the art of the large- microbial cell disruption for intracellular product isolation scale cell disruption technology and disruption methods of will become of increasing importance. potential commercial value. Probably because of the high capital and operating costs of pilot plants for large-scale isolation of intracellular Keywords.. Disruption of microorganisms; cell disintegration; products and the requirement of sizeable teams of scientists intracellular enzymes and technical staff to obtain meaningful biochemical engineering design data, few studies have been published on the subject. 3 This review examines the current state of microbial cell disruption technology from an industrial Introduction applications point of view. -
Creating Nanoparticles with Microfluidizer® High-Shear Fluid Technology
Creating Nanoparticles with Microfluidizer® High-Shear Fluid Technology Yang Su New Technology Manager Microfluidics International Corporation 1 Microfluidics at a Glance • Microfluidics has been located just outside of Boston,Status MA for Update 32 years serving over 2000 customers worldwide. We have sold ~4,000 processors with localized sales and support in 47 countries. • Microfluidizer® high shear fluid processors can produce nanomaterials with a wide variety of multiphase applications. We have vast experience with process development of many different types of applications/formulations. We pride ourselves in our ability to help our customers get the most out of their materials. • Microfluidizer Processors are used for R+D and manufacturing of active pharmaceutical ingredients, vaccines, inkjet inks, coatings, nutraceuticals and cosmetics. 17 of the top 20 pharma companies 8 of the top 10 biotech companies 4 of the top 5 chemical companies ...innovate with Microfluidics technology Tiny Particles, BIG RESULTS 2 What Microfluidics Does Best • Nanoemulsions “The overall satisfaction which we experienced with our • Cell disruption laboratory model Microfluidizer • Protein recovery processor eliminated the need to consider other equipment • Uniform particle size reduction when it was time to scale up to • Nano/microencapsulation production capabilities.” - Amylin Pharmaceuticals • Nanodispersions • Deagglomeration M-110P “plug n’ play” M-110EH-30 M-700 series Fixed-geometry benchtop lab model pilot scale machine production machine interaction -
DIMITAR VALEV: Wastewater Treatment with Algae Doctoral Dissertation, 118 Pp
ANNALES UNIVERSITATIS TURKUENSIS UNIVERSITATIS ANNALES AI 627 AI Dimitar Valev WASTEWATER TREATMENT WITH ALGAE Dimitar Valev Painosalama Oy, Turku, Finland 2020 Finland Turku, Oy, Painosalama ISBN 978-951-29-8094-9 (PRINT) – ISBN 978-951-29-8095-6 (PDF) TURUN YLIOPISTON JULKAISUJA ANNALES UNIVERSITATIS TURKUENSIS ISSN 0082-7002 (Print) SARJA – SER. AI OSA – TOM. 627 | ASTRONOMICA – CHEMICA – PHYSICA – MATHEMATICA | TURKU 2020 ISSN 2343-3175 (Online) WASTEWATER TREATMENT WITH ALGAE Dimitar Valev TURUN YLIOPISTON JULKAISUJA – ANNALES UNIVERSITATIS TURKUENSIS SARJA – SER. AI OSA – TOM. 627 | ASTRONOMICA – CHEMICA – PHYSICA – MATHEMATICA | TURKU 2020 University of Turku Faculty of Science and Engineering Department of Biochemistry / Molecular Plant Biology Doctoral programme in Molecular Life Sciences Supervised by Dr. Esa Tyystjärvi Dr. Taina Tyystjärvi Department of Biochemistry / Department of Biochemistry / Molecular Plant Biology, Molecular Plant Biology, University of Turku, FI-20014 University of Turku, FI-20014 Turku, Finland Turku, Finland Dr. Taras Antal Department of Botany and Plant Ecology Pskov State University Pskov 180000 Russia Reviewed by Professor Amit Bhatnagar Professor Koenraad Muylaert Water Chemistry & Microbiology Laboratory of Aquatic Biology University of Eastern Finland KU Leuven Kuopio, Finland Kortrijk, Belgium Opponent Professor Ondřej Prášil Centre Algatech Institute of Microbiology, The Czech Academy of Sciences Třeboň, Czech Republic The originality of this publication has been checked in accordance with the University of Turku quality assurance system using the Turnitin OriginalityCheck service. ISBN 978-951-29-8094-9 (PRINT) ISBN 978-951-29-8095-6 (PDF) ISSN 0082-7002 (Painettu/Print) ISSN 2343-3175 (Sähköinen/Online) Painosalama Oy, Turku, Finland 2020 UNIVERSITY OF TURKU Faculty of Science and Engineering Department of Biochemistry Molecular Plant Biology DIMITAR VALEV: Wastewater treatment with algae Doctoral Dissertation, 118 pp. -
Comparison of Methods for DNA Extraction from Candida Albicans
Department of Medical Biochemistry and Microbiology Uppsala University Comparison of methods for DNA extraction from Candida albicans Ashraf Dadgar 2006 Department of Clinical Microbiology, Uppsala University Hospital, SE- 751 85 Uppsala Sweden Supervisor: Åsa Innings and Björn Herrmann ABSTRACT Invasive Candida infection is an increasing cause of morbidity and mortality in the immunocompromised patient. Molecular diagnosis based on genomic amplification methods, such as real time PCR, has been reported as an alternative to conventional culture for early detection of invasive candidiasis. The template DNA extraction step has been the major limitation in most reported nucleic acid based assays, due to problems in breaking fungal cell walls and incomplete purification in PCR inhibitor substances. The aim of this study was to compare enzymatic cell wall disruption using recombinant lyticase with mechanical disruption using glass beads. The QIAamp tissue kit was compared with two automated DNA extraction robots, the BioRobot M48 and NucliSens easyMAG, to determine their sensitivity, reliability and duration for DNA release of C. albicans. Mechanical cell wall disruption shortened and facilitated the extraction procedure, but the quantity of released DNA was significantly lower than when enzymatic cell wall disruption was used. Use of robots did not significantly shorten the DNA extraction time, compared with manual DNA extraction. However the NucliSens easyMAG resulted in a higher yield of target DNA compared to the BioRobot M48 and the manual QIAamp tissue kit. KEYWORDS: Candida albicans, candidemia, DNA extraction, cell wall disruption, real time PCR 2 SAMMANFATTNING Invasiva svampinfektioner är ett stort problem hos patienter med dåligt immunförsvar. Förekomst av invasiva svampinfektioner har ökat under senare år och medför hög dödlighet. -
Applied Microbiology and Biotechnology
Applied Microbiology and Biotechnology Re-cultivation of Neochloris oleoabundans in exhausted autotrophic and mixotrophic media: the potential role of polyamines and free fatty acids --Manuscript Draft-- Manuscript Number: AMAB-D-15-00751R1 Full Title: Re-cultivation of Neochloris oleoabundans in exhausted autotrophic and mixotrophic media: the potential role of polyamines and free fatty acids Article Type: Original Article Section/Category: Applied microbial and cell physiology Corresponding Author: Simonetta Pancaldi Italy Corresponding Author Secondary Information: Corresponding Author's Institution: Corresponding Author's Secondary Institution: First Author: Alessandra Sabia First Author Secondary Information: Order of Authors: Alessandra Sabia Costanza Baldisserotto Stefania Biondi Roberta Marchesini Paola Tedeschi Annalisa Maietti Martina Giovanardi Lorenzo Ferroni Simonetta Pancaldi Order of Authors Secondary Information: Funding Information: Abstract: Neochloris oleoabundans (Chlorophyta) is widely considered one of the most promising microalgae for biotechnological applications. However, the large-scale production of microalgae requires large amounts of water. In this perspective, the possibility of using exhausted growth media for the re-cultivation of N. oleoabundans was investigated in order to simultaneously make the cultivation more economically feasible and environmentally sustainable. Experiments were performed by testing the following media: autotrophic exhausted medium (E+) and mixotrophic exhausted medium after cultivation -
Downloaded from Genbank on That Full Plastid Genomes Are Not Sufficient to Reject Al- February 28, 2012
Ruhfel et al. BMC Evolutionary Biology 2014, 14:23 http://www.biomedcentral.com/1471-2148/14/23 RESEARCH ARTICLE Open Access From algae to angiosperms–inferring the phylogeny of green plants (Viridiplantae) from 360 plastid genomes Brad R Ruhfel1*, Matthew A Gitzendanner2,3,4, Pamela S Soltis3,4, Douglas E Soltis2,3,4 and J Gordon Burleigh2,4 Abstract Background: Next-generation sequencing has provided a wealth of plastid genome sequence data from an increasingly diverse set of green plants (Viridiplantae). Although these data have helped resolve the phylogeny of numerous clades (e.g., green algae, angiosperms, and gymnosperms), their utility for inferring relationships across all green plants is uncertain. Viridiplantae originated 700-1500 million years ago and may comprise as many as 500,000 species. This clade represents a major source of photosynthetic carbon and contains an immense diversity of life forms, including some of the smallest and largest eukaryotes. Here we explore the limits and challenges of inferring a comprehensive green plant phylogeny from available complete or nearly complete plastid genome sequence data. Results: We assembled protein-coding sequence data for 78 genes from 360 diverse green plant taxa with complete or nearly complete plastid genome sequences available from GenBank. Phylogenetic analyses of the plastid data recovered well-supported backbone relationships and strong support for relationships that were not observed in previous analyses of major subclades within Viridiplantae. However, there also is evidence of systematic error in some analyses. In several instances we obtained strongly supported but conflicting topologies from analyses of nucleotides versus amino acid characters, and the considerable variation in GC content among lineages and within single genomes affected the phylogenetic placement of several taxa. -
Mixotrophy in Chlorella Sorokiniana – Physiology, Biotechnological Potential and Ecotoxicology
UNIVERSIDADE FEDERAL DE SÃO CARLOS CENTRO DE CIÊNCIAS BIOLÓGICAS E DA SAÚDE PROGRAMA DE PÓS-GRADUAÇÃO EM ECOLOGIA E RECURSOS NATURAIS Mixotrophy in Chlorella sorokiniana – Physiology, Biotechnological Potential and Ecotoxicology ADRIANO EVANDIR MARCHELLO São Carlos – 2017 UNIVERSIDADE FEDERAL DE SÃO CARLOS CENTRO DE CIÊNCIAS BIOLÓGICAS E DA SAÚDE PROGRAMA DE PÓS-GRADUAÇÃO EM ECOLOGIA E RECURSOS NATURAIS ADRIANO EVANDIR MARCHELLO Mixotrophy in Chlorella sorokiniana – Physiology, Biotechnological Potential and Ecotoxicology Orientadora: Profa Dra Ana Teresa Lombardi Tese apresentada ao Programa de Pós- Graduação em Ecologia e Recursos Naturais (PPGERN) como parte dos requisitos para obtenção do título de Doutor em Ecologia e Recursos Naturais. São Carlos – 2017 Dedico este trabalho aos meus pais Adilson e Rosana, minha irmã Amanda e meus sobrinhos Maressa e Lucca. “Só Tu és o Senhor. Fizeste os céus, e os mais altos céus, e tudo o que neles há, a terra e tudo o que nela existe, os mares e tudo o que neles existe. Tu deste vida a todos os seres, e os exércitos dos céus te adoram.” Neemias 9:6 “O que sabemos é uma gota; o que ignoramos é um oceano.” Isaac Newton Agradecimentos Acima de tudo agradeço a DEUS por ter me dado a vida, criado a vida em todas as suas formas e me dado a oportunidade de estudar sua maior criação, a VIDA! Agradeço minha família (meu pai Adilson, minha mãe Rosana, minha irmã Amanda, e meus avós Laurinda in memoriam, Teresinha e João) por sempre ter me apoiado em todo esse percurso. Se cheguei onde estou, com certeza é graças ao incentivo e esforço de cada um deles.