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New Biotechnology

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https://www.readbyqxmd.com/read/29656128/agrobacterium-rhizogenes-mediated-transformation-of-a-dioecious-plant-model-silene-latifolia
#1
Vojtech Hudzieczek, Radim Cegan, Tomas Cermak, Nela Bacovska, Zuzana Machalkova, Karel Dolezal, Lucie Plihalova, Daniel Voytas, Roman Hobza, Boris Vyskot
Silene latifolia serves as a model species to study dioecy, the evolution of sex chromosomes, dosage compensation and sex-determination systems in plants. Currently, no protocol for genetic transformation is available for this species, mainly because S. latifolia is considered recalcitrant to in vitro regeneration and infection with Agrobacterium tumefaciens. Using cytokinins and their synthetic derivatives, we markedly improved the efficiency of regeneration. Several agrobacterial strains were tested for their ability to deliver DNA into S...
April 12, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29635104/generation-and-validation-of-murine-monoclonal-and-camelid-recombinant-single-domain-antibodies-specific-for-human-pancreatic-glycoprotein-2
#2
Anja Schlör, Pamela Holzlöhner, Martin Listek, Cindy Grieß, Monique Butze, Burkhard Micheel, Christian Hentschel, Mandy Sowa, Dirk Roggenbuck, Peter Schierack, Jonas Füner, Erik Schliebs, Alexander Goihl, Dirk Reinhold, Katja Hanack
Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified as a major autoantigenic target in Crohńs disease patients. It was reported recently that a long (GP2a) and a short (GP2b) isoform of GP2 exist and that in the outcome of inflammatory bowel diseases (IBD) GP2-specific autoantibodies probably appear as new serological markers for diagnosis and therapeutic monitoring. To investigate this further and in order to establish diagnostic tools for the discrimination of both GP2 isoforms, a set of different murine monoclonal and camelid recombinant single domain antibodies (camelid VHH) was generated and validated in various enzyme-linked immunosorbent assay (ELISA) formats, immunofluorescence on transgenic cell lines and immunohistochemistry on monkey pancreas tissue sections...
April 7, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29614323/optimization-of-aqueous-two-phase-systems-for-the-production-of-6-aminopenicillanic-acid-in-integrated-microfluidic-reactors-separators
#3
Lucie Vobecká, Alexandr Romanov, Zdeněk Slouka, Pavel Hasal, Michal Přibyl
Aqueous two-phase systems (ATPSs) were screened for the production of 6-aminopenicillanic acid (6-APA) catalyzed by penicillin acylase, followed by the extractive separation of 6-APA from the reaction mixture. The key point of this study was to find an ATPS exhibiting a large difference in the partition coefficients of the biocatalyst and reaction products. Several ATPSs based on polyethylene glycol (PEG)/phosphate, PEG/citrate, and PEG/dextran were tested. We found that an ATPS consisting of 15 wt% of PEG 4000, 10 wt% of phosphates, 75 wt% of water (pH value 8...
March 31, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29574274/pharmacokinetics-of-radiolabeled-dimeric-sdabs-constructs-targeting-human-cd20
#4
Ahmet Krasniqi, Magdalena Bialkowska, Catarina Xavier, Kevin Van der Jeught, Serge Muyldermans, Nick Devoogdt, Matthias D'Huyvetter
Single-domain antibody fragments (sdAbs) are the smallest functional antigen-binding fragments, derived from heavy chain-only camelid antibodies. When designed as radiolabeled monomeric probes for imaging and therapy of cancer, their fast and specific targeting results in high tumor-to-background ratios early after injection. However, their moderate absolute uptake into tumors might not always be sufficient to treat cancerous lesions. We have evaluated the pharmacokinetics of seven constructs derived from a CD20-targeting monomeric sdAb (αCD20)...
March 21, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29550523/microbial-single-cell-analysis-in-picoliter-sized-batch-cultivation-chambers
#5
Eugen Kaganovitch, Xenia Steurer, Deniz Dogan, Christopher Probst, Wolfgang Wiechert, Dietrich Kohlheyer
Microfluidics has enabled various research projects in the field of microbial single-cell analysis. In particular, single-use microfluidic cultivation devices combined with automated time-lapse imaging provide powerful approaches for analyzing microbial phenomena at the single-cell level. High spatiotemporal resolution facilitates individual cell identification and tracking, delivering detailed insights into areas like phenotypic population heterogeneity, which can be highly relevant to the fate of a microbial population and may negatively impact the efficiency of biotechnological fermentations...
March 14, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29530668/cleavage-of-poly-cis-1-4-isoprene-rubber-as-solid-substrate-by-cultures-of-gordonia-polyisoprenivorans
#6
R Andler, S Hiessl, O Yücel, M Tesch, A Steinbüchel
Potential biotechnological recycling processes for rubber products include the bacterial degradation of poly(cis-1,4-isoprene) (IR) in order to achieve its total biodegradation or its biotransformation into useful products. The actinomycete Gordonia polyisoprenivorans strain VH2 catalyzes the degradation of IR and enables its use as a sole carbon source via β-oxidation. The initial cleavage reaction is catalyzed by the extracellular latex clearing protein (Lcp). This dioxygenase is the key enzyme for the formation of oligo(cis-1,4-isoprene) molecules with different lengths, i...
March 9, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29526810/integrated-physical-map-of-bread-wheat-chromosome-arm-7ds-to-facilitate-gene-cloning-and-comparative-studies
#7
Zuzana Tulpová, Ming-Cheng Luo, Helena Toegelová, Paul Visendi, Satomi Hayashi, Petr Vojta, Etienne Paux, Andrzej Kilian, Michaël Abrouk, Jan Bartoš, Marián Hajdúch, Jacqueline Batley, David Edwards, Jaroslav Doležel, Hana Šimková
Bread wheat (Triticum aestivum L.) is a staple food for a significant part of the world's population. The growing demand on its production can be satisfied by improving yield and resistance to biotic and abiotic stress. Knowledge of the genome sequence would aid in discovering genes and QTLs underlying these traits and provide a basis for genomics-assisted breeding. Physical maps and BAC clones associated with them have been valuable resources from which to generate a reference genome of bread wheat and to assist map-based gene cloning...
March 8, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29522860/multi-function-microfluidic-platform-for-sensor-integration
#8
Ana C Fernandes, Daria Semenova, Peter Panjan, Adama M Sesay, Krist V Gernaey, Ulrich Krühne
The limited availability of metabolite-specific sensors for continuous sampling and monitoring is one of the main bottlenecks contributing to failures in bioprocess development. Furthermore, only a limited number of approaches exist to connect currently available measurement systems with high throughput reactor units. This is especially relevant in the biocatalyst screening and characterization stage of process development. In this work, a strategy for sensor integration in microfluidic platforms is demonstrated, to address the need for rapid, cost-effective and high-throughput screening in bioprocesses...
March 6, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29510237/relationship-between-microbial-community-operational-factors-and-ammonia-inhibition-resilience-in-anaerobic-digesters-at-low-and-moderate-ammonia-background-concentrations
#9
Y Lu, R Liaquat, S Astals, P D Jensen, D J Batstone, S Tait
The relationship between anaerobic digestion operational conditions and (i) microbial community, (ii) acetoclastic methanogenic activity and (iii) free ammonia (NH3 ) inhibition resilience was investigated. Thirteen inocula were obtained from full and pilot scale digesters fed with different substrates, digester configurations, operating temperatures and NH3 concentrations (0.1-241 mgN·L-1 ). Substrate type and temperature were the primary factors influencing microbial community composition. Methanogenic activity ranged from 0...
March 3, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29501588/migration-of-blood-cells-and-phospholipid-vesicles-induced-by-concentration-gradients-in-microcavities
#10
Saša Vrhovec Hartman, Bojan Božič, Jure Derganc
Microcavities provide a well-controlled flow-free microenvironment and play an important role in many microfluidic systems, for example as cell-culturing microchambers. Here we show that transient concentration gradients that emerge during diffusive exchange of solutes in microcavities induce passive migration (diffusiophoresis) of blood cells and synthetic phospholipid vesicles. The passive migration is observed in various concentration gradients comprising non-electrolytes and electrolytes, i.e., glucose, sucrose, sodium chloride, potassium chloride, potassium benzoate, and potassium sulfate...
February 28, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29496641/potential-use-of-methane-fermentation-digested-slurry-as-a-low-cost-environmentally-friendly-nutrient-for-bioethanol-production-from-crude-glycerol-by-klebsiella-variicola-tb-83d
#11
Kohei Seta, Toshihiro Suzuki, Keiji Kiyoshi, Toshiya Shigeno, Toshiaki Nakajima-Kambe
A methane fermentation digested slurry (MFDS) was evaluated as a substitute for the commercial nutrient, yeast extract (YE), in ethanol production from glycerol by Klebsiella variicola strain TB-83D. In pH-controlled fed-batch cultures, partial replacement of YE by MFDS did not reduce ethanol productivity significantly. However, non-sterilized MFDS had negative effects on glycerol fermentation by this strain. Although ethanol production decreased when YE was completely replaced by sterilized MFDS, the use of crude glycerol and sterilized MFDS achieved a yield of 14...
February 26, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29477599/rosa-hybrida-orcinol-o-methyl-transferase-mediated-production-of-pterostilbene-in-metabolically-engineered-grapevine-cell-cultures
#12
Ascensión Martínez-Márquez, Jaime A Morante-Carriel, Javier Palazon, Roque Bru-Martínez
Stilbenes are naturally scarce high-added-value plant compounds with chemopreventive, pharmacological and cosmetic properties. Bioproduction strategies include engineering the metabolisms of bacterial, fungal and plant cell systems. Strikingly, one of the most effective strategies consists in the elicitation of wild grapevine cell cultures, which leads to vast stilbene resveratrol accumulation in the extracellular medium. The combination of both cell culture elicitation and metabolic engineering strategies to produce resveratrol analogs proved more efficient for the hydroxylated derivative piceatannol than for the dimethylated derivative pterostilbene, for which human hydroxylase HsCYP1B1- and grapevine O-methyltransferase VvROMT-transformed cell cultures were respectively used...
February 24, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29477600/improved-continuous-fumaric-acid-production-with-immobilised-rhizopus-oryzae-by-implementation-of-a-revised-nitrogen-control-strategy
#13
Andre Naude, Willie Nicol
A novel fermentation system was employed whereby the mycelial mat of Rhizopus oryzae was attached to a polypropylene tube. Batch operation was used for growth, while continuous operation was employed during the fumaric acid production phase. A clear decrease in respiration, fumaric acid (FA) and ethanol production was observed when zero nitrogen was fed in the production phase, with FA productivity decreasing from an initial 0.7 g L-1  h-1 to 0.3 g L-1  h-1 after 150 hours. With the addition of 0...
February 22, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29476816/potential-use-of-ricotta-cheese-whey-for-the-production-of-lactobionic-acid-by-pseudomonas-taetrolens-strains
#14
Stefania De Giorgi, Noura Raddadi, Angelo Fabbri, Tullia Gallina Toschi, Fabio Fava
Lactobionic acid (LBA) is a fine chemical largely applied in the food, chemical, cosmetics and pharmaceutical industries. Here, its production from ricotta cheese whey (RCW), or scotta, the main by-product obtained from ricotta cheese production process and currently employed mainly for cattle feed, was evaluated. Among seven bacterial species tested, only two Pseudomonas taetrolens strains were selected after preliminary screening in shake-flasks. When autoclaved RCW was used, a lactobionic acid titer of 34...
February 21, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29476815/enhanced-bioproduction-of-2-phenylethanol-in-a-biphasic-system-with-rapeseed-oil
#15
Karolina Chreptowicz, Jolanta Mierzejewska
Increasing demand for natural fragrance ingredients and products has led to their global market growth. 2-Phenylethanol (2-PE) is a volatile substance widely used in food and cosmetics manufacturing. It is generally known that yeast can metabolize L-phenylalanine (L-Phe) to produce 2-PE. However, because the product exhibits an inhibitory effect on yeast cells, simple batch cultivation is uneconomic. The aim of this study was to enhance 2-PE productivity using in situ product removal. Here we present a new method of 2-PE production by yeast in a biphasic system with rapeseed oil as the second phase...
February 21, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29474834/non-immunoglobulin-scaffold-proteins-precision-tools-for-studying-protein-protein-interactions-in-cancer
#16
REVIEW
Heather L Martin, Robert Bedford, Sophie J Heseltine, Anna A Tang, Katarzyna Z Haza, Ajinkya Rao, Michael J McPherson, Darren C Tomlinson
Cancer is frequently characterised by dysregulation of the cellular signalling processes that govern proliferation, survival and attachment. Understanding such dysregulation continues to present a challenge given the importance of protein-protein interactions in intracellular processes. Exploring this protein-protein interactome requires novel tools capable of discriminating between highly homologous proteins, individual domains and post-translational modifications. This review examines the potential of scaffold-based binding proteins to fulfil these requirements...
February 20, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29474833/one-step-integrated-clarification-and-purification-of-a-monoclonal-antibody-using-protein-a-mag-sepharose-beads-and-a-cgmp-compliant-high-gradient-magnetic-separator
#17
Moritz Ebeler, Ola Lind, Nils Norrman, Ronnie Palmgren, Matthias Franzreb
Monoclonal antibodies are a dominant component of today's biopharmaceutical market and are typically purified by classical platform processes. However, high costs and rising demands are drivers for the development of new, efficient and flexible integrated purification processes. Currently, high-gradient magnetic separation as a direct capturing tool for protein purification suffers from the lack of suitable GMP-compliant separation equipment for industrial scale. As a solution for this bottleneck, we present a purification process for a monoclonal antibody directly from CHO cell culture by use of protein A-functionalized magnetic particles together with the first pilot-scale GMP-compliant 'rotor-stator' high-gradient magnetic separator...
February 20, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29454893/deciphering-the-growth-pattern-and-phytohormonal-content-in-saskatoon-berry-amelanchier-alnifolia-in-response-to-in-vitro-cytokinin-application
#18
Mack Moyo, Adeyemi O Aremu, Lenka Plačková, Lucie Plíhalová, Aleš Pěnčík, Ondřej Novák, Jan Holub, Karel Doležal, Johannes Van Staden
Clonal propagation plays a critical integral role in the growth and success of a global multi-billion dollar horticulture industry through a constant supply of healthy stock plants. The supply chain depends on continuously improving the micropropagation process, thus, understanding the physiology of in vitro plants remains a core component. We evaluated the influence of exogenously applied cytokinins (CKs, N6 -benzyladenine = BA, isopentenyladenine = iP, meta-topolin = mT, 6-(3-hydroxybenzylamino)-9-(tetrahydropyran-2-yl)purine = mTTHP) in Murashige and Skoog (MS)-supplemented media on organogenic response and accumulation of endogenous CK and indole-3-acetic acid (IAA) metabolites...
February 15, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29452271/biotechnologically-produced-chitosan-for-nanoscale-products-a-legal-analysis
#19
Greet Smets, Patrick Rüdelsheim
Conventionally, chitosans are derived from shrimp and other crustacean shells. Biotechnology offers an alternative route to produce chitosans and more importantly, specific chitosan structures tailored to the needs of a diversity of industries. However, for biotech chitosans and products thereof to be commercialised, legislation should not create a burden. Here, the requirements of the EU regulatory framework have been analysed for the entire chain from research to development and production of several potential applications including nanomaterials...
February 13, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29448030/a-sep-tag-enhances-the-expression-solubility-and-yield-of-recombinant-tev-protease-without-altering-its-activity
#20
Kalpana Nautiyal, Yutaka Kuroda
Tobacco Etch Virus (TEV) protease is used in the purification of recombinant proteins, but its use is often hampered by solubility issues. Here, we report a short, 12-residue solubility enhancing peptide (SEP) tag attached at the C-terminus of TEV (TEV-C9R). We assessed the effects of the C9R tag on the biophysical and biochemical characteristics of TEV. The yield of HPLC purified TEV-C9R expressed in E. coli grown in 200 mL LB or TB media was between 10 and 13 mg, which was up to 6.5 times higher than the yield of the untagged TEV...
February 12, 2018: New Biotechnology
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