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Biotechnology and Bioengineering

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https://www.readbyqxmd.com/read/28092110/simultaneous-detection-of-three-sex-steroid-hormone-classes-using-a-novel-yeast-based-biosensor
#1
Alexandre Chamas, Ha Thi Minh Pham, Martin Jähne, Karina Wettwer, Steffen Uhlig, Kirsten Simon, Almuth Einspanier, Kim Baronian, Gotthard Kunze
A biosensor detecting estrogens, progestogens and androgens in complex samples and in a single step is described. Three Arxula adeninivorans yeast strains were created, each strain producing a different recombinant human hormone receptor and a different fluorescent reporter protein. These strains were then mixed to create G1212/YRC102-hHR-fluo, the biological component of the biosensor. During incubation with G1212/YRC102-hHR-fluo, hormones present in a sample bind to their target receptor, which leads to the production of a specific fluorescent protein...
January 16, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28090632/identification-of-highly-selective-mmp-14-inhibitory-fabs-by-deep-sequencing
#2
Tyler Lopez, Dong Hyun Nam, Evan Kaihara, Zahid Mustafa, Xin Ge
Matrix metalloproteinase (MMP)-14 is an important target for cancer treatment due to its critical roles in tumor invasion and metastasis. Previous failures of all compound-based broad-spectrum MMP inhibitors in clinical trials suggest that selectivity is the key for a successful therapy. With inherent high specificity, monoclonal antibodies (mAbs) therefore arise as attractive inhibitors able to target the particular MMP of interest. As a routine screening method, enzyme-linked immunosorbent assays (ELISA) have been applied to panned phage libraries for the isolation of mAbs inhibiting MMP-14...
January 16, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28067424/human-pathogens-in-plant-biofilms-formation-physiology-and-detection
#3
REVIEW
Eduardo Ximenes, Lori Hoagland, Seockmo Ku, Xuan Li, Michael Ladisch
Fresh produce, viewed as an essential part of a healthy life style, usually consumed in the form of raw or minimally processed fruits and vegetables, is a potentially important source of food-borne disease including human pathogenic bacteria and viruses. These are passed on to the consumer since the bacteria can form biofilms or otherwise populate plant tissues, thereby using plants as vectors to infect animal hosts. The life cycle of the bacteria in plants differs from those in animals or humans and results in altered physiochemical and biological properties (e...
January 9, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28067404/electrochemical-techniques-reveal-that-total-ammonium-stress-increases-electron-flow-to-anode-respiration-in-mixed-species-bacterial-anode-biofilms
#4
Mohamed Mahmoud, Prathap Parameswaran, César I Torres, Bruce E Rittmann
When anode-respiring bacteria (ARB) respire electrons to an anode in microbial electrochemical cells (MXCs), they harvest only a small amount of free energy. This means that ARB must have a high substrate-oxidation rate coupled with a high ratio of electrons used for respiration compared to total electrons removed by substrate utilization. It also means that they are especially susceptible to inhibition that slows anode respiration or lowers their biomass yield. Using several electrochemical techniques, we show that a relatively high total ammonium-nitrogen (TAN) concentration (2...
January 9, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28059445/computational-de-novo-design-of-antibodies-binding-to-a-peptide-with-high-affinity
#5
Venkata Giridhar Poosarla, Tong Li, Boon Chong Goh, Klaus Schulten, Thomas K Wood, Costas D Maranas
Antibody drugs play a critical role in infectious diseases, cancer, autoimmune diseases, and inflammation. However, experimental methods for the generation of therapeutic antibodies such as using immunized mice or directed evolution remain time consuming and cannot target a specific antigen epitope. Here, we describe the application of a computational framework called OptMAVEn combined with molecular dynamics to de novo design antibodies. Our reference system is antibody 2D10, a single-chain antibody (scFv) that recognizes the dodecapeptide DVFYPYPYASGS, a peptide mimic of mannose-containing carbohydrates...
January 6, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28059435/acetoin-production-via-unbalanced-fermentation-in-shewanella-oneidensis
#6
Thea Bursac, Jeffrey A Gralnick, Johannes Gescher
This study describes the realization of an anoxic acetoin production process using the proteobacterium Shewanella oneidensis. Fermentative processes are of high biotechnological relevance since they offer high productivity and a low percentage of substrate consumption for anabolic processes. Nevertheless, the range of compounds that can be produced as sole end product of a fermentative process is limited, since the average oxidation state of substrate and products has to be identical in the absence of an external electron acceptor...
January 6, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28019666/engineering-the-n-terminal-end-of-cela-results-in-improved-performance-and-growth-of-caldicellulosiruptor-bescii-on-crystalline-cellulose
#7
Sun-Ki Kim, Daehwan Chung, Michael E Himmel, Yannick J Bomble, Janet Westpheling
CelA is the most abundant enzyme secreted by Caldicellulosiruptor bescii and has been shown to outperform mixtures of commercially available exo- and endoglucanases in vitro. CelA contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. Here, repeated aspartate residues were introduced into the N-terminal ends of CelA GH9 and GH48 domains to improve secretion efficiency and/or catalytic efficiency of CelA...
December 26, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28019665/the-impact-of-left-ventricular-stretching-in-model-cultivations-with-neonatal-cardiomyocytes-in-a-whole-heart-bioreactor
#8
Jörn Hülsmann, Hug Aubin, Alexander Wehrmann, Artur Lichtenberg, Payam Akhyari
Here, we investigate the impact of integrated three-dimensional (3D) left ventricular (LV) stretching on myocardial maturation in a whole-heart bioreactor setting. Therefore, decellularized rat hearts were selectively repopulated with rodent neonatal cardiomyocytes (5•10(6) cells per heart) and cultured over 5 d. Continuous medium perfusion was maintained through the coronary artery system in a customized whole-heart bioreactor system with or without integrated biomechanical stimulation of LV. 3D repopulation effectiveness and cellular vitality were evaluated by repetitive metabolic WST-1 assays and 3D confocal microscopy analysis through fluorescent staining, also assessing cellular organization...
December 26, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27987323/translation-system-engineering-in-escherichia-coli-enhances-non-canonical-amino-acid-incorporation-into-proteins
#9
Rui Gan, Jessica G Perez, Erik D Carlson, Ioanna Ntai, Farren J Isaacs, Neil L Kelleher, Michael C Jewett
The ability to site-specifically incorporate noncanonical amino acids (ncAAs) into proteins has made possible the study of protein structure and function in fundamentally new ways, as well as the synthesis of unnatural polymers. However, the task of site-specifically incorporating multiple ncAAs into proteins with high purity and yield continues to present a challenge. At the heart of this challenge lies the lower efficiency of engineered orthogonal translation system components compared to their natural counterparts (e...
December 17, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27987304/identification-of-lanthanum-specific-peptides-for-future-recycling-of-rare-earth-elements-from-compact-fluorescent-lamps
#10
Franziska L Lederer, Susan B Curtis, Stefanie Bachmann, W Scott Dunbar, Ross T A MacGillivray
As components of electronic scrap, rare earth minerals are an interesting but little used source of raw materials that are highly important for the recycling industry. Currently, there exists no cost-efficient technology to separate rare earth minerals from an electronic scrap mixture. In this study, phage surface display has been used as a key method to develop peptides with high specificity for particular inorganic targets in electronic scrap. Lanthanum phosphate doped with cerium and terbium as part of the fluorescent phosphors of spent compact fluorescent lamps (CFL) was used as a target material of economic interest to test the suitability of the phage display method to the separation of rare earth minerals...
December 17, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27943258/improving-metabolite-production-in-microbial-cocultures-using-a-spatially-constrained-hydrogel
#11
Matthew J Smith, Matthew B Francis
Microbial cocultures promise the development of more efficient bioproductions. However, the design of obligate mutualisms is complicated when using organisms that possess differing growth rates or incompatible media requirements. In this work, we investigate sucrose production by cscB Synechococcus elongatus PCC 7942 within a polyacrylate hydrogel matrix. This system secretes sucrose only when the hydrogel is spatially constrained, demonstrating a new utilization of hydrogel swelling pressure to control the osmotic strength of a microbial microenvironment...
December 12, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27943249/metabolic-engineering-of-fatty-alcohol-production-in-transgenic-hairy-roots-of-crambe-abyssinica
#12
Magdalena Miklaszewska, Antoni Banaś, Aleksandra Królicka
Biotechnological production of fatty alcohols, important raw materials in the chemical industry, has been receiving considerable attention in recent years. Fatty alcohols are formed by the reduction of fatty acyl-CoAs or fatty acyl-ACPs catalyzed by a fatty acyl reductase (FAR). In this study, we introduced genes encoding FARs from Arabidopsis thaliana (AtFAR5) and Simmondsia chinensis (ScFAR) into Crambe abyssinica hairy roots via Agrobacterium rhizogenes-mediated transformation. The efficiency of the transformation ranged between 30-45%...
December 12, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27943246/producing-desulfurized-biogas-through-removal-of-sulfate-in-the-first-stage-of-a-two-stage-anaerobic-digestion
#13
Yeo-Myeong Yun, Shihwu Sung, Hang-Sik Shin, Jong-In Han, Hyun-Woo Kim, Dong-Hoon Kim
In the present work, a two-stage anaerobic digestion system (TSADS) was newly designed to produce biogas with a greatly reduced H2 S content. The role of first (sulfidogenic)-stage digester was not only acidogenesis but also sulfidogenesis (sulfate reduction to H2 S), which would minimize the input of H2 S-producing source in the followed second (methanogenic)-stage digester. For the coexistence of acidogens and sulfate reducing bacteria (SRB) in the sulfidogenic-stage digester, it was found that pH played a crucial role...
December 12, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27943242/knockout-of-a-difficult-to-remove-cho-host-cell-protein-lipoprotein-lipase-for-improved-polysorbate-stability-in-monoclonal-antibody-formulations
#14
Josephine Chiu, Kristin N Valente, Nicholas E Levy, Lie Min, Abraham M Lenhoff, Kelvin H Lee
While the majority of host cell protein (HCP) impurities are effectively removed in typical downstream purification processes, a small population of HCPs are particularly challenging. Previous studies have identified HCPs that are challenging for a variety of reasons. Lipoprotein lipase (LPL)-a Chinese hamster ovary (CHO) HCP that functions to hydrolyze esters in triglycerides-was one of ten HCPs identified in previous studies as being susceptible to retention in downstream processing. LPL may degrade polysorbate 80 (PS-80) and polysorbate 20 (PS-20) in final product formulations due to the structural similarity between polysorbates and triglycerides...
December 12, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27943241/qsar-models-for-prediction-of-chromatographic-behavior-of-homologous-fab-variants
#15
Julie R Robinson, Hanne Sophie Karkov, James A Woo, Berit Olsen Krogh, Steven M Cramer
While quantitative structure activity relationship (QSAR) models have been employed successfully for the prediction of small model protein chromatographic behavior, there have been no reports to date on the use of this methodology for larger, more complex proteins. Recently our group generated focused libraries of antibody Fab fragment variants with different combinations of surface hydrophobicities and electrostatic potentials, and demonstrated that the unique selectivities of multimodal resins can be exploited to separate these Fab variants...
December 12, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27943233/enhancing-functional-expression-of-codon-optimized-heterologous-enzymes-in-escherichia-coli-bl21-de3-by-selective-introduction-of-synonymous-rare-codons
#16
Chao Zhong, Ping Wei, Yi-Heng Percival Zhang
Rare codon in a heterologous gene may cause premature termination of protein synthesis, misincorporation of amino acids, and/or slow translation of mRNA, decreasing the heterologous protein expression. However, its hypothetical function pertaining to functional protein folding has been barely reported. Here, we investigated the effects of selective introduction of synonymous rare codons (SRCs) to two codon-optimized (i.e., rare codon-free) genes sucrose phosphorylase (SP) gene from Thermoanaerobacterium thermosaccharolyticum and amidohydrolase gene from Streptomyces caatingaensis on their expression levels in Escherichia coli BL21(DE3)...
December 12, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27922181/evaluation-of-gas-and-carbon-transport-in-a-methanogenic-bioelectrochemical-system-bes
#17
Christy M Dykstra, Spyros G Pavlostathis
Bioelectrochemical systems (BESs) may be used to upgrade anaerobic digester biogas by directly converting CO2 to CH4 . The objective of this study was to evaluate gas (N2 , CO2 , CH4 , and H2 ) and carbon transport within a methanogenic BES. Four BES configurations were evaluated: abiotic anode with abiotic cathode (AAn-ACa), bioanode with abiotic cathode (BAn-ACa), abiotic anode with biocathode (AAn-BCa), and bioanode with biocathode (BAn-BCa). Transport of N2 , a gas commonly used for flushing anoxic systems, out of the anode headspace ranged from 3...
December 6, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27922179/a-carbon-dioxide-stripping-model-for-mammalian-cell-culture-in-manufacturing-scale-bioreactors
#18
Zizhuo Xing, Amanda M Lewis, Michael C Borys, Zheng Jian Li
Control of carbon dioxide within the optimum range is important in mammalian bioprocesses at the manufacturing scale in order to ensure robust cell growth, high protein yields, and consistent quality attributes. The majority of bioprocess development work is done in laboratory bioreactors, in which carbon dioxide levels are more easily controlled. Some challenges in carbon dioxide control can present themselves when cell culture processes are scaled up, because carbon dioxide accumulation is a common feature due to longer gas-residence time of mammalian cell culture in large scale bioreactors...
December 6, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27922175/a-framework-to-quantify-karyotype-variation-associated-with-cho-production-instability
#19
Jong Youn Baik, Kelvin H Lee
Chinese hamster ovary (CHO) cells, major mammalian host cells for biomanufacturing of therapeutic proteins, have been extensively investigated to enhance productivity and product quality. However, cell line instability resulting in unexpected changes in productivity or product quality is challenging for biomanufacturing. Based on previous reports about causes and characteristics of production instability, we hypothesized that chromosomal rearrangements due to genomic instability are associated with production instability...
December 6, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27888663/effect-of-structural-carbohydrates-and-lignin-content-on-the-anaerobic-digestion-of-paper-and-paper-board-materials-by-anaerobic-granular-sludge
#20
Jorge Gonzalez-Estrella, Caitlin M Asato, Amber C Jerke, James J Stone, Patrick C Gilcrease
Anaerobic digestion (AD) of lignocellulosic materials is commonly limited by the hydrolysis step. Unlike unprocessed lignocellulosic materials, paper and paper board (PPB) are processed for their fabrication. Such modifications may affect their methane yields and methane production rates. Previous studies have investigated the correlation between lignin and biomethane yields of unprocessed lignocellulosic materials; nevertheless, there is limited knowledge regarding the relationship between the AD kinetic parameters and composition of PPB...
November 26, 2016: Biotechnology and Bioengineering
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