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Biotechnology and Bioengineering

Jie Dong, Yan Chen, Veronica Teixeira Benites, Edward E K Baidoo, Christopher J Petzold, Harry R Beller, Aymerick Eudes, Henrik V Scheller, Paul D Adams, Aindrila Mukhopadhyay, Blake A Simmons, Steven W Singer
Plants are an attractive source of renewable carbon for conversion to biofuels and bio-based chemicals. Conversion strategies often use a fraction of the biomass, focusing on sugars from cellulose and hemicellulose. Strategies that use plant components such as aromatics and amino acids may improve the efficiency of biomass conversion. Pseudomonas putida is a promising host for its ability to metabolize a wide variety of organic compounds. P. putida was engineered to produce methyl ketones, which are promising diesel blendstocks and potential platform chemicals, from glucose and lignin-related aromatics...
April 14, 2019: Biotechnology and Bioengineering
Samuel D Stimple, Sibel Kalyoncu, Alec A Desai, Jesper E Mogensen, Lotte T Spang, Désirée J Asgreen, Arne Staby, Peter M Tessier
Sensitive detection of protein aggregates is important for evaluating the quality of biopharmaceuticals and detecting misfolded proteins in several neurodegenerative diseases. However, it is challenging to detect extremely low concentrations (<10 ppm) of aggregated protein in the presence of high concentrations of natively-folded (soluble) protein. Glucagon, a peptide hormone used in the treatment of extreme hypoglycemia, is aggregation-prone and forms amyloid fibrils. Detection of glucagon fibrils using conformation-specific antibodies is an attractive approach for identifying such aggregates during process and formulation development...
April 14, 2019: Biotechnology and Bioengineering
Christopher T Straub, Piyum A Khatibi, Jonathan K Otten, Michael W W Adams, Robert M Kelly
The extreme thermophile Caldicellulosiruptor bescii solubilizes and metabolizes the carbohydrate content of lignocellulose, a process that ultimately ceases because of biomass recalcitrance, accumulation of fermentation products, inhibition by lignin moieties, and reduction of metabolic activity. Deconstruction of low loadings of lignocellulose (5 g/l), either natural or transgenic, whether unpretreated or subjected to hydrothermal processing, by C. bescii typically results in less than 40% carbohydrate solubilization...
April 14, 2019: Biotechnology and Bioengineering
Hadi Nazem-Bokaee, Dayue Chen, Sean Michael O'Donnell, Andrew L Zydney
Virus filtration remains a critical step in the downstream process for the production of monoclonal antibodies and other mammalian cell-derived biotherapeutics. Recent studies have shown large differences in virus capture behavior of different virus filters, although the origin of these differences is still unclear. The objective of this study was to use confocal and scanning electron microscopy to directly evaluate the capture of virus-size nanoparticles in Planova 20N and BioEX hollow fiber virus filters...
April 14, 2019: Biotechnology and Bioengineering
Lina Apitius, Kristin Rübsam, Christina Jakesch, Felix Jakob, Ulrich Schwaneberg
Accumulation of plastics in the environment became a geological indicator of the Anthropocene era. An effective reduction of long-lasting plastics requires a treatment with microorganisms that release polymer-degrading enzymes. Polymer binding peptides function as adhesion promoters and enable a targeted binding of whole cells to polymer surfaces. An esterase A-based E. coli cell surface display screening system was developed, that enabled directed evolution of polymer binding peptides for improved binding strength to polymers...
April 14, 2019: Biotechnology and Bioengineering
Ghislaine Béhar, Axelle Renodon-Cornière, Stanimir Kambarev, Petar Vukojicic, Nathalie Caroff, Stéphane Corvec, Barbara Mouratou, Frédéric Pecorari
Detection and capture methods using antibodies have been developed to ensure identification of pathogens in biological samples. Though antibodies have many attractive properties, they also have limitations and there are needs to expand the panel of available affinity proteins with different properties. Affitins, that we developed from the Sul7d proteins, are a solid class of affinity proteins which can be used as substitutes to antibodies or to complement them. We report the generation and characterization of anti-bacterial Affitins with high specificity for Staphylococcus aureus...
April 14, 2019: Biotechnology and Bioengineering
Salvatore Petralia, Daniele Motta, Sabrina Conoci
In this paper, a miniaturized biosensor containing 96 silicon micro-chambers electro-loaded with nano-volumes of liquid (EW-Chip) is presented. The liquid electro-loading is achieved by the appropriate modulation of interface properties. The surface chemistries have been studied to guarantee effective interface properties for both ElectroWetting on Dielectric (EWoD) actuaction and biocompatibility versus biochemical reactions. The silicon micro-chambers are 200 nl in volume and are connected to a specific system of electrodes able to deliver liquid sample on each wells...
April 9, 2019: Biotechnology and Bioengineering
Shyam Sundhar Bale, Andrea Manoppo, Rebecca Thompson, Alex Markoski, Jonathan Coppeta, Hesham Azizgolshani, Mingjian Lu, James Gosset, Philip Keegan, Joe Charest
Hepatic in vitro platforms ranging from multi-well cultures to bioreactors and microscale systems have been developed as tools to recapitulate cellular function and responses to aid in drug screening and disease model development. Recent developments in microfabrication techniques and cellular materials enabled fabrication of next-generation, advanced microphysiological systems (MPSs) that aim to capture the cellular complexity and dynamic nature of the organ presenting highly controlled extracellular cues to cells in a physiologically relevant context...
April 9, 2019: Biotechnology and Bioengineering
Nico Betterle, Anastasios Melis
The work aims to convert the secondary slow metabolism of the terpenoid biosynthetic pathway into a primary activity in cyanobacteria, and to generate heterologous products using these photosynthetic microorganisms as cell factories. Case study is the production of the 10-carbon monoterpene β-phellandrene (PHL) in Synechocystis sp. PCC 6803 (Synechocystis). Barriers to this objective include the slow catalytic activity of the terpenoid metabolism enzymes that limits rates and yield of product synthesis and accumulation...
April 9, 2019: Biotechnology and Bioengineering
Matthew Gagnon, Shashikant Nagre, Wenge Wang, Jon Coffman, Gregory W Hiller
A novel, alternative intensified cell culture process comprised of a linked bioreactor system is presented. An N-1 perfusion bioreactor maintained cells in a highly proliferative state and provided a continuous inoculum source to a second bioreactor operating as a continuous-flow stirred-tank reactor (CSTR). An initial study evaluated multiple system steady-states by varying N-1 steady-state viable cell densities, N-1 to CSTR working volume ratios, and CSTR dilution rates. After identifying near optimum system steady-state parameters yielding a relatively high volumetric productivity while efficiently consuming media, a subsequent lab scale experiment demonstrated the start-up and long-term operation of the envisioned manufacturing process for 83 days...
April 5, 2019: Biotechnology and Bioengineering
Chae Rin Park, Won Jong Rhee, Kyu Won Kim, Byeong Hee Hwang
On-site genetic detection needs to develop a sensitive and straightforward biosensor without special equipment which can detect various genetic biomarkers. Hybridization chain reaction amplifying signal isothermally could be considered as a good candidate for on-site detection. Here, we developed a novel genetic biosensor based on enzyme-free dual-amplification of universal hybridization chain reaction (uHCR) and hemin/G-quadruplex horseradish peroxidase (HRP)-mimicking DNAzyme. The uHCR is the strategy which enables simple design for multiple target detection by the introduction of target-specific trigger hairpin without changing the whole system according to a target change...
April 1, 2019: Biotechnology and Bioengineering
Sun Jin Kim, Samuel T Hatch, Andrew S Dixon, Shawn C Owen
Expressing, isolating, and characterizing recombinant proteins is crucial to many disciplines within the biological sciences. Different molecular tagging technologies have been developed to enable each individual step of protein production, from expression through purification and characterization. Monitoring the entire production process requires multiple tags or molecular interactions, because no individual tag has provided the comprehensive breadth of utility. An ideal molecular tag is small and does not interrupt expression, solubility, folding or function of the protein being purified and can be used throughout the production process...
April 1, 2019: Biotechnology and Bioengineering
Zhongmei Liu, Han Zhao, Laichuang Han, Wenjing Cui, Li Zhou, Zhemin Zhou
Nattokinase (NK) is a serine protease of the subtilisin family; as a potent fibrinolytic enzyme, it is potentially useful for thrombosis therapy. For NK to be applied as an oral medicine for the treatment of cardiovascular diseases, it must overcome the extremely acidic environments of the gastrointestinal tract despite its limited acidic stability. In this study, three strategies were adopted to improve the acid resistance of NK: (1) surface charge engineering, (2) sequence alignment, and (3) mutation based on the literature...
April 1, 2019: Biotechnology and Bioengineering
Chengchao Zhu, Jiuzhou Chen, Yu Wang, Lixian Wang, Xuan Guo, Ning Chen, Ping Zheng, Jibin Sun, Yanhe Ma
5-Aminolevulinic acid (ALA) is a value-added compound with potential applications in the fields of agriculture and medicine. Although massive efforts have recently been devoted to building microbial producers of ALA through metabolic engineering, few studies focused on the cellular response and tolerance to ALA. In this study, we demonstrated that ALA caused severe cell damage and morphology change of Escherichia coli via generating reactive oxygen species (ROS), which were further determined to be mainly hydrogen peroxide and superoxide anion radical...
April 1, 2019: Biotechnology and Bioengineering
Ru Zang, Xin Xin, Fengli Zhang, Ding Li, Shang-Tian Yang
Embryonic stem cell test (EST) is the only generally accepted in vitro method for assessing embryotoxicity without animal sacrifice. However, the implementation and application of EST for regulatory embryotoxicity screening are impeded by its technical complexity, long testing period, and limited endpoint data. In this study, a high throughput embryotoxicity screening based on mouse embryonic stem cells (mESCs) expressing enhanced green fluorescent protein (EGFP) driven by a human survivin promoter and a human cytomegalovirus (CMV) promoter, respectively, was developed...
April 1, 2019: Biotechnology and Bioengineering
Dominik Georg Sauer, Michael Melcher, Magdalena Mosor, Nicole Walch, Matthias Berkemeyer, Theresa Scharl-Hirsch, Friedrich Leisch, Alois Jungbauer, Astrid Dürauer
Process analytical technology combines understanding and control of the process with real-time monitoring of critical quality and performance attributes. The goal is to ensure the quality of the final product. Currently, chromatographic processes in biopharmaceutical production are predominantly monitored with UV/Vis absorbance and a direct correlation with purity and quantity is limited. In this study a chromatographic workstation was equipped with additional online sensors, such as multi-angle light scattering, refractive index, attenuated total reflection Fourier-transform infrared, and fluorescence spectroscopy...
April 1, 2019: Biotechnology and Bioengineering
Nathanael D Sallada, Lauren E Harkins, Bryan W Berger
Hydrophobins are small highly surface-active fungal proteins with potential as biosurfactants in a wide array of applications. However, practical implementation of hydrophobins at large scale has been hindered by low recombinant yields. In this study, the effects of increasing hydrophobin gene copy number and overexpressing endoplasmic reticulum resident chaperone proteins Kar2p, Pdi1p, and Ero1p were explored as a means to enhance recombinant yields of the class II hydrophobin HFBI in the eukaryotic expression host Pichia pastoris...
April 1, 2019: Biotechnology and Bioengineering
Jacob Nedergaard Pedersen, Ye Zhou, Zheng Guo, Bianca Pérez
Surface charge engineering has received considerable interest from the scientific and industrial community the last decades. Although it was previously hypothesized that the surface charge-charge interactions were not a fundamental force to determine protein folding and stability, many studies today show that surface charge plays a key role determining protein structure and activity. This review aims to i) highlight the value of surface charged engineering of proteins to improve enzyme stability and activity in aqueous media and in the presence of ionic liquids, and organic solvents, ii) describe the existing approaches (genetic engineering or chemical modifications) for surface charged engineering, and iii) demonstrate the applicability of these surface charged enzymes in biocatalysis...
March 30, 2019: Biotechnology and Bioengineering
Kyoung-Joo Jenny Park, James Kim, Thomas Testoff, Joseph Adams, Miranda Poklar, Maciej Zborowski, Monica Venere, Jeffrey J Chalmers
This study focuses on different iron regulation mechanisms of glioblastoma (GBM) cancer stem-like cells (CSCs) and non-stem tumor cells (NSTCs) using multiple approaches: cell viability, density and magnetophoresis. GBM CSCs and NSTCs were exposed to elevated iron concentration, and their magnetic susceptibility was measured using Single Cell Magnetophoresis (SCM), which tracks the magnetic and settling velocities of thousands of individual cells passing through the magnetic field with a constant energy gradient...
March 24, 2019: Biotechnology and Bioengineering
Maryam Ataeian, Yihua Liu, Karen Andrea Canon-Rubio, Michael Nightingale, Marc Strous, Agasteswar Vadlamani
Bioenergy with carbon capture and storage (BECCS) is recognized as a potential negative emission technology, needed to keep global warming within safe limits. With current technologies, large-scale implementation of BECCS would compromise food production. Bioenergy derived from phototrophic microorganisms, with direct capture of CO2 from air, could overcome this challenge and become a sustainable way to realize BECCS. Here we present an alkaline capture and conversion system that combines high atmospheric CO2 transfer rates with high and robust phototrophic biomass productivity (15...
March 24, 2019: Biotechnology and Bioengineering
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