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https://www.readbyqxmd.com/read/27933725/cellular-engineering-for-therapeutic-protein-production-product-quality-host-modification-and-process-improvement
#1
REVIEW
Evan Wells, Anne Skaja Robinson
Recombinant proteins offer many therapeutic advantages unavailable in traditional small molecule drugs, but the need for cellular versus chemical synthesis complicates production. Avenues for producing therapeutic biologics are continuously expanding, and developments in biochemistry, cell biology, and bioengineering fuel new discoveries that promise safer, more efficient, and cheaper drugs for consumers. Numerous approaches to express recombinant proteins exist, but Escherichia coli, Saccharomyces cerevisiae, and mammalian systems (e...
December 9, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27918147/genome-analysis-of-a-hyper-acetone-butanol-ethanol-abe-producing-clostridium-acetobutylicum-bkm19
#2
Changhee Cho, Donghui Choe, Yu-Sin Jang, Kyung-Jin Kim, Won Jun Kim, Byung-Kwan Cho, E Terry Papoutsakis, George N Bennett, Do Young Seung, Sang Yup Lee
We previously reported development of a hyper acetone-butanol-ethanol (ABE) producing Clostridium acetobutylicum BKM19 strain capable of producing 30.5% more total solvent by random mutagenesis of its parental strain PJC4BK, which is a buk mutant C. acetobutylicum ATCC 824 strain. Here BKM19 and PJC4BK strains were re-sequenced by a high-throughput sequencing technique to understand the mutations responsible for enhanced solvent production. In comparison with the C. acetobutylicum PJC4BK, 13 single nucleotide variants (SNVs), 1 deletion and 1 back mutation SNV were identified in the C...
December 5, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27906513/smartphone-based-portable-wireless-optical-system-for-the-detection-of-target-analytes
#3
Shreedhar Gautam, Bhagwan S Batule, Hyo Yong Kim, Ki Soo Park, Hyun Gyu Park
Rapid and accurate on-site wireless measurement of hazardous molecules or biomarkers is one of the biggest challenges in nanobiotechnology. We herein describe a novel smartphone-based Portable and Wireless Optical System (PAWS) for rapid, quantitative, and on-site analysis of target analytes. As a proof-of-concept, we employed gold nanoparticles (GNP) and an enzyme, horse radish peroxidase (HRP), to generate colorimetric signals in response to two model target molecules, melamine and hydrogen peroxide, respectively...
December 1, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27906504/combination-of-two-epitope-identification-techniques-enables-the-rational-design-of-soy-allergen-gly-m-4-mutants
#4
Heide Havenith, Karolin Kern, Paul Rautenberger, Holger Spiegel, Michael Szardenings, Elke Ueberham, Jörg Lehmann, Matthias Buntru, Simon Vogel, Regina Treudler, Rainer Fischer, Stefan Schillberg
BACKGROUND: Detailed IgE-binding epitope analysis is a key requirement for the understanding and development of diagnostic and therapeutic agents to address food allergies. METHODS: We combined an IgE-specific linear peptide microarray with random phage peptide display for the high-resolution mapping of IgE-binding epitopes of the major soybean allergen Gly m 4, which is a homologue to the birch pollen allergen Bet v1. RESULTS: Three epitopes were identified and mapped to a resolution of four key amino acids, allowing the rational design and the production of three Gly m 4 mutants with the aim to abolish or reduce the binding of epitope-specific IgE...
December 1, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27906496/aerobic-expression-of-vitreoscilla-hemoglobin-improves-the-growth-performance-of-cho-k1-cells
#5
Mariana Juárez, Claudia H González-De la Rosa, Elisa Memún, Juan-Carlos Sigala, Alvaro R Lara
Inefficient carbon metabolism is a relevant issue during the culture of mammalian cells for the production of biopharmaceuticals. Therefore, cell engineering strategies to improve the metabolic and growth performance of cell lines are needed. The expression of Vitreoscilla stercoraria hemoglobin (VHb) has been shown to significantly reduce overflow metabolism and improve the aerobic growth of bacteria. However, the effects of VHb on mammalian cells have been rarely studied. Here, the impact of VHb on growth and lactate accumulation during CHO-K1 cell culture was investigated...
December 1, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27901315/unveiling-the-30-year-mystery-of-polyhydroxyalkanoate-pha-synthase
#6
George Guo-Qiang Chen
No abstract text is available yet for this article.
November 30, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27897385/reconstruction-of-biological-pathways-and-metabolic-networks-from-in-silico-labeled-metabolites
#7
Noushin Hadadi, Jasmin Hafner, Keng Cher Soh, Vassily Hatzimanikatis
Reaction atom mappings track the positional changes of all of the atoms between the substrates and the products as they undergo the biochemical transformation. However, information on atom transitions in the context of metabolic pathways is not widely available in the literature. The understanding of metabolic pathways at the atomic level is of great importance as it can deconvolute the overlapping catabolic/anabolic pathways resulting in the observed metabolic phenotype. The automated identification of atom transitions within a metabolic network is a very challenging task since the degree of complexity of metabolic networks dramatically increases when we transit from metabolite-level studies to atom-level studies...
November 29, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27883271/high-throughput-downstream-process-development-for-cell-based-products-using-aqueous-two-phase-systems-atps-a-case-study
#8
Sarah Zimmermann, Christian Scheeder, Philipp K Zimmermann, Are Bogsnes, Mattias Hansson, Arne Staby, Jürgen Hubbuch
The availability of preparative-scale downstream processing strategies for cell-based products presents a critical juncture between fundamental research and clinical development. Aqueous two-phase systems (ATPS) present a gentle, scalable, label-free, and cost-effective method for cell purification, and are thus a promising tool for downstream processing of cell-based therapeutics. Here, we report the application of a previously developed robotic screening platform that enables high-throughput cell partitioning analysis in ATPS...
November 24, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27878965/a-simplified-procedure-for-antibody-engineering-by-yeast-surface-display-coupling-display-levels-and-target-binding-by-ribosomal-skipping
#9
Julius Grzeschik, Steffen C Hinz, Doreen Könning, Thomas Pirzer, Stefan Becker, Stefan Zielonka, Harald Kolmar
Yeast surface display is a valuable, widely used method for protein engineering. However, current yeast display applications rely on the staining of epitope tags in order to verify full-length presentation of the protein of interest on the cell surface. We aimed at developing a modified yeast display approach that relies on ribosomal skipping, thereby enabling the translation of two proteins from one open reading frame and, in that manner, generating an intracellular fluorescence signal. This improved setup is based on a 2A sequence that is encoded between the protein to be displayed and a gene for green fluorescent protein (GFP)...
November 23, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27860405/flow-cytometric-screening-of-aggregation-inhibitors-using-a-fluorescence-assisted-intracellular-method
#10
Hanna Lindberg, Lisa Sandersjöö, Sebastian W Meister, Mathias Uhlén, John Löfblom, Stefan Ståhl
Aggregation of misfolded peptides and proteins is a key event in several neurodegenerative diseases. Suggested treatments of such disorders aim to inhibit the initial aggregation process. Here, we have developed an intracellular, function-based screening method, intended for isolation of aggregation-inhibitors from combinatorial protein libraries by flow-cytometric cell sorting. The method is based on fusion of aggregation-prone peptides to a fluorescent protein, functioning as a solubility reporter. Co-expression of a protein-based aggregation-inhibitor should prevent aggregation and thus increase the whole-cell fluorescence...
November 17, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27860394/simultaneous-conversion-of-free-fatty-acids-and-triglycerides-to-biodiesel-by-immobilized-aspergillus-oryzae-expressing-fusarium-heterosporum-lipase
#11
Jerome Amoah, Emmanuel Quayson, Shinji Hama, Ayumi Yoshida, Tomohisa Hasunuma, Chiaki Ogino, Akihiko Kondo
The presence of high levels of free fatty acids in oil is a barrier to one-step biodiesel production. Undesirable soaps are formed during conventional chemical methods, and enzyme deactivation occurs when enzymatic methods are used. This work investigates an efficient technique to simultaneously convert a mixture of free fatty acids and triglycerides. A partial soybean hydrolysate containing 73.04% free fatty acids and 24.81% triglycerides was used as a substrate for the enzymatic production of fatty acid methyl ester (FAME)...
November 17, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27860290/predictive-glycoengineering-of-biosimilars-using-a-markov-chain-glycosylation-model
#12
Philipp N Spahn, Anders H Hansen, Stefan Kol, Bjørn G Voldborg, Nathan E Lewis
Biosimilar drugs must closely resemble the pharmacological attributes of innovator products to ensure safetyand efficacy to obtain regulatory approval. Glycosylation is one critical quality attribute that must be matched, but it is inherently difficult to control due to the complexity of its biogenesis. This usually implies that costly and time-consuming experimentation is required for clone identification and optimization of biosimilar glycosylation. Here, we describe a computational method that utilizes a Markov model of glycosylation to predict optimal glycoengineering strategies to obtain a specific glycosylation profile with desired properties...
November 17, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27863041/immobilized-hematopoietic-growth-factors-onto-magnetic-particles-offer-a-scalable-strategy-for-cell-therapy-manufacturing-in-suspension-cultures
#13
Matthew J Worrallo, Rebecca L L Moore, Katie E Glen, Robert J Thomas
Hematopoietic therapies require high cell dosages and precise phenotype control for clinical success; scalable manufacturing processes therefore need to be economic and controllable, in particular with respect to culture medium and growth factor (GF) strategy. The aim of this work was to demonstrate the biological function, and integration within scalable systems, of a highly controllable immobilized growth factor (iGF) approach. GFs were biotinylated and attached to streptavidin coated magnetic particles. GF concentration during biotinylation, GF-biotin ratio, and GF lysine content were shown to control iGF surface concentration and enable predictable co-presentation of multiple GF on a single bead...
November 11, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27809410/systems-biology-for-understanding-and-engineering-of-heterotrophic-oleaginous-microorganisms
#14
REVIEW
Beom Gi Park, Minsuk Kim, Joonwon Kim, Heewang Yoo, Byung-Gee Kim
Heterotrophic oleaginous microorganisms continue to draw interest as they can accumulate a large amount of lipids which is a promising feedstock for the production of biofuels and oleochemicals. Nutrient limitation, especially nitrogen limitation, is known to effectively trigger the lipid production in these microorganisms. For the aim of developing improved strains, the mechanisms behind the lipid production have been studied for a long time. Nowadays, system-level understanding of their metabolism and associated metabolic switches is attainable with modern systems biology tools...
November 3, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27808482/crystal-structure-of-ralstonia-eutropha-polyhydroxyalkanoate-synthase-c-terminal-domain-and-reaction-mechanisms
#15
Jieun Kim, Yeo-Jin Kim, So Young Choi, Sang Yup Lee, Kyung-Jin Kim
Polyhydroxyalkanoates (PHAs) are natural polyesters synthesized by numerous microorganisms as energy and reducing power storage materials, and have attracted much attention as substitutes for petroleum-based plastics. Here, we report the first crystal structure of Ralstonia eutropha PHA synthase at 1.8 Å resolution and structure-based mechanisms for PHA polymerization. RePhaC1 contains two distinct domains, the N-terminal (RePhaC1ND ) and C-terminal domains (RePhaC1CD ), and exists as a dimer. RePhaC1CD catalyzes polymerization via non-processive ping-pong mechanism using a Cys-His-Asp catalytic triad...
November 3, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27808475/structure-and-function-of-the-n-terminal-domain-of-ralstonia-eutropha-polyhydroxyalkanoate-synthase-and-the-proposed-structure-and-mechanisms-of-the-whole-enzyme
#16
Yeo-Jin Kim, So Young Choi, Jieun Kim, Kyeong Sik Jin, Sang Yup Lee, Kyung-Jin Kim
Polyhydroxyalkanoates (PHAs) are natural polyesters synthesized by numerous microorganisms as energy and reducing power storage materials, and have attracted much attention as substitutes for petroleum-based plastics. In an accompanying paper, the authors reported the crystal structure of the C-terminal domain of Ralstonia eutropha PHA synthase (PhaC1). Here, the authors report the 3D reconstructed model of full-length of R. eutropha PhaC1 (RePhaC1F ) by small angle X-ray scattering (SAXS) analysis. The catalytic C-terminal domain of RePhaC1 (RePhaC1CD ) dimer is located at the center of RePhaC1F , and the N-terminal domain of RePhaC1 (RePhaC1ND ) is located opposite the dimerization subdomain of RePhaC1CD , indicating that RePhaC1ND is not directly involved in the enzyme catalysis...
November 3, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27797150/the-potential-of-nanoparticles-in-stem-cell-differentiation-and-further-therapeutic-applications
#17
REVIEW
Ahmed Abdal Dayem, Hye Yeon Choi, Gwang-Mo Yang, Kyeongseok Kim, Subbroto Kumar Saha, Jin-Hoi Kim, Ssang-Goo Cho
Tissue regeneration could offer therapeutic advantages for individuals experiencing organ or tissue damage. Recently, advances in nanotechnology have provided various nanomaterials, with a wide range of applications, for modulating stem cell behavior and for further therapeutic applications in tissue regeneration. Defects in cell proliferation and differentiation, a low mechanical strength of scaffolds, and inefficient production of factors that are essential for stem cell differentiation are the current challenges in tissue regeneration...
October 31, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27787955/nanomaterials-based-biosensors-for-detection-of-microorganisms-and-microbial-toxins
#18
REVIEW
Laura Sutarlie, Sian Yang Ow, Xiaodi Su
Detection of microorganisms and microbial toxins is important for health and safety. Due to their unique physical and chemical properties, nanomaterials have been extensively used to develop biosensors for rapid detection of microorganisms with microbial cells and toxins as target analytes. In this paper, the design principles of nanomaterials-based biosensors for four selected analyte categories (bacteria cells, toxins, mycotoxins, and protozoa cells), closely associated with the target analytes' properties is reviewed...
October 27, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27783465/protease-substrate-profiling-using-bacterial-display-of-self-blocking-affinity-proteins-and-flow-cytometric-sorting
#19
Lisa Sandersjöö, Andreas Jonsson, John Löfblom
Proteases are involved in fundamental biological processes and are important tools in both biotechnological and biomedical research. An important property of proteases is to discriminate among potential substrates. Here, we present a new method for substrate profiling of proteases. The substrates are displayed between two anti-idiotypic affinity domains on the Gram-positive bacterium Staphylococcus carnosus. The first domain functions as a reporter tag and has affinity for a labeled reporter protein, whereas the second domain blocks the reporter tag from interacting with the reporter protein...
October 26, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27778472/impedance-technology-reduces-the-enumeration-time-of-brettanomyces-yeast-during-beer-fermentation
#20
Sanelle van Wyk, Filipa V M Silva
Brettanomyces yeasts are increasingly being used to produce lambic style beers and craft beers with unique flavours. Currently, the industry monitors Brettanomyces bruxellensis using time consuming plate counting. B. bruxellensis is a fastidious slow growing organism, requiring 5 days of incubation at 30 °C for visible growth on agar plates. Thus, a need exists to develop a quicker, feasible method to enumerate this yeast. The aim of this study was therefore to determine the feasibility of using the 'direct' and 'indirect' impedance methods for the enumeration of B...
October 25, 2016: Biotechnology Journal
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