journal
MENU ▼
Read by QxMD icon Read
search

Biotechnology Journal

journal
https://www.readbyqxmd.com/read/28301074/high-throughput-inclusion-body-sizing-nano-particle-tracking-analysis
#1
Wieland N Reichelt, Andreas Kaineder, Markus Brillmann, Lukas Neutsch, Alexander Taschauer, Hans Lohninger, Christoph Herwig
The expression of pharmaceutical relevant proteins in E. coli frequently triggers inclusion body (IB) formation caused by protein aggregation. In literature, substantial effort has been invested into the quantification of IB size. But up to this date particle based methods to analyze physical IB properties of representative numbers of IBs lack sensitivity and/or lack orthogonal verification. Using high pressure freezing and automated freeze substitution for transmission electron microscopy (TEM) the cytosolic inclusion body structure was preserved within the cells...
March 16, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28299901/dual-display-of-proteins-on-the-yeast-cell-surface-simplifies-quantification-of-binding-interactions-and-enzymatic-bioconjugation-reactions
#2
Sungwon Lim, Jeff E Glasgow, Maria Filsinger Interrante, Erica M Storm, Jennifer R Cochran
Yeast surface display, a well-established technology for protein analysis and engineering, involves expressing a protein of interest as a genetic fusion to either the N- or C-terminus of the yeast Aga2p mating protein. Historically, yeast-displayed protein variants are flanked by peptide epitope tags that enable flow cytometric measurement of construct expression using fluorescent primary or secondary antibodies. Here, we built upon this technology to develop a new yeast display strategy that comprises fusion of two different proteins to Aga2p, one to the N-terminus and one to the C-terminus...
March 16, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28296345/a-cleavable-self-assembling-tag-strategy-for-preparing-proteins-and-peptides-with-an-authentic-n-terminus
#3
Qing Zhao, Bihong Zhou, Xianxing Gao, Lei Xing, Xu Wang, Zhanglin Lin
Recombinant protein expression and purification remains a central need for biotechnology. Herein, we report a streamlined protein and peptide purification strategy using short self-assembling peptides and a C-terminal cleavage intein. In this strategy, the fusion protein is first expressed as an aggregate induced by the self-assembling peptide. Upon simple separation, the target protein or peptide with an authentic N-terminus is then released in the solution by intein-mediated cleavage. Different combinations of four self-assembling peptides (ELK16, L6 KD, FK and FR) with three inteins (Sce VMA, Mtu ΔI-CM and Ssp DnaB) were explored...
March 15, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28294579/experimental-validation-of-in-silico-estimated-biomass-yields-of-pseudomonas-putida-kt2440
#4
Sarah Beate Hintermayer, Dirk Weuster-Botz
Pseudomonas putida is rapidly becoming a microbial cell platform for biotechnological applications. In order to understand genotype-phenotype relationships genome scale models represent helpful tools. However, the validation of in silico predictions of genome scale models is a task that is rarely performed. In this study the theoretical biomass yields of Pseudomonas putida KT2440 were estimated for 57 different carbon sources based on a genome scale stoichiometric model applying flux balance analysis. The batch growth of P...
March 14, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28294544/identification-of-acetylcholinesterase-inhibitors-using-homogenous-cell-based-assays-in-quantitative-high-throughput-screening-platforms
#5
Shuaizhang Li, Ruili Huang, Samuel Solomon, Yitong Liu, Bin Zhao, Michael F Santillo, Menghang Xia
Acetylcholinesterase (AChE) is an enzyme responsible for metabolism of acetylcholine, a neurotransmitter associated with muscle movement, cognition, and other neurobiological processes. Inhibition of AChE activity can serve as a therapeutic mechanism, but also cause adverse health effects and neurotoxicity. In order to efficiently identify AChE inhibitors from large compound libraries, homogenous cell-based assays in high-throughput screening platforms are needed. In this study, a fluorescent method using Amplex Red (10-acetyl-3,7-dihydroxyphenoxazine) and the Ellman absorbance method were both developed in a homogenous format using a human neuroblastoma cell line (SH-SY5Y)...
March 14, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28296355/biosensor-enabled-directed-evolution-to-improve-muconic-acid-production-in-saccharomyces-cerevisiae
#6
John M Leavitt, James M Wagner, Cuong Chi Tu, Alice Tong, Yanyi Liu, Hal S Alper
Muconic acid is a valuable platform chemical with potential applications in the production of polymers such as nylon and polyethylene terephthalate (PET). The conjugate base, muconate, was previously biosynthesized in the bacterial host Escherichia coli. Significant pathway engineering led to the first reported instance of rationally engineered production of muconic acid in the yeast Saccharomyces cerevisiae. To further increase muconic acid production in this host, we employed an adaptive laboratory evolution (ALE) strategy to complement rational metabolic engineering...
March 10, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28296342/delivery-of-growth-factor-based-therapeutics-in-vascular-diseases-challenges-and-strategies
#7
REVIEW
He-Lin Xu, Wen-Ze Yu, Cui-Tao Lu, Xiao-Kun Li, Ying-Zheng Zhao
Either cardiovascular or peripheral vascular diseases have become the major cause of morbidity and mortality worldwide. Recently, growth factors therapeutics, whatever administrated in form of exogenous growth factors or their relevant genes have been discovered to be an effective strategy for the prevention and therapy of vascular diseases, because of their promoting angiogenesis. Besides, as an alternative, stem cell-based therapy has been also developed in view of their paracrine-mediated effect or ability of differentiation toward angiogenesis-related cells under assistance of growth factors...
March 10, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28294564/cultivation-based-strategies-to-find-efficient-marine-biocatalysts
#8
Carlos J C Rodrigues, Ricardo F S Pereira, Pedro Fernandes, Joaquim M S Cabral, Carla C C R de Carvalho
Marine bacteria have evolved to survive in the marine environment by using unique physiological, biochemical and metabolic features and the ability to produce enzymes and compounds which may have commercial value. The Azores archipelago presents several ecosystems with strong volcanic activity where bacteria thrive under e.g. high temperatures. In this study, samples collected in the island of São Miguel were screened for biocatalysts possessing e.g. lipase, esterase, amylase, and inulinase activities. After isolation of several hundred bacterial strains, high throughput screening methods allowed the fast identification of biocatalysts...
March 10, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28263439/protein-refolding-is-improved-by-adding-nonionic-polyethylene-glycol-monooleyl-ethers-with-various-polyethylene-glycol-lengths
#9
Etsushi Yamamoto, Satoshi Yamaguchi, Teruyuki Nagamune
Protein refolding from bacterial inclusion bodies is a crucial step for the production of recombinant proteins, but the refolding step often results in significantly lower yields due to aggregation. To prevent aggregation, chemical additives are often used. However, the ability of additives to effectively increase refolding yields are protein dependent, and therefore, it is important to understand the manner in which the substructures of additives confer suitable properties on protein refolding. We focused attention on nonionic detergents, the polyethylene glycol monooleyl ether (PGME) series, and systematically studied the influence of two to 90 polyethylene glycol (PEG) lengths of PGMEs on the refolding of pig muscle lactate dehydrogenase (LDH), hen egg white lysozyme, and yeast α-glucosidase...
March 6, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28256799/prospects-and-challenges-for-the-recovery-of-2-butanol-produced-by-vacuum-fermentation-a-techno-economic-analysis
#10
Joana P C Pereira, Gustavo Lopez-Gomez, Noelia G Reyes, Luuk A M van der Wielen, Adrie J J Straathof
The conceptual design of a bio-based process for 2-butanol production is presented for the first time. Considering a hypothetical efficient producing strain, a vacuum fermentation is proposed to alleviate product toxicity, but the main challenge is the energy-efficient product recovery from the vapor. Three downstream scenarios were examined for this purpose: 1) multi-stage vapor recompression; 2) temperature swing adsorption; and 3) vapor absorption. The processes were simulated using Aspen Plus, considering a production capacity of 101 kton/yr...
March 3, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28233468/an-automated-laboratory-scale-methodology-for-the-generation-of-sheared-mammalian-cell-culture-samples
#11
Adrian Joseph, Stephen Goldrick, Michael Mollet, Richard Turner, Jean Bender, David Gruber, Suzanne S Farid, Nigel Titchener-Hooker
Continuous disk-stack centrifugation is typically used for the removal of cells and cellular debris from mammalian cell culture broths at manufacturing-scale. The use of scale-down methods to characterise disk-stack centrifugation performance enables substantial reductions in material requirements and allows a much wider design space to be tested than is currently possible at pilot-scale. The process of scaling down centrifugation has historically been challenging due to the difficulties in mimicking the Energy Dissipation Rates (EDRs) in typical machines...
February 24, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28230321/disruption-of-genes-involved-in-corvet-complex-leads-to-enhanced-secretion-of-heterologous-carboxylesterase-only-in-protease-deficient-pichia-pastoris
#12
Lukas Marsalek, Clemens Gruber, Friedrich Altmann, Markus Aleschko, Diethard Mattanovich, Brigitte Gasser, Verena Puxbaum
The methylotrophic yeast Pichia pastoris (Komagataella spp.) is a popular microbial host for the production of recombinant proteins. Previous studies have shown that mis-sorting to the vacuole can be a bottleneck during production of recombinant secretory proteins in yeast, however, no information was available for P. pastoris. In this work we have therefore generated vps (vacuolar protein sorting) mutant strains disrupted in genes involved in the CORVET tethering complex at the early stages of endosomal sorting...
February 23, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28221723/animal-component-free-agrobacterium-tumefaciens-cultivation-media-for-better-gmp-compliance-increases-biomass-yield-and-pharmaceutical-protein-expression-in-nicotiana-benthamiana
#13
Marcel Houdelet, Anna Galinski, Tanja Holland, Kathrin Wenzel, Stefan Schillberg, Johannes Felix Buyel
Transient expression systems allow the rapid production of recombinant proteins in plants. Such systems can be scaled up to several hundred kilograms of biomass, making them suitable for the production of pharmaceutical proteins required at short notice, such as emergency vaccines. However, large-scale transient expression requires the production of recombinant Agrobacterium tumefaciens strains with the capacity for efficient gene transfer to plant cells. The complex media often used for the cultivation of this species typically include animal-derived ingredients that can contain human pathogens, thus conflicting with the requirements of good manufacturing practice (GMP)...
February 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28220995/development-of-hydrogels-for-regenerative-engineering
#14
REVIEW
Xiaofei Guan, Meltem Avci-Adali, Emine Alarçin, Hao Cheng, Sara Saheb Kashaf, Yuxiao Li, Aditya Chawla, Hae Lin Jang, Ali Khademhosseini
The aim of regenerative engineering is to restore complex tissues and biological systems through convergence in the fields of advanced biomaterials, stem cell science, and developmental biology. Hydrogels are one of the most attractive biomaterials for regenerative engineering, since they can be engineered into tissue mimetic 3D scaffolds to support cell growth due to their similarity to native extracellular matrix. Advanced nano- and micro-technologies have dramatically increased the ability to control properties and functionalities of hydrogel materials by facilitating biomimetic fabrication of more sophisticated compositions and architectures, thus extending our understanding of cell-matrix interactions at the nanoscale...
February 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28217906/production-of-taxadiene-by-engineering-of-mevalonate-pathway-in-escherichia-coli-and-endophytic-fungus-alternaria-alternata-tpf6
#15
Guangkai Bian, Yujie Yuan, Hui Tao, Xiaofei Shi, Xiaofang Zhong, Yichao Han, Shuai Fu, Chengxiang Fang, Zixin Deng, Tiangang Liu
Taxol (paclitaxel) is a diterpenoid compound with significant and extensive applications in the treatment of cancer. The production of Taxol and relevant intermediates by engineered microbes is an attractive alternative to the semichemical synthesis of Taxol. In this study, based on a previously developed platform, we first established taxadiene production in mutant E. coli T2 and T4 by engineering of the mevalonate (MVA) pathway. We then developed an Agrobacterium tumefaciens-mediated transformation (ATMT) method and verified the strength of heterologous promoters in Alternaria alternata TPF6...
February 20, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28177193/manufacturing-of-recombinant-adeno-associated-viruses-using-mammalian-expression-platforms
#16
REVIEW
Marc-André Robert, Parminder S Chahal, Alexandre Audy, Amine Kamen, Rénald Gilbert, Bruno Gaillet
Manufacturing practices for recombinant adeno-associated viruses (AAV) have improved in the last decade through the development of new platforms in conjunction with better production and purification methods. In this review, we discuss the advantages and limitations of the most popular systems and methods employed with mammalian cell platforms. Methods and systems such as transient transfection, packaging and producer cells and adenovirus and herpes simplex virus are described. In terms of best production yields, they are comparable with about 10(4) -10(5) vector genomes produced per cell but transient transfection of HEK293 cells is by far the most commonly used...
February 8, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28169491/fermentative-production-of-l-pipecolic-acid-from-glucose-and-alternative-carbon-sources
#17
Fernando Pérez-García, Joe Max Risse, Karl Friehs, Volker F Wendisch
Corynebacterium glutamicum is used for the million-ton scale production of amino acids and has recently been engineered for production of the cyclic non-proteinogenic amino acid L-pipecolic acid (L-PA). In this synthetic pathway L-lysine was converted to L-PA by oxidative deamination, dehydration and reduction by L-lysine 6-dehydrogenase (deaminating) from Silicibacter pomeroyi and pyrroline 5-carboxylate reductase from C. glutamicum. However, production of L-PA occurred as by-product of L-lysine production only...
February 7, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28166394/exosome-separation-using-microfluidic-systems-size-based-immunoaffinity-based-and-dynamic-methodologies
#18
REVIEW
Fang Yang, Xiangzhi Liao, Yuan Tian, Guiying Li
Exosomes, nanovesicles secreted by most types of cells, exist in virtually all bodily fluids. Their rich nucleic acid and protein content make them potentially valuable biomarkers for noninvasive molecular diagnostics. They also show promise, after further development, to serve as a drug delivery system. Unfortunately, existing exosome separation technologies, such as ultracentrifugation and methods incorporating magnetic beads, are time-consuming, laborious and separate only exosomes of low purity. Thus, a more effective separation method is highly desirable...
February 6, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28146316/mir-92a-enhances-recombinant-protein-productivity-in-cho-cells-by-increasing-intracellular-cholesterol-levels
#19
Wan Ping Loh, Yuansheng Yang, Kong Peng Lam
MicroRNAs (miRNAs) have emerged as promising targets for engineering of CHO cell factories to enhance recombinant protein productivity. Manipulation of miRNA levels in CHO cells have been shown to improve product yield by increasing proliferation and specific productivity (qP), resisting apoptosis and enhancing oxidative metabolism. We previously demonstrated that over-expressing miR-92a results in increases in qP and titer of CHO-IgG cells. However, the mechanisms by which miR-92a enhances qP in CHO cells are still uninvestigated...
February 1, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28139886/genomic-integration-of-the-full-length-dystrophin-coding-sequence-in-duchenne-muscular-dystrophy-induced-pluripotent-stem-cells
#20
Alfonso P Farruggio, Mital S Bhakta, Haley du Bois, Julia Ma, Michele P Calos
We developed plasmid vectors that express the full-length human dystrophin coding sequence in human cells. Dystrophin, the protein mutated in Duchenne muscular dystrophy, is extraordinarily large, providing challenges for cloning and plasmid production in E. coli. We expressed dystrophin from the strong, widely expressed CAG promoter, along with co-transcribed luciferase and mCherry marker genes useful for tracking plasmid expression. Introns were added at the 3' and 5' ends of the dystrophin sequence to prevent translation in E...
January 31, 2017: Biotechnology Journal
journal
journal
41277
1
2
Fetch more papers »
Fetching more papers... Fetching...
Read by QxMD. Sign in or create an account to discover new knowledge that matter to you.
Remove bar
Read by QxMD icon Read
×

Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"