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https://www.readbyqxmd.com/read/27887723/synergistic-effect-of-cavitation-and-agitation-on-protein-aggregation
#1
Tetsuo Torisu, Takahiro Maruno, Yoshinori Hamaji, Tadayasu Ohkubo, Susumu Uchiyama
It was recently reported that dropping induces protein aggregation due to the occurrence of cavitation. Agitation also causes protein aggregation. In this study, vials filled with antibody solution were subjected to a cycle of dropping and shaking using the friability testing apparatus to examine the combined effect of cavitation and agitation on protein aggregation. A cycle of dropping and shaking generated a massive amount of subvisible particles. Comparison of aggregation rate at different fill volumes indicated that shaking plays an important role in protein aggregation due to combination stress...
November 22, 2016: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/27864159/high-level-expression-and-efficient-refolding-of-therapeutically-important-recombinant-human-interleukin-3-hil-3-in-e-%C3%A2-coli
#2
Vikas Kumar Dagar, Adivitiya, Yogender Pal Khasa
Human interleukin-3 (hIL-3) is a pleiotropic cytokine that stimulates the differentiation and proliferation of multipotent hematopoietic cells thus making it a therapeutically important molecule. In this study, its poor expression yield was improved by addressing various upstream bottlenecks in E. coli heterologous system. The codon-optimized hIL-3 gene was cloned under various signal sequences and solubility enhancer fusion tags for its hyper-expression under a strong T7 promoter. The optimization of shake flask expression studies resulted in a hIL-3 protein concentration of 225 mg/L in the form of inclusion bodies (IBs)...
November 15, 2016: Protein Expression and Purification
https://www.readbyqxmd.com/read/27836963/phosphorylated-groucho-delays-differentiation-in-the-follicle-stem-cell-lineage-by-providing-a-molecular-memory-of-egfr-signaling-in-the-niche
#3
Michael J Johnston, Shaked Bar-Cohen, Ze'ev Paroush, Todd G Nystul
In the epithelial follicle stem cells (FSCs) of the Drosophila ovary, Epidermal Growth Factor Receptor (EGFR) signaling promotes self-renewal whereas Notch signaling promotes differentiation of the prefollicle cell (pFC) daughters. We have identified two proteins, Six4 and Groucho (Gro), that link the activity of these two pathways to regulate the earliest cell fate decision in the FSC lineage. Our data indicate that Six4 and Gro promote differentiation toward the polar cell fate by promoting Notch pathway activity...
November 11, 2016: Development
https://www.readbyqxmd.com/read/27825486/high-level-expression-of-a-zen-detoxifying-gene-by-codon-optimization-and-biobrick-in-pichia-pastoris
#4
La Xiang, Qinhong Wang, Yuling Zhou, Lifeng Yin, Guimin Zhang, Yanhe Ma
The mycotoxin zearalenone (ZEN) can be degraded by a lactone hydrolase ZHD, which was derived from Gliocladium roseum. Here, based on the native ZHD encoding gene zhd101, a codon optimized zhd gene was synthesized, which was used for high expression of ZHD in Pichia pastoris GS115. Meanwhile, to further improve the expression of recombinant ZHD, the plasmids containing 1 to 4 copies of the zhd expression cassette were constructed, respectively, using the biobrick method. The protein expression in the recombinant P...
December 2016: Microbiological Research
https://www.readbyqxmd.com/read/27790212/the-influence-of-sporulation-conditions-on-the-spore-coat-protein-composition-of-bacillus-subtilis-spores
#5
Wishwas R Abhyankar, Kiki Kamphorst, Bhagyashree N Swarge, Henk van Veen, Nicole N van der Wel, Stanley Brul, Chris G de Koster, Leo J de Koning
Spores are of high interest to the food and health sectors because of their extreme resistance to harsh conditions, especially against heat. Earlier research has shown that spores prepared on solid agar plates have a higher heat resistance than those prepared under a liquid medium condition. It has also been shown that the more mature a spore is, the higher is its heat resistance most likely mediated, at least in part, by the progressive cross-linking of coat proteins. The current study for the first time assesses, at the proteomic level, the effect of two commonly used sporulation conditions on spore protein presence...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27788200/physiological-morphological-and-behavioural-responses-of-self-feeding-precocial-chicks-copying-with-contrasting-levels-of-water-salinity-during-development
#6
Afonso R Rocha, Rita Silva, Auxiliadora Villegas, Juan M Sánchez-Guzmán, Jaime A Ramos, José A Masero
Combined physiological and behavioural responses to salt loads during development have rarely been studied in air-breathing vertebrates able to inhabit hypersaline habitats, but they may be of particular importance in understanding, for example, the differences among species in patterns of habitat use or ontogenetic diet switches. Here, we compared the physiological and behavioural responses of self-feeding precocial chicks developed in contrasting levels of water salinity. The model species was the Black-winged Stilt (Himantopus himantopus) a precocial shorebird that breeds in a range of habitats from freshwater to hypersaline wetlands...
2016: PloS One
https://www.readbyqxmd.com/read/27771392/high-throughput-purification-of-recombinant-proteins-using-self-cleaving-intein-tags
#7
M J Coolbaugh, M J Shakalli Tang, D W Wood
High throughput methods for recombinant protein production using E. coli typically involve the use of affinity tags for simple purification of the protein of interest. One drawback of these techniques is the occasional need for tag removal before study, which can be hard to predict. In this work, we demonstrate two high throughput purification methods for untagged protein targets based on simple and cost-effective self-cleaving intein tags. Two model proteins, E. coli beta-galactosidase (βGal) and superfolder green fluorescent protein (sfGFP), were purified using self-cleaving versions of the conventional chitin-binding domain (CBD) affinity tag and the nonchromatographic elastin-like-polypeptide (ELP) precipitation tag in a 96-well filter plate format...
October 19, 2016: Analytical Biochemistry
https://www.readbyqxmd.com/read/27746699/morphine-dependence-is-attenuated-by-red-ginseng-extract-and-ginsenosides-rh2-rg3-and-compound-k
#8
Taddesse Yayeh, Kyunghwa Yun, Soyong Jang, Seikwan Oh
BACKGROUND: Red ginseng and ginsenosides have shown plethoric effects against various ailments. However, little is known regarding the effect of red ginseng on morphine-induced dependence and tolerance. We therefore investigated the effect of red ginseng extract (RGE) and biotransformed ginsenosides Rh2, Rg3, and compound K on morphine-induced dependence in mice and rats. METHODS: While mice were pretreated with RGE and then morphine was injected intraperitoneally, rats were infused with ginsenosides and morphine intracranially for 7 days...
October 2016: Journal of Ginseng Research
https://www.readbyqxmd.com/read/27744524/production-of-laccase-by-recombinant-yarrowia-lipolytica-from-molasses-bioprocess-development-using-statistical-modeling-and-increase-productivity-in-shake-flask-and-bioreactor-cultures
#9
Farshad Darvishi, Marzieh Moradi, Catherine Madzak, Claude Jolivalt
Laccases are used in numerous applications, from green degradation of various xenobiotic compounds, waste detoxification, textile dye bleaching, and delignification of lignocellulose materials to biofuel production. In this study, the recombinant Yarrowia lipolytica YL4 strain carrying the white-rot fungus Trametes versicolor laccase IIIb gene was used for laccase production from beet molasses as an agro-industrial residue. Response surface methodology was used to statistical optimization of the production of laccase by Y...
October 15, 2016: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/27744242/mycothermus-thermophilus-syn-scytalidium-thermophilum-repertoire-of-a-diverse-array-of-efficient-cellulases-and-hemicellulases-in-the-secretome-revealed
#10
Neha Basotra, Baljit Kaur, Marcos Di Falco, Adrian Tsang, Bhupinder Singh Chadha
Mycothermus thermophilus (Syn. Scytalidium thermophilum/Humicola insolens), a thermophilic fungus, is being reported to produce appreciable titers of cellulases and hemicellulases during shake flask culturing on cellulose/wheat-bran/rice straw based production medium. The sequential and differential expression profile of endoglucanases, β-glucosidases, cellobiohydrolases and xylanases using zymography was studied. Mass spectrometry analysis of secretome (Q-TOF LC/MS) revealed a total of 240 proteins with 92 CAZymes of which 62 glycosyl hydrolases belonging to 30 different families were present...
October 8, 2016: Bioresource Technology
https://www.readbyqxmd.com/read/27737615/kinetic-characterization-and-fed-batch-fermentation-for-maximal-simultaneous-production-of-esterase-and-protease-from-lysinibacillus-fusiformis-au01
#11
K Divakar, M Suryia Prabha, G Nandhinidevi, P Gautam
The simultaneous production of intracellular esterase and extracellular protease from the strain Lysinibacillus fusiformis AU01 was studied in detail. The production was performed both under batch and fed-batch modes. The maximum yield of intracellular esterase and protease was obtained under full oxygen saturation at the beginning of the fermentation. The data were fitted to the Luedeking-Piret model and it was shown that the enzyme (both esterase and protease) production was growth associated. A decrease in intracellular esterase and increase in the extracellular esterase were observed during late stationary phase...
October 13, 2016: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/27735933/the-real-time-quaking-induced-conversion-assay-for-detection-of-human-prion-disease-and-study-of-other-protein-misfolding-diseases
#12
Matthias Schmitz, Maria Cramm, Franc Llorens, Dominik Müller-Cramm, Steven Collins, Ryuichiro Atarashi, Katsuya Satoh, Christina D Orrù, Bradley R Groveman, Saima Zafar, Walter J Schulz-Schaeffer, Byron Caughey, Inga Zerr
The development and adaption of in vitro misfolded protein amplification systems has been a major innovation in the detection of abnormally folded prion protein scrapie (PrP(Sc)) in human brain and cerebrospinal fluid (CSF) samples. Herein, we describe a fast and efficient protein amplification technique, real-time quaking-induced conversion (RT-QuIC), for the detection of a PrP(Sc) seed in human brain and CSF. In contrast to other in vitro misfolded protein amplification assays-such as protein misfolding cyclic amplification (PMCA)-which are based on sonication, the RT-QuIC technique is based on prion seed-induced misfolding and aggregation of recombinant prion protein substrate, accelerated by alternating cycles of shaking and rest in fluorescence plate readers...
November 2016: Nature Protocols
https://www.readbyqxmd.com/read/27718028/measuring-experimental-cyclohexane-water-distribution-coefficients-for-the-sampl5-challenge
#13
Ariën S Rustenburg, Justin Dancer, Baiwei Lin, Jianwen A Feng, Daniel F Ortwine, David L Mobley, John D Chodera
Small molecule distribution coefficients between immiscible nonaqueuous and aqueous phases-such as cyclohexane and water-measure the degree to which small molecules prefer one phase over another at a given pH. As distribution coefficients capture both thermodynamic effects (the free energy of transfer between phases) and chemical effects (protonation state and tautomer effects in aqueous solution), they provide an exacting test of the thermodynamic and chemical accuracy of physical models without the long correlation times inherent to the prediction of more complex properties of relevance to drug discovery, such as protein-ligand binding affinities...
October 7, 2016: Journal of Computer-aided Molecular Design
https://www.readbyqxmd.com/read/27690107/modeling-fragile-x-syndrome-using-human-pluripotent-stem-cells
#14
Hagar Mor-Shaked, Rachel Eiges
Fragile X syndrome (FXS) is the most common heritable form of cognitive impairment. It results from a loss-of-function mutation by a CGG repeat expansion at the 5' untranslated region of the X-linked fragile X mental retardation 1 (FMR1) gene. Expansion of the CGG repeats beyond 200 copies results in protein deficiency by leading to aberrant methylation of the FMR1 promoter and the switch from active to repressive histone modifications. Additionally, the CGGs become increasingly unstable, resulting in high degree of variation in expansion size between and within tissues of affected individuals...
2016: Genes
https://www.readbyqxmd.com/read/27684722/fish-based-analysis-of-clonally-derived-cho-cell-populations-reveals-high-probability-for-transgene-integration-in-a-terminal-region-of-chromosome-1-1q13
#15
Shengwei Li, Xiaoping Gao, Rui Peng, Sheng Zhang, Wei Fu, Fangdong Zou
A basic goal in the development of recombinant proteins is the generation of cell lines that express the desired protein stably over many generations. Here, we constructed engineered Chinese hamster ovary cell lines (CHO-S) with a pCHO-hVR1 vector that carried an extracellular domain of a VEGF receptor (VR) fusion gene. Forty-five clones with high hVR1 expression were selected for karyotype analysis. Using fluorescence in situ hybridization (FISH) and G-banding, we found that pCHO-hVR1 was integrated into three chromosomes, including chromosomes 1, Z3 and Z4...
2016: PloS One
https://www.readbyqxmd.com/read/27668880/valorization-of-sugar-to-ethanol-process-waste-vinasse-a-novel-biorefinery-approach-using-edible-ascomycetes-filamentous-fungi
#16
Ramkumar B Nair, Mohammad J Taherzadeh
The aim of the present work was to study the integration of edible ascomycetes filamentous fungi into the existing sugar- or molasses-to-ethanol processes, to grow on vinasse or stillage and produce ethanol and protein-rich fungal biomass. Two fungal strains, Neurospora intermedia and Aspergillus oryzae were examined in shake flasks and airlift-bioreactors, resulting in reduction of vinasse COD by 34% and viscosity by 21%. Utilization of glycerol and sugars were observed, yielding 202.4 or 222.8g dry fungal biomass of N...
December 2016: Bioresource Technology
https://www.readbyqxmd.com/read/27663383/a-comprehensive-evaluation-of-nanoparticle-tracking-analysis-nanosight-for-characterization-of-proteinaceous-submicron-particles
#17
Xinsheng Tian, M Reza Nejadnik, Dorrit Baunsgaard, Anette Henriksen, Christian Rischel, Wim Jiskoot
Nanoparticle tracking analysis (NTA) has attracted great interest for application in the field of submicron particle characterization for biopharmaceuticals. It has the virtue of direct sample visualization and particle-by-particle tracking, but the complexity of method development has limited its routine applicability. We systematically evaluated data collection and processing parameters as well as sample handling methods using shake-stressed protein samples. The camera shutter and gain were identified as the key factors influencing NTA results...
November 2016: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/27652061/role-of-ligninolytic-enzymes-of-white-rot-fungi-pleurotus-spp-grown-with-azo-dyes
#18
Prashant D Kunjadia, Gaurav V Sanghvi, Anju P Kunjadia, Pratap N Mukhopadhyay, Gaurav S Dave
BACKGROUND: Total three Pleurotus species (P. ostreatus, P. sapidus, P. florida) was compared for ligninolytic enzyme production grown with Coralene Golden Yellow, Coralene Navy Blue and Coralene Dark Red azo dyes in liquid medium under shaking condition. RESULTS: The biodegradation competency varied from species to species and it was found that P. ostreatus, P. sapidus and P. florida to 20 ppm dye concentration shows 88, 92 and 98 % decolorization, respectively for all three dyes...
2016: SpringerPlus
https://www.readbyqxmd.com/read/27630761/applying-central-composite-design-and-response-surface-methodology-to-optimize-growth-and-biomass-production-of-haemophilus-influenzae-type-b
#19
Seyed Bahman Momen, Seyed Davar Siadat, Neda Akbari, Bijan Ranjbar, Khosro Khajeh
BACKGROUND: Haemophilus influenzae type b (Hib) is the leading cause of bacterial meningitis, otitis media, pneumonia, cellulitis, bacteremia, and septic arthritis in infants and young children. The Hib capsule contains the major virulence factor, and is composed of polyribosyl ribitol phosphate (PRP) that can induce immune system response. Vaccines consisting of Hib capsular polysaccharide (PRP) conjugated to a carrier protein are effective in the prevention of the infections. However, due to costly processes in PRP production, these vaccines are too expensive...
June 2016: Jundishapur Journal of Microbiology
https://www.readbyqxmd.com/read/27542751/heterologous-expression-of-codon-optimized-trichoderma-reesei-cel6a-in-pichia-pastoris
#20
Fubao Fuelbiol Sun, Renhui Bai, Huimin Yang, Fei Wang, Jing He, Chundi Wang, Maobing Tu
The Cel6A deficiency has become one of the limiting factors for cellulose saccharification in biochemical conversion of cellulosic biomass to fuels and chemicals. The work attempted to use codon optimization to enhance Trichoderma reesei Cel6A expression in Pichia pastoris. Two recombinants P. pastoris GS115 containing AOX1 and GAP promotors were successfully constructed, respectively. The optimal temperatures and pHs of the expressed Cel6A from two recombinants were consistent with each other, were also in the extremely similar range to that reported on the native Cel6A from T...
October 2016: Enzyme and Microbial Technology
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