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https://www.readbyqxmd.com/read/28068309/biomimetic-spinning-of-artificial-spider-silk-from-a-chimeric-minispidroin
#1
Marlene Andersson, Qiupin Jia, Ana Abella, Xiau-Yeen Lee, Michael Landreh, Pasi Purhonen, Hans Hebert, Maria Tenje, Carol V Robinson, Qing Meng, Gustavo R Plaza, Jan Johansson, Anna Rising
Herein we present a chimeric recombinant spider silk protein (spidroin) whose aqueous solubility equals that of native spider silk dope and a spinning device that is based solely on aqueous buffers, shear forces and lowered pH. The process recapitulates the complex molecular mechanisms that dictate native spider silk spinning and is highly efficient; spidroin from one liter of bacterial shake-flask culture is enough to spin a kilometer of the hitherto toughest as-spun artificial spider silk fiber.
January 9, 2017: Nature Chemical Biology
https://www.readbyqxmd.com/read/28039549/expression-of-talaromyces-thermophilus-lipase-gene-in-trichoderma-reesei-by-homologous-recombination-at-the-cbh1-locus
#2
Xu Zhang, Liming Xia
CBH1 (cellobiohydrolase) comprises the majority of secreted proteins by Trichoderma reesei. For expression of Talaromyces thermophilus lipase gene in T. reesei, a self-designed CBH1 promoter was applied to drive the lipase gene expression cassette which was bracketed by flanking sequences of cbh1 gene for homologous recombination. Protoplast and Agrobacterium-mediated plasmid transformations were performed and compared, resultantly, transformation mediated by Agrobacterium was overall proved to be more efficient...
December 30, 2016: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28012242/a-12-week-intervention-with-nonivamide-a-trpv1-agonist-prevents-a-dietary-induced-body-fat-gain-and-increases-peripheral-serotonin-in-moderately-overweight-subjects
#3
Christina M Hochkogler, Barbara Lieder, Petra Rust, David Berry, Samuel M Meier, Marc Pignitter, Alessandra Riva, Alina Leitinger, Anne Bruk, Simone Wagner, Joachim Hans, Sabine Widder, Jakob P Ley, Gerhard E Krammer, Veronika Somoza
SCOPE: A bolus administration of 0.15 mg nonivamide has previously been demonstrated to reduce energy intake in moderately overweight men. This 12-week intervention investigated whether a daily consumption of nonivamide in a protein-based product formulation promotes a reduction in body weight in healthy overweight subjects, and affects outcome measures associated with mechanisms regulating food intake, e.g. plasma concentrations of an-/orexigenic hormones, energy substrates as well as changes in fecal microbiota ...
December 24, 2016: Molecular Nutrition & Food Research
https://www.readbyqxmd.com/read/28005561/risk-factors-for-collegiate-swimmers-hospitalized-with-exertional-rhabdomyolysis
#4
Margo R Stanfa, Navjeet N Silles, Amanda Cooper, Sara Arena, Kristin Landis-Piwowar, Carrie Aprik, Tamara Hew-Butler
OBJECTIVE: To identify midseason risk factors for symptomatic exertional rhabdomyolysis (sER) in swimmers after a novel upper body workout. DESIGN: Retrospective (1) survey and (2) analyses of observational laboratory data conducted over a 16-week training period, 2 months before sER. SETTING: Midwest University. PARTICIPANTS: Thirty-four collegiate swimmers. INDEPENDENT VARIABLES: (1) Motivation, symptoms, and supplements for survey variables...
January 2017: Clinical Journal of Sport Medicine: Official Journal of the Canadian Academy of Sport Medicine
https://www.readbyqxmd.com/read/27991937/silver-ions-as-em-marker-of-congo-red-ligation-sites-in-amyloids-and-amyloid-like-aggregates
#5
Janina Rybarska, Leszek Konieczny, Anna Jagusiak, Katarzyna Chłopaś, Grzegorz Zemanek, Barbara Piekarska, Barabara Stopa, Piotr Piwowar, Olga Woźnicka, Irena Roterman
Congo red (CR) is a known selective amyloid ligand. The focus of our work is identification (by EM imaging) of dye binding sites and their distribution in amyloids and amyloid-like aggregates formed in vitro. In order to produce the required contrast, CR has been indirectly combined with metal via including Titan yellow (TY) by intercalation which exhibits a relatively strong affinity for silver ions. The resulting combined ligand retains its ability to bind to proteins (which it owes to CR) and can easily be detected in EM studies thanks to TY...
December 19, 2016: Acta Biochimica Polonica
https://www.readbyqxmd.com/read/27939273/dimerization-of-tyr136cys-alpha-synuclein-prevents-amyloid-transformation-of-wild-type-alpha-synuclein
#6
K V Barinova, M L Kuravsky, A M Arutyunyan, M V Serebryakova, E V Schmalhausen, V I Muronetz
Expression of human alpha-synuclein in E. coli cells is known to result in a mixture of the wild type alpha-synuclein and the protein containing Tyr136Cys substitution due to the translational error. The amount of Cys136 alpha-synuclein (Cys136-AS) may reach approximately 50% of the recombinant protein. The wild-type and Cys136-containing fractions of alpha-synuclein were separated using thiol-Sepharose, and their properties were investigated. In the absence of reducing agents, Cys136-AS forms dimers due to the disulfide bonding...
December 6, 2016: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/27887723/synergistic-effect-of-cavitation-and-agitation-on-protein-aggregation
#7
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...
February 2017: 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
#8
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
#9
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 towards the polar cell fate by promoting Notch pathway activity...
December 15, 2016: Development
https://www.readbyqxmd.com/read/27825486/high-level-expression-of-a-zen-detoxifying-gene-by-codon-optimization-and-biobrick-in-pichia-pastoris
#10
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
#11
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
#12
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
#13
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
#14
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
#15
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
#16
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...
December 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
#17
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
#18
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
#19
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...
November 2016: Journal of Computer-aided Molecular Design
https://www.readbyqxmd.com/read/27690107/modeling-fragile-x-syndrome-using-human-pluripotent-stem-cells
#20
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
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