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https://www.readbyqxmd.com/read/28219382/high-level-extracellular-protein-production-in-bacillus-subtilis-using-an-optimized-dual-promoter-expression-system
#1
Kang Zhang, Lingqia Su, Xuguo Duan, Lina Liu, Jing Wu
BACKGROUND: We recently constructed a Bacillus subtilis strain (CCTCC M 2016536) from which we had deleted the srfC, spoIIAC, nprE, aprE and amyE genes. This strain is capable of robust recombinant protein production and amenable to high-cell-density fermentation. Because the promoter is among the factors that influence the production of target proteins, optimization of the initial promoter, PamyQ from Bacillus amyloliquefaciens, should improve protein expression using this strain. This study was undertaken to develop a new, high-level expression system in B...
February 20, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28197999/parallel-experimental-design-and-multivariate-analysis-provides-efficient-screening-of-cell-culture-media-supplements-to-improve-biosimilar-product-quality
#2
David Brühlmann, Michael Sokolov, Alessandro Butté, Markus Sauer, Jürgen Hemberger, Jonathan Souquet, Hervé Broly, Martin Jordan
Rational and high-throughput optimization of mammalian cell culture media has a great potential to modulate recombinant protein product quality. We present a process design method based on parallel design-of-experiment (DoE) of CHO fed-batch cultures in 96-deepwell plates to modulate monoclonal antibody (mAb) glycosylation using medium supplements. To reduce risk of losing valuable information in an intricate joint screening, 17 compounds were separated into five different groups, considering their mode of biological action...
February 15, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28188692/bioreactor-scale-up-and-protein-product-quality-characterization-of-piggybac-transposon-derived-cho-pools
#3
Yashas Rajendra, Sowmya Balasubramanian, Robert B Peery, James R Swartling, Neil A McCracken, Dawn L Norris, Christopher C Frye, Gavin C Barnard
Chinese hamster ovary (CHO) cells remain the most popular host for the production of biopharmaceutical drugs, particularly monoclonal antibodies (mAbs), bispecific antibodies and Fc-fusion proteins. Creating and characterizing the stable CHO clonally-derived cell lines (CDCLs) needed to manufacture these therapeutic proteins is a lengthy and laborious process. Therefore, CHO pools have increasingly been used to rapidly produce protein to support and enable preclinical drug development. We recently described the generation of CHO pools yielding mAb titers as high as 7...
February 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28147314/the-antiangiogenic-role-of-the-pro-inflammatory-cytokine-interleukin-31
#4
Shiri Davidi, Ella Fremder, Tal Kan, Ziv Raviv, Michael Timaner, Nathan Karin, Dov Hershkovitz, Ami Arohneim, Yuval Shaked
Pro-inflammatory cytokines in the tumor microenvironment are known for their ability to either inhibit or promote cancer progression. Here we evaluated the role of Interleukin-31 (IL31), a protein belonging to the pro-inflammatory IL-6 cytokine family which has been characterized in autoimmune disease, in tumorigenesis. We show that IL31 and its receptor, IL31RA, are highly expressed in various human and mouse cancer cell lines, as well as in tumor specimens from cancer patients. MC38 murine colon carcinoma cells depleted of IL31 exhibit an increase in invasive and migratory properties in vitro, effects that are reversed by supplementing the cells with exogenous IL31...
January 27, 2017: Oncotarget
https://www.readbyqxmd.com/read/28140467/a-survey-of-commercially-available-isomaltooligosaccharide-based-food-ingredients
#5
Lee R Madsen, Sarah Stanley, Peter Swann, Jack Oswald
Isomaltooligosaccharides (IMOs) are included in many commercially available food products including protein/fiber bars, shakes, and other dietary supplements. Marketed as "high fiber," "prebiotic soluble fiber," and/or as a "low-calorie, low glycemic sweetener," IMO may be present in significant amounts, for example, more than 15 g/item or serving. Herein, high-pressure anion exchange chromatography with pulsed amperometric detection and high-pressure liquid chromatography with differential refractive index detection are used to compare 7 commercially available IMO-containing bulk food ingredients...
January 31, 2017: Journal of Food Science
https://www.readbyqxmd.com/read/28117940/peripheral-blood-gene-expression-changes-associated-with-primary-graft-dysfunction-after-lung-transplantation
#6
J M Diamond, E Cantu, M K Porteous, Y Suzuki, K C Meyer, D J Lederer, R K Milewski, S Arcasoy, F D'Ovidio, M Bacchetta, J R Sonett, G Singh, J Costa, J W Tobias, H Rodriguez, V M Van Deerlin, K M Olthoff, A Shaked, B-L Chang, J D Christie
Recipient responses to primary graft dysfunction (PGD) after lung transplantation may have important implications to the fate of the allograft. We therefore evaluated longitudinal differences in peripheral blood gene expression in subjects with PGD. RNA expression was measured throughout the first transplant year in 106 subjects enrolled in the Clinical Trials in Organ Transplantation-03 study using a panel of 100 hypothesis-driven genes. PGD was defined as grade 3 in the first 72 posttransplant hours. Eighteen genes were differentially expressed over the first year based on PGD development, with significant representation from innate and adaptive immunity genes, with most differences identified very early after transplant...
January 24, 2017: American Journal of Transplantation
https://www.readbyqxmd.com/read/28115011/recombinant-production-of-the-antibody-fragment-d1-3-scfv-with-different-bacillus-strains
#7
Antonia Lakowitz, Rainer Krull, Rebekka Biedendieck
BACKGROUND: Different strains of the genus Bacillus are versatile candidates for the industrial production and secretion of heterologous proteins. They can be cultivated quite easily, show high growth rates and are usually non-pathogenic and free of endo- and exotoxins. They have the ability to secrete proteins with high efficiency into the growth medium, which allows cost-effective downstream purification processing. Some of the most interesting and challenging heterologous proteins are recombinant antibodies and antibody fragments...
January 23, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28100236/human-%C3%AE-defensin-2-production-from-s-cerevisiae-using-the-repressible-met17-promoter
#8
Thea S B Møller, Joanna Hay, Malcolm J Saxton, Karen Bunting, Evamaria I Petersen, Søren Kjærulff, Christopher J A Finnis
BACKGROUND: Baker's yeast Saccharomyces cerevisiae is a proven host for the commercial production of recombinant biopharmaceutical proteins. For the manufacture of heterologous proteins with activities deleterious to the host it can be desirable to minimise production during the growth phase and induce production late in the exponential phase. Protein expression by regulated promoter systems offers the possibility of improving productivity in this way by separating the recombinant protein production phase from the yeast growth phase...
January 18, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28068309/biomimetic-spinning-of-artificial-spider-silk-from-a-chimeric-minispidroin
#9
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.
March 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
#10
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
#11
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
#12
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
#13
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
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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