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https://www.readbyqxmd.com/read/28330255/ascorbyl-palmitate-synthesis-in-an-organic-solvent-system-using-a-celite-immobilized-commercial-lipase-lipolase-100l
#1
Shivika Sharma, Kriti Kanwar, Shamsher S Kanwar
Ascorbyl palmitate was synthesized using a Celite-immobilized commercial lipase (Lipolase 100L) in dimethylsulfoxide (DMSO) as an organic solvent system. Lipase immobilized by surface adsorption onto Celite 545 matrix and subsequently exposed to 1 % glutaraldehyde showed 75 % binding of protein. The Celite-bound lipase was optimally active at 75 °C and pH 8.5 under shaking and showed maximum hydrolytic activity toward p-NPP as a substrate. The bound lipase was found to be stimulated only in the presence of Al(3+) and EDTA...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28323213/oral-delivery-of-bacillus-subtilis-spores-expressing-cysteine-protease-of-clonorchis-sinensis-to-grass-carp-ctenopharyngodon-idellus-induces-immune-responses-and-has-no-damage-on-liver-and-intestine-function
#2
Zeli Tang, Hengchang Sun, TingJin Chen, Zhipeng Lin, Hongye Jiang, Xinyi Zhou, Cunbin Shi, Houjun Pan, Ouqin Chang, Pengli Ren, Jinyun Yu, Xuerong Li, Jin Xu, Yan Huang, Xinbing Yu
Clonorchis sinensis (C. sinensis) is a fish-borne trematode. Human can be infected by ingestion of C. sinensis metacercariae parasitized in grass carp (Ctenopharyngodon idella). For induction of effective oral immune responses, spores of Bacillus subtilis (B. subtilis) WB600 were utilized as vehicle to delivery CsCP (cysteine protease of C. sinensis) cooperated with CotC (B.s-CotC-CP), one of coat proteins, to the gastrointestinal tract. After routine culture of 8-12 h in LB medium, B. subtilis containing CotC-CsCP was transferred into the sporulation culture medium...
March 17, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28321795/phytase-production-by-aspergillus-niger-ncim-563-for-a-novel-application-to-degrade-organophosphorus-pesticides
#3
Parin C Shah, V Ravi Kumar, Syed G Dastager, Jayant M Khire
The production of phytase using Aspergillus niger NCIM 563 under submerged fermentation conditions was studied using protein rich chickpea flour as substrate. Employing a hybrid statistical media optimization strategy of Plackett-Burman and Box-Behnken experimental designs in shake-flasks gave an increased phytase activity from an initial 66 IU/mL in 216 h to 160 IU/mL in a reduced time of 132 h. Productivity, thus increased by 3.97 times from 7.3 to 29 IU/mL/day. Using the optimized media, the production was successfully scaled-up further and improved up to 164 IU/mL in 96 h by studies carried out employing 2 and 10-L fermenters...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28318397/baiba-does-not-regulate-ucp-3-expression-in-human-skeletal-muscle-as-a-response-to-aerobic-exercise
#4
Flor E Morales, Jeffrey S Forsse, Thomas L Andre, Sarah K McKinley-Barnard, Paul S Hwang, Ian G Anthony, Grant M Tinsley, Mike Spillane, Peter W Grandjean, Alejandro Ramirez, Darryn S Willoughby
OBJECTIVE: β-Aminoisobutyric acid (BAIBA) has shown to modulate uncoupling protein (UCP)-1 expression, which is mainly expressed in white adipose tissue; however, no studies to date have analyzed its potential effect on the main uncoupling protein of skeletal muscle, UCP-3. The main goal of this study was to assess the potential effect of acute aerobic exercise on serum BAIBA and skeletal muscle UCP-3. The secondary goal was to assess the potential involvement of the transcription factors proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and peroxisome proliferator-activated receptor alpha (PPARα), as well as free fatty acids (FFAs) in UCP-3 expression...
March 20, 2017: Journal of the American College of Nutrition
https://www.readbyqxmd.com/read/28315397/efficient-extracellular-expression-of-transpeptidase-sortase-a-in-pichia-pastoris
#5
Xinrui Zhao, Haofei Hong, Zhimeng Wu
In order to achieve efficient extracellular expression of Sortase A (SrtA), various strategies in Pichia pastoris system were applied in this study. Among different constructed recombinant strains, the SMD1168 strain integrated 5.7 copies of srtA gene under control of AOX1 promoter was proved to be the best strain for the extracellular SrtA expression. After the optimization of fermentation conditions (induction 72 h at 28 °C, initial pH 6.0, supplemented with 1.5% methanol), the highest yield and activity of extracellular SrtA reached 97...
March 15, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28302114/implications-of-evolutionary-engineering-for-growth-and-recombinant-protein-production-in-methanol-based-growth-media-in-the-yeast-pichia-pastoris
#6
Josef W Moser, Roland Prielhofer, Samuel M Gerner, Alexandra B Graf, Iain B H Wilson, Diethard Mattanovich, Martin Dragosits
BACKGROUND: Pichia pastoris is a widely used eukaryotic expression host for recombinant protein production. Adaptive laboratory evolution (ALE) has been applied in a wide range of studies in order to improve strains for biotechnological purposes. In this context, the impact of long-term carbon source adaptation in P. pastoris has not been addressed so far. Thus, we performed a pilot experiment in order to analyze the applicability and potential benefits of ALE towards improved growth and recombinant protein production in P...
March 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28296355/biosensor-enabled-directed-evolution-to-improve-muconic-acid-production-in-saccharomyces-cerevisiae
#7
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/28291648/combining-stable-insect-cell-lines-with-baculovirus-mediated-expression-for-multi-ha-influenza-vlp-production
#8
Daniela P Sequeira, Ricardo Correia, Manuel J T Carrondo, António Roldão, Ana P Teixeira, Paula M Alves
Safer and broadly protective vaccines are needed to cope with the continuous evolution of circulating influenza virus strains and promising approaches based on the expression of multiple hemagglutinins (HA) in a virus-like particle (VLP) have been proposed. However, expression of multiple genes in the same vector can lead to its instability due to tandem repetition of similar sequences. By combining stable with transient expression systems we can rationally distribute the number of genes to be expressed per platform and thus mitigate this risk...
March 10, 2017: Vaccine
https://www.readbyqxmd.com/read/28291518/a-novel-expression-system-for-lytic-polysaccharide-monooxygenases
#9
Gaston Courtade, Simone Balzer Le, Gerd Inger Sætrom, Trygve Brautaset, Finn L Aachmann
Lytic polysaccharide monooxygenases (LPMOs) are key enzymatic players of lignocellulosic biomass degradation processes. As such, they have been introduced in cellulolytic cocktails for more efficient and less expensive lignocellulose saccharification. The recombinant production of LPMOs in bacteria for scientific investigations using vectors typically based on the T7 and lacUV5 promoters has been hampered by low yields. Reasons for this have been catabolite repression when producing the proteins in defined media with glucose as the sole carbon source, as well as the lack of an inducible expression system that allows controlled production of LPMOs that are correctly processed during translocation to the periplasmic space...
February 14, 2017: Carbohydrate Research
https://www.readbyqxmd.com/read/28284309/alleviation-of-metabolic-bottleneck-by-combinatorial-engineering-enhanced-astaxanthin-synthesis-in-saccharomyces-cerevisiae
#10
Pingping Zhou, Wenping Xie, Aipeng Li, Fan Wang, Zhen Yao, Qi Bian, Yongqiang Zhu, Hongwei Yu, Lidan Ye
Highly efficient biosynthesis of the commercially valuable carotenoid astaxanthin by microbial cells is an attractive alternative to chemical synthesis and microalgae extraction. With the goal of enhancing heterologous astaxanthin production in Saccharomyces cerevisiae, metabolic engineering and protein engineering were integrated to improve both the expression and activity of rate-limiting enzymes. Firstly, to increase the supply of β-carotene as a key precursor for astaxanthin, a positive mutant of GGPP synthase (CrtE03M) was overexpressed together with three other rate-limiting enzymes tHMG1, CrtI and CrtYB...
May 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/28280340/wheat-germ-agglutinin-conjugated-liposomes-incorporated-with-cardiolipin-to-improve-neuronal-survival-in-alzheimer-s-disease-treatment
#11
Yung-Chih Kuo, Che-Yu Lin, Jay-Shake Li, Yung-I Lou
Curcumin (CRM) and nerve growth factor (NGF) were entrapped in liposomes (LIP) with surface wheat germ agglutinin (WGA) to downregulate the phosphorylation of kinases in Alzheimer's disease (AD) therapy. Cardiolipin (CL)-conjugated LIP carrying CRM (CRM-CL/LIP) and also carrying NGF (NGF-CL/LIP) were used with AD models of SK-N-MC cells and Wistar rats after an insult with β-amyloid peptide (Aβ). We found that CRM-CL/LIP inhibited the expression of phosphorylated p38 (p-p38), phosphorylated c-Jun N-terminal kinase (p-JNK), and p-tau protein at serine 202 and prevented neurodegeneration of SK-N-MC cells...
2017: International Journal of Nanomedicine
https://www.readbyqxmd.com/read/28219382/high-level-extracellular-protein-production-in-bacillus-subtilis-using-an-optimized-dual-promoter-expression-system
#12
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
#13
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
#14
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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