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https://www.readbyqxmd.com/read/29331528/expression-of-a-thermotolerant-laccase-from-pycnoporus-sanguineus-in-trichoderma-reesei-and-its-application-in-the-degradation-of-bisphenol-a
#1
Jie Zhao, Shenquan Zeng, Ying Xia, Liming Xia
The laccase gene from Pycnoporus sanguineus was cloned and inserted between the strong Pcbh1 promoter and the Tcbh1 terminator from Trichoderma reesei to form the recombinant plasmid pCH-lac. Using Agrobacterium-mediated technique, the pCH-lac was integrated into the chromosomes of T. reesei. Twenty positive transformants were obtained by employing hygromycin B as a selective agent. PCR was used to confirm that the laccase gene was integrated into the chromosomal DNA of T. reesei. Laccase production by recombinant transformants was performed in shaking flasks, and the activity of laccase reached 8...
January 10, 2018: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/29314728/online-measurement-of-the-respiratory-activity-in-shake-flasks-enables-the-identification-of-cultivation-phases-and-patterns-indicating-recombinant-protein-production-in-various-e-coli-host-strains
#2
Nina Ihling, Natalie Bittner, Sylvia Diederichs, Maximilian Schelden, Anna Korona, Georg Theo Höfler, Alexander Fulton, Karl-Erich Jaeger, Kohsuke Honda, Hisao Ohtake, Jochen Büchs
Escherichia coli is commonly used for recombinant protein production with many available host strains. Screening experiments are often performed in batch mode using shake flasks and evaluating only the final product concentration. This conventional approach carries the risk of missing the best strain due to limited monitoring capabilities. Thus, this study focuses on investigating the general suitability of online respiration measurement for selecting expression hosts for heterologous protein production. The oxygen transfer rate (OTR) for different T7-RNA polymerase-dependent Escherichia coli expression strains was compared under inducing and non-inducing conditions...
January 5, 2018: Biotechnology Progress
https://www.readbyqxmd.com/read/29311640/a-specific-form-of-prefibrillar-aggregates-that-functions-as-a-precursor-of-amyloid-nucleation
#3
Naoki Yamamoto, Shoko Tsuhara, Atsuo Tamura, Eri Chatani
Non-fibrillar protein aggregates that appear in the earlier stages of amyloid fibril formation are sometimes considered to play a key role in amyloid nucleation; however, the structural features of these aggregates currently remain unclear. We herein identified a characteristic pathway of fibril formation by human insulin B chain, in which two major species of prefibrillar aggregates were identified. Based on the time-resolved tracking of this pathway with far-UV circular dichroism (CD) spectroscopy, dynamic light scattering (DLS), and 1H-NMR spectroscopy, the first prefibrillar aggregate with a hydrodynamic diameter of approximately 70 nm accumulated concomitantly with the formation of a β-sheet structure, and the size further evolved to 130 nm with an additional structural development...
January 8, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29308528/biosynthesis-of-flavone-c-glucosides-in-engineered-escherichia-coli
#4
Anil Shrestha, Ramesh Prasad Pandey, Dipesh Dhakal, Prakash Parajuli, Jae Kyung Sohng
Two plant-originated C-glucosyltransferases (CGTs) UGT708D1 from Glycine max and GtUF6CGT1 from Gentiana triflora were accessed for glucosylation of selected flavones chrysin and luteolin. Uridine diphosphate (UDP)-glucose pool was enhanced in Escherichia coli cell cytosol by introducing heterologous UDP-glucose biosynthetic genes, i.e., glucokinase (glk), phosphoglucomutase (pgm2), and glucose 1-phosphate uridylyltransferase (galU), along with glucose facilitator diffusion protein from (glf) from different organisms, in a multi-monocistronic vector with individual T7 promoter, ribosome binding site, and terminator for each gene...
January 7, 2018: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29300306/uptake-of-tyrosine-amino-acid-on-nano-graphene-oxide
#5
Hossam M Nassef, Mohamed Hagar, Zeiad Malek, Abdelhameed M Othman
Graphene oxide (GO) is emerging as a promising nanomaterial with potential application in the detection and analysis of amino acids, DNA, enzymes, and proteins in biological fluid samples. So, the reaction of GO with amino acids should be characterized and determined before using it in biosensing methods and devices. In this study, the reaction of tyrosine amino acid (Tyr) with GO was characterized using FT-IR, UV-vis spectrophotometry, and scanning electron microscopy (SEM) before its use. The optimum conditions for GO's interaction with Tyr amino acid have been studied under variable conditions...
January 4, 2018: Materials
https://www.readbyqxmd.com/read/29288112/the-genetics-of-parkinson-disease
#6
REVIEW
Hao Deng, Peng Wang, Joseph Jankovic
About 15% of patients with Parkinson disease (PD) have family history and 5-10% have a monogenic form of the disease with Mendelian inheritance. To date, at least 23 loci and 19 disease-causing genes for parkinsonism have been found, but many more genetic risk loci and variants for sporadic PD phenotype have been identified in various association studies. Investigating the mutated protein products has uncovered potential pathogenic pathways that provide insights into mechanisms of neurodegeneration in familial and sporadic PD...
December 26, 2017: Ageing Research Reviews
https://www.readbyqxmd.com/read/29288093/notorious-but-not-understood-how-liquid-air-interfacial-stress-triggers-protein-aggregation
#7
Ellen Koepf, Simon Eisele, Rudolf Schroeder, Gerald Brezesinski, Wolfgang Friess
Protein aggregation is a major challenge in the development of biopharmaceuticals. As the pathways of aggregation are manifold, good understanding of the mechanisms behind is essential. Particularly, the presence of liquid-air interfaces has been identified to trigger the formation of large protein particles. Investigations of two monoclonal antibodies (IgGs) at the liquid-air interface exhibited the formation of a highly compressible film. An inhomogeneous protein distribution across the interface with areas of increased packing density was discovered by Brewster-Angle microscopy...
December 26, 2017: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/29282592/systematic-exploration-of-multiple-drug-binding-sites
#8
Mónika Bálint, Norbert Jeszenői, István Horváth, David van der Spoel, Csaba Hetényi
BACKGROUND: Targets with multiple (prerequisite or allosteric) binding sites have an increasing importance in drug design. Experimental determination of atomic resolution structures of ligands weakly bound to multiple binding sites is often challenging. Blind docking has been widely used for fast mapping of the entire target surface for multiple binding sites. Reliability of blind docking is limited by approximations of hydration models, simplified handling of molecular flexibility, and imperfect search algorithms...
December 28, 2017: Journal of Cheminformatics
https://www.readbyqxmd.com/read/29282165/in-vitro-effects-of-sports-and-energy-drinks-on-streptococcus-mutans-biofilm-formation-and-metabolic-activity
#9
LaQuia A Vinson, Amy K Goodlett, Ruijie Huang, George J Eckert, Richard L Gregory
PURPOSE: Sports and energy drinks are being increasingly consumed and contain large amounts of sugars, which are known to increase Streptococcus mutans biofilm formation and metabolic activity. The purpose of this in vitro study was to investigate the effects of sports and energy drinks on S. mutans biofilm formation and metabolic activity. METHODS: S. mutans UA159 was cultured with and without a dilution (1:3 ratio) of a variety of sports and energy drinks in bacterial media for 24 hours...
September 15, 2017: Journal of Dentistry for Children
https://www.readbyqxmd.com/read/29228335/gram-scale-production-of-recombinant-microbial-enzymes-in-shake-flasks
#10
Lukas Chrast, Radka Chaloupkova, Jiri Damborsky
Heterologous production of recombinant proteins is a cornerstone of microbiological and biochemical research as well as various biotechnological processes. Yields and quality of produced proteins have a tremendous impact on structural and enzymology studies, development of new biopharmaceuticals and establishing new biocatalytic processes. Majority of current protocols for recombinant protein expression in Escherichia coli exploit batch cultures with complex media, often providing low yields of the target protein due to oxygen transfer limitation, rapid depletion of carbon sources, and pH changes during the cultivation...
December 5, 2017: FEMS Microbiology Letters
https://www.readbyqxmd.com/read/29226613/large-scale-culture-of-a-megakaryocytic-progenitor-cell-line-with-a-single-use-bioreactor-system
#11
Retno Wahyu Nurhayati, Yoshihiro Ojima, Takeaki Dohda, Masahiro Kino-Oka
The increasing application of regenerative medicine has generated a growing demand for stem cells and their derivatives. Single-use bioreactors offer an attractive platform for stem cell expansion owing to their scalability for large-scale production and feasibility of meeting clinical-grade standards. The current work evaluated the capacity of a single-use bioreactor system (1 L working volume) for expanding Meg01 cells, a megakaryocytic (MK) progenitor cell line. Oxygen supply was provided by surface aeration to minimize foaming and orbital shaking was used to promote oxygen transfer...
December 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/29215267/new-structural-insights-into-formation-of-the-key-actin-regulating-wip-wasp-complex-determined-by-nmr-and-molecular-imaging
#12
Adi Halle-Bikovski, Sophia Fried, Eva Rozentur-Shkop, Guy Biber, Hadassa Shaked, Noah Joseph, Mira Barda-Saad, Jordan H Chill
Wiskott-Aldrich syndrome protein (WASp) is exclusively expressed in hematopoietic cells and responsible for actin-dependent processes, including cellular activation, migration, and invasiveness. The C-terminal domain of WASp-Interacting Protein (WIP) binds to WASp and regulates its activity by shielding it from degradation in a phosphorylation dependent manner as we previously demonstrated. Mutations in the WAS-encoding gene lead to the primary immunodeficiencies Wiskott-Aldrich syndrome (WAS) and X-linked thrombocytopenia (XLT)...
December 7, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/29183374/optimizing-recombinant-protein-expression-via-automated-induction-profiling-in-microtiter-plates-at-different-temperatures
#13
Martina Mühlmann, Eva Forsten, Saskia Noack, Jochen Büchs
BACKGROUND: Escherichia coli (E. coli) is the most abundant expression host for recombinant proteins. The production efficiency is dependent on a multitude of parameters. Therefore, high-throughput applications have become an increasingly frequent technique to investigate the main factors. Within this study, the effects of temperature, induction time and inducer concentration on the metabolic state and the product formation were extensively examined. Induction profiling of E. coli Tuner(DE3) pRhotHi-2-EcFbFP was performed in 48-well Flowerplates and standard 96-well plates using a robotic platform...
November 28, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/29175761/an-improved-protocol-for-extracting-extracellular-polymeric-substances-from-granular-filter-media
#14
Sarah E Keithley, Mary Jo Kirisits
Extracellular polymeric substances (EPS) are an important parameter in drinking-water biofilters, and, to date, this is the first study to compare protocols from the literature for extracting EPS from granular filter media. Five extraction protocols were compared, and one was improved by varying the type of initial physical treatment and the shaking intensity, temperature, and time of incubation. Extracting EPS from granular filter media in triplicate by combining 2 g (wet weight) of filter media with 10 mL of extraction buffer (10 mM Tris, 10 mM EDTA, 2...
November 8, 2017: Water Research
https://www.readbyqxmd.com/read/29159244/clinical-pathological-and-molecular-data-concerning-coenurus-cerebralis-in-sheep-in-egypt
#15
Said Amer, Ahmed ElKhatam, Yasuhiro Fukuda, Lamia I Bakr, Shereif Zidan, Ahmed Elsify, Mostafa A Mohamed, Chika Tada, Yutaka Nakai
This article contains information related to a recent study "Prevalence and Identity of Taenia multiceps cysts "Coenurus cerebralis" in Sheep in Egypt" (Amer et al., 2017) [1]. Specifically, affected sheep showed neurological disorders manifested as depression, head shaking and circling, altered head position, incoordination and paralysis in some cases. Brain-derived cysts were molecularly identified by PCR-sequence analysis at mitochondrial 12S rRNA gene marker. Cyst-induced pathological changes included degenerative changes and demyelination in brain tissue, infiltration of lymphocytes and histiocytes...
February 2018: Data in Brief
https://www.readbyqxmd.com/read/29150912/sustaining-an-efficient-and-effective-cho-cell-line-development-platform-by-incorporation-of-24-deep-well-plate-screening-and-multivariate-analysis
#16
Alessandro Mora, Sheng Sam Zhang, Gerald Carson, Bernard Nabiswa, Patrick Hossler, Seongkyu Yoon
Efficient and effective cell line screening is paramount toward a successful biomanufacturing program. Here we describe the implementation of 24-deep well plate (24-DWP) screening of CHO lines as part of the cell line development platform at AbbVie. Incorporation of this approach accelerated the identification of the best candidate lines for process development. In an effort to quantify and predict process performance comparability, we compared cell culture performance in and in shake flasks, for a panel of Chinese Hamster Ovary cell lines expressing a monoclonal antibody...
November 18, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/29137641/improving-e-coli-growth-performance-by-manipulating-small-rna-expression
#17
REVIEW
Alejandro Negrete, Joseph Shiloach
Efficient growth of E. coli, especially for production of recombinant proteins, has been a challenge for the biotechnological industry since the early 1970s. By employing multiple approaches, such as different media composition, various growth strategies and specific genetic manipulations, it is now possible to grow bacteria to concentrations exceeding 100 g/L and to achieve high concentrations of recombinant proteins. Although the growth conditions are carefully monitored and maintained, it is likely that during the growth process cells are exposed to periodic stress conditions, created by fluctuations in pH, dissolved oxygen, temperature, glucose, and salt concentration...
November 14, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/29113894/combined-effect-of-gene-dosage-and-process-optimization-strategies-on-high-level-production-of-recombinant-human-interleukin-3-hil-3-in-pichia-pastoris-fed-batch-culture
#18
Vikas Kumar Dagar, Yogender Pal Khasa
In this work, the combined effects of gene dosage and process optimization strategies were studied to achieve higher hIL-3 expression in Pichia system. The in-vitro multimerization method was used to generate various Pichia X-33 transformants having multi-copy expression cassettes. The quantitative polymerase chain reaction (qPCR) strategy was used to further confirm the genome integration of hIL-3 expression cassette. From shake flask expression studies, the recombinant hIL-3 concentration in culture supernatant increased upto 8 copies to a level of 310mg/L, thereafter a considerably lower expression was observed...
November 4, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29110014/tmao-mediates-effective-attraction-between-lipid-membranes-by-partitioning-unevenly-between-bulk-and-lipid-domains
#19
Shahar Sukenik, Shaked Dunsky, Avishai Barnoy, Ilan Shumilin, Daniel Harries
Under environmental duress, many organisms accumulate large amounts of osmolytes - molecularly small organic solutes. Osmolytes are known to counteract stress, driving proteins to their compact native states by their exclusion from protein surfaces. In contrast, the effect of osmolytes on lipid membranes is poorly understood and widely debated. Many fully membrane-permeable osmolytes exert an apparent attractive force between lipid membranes, yet all proposed models fail to fully account for the origin of this force...
November 7, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/29078278/overcoming-factors-limiting-high-solids-fermentation-of-lignocellulosic-biomass-to-ethanol
#20
Thanh Yen Nguyen, Charles M Cai, Rajeev Kumar, Charles E Wyman
Simultaneous saccharification and fermentation (SSF) of solid biomass can reduce the complexity and improve the economics of lignocellulosic ethanol production by consolidating process steps and reducing end-product inhibition of enzymes compared with separate hydrolysis and fermentation (SHF). However, a long-standing limitation of SSF has been too low ethanol yields at the high-solids loading of biomass needed during fermentation to realize sufficiently high ethanol titers favorable for more economical ethanol recovery...
October 31, 2017: Proceedings of the National Academy of Sciences of the United States of America
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