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https://www.readbyqxmd.com/read/28535614/elucidating-bottlenecks-of-the-efficient-preparation-of-an-ab5-hexamer-mucosal-adjuvant-protein-ltm-by-genetic-engineering
#1
Di Liu, Fabiao Hu, Wenpeng Wang, Dong Wu, Xiujuan He, Wenyun Zheng, Haipeng Liu, Xingyuan Ma
Escherichia coli heat-labile enterotoxin (LT) and its non-toxic mutant (LTm) are well-known powerful mucosal adjuvants and immunogens. However, the yields of these adjuvants from genetically engineered strains remain at extremely low levels, thereby hindering their extensive application in fundamental and clinical research. Therefore, efficient production of these adjuvant proteins from genetically engineered microbes is a huge challenge in the field of molecular biology. In order to explore expression bottlenecks of LTm in E...
May 24, 2017: Journal of Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28535612/efficient-whole-cell-biocatalyst-for-2-3-butanediol-acetoin-production-with-nadh-nad-regeneration-system-in-engineered-serratia-marcescens
#2
Rao Ben, Truong Ngoc Tu, Fan Ji Ying, Sun Jian An, Yaling Shen
Serratia marcescens is reported to possess the potential for industrial 2,3-butanediol or acetoin production by fermentation. But 2,3-butanediol or acetoin are always co-produced and this may make purification process difficult. So biocatalytic technologies may be the appropriate production methods. In this study, we developed an auto-inducing expression system based on pET system and swr quorum sensing system by using S. marcescens. By using this system, S. marcescens could be engineered as whole-cell biocatalyst to obtain 2,3-butanediol or acetoin...
May 24, 2017: Journal of Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28535034/p450-mediated-non-natural-cyclopropanation-of-dehydro-alanine-containing-thiopeptides
#3
Joshua G Gober, Swapnil V Ghodge, Jonathan W Bogart, Walter J Wever, Richard R Watkins, Eric M Brustad, Albert A Bowers
Thiopeptides are a growing class of ribosomally synthesized and posttranslationally modified peptide (RiPP) natural products. Many biosynthetic enzymes for RiPPs, especially thiopeptides, are promiscuous and can accept a wide range of peptide substrates with different amino acid sequence; thus, these enzymes have been used as tools to generate new natural product derivatives. Here, we explore an alternative route to molecular complexity by engineering thiopeptide tailoring enzymes to do new or non-native chemistry...
May 23, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28534616/microbes-and-the-next-nitrogen-revolution
#4
Ilje Pikaar, Silvio Matassa, Korneel Rabaey, Benjamin Leon Bodirsky, Alexander Popp, Mario Herrero, Willy Verstraete
The Haber Bosch process is among the greatest inventions of the 20th century. It provided agriculture with reactive nitrogen and ultimately mankind with nourishment for a population of 7 billion people. However, the present agricultural practice of growing crops for animal production and human food constitutes a major threat to the sustainability of the planet in terms of reactive nitrogen pollution. In view of the shortage of directly feasible and cost-effective measures to avoid these planetary nitrogen burdens and the necessity to remediate this problem, we foresee the absolute need for, and expect, a revolution in the use of microbes as a source of protein...
May 23, 2017: Environmental Science & Technology
https://www.readbyqxmd.com/read/28534229/production-of-a-gonadotropin-releasing-hormone-2-receptor-knockdown-gnrhr2-kd-swine-line
#5
A T Desaulniers, R A Cederberg, G A Mills, C A Lents, B R White
Swine are the only livestock species that produce both the second mammalian isoform of gonadotropin-releasing hormone (GNRH2) and its receptor (GNRHR2). Previously, we reported that GNRH2 and GNRHR2 mediate LH-independent testosterone secretion from porcine testes. To further explore this ligand-receptor complex, a pig model with reduced GNRHR2 expression was developed. Small hairpin RNA sequences targeting porcine GNRHR2 were subcloned into a lentiviral-based vector, lentiviral particles were generated and microinjected into the perivitelline space of zygotes, and embryos were transferred into a recipient...
May 22, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28534111/overexpression-of-ppc-or-deletion-of-mdh-for-improving-production-of-%C3%AE-aminobutyric-acid-in-recombinant-corynebacterium-glutamicum
#6
Feng Shi, Ming Zhang, Yongfu Li
L-Glutamate decarboxylase (GAD) transforms L-glutamate into γ-aminobutyric acid (GABA). Corynebacterium glutamicum that expresses exogenous GAD gene(s) can synthesize GABA from its own produced L-glutamate. To enhance GABA production in recombinant C. glutamicum strain SH, metabolic engineering strategies were used to improve the supply of the GABA precursor, L-glutamate. Five new strains were constructed here. First, the ppc gene was coexpressed with two GAD genes (gadB1 and gadB2). Then, the mdh gene was deleted in C...
June 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28533816/conferring-cellulose-degrading-ability-to-yarrowia-lipolytica-to-facilitate-a-consolidated-bioprocessing-approach
#7
Zhong-Peng Guo, Sophie Duquesne, Sophie Bozonnet, Gianluca Cioci, Jean-Marc Nicaud, Alain Marty, Michael Joseph O'Donohue
BACKGROUND: Yarrowia lipolytica, one of the most widely studied "nonconventional" oleaginous yeast species, is unable to grow on cellulose. Recently, we identified and overexpressed two endogenous β-glucosidases in Y. lipolytica, thus enabling this yeast to use cello-oligosaccharides as a carbon source for growth. Using this engineered yeast platform, we have now gone further toward building a fully cellulolytic Y. lipolytica for use in consolidated bioprocessing of cellulose. RESULTS: Initially, different essential enzyme components of a cellulase cocktail (i...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/28533772/endotoxin-contamination-in-nanomaterials-leads-to-the-misinterpretation-of-immunosafety-results
#8
Yang Li, Mayumi Fujita, Diana Boraschi
Given the presence of engineered nanomaterials in consumers' products and their application in nanomedicine, nanosafety assessment is becoming increasingly important. In particular, immunosafety aspects are being actively investigated. In nanomaterial immunosafety testing strategies, it is important to consider that nanomaterials and nanoparticles are very easy to become contaminated with endotoxin, which is a widespread contaminant coming from the Gram-negative bacterial cell membrane. Because of the potent inflammatory activity of endotoxin, contaminated nanomaterials can show inflammatory/toxic effects due to endotoxin, which may mask or misidentify the real biological effects (or lack thereof) of nanomaterials...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28533324/crispr-cas9-genetic-modification-of-cyp3a5-3-in-huh-7-human-hepatocyte-cell-line-leads-to-cell-lines-with-increased-midazolam-and-tacrolimus-metabolism
#9
Casey R Dorr, Rory P Remmel, Amutha Muthusamy, James Fisher, Branden Moriarity, Kazuto Yasuda, Baolin Wu, Weihua Guan, Erin G Schuetz, William S Oetting, Pamala A Jacobson, Ajay K Israni
CRISPR/Cas9 engineering of the CYP3A5 *3 locus (rs776746) in human liver cell line HuH-7 (CYP3A5 *3/*3) led to three CYP3A5 *1 cell lines by deletion of the exon 3B splice junction or point mutation. Cell lines CYP3A5 *1/*3 sd (single deletion), CYP3A5 *1/*1 dd (double deletion) or CYP3A5 *1/*3 pm (point mutation) expressed the CYP3A5 *1 mRNA, had elevated CYP3A5 mRNA (p<0.0005 for all engineered cell lines) and protein expression compared with HuH-7. In metabolism assays, HuH-7 had less tacrolimus (Tac) (all p-values < 0...
May 22, 2017: Drug Metabolism and Disposition: the Biological Fate of Chemicals
https://www.readbyqxmd.com/read/28533309/aryl-hydrocarbon-receptor-inhibition-promotes-hemato-lymphoid-development-from-human-pluripotent-stem-cells
#10
Mathew G Angelos, Paige N Ruh, Beau R Webber, Robert H Blum, Caitlin D Ryan, Laura Bendzick, Seonhui Shim, Ashley M Yingst, Dejene M Tufa, Michael R Verneris, Dan S Kaufman
The aryl hydrocarbon receptor (AHR) plays an important physiological role in hematopoiesis. AHR is highly expressed in hematopoietic stem/progenitor cells (HSPCs) and inhibition of AHR results in a marked expansion of human umbilical cord blood-derived HSPCs following cytokine stimulation. It is unknown whether AHR also contributes earlier in human hematopoietic development. To model hematopoiesis, human embryonic stem cells (hESCs) were allowed to differentiate in defined conditions in the presence of the AHR antagonist StemReginin-1 (SR-1) or AHR agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)...
May 22, 2017: Blood
https://www.readbyqxmd.com/read/28533066/an-efficient-tool-for-metabolic-pathway-construction-and-gene-integration-for-aspergillus-niger
#11
Parveen Sarkari, Hans Marx, Marzena L Blumhoff, Diethard Mattanovich, Michael Sauer, Matthias G Steiger
Metabolic engineering requires functional genetic tools for easy and quick generation of multiple pathway variants. A genetic engineering toolbox for A. niger is presented, which facilitates the generation of strains carrying heterologous expression cassettes at a defined genetic locus. The system is compatible with Golden Gate cloning, which facilitates the DNA construction process and provides high design flexibility. The integration process is mediated by a CRISPR/Cas9 strategy involving the cutting of both the genetic integration locus (pyrG) as well as the integrating plasmid...
May 4, 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28532902/adipose-derived-delivery-vehicle-for-encapsulated-adipogenic-factors
#12
Christopher M Mahoney, Arta Kelmindi-Doko, Malik J Snowden, J Peter Rubin, Kacey G Marra
Hydrogels derived from adipose tissue extracellular matrix (AdECM) have shown potential in the ability to generate new adipose tissue in vivo. To further enhance adipogenesis, a composite adipose derived delivery system (CADDS) containing single- and double-walled dexamethasone encapsulated microspheres (SW and DW Dex MS) has been developed. Previously, our laboratory has published the use of Dex MS as an additive to enhance adipogenesis and angiogenesis in adipose tissue grafts. In the current work, AdECM and CADDS are extensively characterized, in addition to conducting in vitro cell culture analysis...
May 19, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28532676/novel-strategies-for-the-formulation-and-processing-of-poorly-water-soluble-drugs
#13
REVIEW
Katrin Göke, Thomas Lorenz, Alexandros Repanas, Frederic Schneider, Denise Steiner, Knut Baumann, Heike Bunjes, Andreas Dietzel, Jan H Finke, Birgit Glasmacher, Arno Kwade
Low aqueous solubility of active pharmaceutical ingredients presents a serious challenge in the development process of new drug products. This article provides an overview on some of the current approaches for the formulation of poorly water-soluble drugs with a special focus on strategies pursued at the Center of Pharmaceutical Engineering of the TU Braunschweig. These comprise formulation in lipid-based colloidal drug delivery systems and experimental as well as computational approaches towards the efficient identification of the most suitable carrier systems...
May 19, 2017: European Journal of Pharmaceutics and Biopharmaceutics
https://www.readbyqxmd.com/read/28532663/recent-developments-in-drug-eluting-devices-with-tailored-interfacial-properties
#14
REVIEW
Eva Sanchez-Rexach, Emilio Meaurio, Jose-Ramon Sarasua
Drug eluting devices have greatly evolved during past years to become fundamental products of great marketing importance in the biomedical field. There is currently a large diversity of highly specialized devices for specific applications, making the development of these devices an exciting field of research. The replacement of the former bare metal devices by devices loaded with drugs allowed the sustained and controlled release of drugs, to achieve the desired local therapeutic concentration of drug. The newer devices have been "engineered" with surfaces containing micro- and nanoscale features in a well-controlled manner, that have shown to significantly affect cellular and subcellular function of various biological systems...
May 4, 2017: Advances in Colloid and Interface Science
https://www.readbyqxmd.com/read/28532507/isoprenoid-biosynthesis-in-dandelion-latex-is-enhanced-by-the-overexpression-of-three-key-enzymes-involved-in-the-mevalonate-pathway
#15
Katharina M Pütter, Nicole van Deenen, Kristina Unland, Dirk Prüfer, Christian Schulze Gronover
BACKGROUND: Latex from the dandelion species Taraxacum brevicorniculatum contains many high-value isoprenoid end products, e.g. triterpenes and polyisoprenes such as natural rubber. The isopentenyl pyrophosphate units required as precursors for these isoprenoids are provided by the mevalonate (MVA) pathway. The key enzyme in this pathway is 3-hydroxy-methyl-glutaryl-CoA reductase (HMGR) and its activity has been thoroughly characterized in many plant species including dandelion. However, two enzymes acting upstream of HMGR have not been characterized in dandelion latex: ATP citrate lyase (ACL), which provides the acetyl-CoA utilized in the MVA pathway, and acetoacetyl-CoA thiolase (AACT), which catalyzes the first step in the pathway to produce acetoacetyl-CoA...
May 22, 2017: BMC Plant Biology
https://www.readbyqxmd.com/read/28532497/improving-the-production-of-22-hydroxy-23-24-bisnorchol-4-ene-3-one-from-sterols-in-mycobacterium-neoaurum-by-increasing-cell-permeability-and-modifying-multiple-genes
#16
Liang-Bin Xiong, Hao-Hao Liu, Li-Qin Xu, Wan-Ju Sun, Feng-Qing Wang, Dong-Zhi Wei
BACKGROUND: The strategy of modifying the sterol catabolism pathway in mycobacteria has been adopted to produce steroidal pharmaceutical intermediates, such as 22-hydroxy-23,24-bisnorchol-4-ene-3-one (4-HBC), which is used to synthesize various steroids in the industry. However, the productivity is not desirable due to some inherent problems, including the unsatisfactory uptake rate and the low metabolic efficiency of sterols. The compact cell envelope of mycobacteria is a main barrier for the uptake of sterols...
May 22, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28532451/enhancing-poly-%C3%AE-glutamic-acid-production-in-bacillus-amyloliquefaciens-by-introducing-the-glutamate-synthesis-features-from-corynebacterium-glutamicum
#17
Jun Feng, Yufen Quan, Yanyan Gu, Fenghong Liu, Xiaozhong Huang, Haosheng Shen, Yulei Dang, Mingfeng Cao, Weixia Gao, Xiaoyun Lu, Yi Wang, Cunjiang Song, Shufang Wang
BACKGROUND: Poly-γ-glutamic acid (γ-PGA) is a valuable polymer with glutamate as its sole precursor. Enhancement of the intracellular glutamate synthesis is a very important strategy for the improvement of γ-PGA production, especially for those glutamate-independent γ-PGA producing strains. Corynebacterium glutamicum has long been used for industrial glutamate production and it exhibits some unique features for glutamate synthesis; therefore introduction of these metabolic characters into the γ-PGA producing strain might lead to increased intracellular glutamate availability, and thus ultimate γ-PGA production...
May 22, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28532300/engineered-polymeric-amphiphiles-self-assembling-into-nanostructures-and-acting-as-efficient-gene-and-drug-carriers
#18
Ruby Bansal, Pradeep Kumar
Nonviral gene delivery systems are finding widespread use due to their safety, rapid and economical production, and ease of modification. In this work, series of N-alkyl-substituted linear polyethylenimine (CP) polymers have been synthesized, characterized, and investigated about how degree of substitution (hydrophobic-hydrophilic balance) (i.e. N-alkylation) influenced the transfection efficiency. Mobility shift assay demonstrated efficient binding of plasmid DNA (pDNA). Transfection efficiency and cytotoxicity of CP polymers were assessed in vitro, which revealed that all the formulations exhibited higher transfection activity than linear polyethylenimine (lPEI) and commercial transfection reagents, Lipofectamine and Superfect, with negligible toxicity (MTT assay)...
January 1, 2017: Journal of Biomaterials Applications
https://www.readbyqxmd.com/read/28532245/applicability-of-ribosome-engineering-to-vitamin-b12-production-by-propionibacterium-shermanii
#19
Yukinori Tanaka, Ken Kasahara, Masumi Izawa, Kozo Ochi
Ribosome engineering has been widely utilized for strain improvement, especially for the activation of bacterial secondary metabolism. This study assessed ribosome engineering technology to modulate primary metabolism, taking vitamin B12 production as a representative example. The introduction into Propionibacterium shermanii of mutations conferring resistance to rifampicin, gentamicin, and erythromycin, respectively, increased per cell production (μg/L/OD600) of vitamin B12 5.2-fold, although net production (μg/L) was unchanged, as the cell mass of the mutants was reduced...
May 22, 2017: Bioscience, Biotechnology, and Biochemistry
https://www.readbyqxmd.com/read/28532182/tackling-a-novel-lethal-virus-a-focus-on-h7n9-vaccine-development
#20
Irina Isakova-Sivak, Larisa Rudenko
Avian-origin H7N9 influenza viruses first detected in humans in China in 2013 continue to cause severe human infections with a mortality rate close to 40%. These viruses are acknowledged as the subtype most likely to cause the next influenza pandemic. Areas covered: Here we review published data on the development of H7N9 influenza vaccine candidates and their evaluation in preclinical and clinical trials identified on PubMed database with the term "H7N9 influenza vaccine". In addition, a search with the same term was done on ClinicalTrials...
May 22, 2017: Expert Review of Vaccines
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