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https://www.readbyqxmd.com/read/29777485/co-suppression-of-atmips-demonstrates-cooperation-of-mips1-mips2-and-mips3-in-maintaining-myo-inositol-synthesis
#1
C M Fleet, J Y Yen, E A Hill, G E Gillaspy
Co-suppressed MIPS2 transgenic lines allow bypass of the embryo lethal phenotype of the previously published triple knock-out and demonstrate the effects of MIPS on later stages of development. Regulation of inositol production is of interest broadly for its effects on plant growth and development. The enzyme L-myo-inositol 1-phosphate synthase (MIPS, also known as IPS) isomerizes D-glucose-6-P to D-inositol 3-P, and this is the rate-limiting step in inositol production. In Arabidopsis thaliana, the MIPS enzyme is encoded by three different genes, (AtMIPS1, AtMIPS2 and AtMIPS3), each of which has been shown to produce proteins with biochemically similar properties but differential expression patterns...
May 18, 2018: Plant Molecular Biology
https://www.readbyqxmd.com/read/29777103/bacterial-encapsulins-as-orthogonal-compartments-for-mammalian-cell-engineering
#2
Felix Sigmund, Christoph Massner, Philipp Erdmann, Anja Stelzl, Hannes Rolbieski, Mitul Desai, Sarah Bricault, Tobias P Wörner, Joost Snijder, Arie Geerlof, Helmut Fuchs, Martin Hrabĕ de Angelis, Albert J R Heck, Alan Jasanoff, Vasilis Ntziachristos, Jürgen Plitzko, Gil G Westmeyer
We genetically controlled compartmentalization in eukaryotic cells by heterologous expression of bacterial encapsulin shell and cargo proteins to engineer enclosed enzymatic reactions and size-constrained metal biomineralization. The shell protein (EncA) from Myxococcus xanthus auto-assembles into nanocompartments inside mammalian cells to which sets of native (EncB,C,D) and engineered cargo proteins self-target enabling localized bimolecular fluorescence and enzyme complementation. Encapsulation of the enzyme tyrosinase leads to the confinement of toxic melanin production for robust detection via multispectral optoacoustic tomography (MSOT)...
May 18, 2018: Nature Communications
https://www.readbyqxmd.com/read/29776931/shuffling-the-neutral-drift-of-unspecific-peroxygenase-in-saccharomyces-cerevisiae
#3
Javier Martin-Diaz, Carmen Paret, Eva García-Ruiz, Patricia Molina-Espeja, Miguel Alcalde
Unspecific peroxygenase (UPO) is a highly promiscuous biocatalyst and its selective mono(per)oxygenase activity makes it useful for many synthetic chemistry applications. Among the broad repertory of library creation methods for directed enzyme evolution, genetic drift allows neutral mutations to be accumulated gradually within a polymorphic network of variants. In this study, we conducted a campaign of genetic drift with UPO in Saccharomyces cerevisiae so that neutral mutations were simply added and recombined in vivo With low mutational loading and an activity threshold of 45% of the parent's native function, mutant libraries enriched in folded and active UPO variants were generated...
May 18, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29775852/biomimetic-magnetoelectric-nanocrystals-synthesized-by-polymerization-of-heme-as-advanced-nanomaterials-for-biosensing-application
#4
Jeong Eun Hyeon, Da Woon Jeong, Young Jin Ko, Seung Wook Kim, Chulhwan Park, Sung Ok Han
Regardless of the malaria disease risk, the malaria parasite Plasmodium falciparum has an interesting mechanism. During its growth within the red blood cell, toxic free heme is converted into an insoluble crystalline form called the malaria pigment, or hemozoin. In particular, natural hemozoin nanocrystals can provide multiple applications in biosensing fields for health care and diagnosis as similar to artificial metal nanoparticles. In this study, the heme was biologically synthesized and polymerized by Corynebacterium glutamicum and final polymer was applied as a biomimetic conductive biopolymer...
May 10, 2018: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/29773713/a-thumbwheel-mechanism-for-apoa1-activation-of-lcat-activity-in-hdl
#5
Allison L Cooke, Jamie Morris, John T Melchior, Scott E Street, W Gray Jerome, Rong Huang, Andrew B Herr, Loren E Smith, Jere P Segrest, Alan T Remaley, Amy S Shah, Thomas B Thompson, W Sean Davidson
APOA1 is the most abundant protein in HDL. It modulates interactions that affect HDLs cardioprotective functions, in part via its activation of the enzyme LCAT. On nascent, discoidal HDL, APOA1 comprises 10 alpha-helical repeats arranged in an anti-parallel, stacked-ring structure that encapsulates a lipid bilayer. Previous chemical cross-linking studies suggested that these APOA1 rings can adopt at least two different orientations, or registries, with respect to each other; however, the functional impact of these structural changes is unknown...
May 17, 2018: Journal of Lipid Research
https://www.readbyqxmd.com/read/29773424/a-device-for-high-throughput-monitoring-of-degradation-in-soft-tissue-samples
#6
D S Tzeranis, I Panagiotopoulos, S Gkouma, G Kanakaris, N Georgiou, N Vaindirlis, G Vasileiou, M Neidlin, A Gkousioudi, V Spitas, G A Macheras, L G Alexopoulos
This work describes the design and validation of a novel device, the High-Throughput Degradation Monitoring Device (HDD), for monitoring the degradation of 24 soft tissue samples over incubation periods of several days inside a cell culture incubator. The device quantifies sample degradation by monitoring its deformation induced by a static gravity load. Initial instrument design and experimental protocol development focused on quantifying cartilage degeneration. Characterization of measurement errors, caused mainly by thermal transients and by translating the instrument sensor, demonstrated that HDD can quantify sample degradation with <6 μm precision and <10 μm temperature-induced errors...
May 3, 2018: Journal of Biomechanics
https://www.readbyqxmd.com/read/29773074/enhanced-cellulase-production-in-trichoderma-reesei-rut-c30-via-constitution-of-minimal-transcriptional-activators
#7
Jiajia Zhang, Guoxiu Zhang, Wei Wang, Wei Wang, Dongzhi Wei
BACKGROUND: Cellulase can convert lignocellulosic feedstocks into fermentable sugars, which can be used for the industrial production of biofuels and chemicals. The high cost of cellulase production remains a challenge for lignocellulose breakdown. Trichoderma reesei RUT C30 serves as a well-known industrial workhorse for cellulase production. Therefore, the enhancement of cellulase production by T. reesei RUT C30 is of great importance. RESULTS: Two sets of novel minimal transcriptional activators (DBDace2 -VP16 and DBDcre1 -VP16) were designed and expressed in T...
May 17, 2018: Microbial Cell Factories
https://www.readbyqxmd.com/read/29771352/de-novo-biosynthesis-of-anthocyanins-in-saccharomyces-cerevisiae
#8
Michael Eichenberger, Anders Hansson, David Fischer, Lara Dürr, Michael Naesby
Anthocyanins (ACNs) are plant secondary metabolites responsible for most of the red, purple and blue colors of flowers, fruits and vegetables. They are increasingly used in the food and beverage industry as natural alternative to artificial colorants. Production of these compounds by fermentation of microorganisms would provide an attractive alternative. In this study, Saccharomyces cerevisiae was engineered for de novo production of the three basic anthocyanins, as well as the three main trans-flavan-3-ols...
June 1, 2018: FEMS Yeast Research
https://www.readbyqxmd.com/read/29770981/a-dehydrogenase-dual-hydrogen-abstraction-mechanism-promotes-estrogen-biosynthesis-can-we-expand-the-functional-annotation-of-the-aromatase-enzyme
#9
Angelo Spinello, Matic Pavlin, Lorenzo Casalino, Alessandra Magistrato
Cytochromes P450 (CYP450) are involved in the metabolism of exogenous compounds and in the synthesis of signaling molecules. Among the latter, human aromatase (HA) promotes estrogen biosynthesis, being a key pharmacological target against breast cancers. After decades of debate, the interest in gaining a comprehensive picture of HA catalysis has been renewed by the recent discovery that Compound I (Cpd I) is the reactive species of the peculiar aromatization step. Here, we provide for the first time a complete atomic-level picture of all controversial steps of estrogen biosynthesis...
May 17, 2018: Chemistry: a European Journal
https://www.readbyqxmd.com/read/29770379/the-limits-to-biocatalysis-pushing-the-envelope
#10
Roger A Sheldon, Dean Brady
In the period 1985 to 1995 applications of biocatalysis, driven by the need for more sustainable manufacture of chemicals and catalytic, (enantio)selective methods for the synthesis of pharmaceutical intermediates, largely involved the available hydrolases. This was followed, in the next two decades, by revolutionary developments in protein engineering and directed evolution for the optimisation of enzyme function and performance that totally changed the biocatalysis landscape. In the same period, metabolic engineering and synthetic biology revolutionised the use of whole cell biocatalysis in the synthesis of commodity chemicals by fermentation...
May 17, 2018: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/29769672/pancreas-regeneration
#11
REVIEW
Qiao Zhou, Douglas A Melton
The pancreas is made from two distinct components: the exocrine pancreas, a reservoir of digestive enzymes, and the endocrine islets, the source of the vital metabolic hormone insulin. Human islets possess limited regenerative ability; loss of islet β-cells in diseases such as type 1 diabetes requires therapeutic intervention. The leading strategy for restoration of β-cell mass is through the generation and transplantation of new β-cells derived from human pluripotent stem cells. Other approaches include stimulating endogenous β-cell proliferation, reprogramming non-β-cells to β-like cells, and harvesting islets from genetically engineered animals...
May 2018: Nature
https://www.readbyqxmd.com/read/29769320/o-glcnac-transferase-missense-mutations-linked-to-x-linked-intellectual-disability-deregulate-genes-involved-in-cell-fate-determination-and-signaling
#12
Nithya Selvan, Stephan George, Fatema J Serajee, Marie Shaw, Lynne Hobson, Vera M Kalscheuer, Nripesh Prasad, Shawn E Levy, Juliet Taylor, Salim Afitmos, Charles E Schwartz, Ahm M Huq, Jozef Gecz, Lance Wells
It is estimated that ~1% of the world's population has intellectual disability, with males affected more often than females. OGT is an X-linked gene encoding for the enzyme O-GlcNAc transferase (OGT), which carries out the reversible addition of N-Acetylglucosamine (GlcNAc) to Ser/Thr residues of its intracellular substrates. Three missense mutations in the tetratricopeptide (TPR) repeats of OGT have recently been reported to cause X-linked Intellectual Disability (XLID). Here we report the discovery of two additional novel missense mutations (c...
May 16, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29766662/bioproduction-of-benzylamine-from-renewable-feedstocks-via-a-9-step-artificial-enzyme-cascade-and-engineered-metabolic-pathways
#13
Yi Zhou, Shuke Wu, Jiwei Mao, Zhi Li
Production of chemicals from renewable feedstocks has been an important task for sustainable chemical industry. While microbial fermentation has been widely employed to produce many biochemicals, it is still very challenging to access non-natural chemicals. Herein, we developed two methods (biotransformation and fermentation) for the first bio-derived synthesis of benzylamine, a commodity non-natural amine with broad applications. At first, a 9-step artificial enzyme cascade was designed by biocatalytic retrosynthesis analysis and engineered in recombinant E...
May 15, 2018: ChemSusChem
https://www.readbyqxmd.com/read/29765029/the-crispr-tool-kit-for-genome-editing-and-beyond
#14
REVIEW
Mazhar Adli
CRISPR is becoming an indispensable tool in biological research. Once known as the bacterial immune system against invading viruses, the programmable capacity of the Cas9 enzyme is now revolutionizing diverse fields of medical research, biotechnology, and agriculture. CRISPR-Cas9 is no longer just a gene-editing tool; the application areas of catalytically impaired inactive Cas9, including gene regulation, epigenetic editing, chromatin engineering, and imaging, now exceed the gene-editing functionality of WT Cas9...
May 15, 2018: Nature Communications
https://www.readbyqxmd.com/read/29763548/zein-increases-the-cytoaffinity-and-biodegradability-of-scaffolds-3d-printed-with-zein-and-poly-%C3%AE%C2%B5-caprolactone-composite-ink
#15
Linzhi Jing, Xiang Wang, Hang Liu, Yuyun Lu, Jinsong Bian, Jie Sun, Dejian Huang
Electrohydrodynamic printing (EHDP) has attracted extensive interests as a powerful technology to fabricate micro to nanoscale fibrous scaffolds in a custom-tailored manner for biomedical applications. A few synthetic biopolymer inks are applicable to this EHDP technology, but the fabricated scaffolds suffered from low mechanical strength, biocompatibility and biodegradability. In this study, a series of poly(ε-caprolactone)(PCL)/zein composite inks were developed and their printability was examined on a solution-based EHDP system for scaffold fabrication...
May 15, 2018: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29760773/high-oleate-yeast-oil-without-polyunsaturated-fatty-acids
#16
Vasiliki Tsakraklides, Annapurna Kamineni, Andrew L Consiglio, Kyle MacEwen, Jonathan Friedlander, Hannah G Blitzblau, Maureen A Hamilton, Donald V Crabtree, Austin Su, Jonathan Afshar, John E Sullivan, W Greg LaTouf, Colin R South, Emily H Greenhagen, A Joe Shaw, Elena E Brevnova
Background: Oleate-enriched triacylglycerides are well-suited for lubricant applications that require high oxidative stability. Fatty acid carbon chain length and degree of desaturation are key determinants of triacylglyceride properties and the ability to manipulate fatty acid composition in living organisms is critical to developing a source of bio-based oil tailored to meet specific application requirements. Results: We sought to engineer the oleaginous yeast Yarrowia lipolytica for production of high-oleate triacylglyceride oil...
2018: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/29760099/hydrogel-delivery-of-lysostaphin-eliminates-orthopedic-implant-infection-by-staphylococcus-aureus-and-supports-fracture-healing
#17
Christopher T Johnson, James A Wroe, Rachit Agarwal, Karen E Martin, Robert E Guldberg, Rodney M Donlan, Lars F Westblade, Andrés J García
Orthopedic implant infections are a significant clinical problem, with current therapies limited to surgical debridement and systemic antibiotic regimens. Lysostaphin is a bacteriolytic enzyme with high antistaphylococcal activity. We engineered a lysostaphin-delivering injectable PEG hydrogel to treat Staphylococcus aureus infections in bone fractures. The injectable hydrogel formulation adheres to exposed tissue and fracture surfaces, ensuring efficient, local delivery of lysostaphin. Lysostaphin encapsulation within this synthetic hydrogel maintained enzyme stability and activity...
May 14, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29760092/noncatalytic-chalcone-isomerase-fold-proteins-in-humulus-lupulus-are-auxiliary-components-in-prenylated-flavonoid-biosynthesis
#18
Zhaonan Ban, Hao Qin, Andrew J Mitchell, Baoxiu Liu, Fengxia Zhang, Jing-Ke Weng, Richard A Dixon, Guodong Wang
Xanthohumol (XN) and demethylxanthohumol (DMX) are specialized prenylated chalconoids with multiple pharmaceutical applications that accumulate to high levels in the glandular trichomes of hops ( Humulus lupulus L.). Although all structural enzymes in the XN pathway have been functionally identified, biochemical mechanisms underlying highly efficient production of XN have not been fully resolved. In this study, we characterized two noncatalytic chalcone isomerase (CHI)-like proteins (designated as HlCHIL1 and HlCHIL2) using engineered yeast harboring all genes required for DMX production...
May 14, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29758351/development-of-microreactors-with-surface-immobilized-biocatalysts-for-continuous-transamination
#19
Nataša Miložič, Gorazd Stojkovič, Andreas Vogel, Dominique Bouwes, Polona Žnidaršič-Plazl
The industrial importance of optically pure compounds has thrown a spotlight on ω-transaminases that have shown a high potential for the synthesis of bioactive compounds with a chiral amine moiety. The implementation of biocatalysts in industrial processes relies strongly on fast and cost effective process development, including selection of a biocatalyst form and the strategy for its immobilization. Here, microscale reactors with selected surface-immobilized amine-transaminase (ATA) in various forms are described as platforms for high-throughput process development...
May 11, 2018: New Biotechnology
https://www.readbyqxmd.com/read/29756770/versatile-high-throughput-fluorescence-assay-for-monitoring-cas9-activity
#20
Kyle Seamon, Yooli K Light, Edwin Saada, Joseph S Schoeniger, Brooke Harmon
The RNA-guided DNA nuclease Cas9 is now widely used for the targeted modification of genomes of human cells and various organisms. Despite the extensive use of CRISPR systems for genome engineering and the rapid discovery and engineering of new CRISPR-associated nucleases, there are no high-throughput assays for measuring enzymatic activity. The current laboratory and future therapeutic uses of CRISPR technology have a significant risk of accidental exposure or clinical off-target effects, under-scoring the need for therapeutically-effective inhibitors of Cas9...
May 14, 2018: Analytical Chemistry
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