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yeast glycosylation

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https://www.readbyqxmd.com/read/28512129/mammalian-o-mannosylation-of-cadherins-and-plexins-is-independent-of-protein-o-mannosyltransferase-1-and-2
#1
Ida Signe Bohse Larsen, Yoshiki Narimatsu, Hiren Jitendra Joshi, Zhang Yang, Oliver J Harrison, Julia Brasch, Lawrence Shapiro, Barry Honig, Sergey Y Vakhrushev, Henrik Clausen, Adnan Halim
Protein O-mannosylation is found in yeast and metazoans and a family of conserved orthologous protein O-mannosyltransferases is believed to initiate this important post-translational modification. We recently discovered that the cadherin superfamily carries O-linked mannose (O-Man) glycans at highly conserved residues in specific extracellular cadherin domains, and it was suggested that the function of E-cadherin was dependent on the O-Man glycans. Deficiencies in enzymes catalyzing O-Man biosynthesis, including the two human protein O-mannosyltransferases, POMT1 and POMT2, underlie a subgroup of congenital muscular dystrophies (CMD) designated α-dystroglycanopathies, because deficient O-Man glycosylation of -dystroglycan disrupts laminin interaction with -dystroglycan and the extracellular matrix...
May 16, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28512128/refined-topology-model-of-the-stt3-sttp3-protein-subunit-of-the-oligosaccharyl-transferase-complex
#2
Patricia Lara, Karin Öjemalm, Johannes Reithinger, Aurora Holgado, You Maojun, Abdessalem Hammed, Daniel Mattle, Hyun Kim, IngMarie Nilsson
The oligosaccharyl transferase (OST) complex, localized in the endoplasmic reticulum (ER) of eukaryotic cells, is responsible for the N-linked glycosylation of numerous protein substrates. The membrane protein STT3 is a highly conserved part of the OST, and likely contains the active site of the complex. However, understanding the catalytic determinants of this system has been challenging, in part because of a discrepancy in the structural topology of the bacterial vs eukaryotic proteins and incomplete information about the mechanism of membrane integration...
May 16, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28500401/sources-of-%C3%AE-galactosidase-and-its-applications-in-food-industry
#3
REVIEW
Shaima Saqib, Attiya Akram, Sobia Ahsan Halim, Raazia Tassaduq
The enzyme β-galactosidases have been isolated from various sources such as bacteria, fungi, yeast, vegetables, and recombinant sources. This enzyme holds importance due to its wide applications in food industries to manufacture lactose-hydrolyzed products for lactose-intolerant people and the formation of glycosylated products. Absorption of undigested lactose in small intestine requires the activity of this enzyme; hence, the deficiency of this enzyme leads to lactose intolerance. Lactose intolerance affects around 70% of world's adult population, while the prevalence rate of lactose intolerance is 60% in Pakistan...
May 2017: 3 Biotech
https://www.readbyqxmd.com/read/28499956/group-x-hybrid-histidine-kinase-chk1-is-dispensable-for-stress-adaptation-host-pathogen-interactions-and-virulence-in-the-opportunistic-yeast-candida-guilliermondii
#4
María J Navarro-Arias, Karine Dementhon, Tatiana A Defosse, Emilien Foureau, Vincent Courdavault, Marc Clastre, Solène Le Gal, Gilles Nevez, Yohann Le Govic, Jean-Philippe Bouchara, Nathalie Giglioli-Guivarc'h, Thierry Noël, Hector M Mora-Montes, Nicolas Papon
Hybrid histidine kinases (HHKs) progressively emerge as prominent sensing proteins in the fungal kingdom and as ideal targets for future therapeutics. The group X HHK is of major interest, since it was demonstrated to play an important role in stress adaptation, host-pathogen interactions and virulence in some yeast and mold models, and particularly Chk1, that corresponds to the sole group X HHK in Candida albicans. In the present work, we investigated the role of Chk1 in the low-virulence species Candida guilliermondii, in order to gain insight into putative conservation of the role of group X HHK in opportunistic yeasts...
May 9, 2017: Research in Microbiology
https://www.readbyqxmd.com/read/28470614/not-limited-to-e-coli-versatile-expression-vectors-for-mammalian-protein-expression
#5
Katharina Karste, Maren Bleckmann, Joop van den Heuvel
Recombinant protein expression is not limited to E. coli or other prokaryotic systems. It is inevitable to use eukaryotic systems in order to express challenging mammalian proteins. Eukaryotic systems are able to perform complex posttranslational modifications like protein processing, phosphorylation, glycosylation, which are essential for stability and functionality of many proteins. Different eukaryotic protein expression systems employing yeast, insect, or mammalian cell lines are established with each having its own advantages and disadvantages...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28467350/chemical-profile-and-antioxidant-anti-inflammatory-antimutagenic-and-antimicrobial-activities-of-geopropolis-from-the-stingless-bee-melipona-orbignyi
#6
Helder Freitas Dos Santos, Jaqueline Ferreira Campos, Cintia Miranda Dos Santos, José Benedito Perrella Balestieri, Denise Brentan Silva, Carlos Alexandre Carollo, Kely de Picoli Souza, Leticia Miranda Estevinho, Edson Lucas Dos Santos
Geopropolis is a resin mixed with mud, produced only by stingless bees. Despite being popularly known for its medicinal properties, few scientific studies have proven its biological activities. In this context, the objective of this study was to determine the chemical composition and antioxidant, anti-inflammatory, antimutagenic and antimicrobial activities of the Melipona orbignyi geopropolis. The hydroalcoholic extract of geopropolis (HEGP) was prepared and its chemical composition determined by high performance liquid chromatography coupled to diode array detector and mass spectrometry (HPLC-DAD-MS)...
May 3, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28446609/glycosylation-of-the-core-of-the-hiv-1-envelope-subunit-protein-gp120-is-not-required-for-native-trimer-formation-or-viral-infectivity
#7
Ujjwal Rathore, Piyali Saha, Sannula Kesavardhana, Aditya Arun Kumar, Rohini Datta, Sivasankar Devanarayanan, Raksha Das, John R Mascola, Raghavan Varadarajan
The gp120 subunit of HIV-1 envelope (Env) protein is heavily glycosylated at approximately 25 glycosylation sites, of which ~7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. Glycans partially shield Env from recognition by the host immune system and also are believed to be indispensable for proper folding of gp120 and for viral infectivity. Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or to alanines...
April 26, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28433761/a-glycoproteomic-approach-reveals-that-the-s-layer-glycoprotein-of-lactobacillus-kefiri-cidca-83111-is-o-and-n-glycosylated
#8
Gustavo J Cavallero, Mariano Malamud, Adriana C Casabuono, M de Los Ángeles Serradell, Alicia S Couto
In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid...
April 19, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/28411481/a-re-evaluation-of-the-final-step-of-vanillin-biosynthesis-in-the-orchid-vanilla-planifolia
#9
Hailian Yang, Jaime Barros-Rios, Galina Kourteva, Xiaolan Rao, Fang Chen, Hui Shen, Chenggang Liu, Andrzej Podstolski, Faith Belanger, Daphna Havkin-Frenkel, Richard A Dixon
A recent publication describes an enzyme from the vanilla orchid Vanilla planifolia with the ability to convert ferulic acid directly to vanillin. The authors propose that this represents the final step in the biosynthesis of vanillin, which is then converted to its storage form, glucovanillin, by glycosylation. The existence of such a "vanillin synthase" could enable biotechnological production of vanillin from ferulic acid using a "natural" vanilla enzyme. The proposed vanillin synthase exhibits high identity to cysteine proteases, and is identical at the protein sequence level to a protein identified in 2003 as being associated with the conversion of 4-coumaric acid to 4-hydroxybenzaldehyde...
July 2017: Phytochemistry
https://www.readbyqxmd.com/read/28409466/the-use-of-glycosylation-tags-as-reporters-for-protein-entry-into-the-endoplasmic-reticulum-in-yeast-and-mammalian-cells
#10
Judith Buentzel, Sven Thoms
N-glycosylation is a process occurring in the Endoplasmic Reticulum (ER) in nearly every organism. Proteins containing a glycosylation site are quickly glycosylated by oligosaccharyltransferases once the glycosylation site is exposed to the ER lumen. The oligosaccharide tree is then modified and proteins are targeted to specific organelles or subcompartments. For a long time peroxisomal membrane proteins (PMP) were thought to be targeted directly to the peroxisome. However, in the course of recent years, several PMPs were found to be targeted via the ER...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28407119/a-barley-udp-glucosyltransferase-inactivates-nivalenol-and-provides-fusarium-head-blight-resistance-in-transgenic-wheat
#11
Xin Li, Herbert Michlmayr, Wolfgang Schweiger, Alexandra Malachova, Sanghyun Shin, Yadong Huang, Yanhong Dong, Gerlinde Wiesenberger, Susan McCormick, Marc Lemmens, Philipp Fruhmann, Christian Hametner, Franz Berthiller, Gerhard Adam, Gary J Muehlbauer
Fusarium Head Blight is a disease of cereal crops that causes severe yield losses and mycotoxin contamination of grain. The main causal pathogen, Fusarium graminearum, produces the trichothecene toxins deoxynivalenol or nivalenol as virulence factors. Nivalenol-producing isolates are most prevalent in Asia but co-exist with deoxynivalenol producers in lower frequency in North America and Europe. Previous studies identified a barley UDP-glucosyltransferase, HvUGT13248, that efficiently detoxifies deoxynivalenol, and when expressed in transgenic wheat results in high levels of type II resistance against deoxynivalenol-producing F...
April 12, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/28376863/pichia-pastoris-versus-saccharomyces-cerevisiae-a-case-study-on-the-recombinant-production-of-human-granulocyte-macrophage-colony-stimulating-factor
#12
Anh-Minh Tran, Thanh-Thao Nguyen, Cong-Thuan Nguyen, Xuan-Mai Huynh-Thi, Cao-Tri Nguyen, Minh-Thuong Trinh, Linh-Thuoc Tran, Stephanie P Cartwright, Roslyn M Bill, Hieu Tran-Van
BACKGROUND: Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) is a glycoprotein that has been approved by the FDA for the treatment of neutropenia and leukemia in combination with chemotherapies. Recombinant hGM-CSF is produced industrially using the baker's yeast, Saccharomyces cerevisiae, by large-scale fermentation. The methylotrophic yeast, Pichia pastoris, has emerged as an alternative host cell system due to its shorter and less immunogenic glycosylation pattern together with higher cell density growth and higher secreted protein yield than S...
April 4, 2017: BMC Research Notes
https://www.readbyqxmd.com/read/28366604/single-n277a-substitution-in-c2-of-simian-immunodeficiency-virus-envelope-influences-vaccine-elicited-cd4i-neutralizing-and-anti-v2-antibody-responses
#13
Xian Tang, Jia Guo, Lin Cheng, Caijun Sun, Li Liu, Teng Zuo, Hui Wang, Ling Chen, Linqi Zhang, Zhiwei Chen
An effective HIV vaccine remains elusive, and immunogens capable of eliciting protective host humoral immunity have not yet been identified. Although HIV/SIV infections result in the abundant production of CD4-induced (CD4i) antibodies (Abs), these Abs are not protective due to steric restrictions following gp120 binding to CD4 on target cells. Here we report that both DNA- and vaccinia-based vaccines encoding SIVmac239 gp160 readily elicited high levels of CD4i Abs in experimental animals. We identified a highly conserved N-linked glycosylation site N277 in the C2 region which strongly affected the immunogenicity of the CD4i Ab domain...
May 2, 2017: Vaccine
https://www.readbyqxmd.com/read/28356218/efficient-antibody-production-in-the-methylotrophic-yeast-ogataea-minuta-by%C3%A2-overexpression-of-chaperones
#14
Takeshi Suzuki, Satoshi Baba, Minako Ono, Koichi Nonaka, Kimihisa Ichikawa, Masayuki Yabuta, Rie Ito, Yasunori Chiba
A production system for a therapeutic monoclonal antibody was developed using the methylotrophic yeast Ogataea minuta IFO10746. The genetically engineered O. minuta secreted a detectable amount of anti-TRAIL receptor antibody into the culture supernatant, and the secreted antibody was purified by multiple column chromatography steps. In the purification process, both fully and partially assembled antibodies were detected and isolated. The fully assembled antibody from O. minuta showed almost the same biological activity as that derived from mammalian cells despite the distinct glycosylation profile, whereas the partially assembled antibody showed no cytotoxic activity...
March 26, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28348446/exploring-the-genomic-diversity-of-black-yeasts-and-relatives-chaetothyriales-ascomycota
#15
M M Teixeira, L F Moreno, B J Stielow, A Muszewska, M Hainaut, L Gonzaga, A Abouelleil, J S L Patané, M Priest, R Souza, S Young, K S Ferreira, Q Zeng, M M L da Cunha, A Gladki, B Barker, V A Vicente, E M de Souza, S Almeida, B Henrissat, A T R Vasconcelos, S Deng, H Voglmayr, T A A Moussa, A Gorbushina, M S S Felipe, C A Cuomo, G Sybren de Hoog
The order Chaetothyriales (Pezizomycotina, Ascomycetes) harbours obligatorily melanised fungi and includes numerous etiologic agents of chromoblastomycosis, phaeohyphomycosis and other diseases of vertebrate hosts. Diseases range from mild cutaneous to fatal cerebral or disseminated infections and affect humans and cold-blooded animals globally. In addition, Chaetothyriales comprise species with aquatic, rock-inhabiting, ant-associated, and mycoparasitic life-styles, as well as species that tolerate toxic compounds, suggesting a high degree of versatile extremotolerance...
March 2017: Studies in Mycology
https://www.readbyqxmd.com/read/28330196/cloning-and-expression-of-saccharomyces-cerevisiae-suc2-gene-in-yeast-platform-and-characterization-of-recombinant-enzyme-biochemical-properties
#16
Nooshin Mohandesi, Seyed Omid Ranaei Siadat, Kamahldin Haghbeen, Ardeshir Hesampour
Invertase (EC.3.2.1.26) catalyzes the hydrolysis of sucrose to an equimolar mixture of D-glucose and D-fructose which is of interest for various industrial applications. In this research, Saccharomyces cerevisiae invertase gene (SUC2) was optimized based on Pichia pastoris codon preference. The synthetic gene was introduced into the methylotrophic yeast Pichia pastoris under the control of the inducible AOX1 promoter. High level of the extracellular recombinant invertase (R-inv) production was achieved via methanol induction for 4 days and purified by His-Tag affinity chromatography which appeared to be a mixture of glycosylated proteins with various sizes of 85-95 kDa on SDS-PAGE...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28323393/evaluation-of-antidiabetic-activity-of-biologically-synthesized-silver-nanoparticles-agnps-using-pouteria-sapota-on-stz-induced-diabetic-rats
#17
Sathya Prabhu, Shanmugam Vinodhini, Chakravarthy Elenchezhiyan, Devi Rajeswari
BACKGROUND: Medicinal plants and green synthesis of silver nanoparticles (AgNPs) have proven to be potent source effective agents in the treatment of diabetes mellitus. The present study is focused on the green synthesis of AgNPs from aqueous leaf extract of Pouteria sapota (P. sapota) in order to evaluate the in-vitro and in-vivo anti-diabetic property of aqueous extracts and their synthesized AgNPs. METHODS: AgNPs were biologically synthesized under ambient conditions from an aqueous leaf extract of P...
March 21, 2017: Journal of Diabetes
https://www.readbyqxmd.com/read/28299731/heterologous-expression-of-the-astacin-protease-meprin-%C3%AE-in-pichia-pastoris
#18
Dagmar Schlenzig, Stephan Schilling
Meprins are zinc-dependent proteases of the metzincin superfamily of metalloproteases. The enzymes are extracellular multi-domain proteins which are stabilized by disulfide bridges, dimerization, and glycosylation. Due to their complex structure, recombinant expression was first established in mammalian and insect cells. However, these methods have several disadvantages such as high costs and the low yields. For this reason, yeast is often considered a preferable expression system. Here, we describe the manipulation and secretory expression of human meprin β in the methylotrophic yeast P...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28288897/efficient-production-of-glycosylated-cypridina-luciferase-using-plant-cells
#19
Yasuo Mitani, Yoshimi Oshima, Nobutaka Mitsuda, Azusa Tomioka, Masako Sukegawa, Mika Fujita, Hiroyuki Kaji, Yoshihiro Ohmiya
Cypridina noctiluca luciferase has been utilized for biochemical and molecular biological applications, including bioluminescent enzyme immunoassays, far-red luminescence imaging, and high-throughput reporter assays. Some of these applications require a large amount of purified luciferase. However, conventional protein expression systems are not capable of producing sufficient quantities of protein with a high quality and purity without laborious and costly purification processes. To improve the productivity and expand the breadth of possibilities for Cypridina luciferase applications, we employed a variety of secretion expression systems, including yeast, mammalian cells, and silk worms...
March 11, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28277249/glycan-based-cell-targeting-to-modulate-immune-responses
#20
REVIEW
Timo Johannssen, Bernd Lepenies
Glycosylation is an integral post-translational modification present in more than half of all eukaryotic proteins. It affects key protein functions, including folding, stability, and immunogenicity. Glycoengineering approaches, such as the use of bacterial N-glycosylation systems, or expression systems, including yeasts, insect cells, and mammalian cells, have enabled access to defined and homogenous glycoproteins. Given that glycan structures on proteins can be recognized by host lectin receptors, they may facilitate cell-specific targeting and immune modulation...
April 2017: Trends in Biotechnology
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